%0 Journal Article %J Sci Rep %D 2015 %T Identification of genetic loci that control mammary tumor susceptibility through the host microenvironment. %A Zhang, Pengju %A Lo, Alvin %A Huang, Yurong %A Huang, Ge %A Liang, Guozhou %A Mott, Joni %A Karpen, Gary H %A Blakely, Eleanor A %A Bissell, Mina J %A Barcellos-Hoff, Mary Helen %A A Snijders %A Jiang-Hua Mao %K Animals %K Breast Neoplasms %K Cell Line, Tumor %K Cytokines %K Female %K Genetic Predisposition to Disease %K Mice %K Mice, Inbred BALB C %K Neoplasms, Radiation-Induced %K Quantitative Trait Loci %K Risk Factors %K Transforming Growth Factor beta1 %K Tumor Microenvironment %X

The interplay between host genetics, tumor microenvironment and environmental exposure in cancer susceptibility remains poorly understood. Here we assessed the genetic control of stromal mediation of mammary tumor susceptibility to low dose ionizing radiation (LDIR) using backcrossed F1 into BALB/c (F1Bx) between cancer susceptible (BALB/c) and resistant (SPRET/EiJ) mouse strains. Tumor formation was evaluated after transplantation of non-irradiated Trp53-/- BALB/c mammary gland fragments into cleared fat pads of F1Bx hosts. Genome-wide linkage analysis revealed 2 genetic loci that constitute the baseline susceptibility via host microenvironment. However, once challenged with LDIR, we discovered 13 additional loci that were enriched for genes involved in cytokines, including TGFβ1 signaling. Surprisingly, LDIR-treated F1Bx cohort significantly reduced incidence of mammary tumors from Trp53-/- fragments as well as prolonged tumor latency, compared to sham-treated controls. We demonstrated further that plasma levels of specific cytokines were significantly correlated with tumor latency. Using an ex vivo 3-D assay, we confirmed TGFβ1 as a strong candidate for reduced mammary invasion in SPRET/EiJ, which could explain resistance of this strain to mammary cancer risk following LDIR. Our results open possible new avenues to understand mechanisms of genes operating via the stroma that affect cancer risk from external environmental exposures.

%B Sci Rep %V 5 %P 8919 %8 2015 %G eng %1 http://www.ncbi.nlm.nih.gov/pubmed/25747469?dopt=Abstract %R 10.1038/srep08919 %0 Journal Article %J Oncogene %D 2015 %T A new role of SNAI2 in postlactational involution of the mammary gland links it to luminal breast cancer development. %A Castillo-Lluva, S %A Hontecillas-Prieto, L %A Blanco-Gómez, A %A Del Mar Sáez-Freire, M %A García-Cenador, B %A García-Criado, J %A Pérez-Andrés, M %A Orfao, A %A Cañamero, M %A Jiang-Hua Mao %A Gridley, T %A Castellanos-Martín, A %A Perez-Losada, J %K Animals %K Apoptosis %K Breast Neoplasms %K Carcinogenesis %K Carrier Proteins %K Cell Line, Tumor %K Cell Proliferation %K Female %K Gene Expression Regulation, Neoplastic %K Humans %K Lactation %K Lung Neoplasms %K Mammary Glands, Animal %K Mice %K Mice, Knockout %K Pregnancy %K Proto-Oncogene Proteins c-akt %K STAT3 Transcription Factor %K Transcription Factors %X

Breast cancer is a major cause of mortality in women. The transcription factor SNAI2 has been implicated in the pathogenesis of several types of cancer, including breast cancer of basal origin. Here we show that SNAI2 is also important in the development of breast cancer of luminal origin in MMTV-ErbB2 mice. SNAI2 deficiency leads to longer latency and fewer luminal tumors, both of these being characteristics of pretumoral origin. These effects were associated with reduced proliferation and a decreased ability to generate mammospheres in normal mammary glands. However, the capacity to metastasize was not modified. Under conditions of increased ERBB2 oncogenic activity after pregnancy plus SNAI2 deficiency, both pretumoral defects-latency and tumor load-were compensated. However, the incidence of lung metastases was dramatically reduced. Furthermore, SNAI2 was required for proper postlactational involution of the breast. At 3 days post lactational involution, the mammary glands of Snai2-deficient mice exhibited lower levels of pSTAT3 and higher levels of pAKT1, resulting in decreased apoptosis. Abundant noninvoluted ducts were still present at 30 days post lactation, with a greater number of residual ERBB2+ cells. These results suggest that this defect in involution leads to an increase in the number of susceptible target cells for transformation, to the recovery of the capacity to generate mammospheres and to an increase in the number of tumors. Our work demonstrates the participation of SNAI2 in the pathogenesis of luminal breast cancer, and reveals an unexpected connection between the processes of postlactational involution and breast tumorigenesis in Snai2-null mutant mice.

%B Oncogene %V 34 %P 4777-90 %8 2015 Sep 3 %G eng %N 36 %1 http://www.ncbi.nlm.nih.gov/pubmed/26096931?dopt=Abstract %R 10.1038/onc.2015.224 %0 Journal Article %J Genome Biol %D 2015 %T Unraveling heterogeneous susceptibility and the evolution of breast cancer using a systems biology approach. %A Castellanos-Martín, Andrés %A Castillo-Lluva, Sonia %A Sáez-Freire, María Del Mar %A Blanco-Gómez, Adrián %A Hontecillas-Prieto, Lourdes %A Patino-Alonso, Carmen %A Galindo-Villardon, Purificación %A Pérez Del Villar, Luis %A Martín-Seisdedos, Carmen %A Isidoro-Garcia, María %A Abad-Hernández, María Del Mar %A Cruz-Hernández, Juan Jesús %A Rodríguez-Sánchez, César Augusto %A González-Sarmiento, Rogelio %A Alonso-López, Diego %A De Las Rivas, Javier %A García-Cenador, Begoña %A García-Criado, Javier %A Lee, Do Yup %A Bowen, Benjamin %A Reindl, Wolfgang %A Northen, Trent %A Jiang-Hua Mao %A Perez-Losada, Jesus %K Animals %K Breast Neoplasms %K Disease Progression %K Female %K Gene Expression Regulation, Neoplastic %K Humans %K MAP Kinase Signaling System %K Mice %K Models, Genetic %K Neoplasm Metastasis %K Proto-Oncogene Proteins c-akt %K Receptor, ErbB-2 %K Systems Biology %X

BACKGROUND: An essential question in cancer is why individuals with the same disease have different clinical outcomes. Progress toward a more personalized medicine in cancer patients requires taking into account the underlying heterogeneity at different molecular levels.

RESULTS: Here, we present a model in which there are complex interactions at different cellular and systemic levels that account for the heterogeneity of susceptibility to and evolution of ERBB2-positive breast cancers. Our model is based on our analyses of a cohort of mice that are characterized by heterogeneous susceptibility to ERBB2-positive breast cancers. Our analysis reveals that there are similarities between ERBB2 tumors in humans and those of backcross mice at clinical, genomic, expression, and signaling levels. We also show that mice that have tumors with intrinsically high levels of active AKT and ERK are more resistant to tumor metastasis. Our findings suggest for the first time that a site-specific phosphorylation at the serine 473 residue of AKT1 modifies the capacity for tumors to disseminate. Finally, we present two predictive models that can explain the heterogeneous behavior of the disease in the mouse population when we consider simultaneously certain genetic markers, liver cell signaling and serum biomarkers that are identified before the onset of the disease.

CONCLUSIONS: Considering simultaneously tumor pathophenotypes and several molecular levels, we show the heterogeneous behavior of ERBB2-positive breast cancer in terms of disease progression. This and similar studies should help to better understand disease variability in patient populations.

%B Genome Biol %V 16 %P 40 %8 2015 %G eng %1 http://www.ncbi.nlm.nih.gov/pubmed/25853295?dopt=Abstract %R 10.1186/s13059-015-0599-z %0 Journal Article %J Cancer Res %D 2014 %T CUL4A induces epithelial-mesenchymal transition and promotes cancer metastasis by regulating ZEB1 expression. %A Wang, Yunshan %A Wen, Mingxin %A Kwon, Yongwon %A Xu, Yangyang %A Liu, Yueyong %A Zhang, Pengju %A He, Xiuquan %A Wang, Qin %A Huang, Yurong %A Jen, Kuang-Yu %A LaBarge, Mark A %A You, Liang %A Kogan, Scott C %A Gray, Joe W %A Jiang-Hua Mao %A Wei, Guangwei %K Animals %K Breast %K Breast Neoplasms %K Cell Line, Tumor %K Cell Proliferation %K Cullin Proteins %K Epithelial-Mesenchymal Transition %K Female %K Gene Expression Regulation, Neoplastic %K Histones %K Homeodomain Proteins %K Humans %K Kruppel-Like Transcription Factors %K Mammary Neoplasms, Experimental %K Mice %K Mice, Nude %K Neoplasm Metastasis %K Neoplasm Transplantation %K Promoter Regions, Genetic %K Signal Transduction %K Transcription Factors %X

The ubiquitin ligase CUL4A has been implicated in tumorigenesis, but its contributions to progression and metastasis have not been evaluated. Here, we show that CUL4A is elevated in breast cancer as well as in ovarian, gastric, and colorectal tumors in which its expression level correlates positively with distant metastasis. CUL4A overexpression in normal or malignant human mammary epithelial cells increased their neoplastic properties in vitro and in vivo, markedly increasing epithelial-mesenchymal transition (EMT) and the metastatic capacity of malignant cells. In contrast, silencing CUL4A in aggressive breast cancer cells inhibited these processes. Mechanistically, we found that CUL4A modulated histone H3K4me3 at the promoter of the EMT regulatory gene ZEB1 in a manner associated with its transcription. ZEB1 silencing blocked CUL4A-driven proliferation, EMT, tumorigenesis, and metastasis. Furthermore, in human breast cancers, ZEB1 expression correlated positively with CUL4A expression and distant metastasis. Taken together, our findings reveal a pivotal role of CUL4A in regulating the metastatic behavior of breast cancer cells.

%B Cancer Res %V 74 %P 520-31 %8 2014 Jan 15 %G eng %N 2 %1 http://www.ncbi.nlm.nih.gov/pubmed/24305877?dopt=Abstract %R 10.1158/0008-5472.CAN-13-2182 %0 Journal Article %J Mol Cancer %D 2014 %T CUL4A overexpression enhances lung tumor growth and sensitizes lung cancer cells to erlotinib via transcriptional regulation of EGFR. %A Wang, Yunshan %A Zhang, Pengju %A Liu, Ziming %A Wang, Qin %A Wen, Mingxin %A Wang, Yuli %A Yuan, Hongtu %A Jiang-Hua Mao %A Wei, Guangwei %K Apoptosis %K Carcinoma, Non-Small-Cell Lung %K Cell Line, Tumor %K Cell Proliferation %K Cullin Proteins %K Erlotinib Hydrochloride %K Female %K Humans %K Lung Neoplasms %K Male %K Middle Aged %K Protein Kinase Inhibitors %K Quinazolines %K Receptor, Epidermal Growth Factor %K Transcription, Genetic %X

BACKGROUND: CUL4A has been proposed as oncogene in several types of human cancer, but its clinical significance and functional role in human non-small cell lung cancer (NSCLC) remain unclear.

METHODS: Expression level of CUL4A was examined by RT-PCR and Western blot. Forced expression of CUL4A was mediated by retroviruses, and CUL4A silencing by shRNAs expressing lentiviruses. Growth capacity of lung cancer cells was measured by MTT in vitro and tumorigenesis in vivo, respectively.

RESULTS: We found that CUL4A was highly expressed in human lung cancer tissues and lung cancer cell lines, and this elevated expression positively correlated with disease progression and prognosis. Overexpression of CUL4A in human lung cancer cell lines increased cell proliferation, inhibited apoptosis, and subsequently conferred resistance to chemotherapy. On other hand, silencing CUL4A expression in NSCLC cells reduced proliferation, promoted apoptosis and resulted in tumor growth inhibition in cancer xenograft model. Mechanistically, we revealed CUL4A regulated EGFR transcriptional expression and activation, and subsequently activated AKT. Targeted inhibition of EGFR activity blocked these CUL4A induced oncogenic activities.

CONCLUSIONS: Our results highlight the significance of CUL4A in NSCLC and suggest that CUL4A could be a promising therapy target and a potential biomarker for prognosis and EGFR target therapy in NSCLC patients.

%B Mol Cancer %V 13 %P 252 %8 2014 %G eng %1 http://www.ncbi.nlm.nih.gov/pubmed/25413624?dopt=Abstract %R 10.1186/1476-4598-13-252 %0 Journal Article %J Cancer Res %D 2014 %T Densely ionizing radiation acts via the microenvironment to promote aggressive Trp53-null mammary carcinomas. %A Illa-Bochaca, Irineu %A Ouyang, Haoxu %A Tang, Jonathan %A Sebastiano, Christopher %A Jiang-Hua Mao %A Costes, Sylvain V %A Demaria, Sandra %A Barcellos-Hoff, Mary Helen %K Animals %K Biomarkers, Tumor %K Cadherins %K Female %K Gene Expression Profiling %K Keratins %K Mammary Neoplasms, Experimental %K Mice %K Mice, Inbred BALB C %K Proto-Oncogene Proteins p21(ras) %K Radiation, Ionizing %K Receptor, ErbB-2 %K Receptors, Estrogen %K Receptors, Notch %K Stem Cells %K Tumor Microenvironment %K Tumor Suppressor Protein p53 %X

Densely ionizing radiation, which is present in the space radiation environment and used in radiation oncology, has potentially greater carcinogenic effect compared with sparsely ionizing radiation that is prevalent on earth. Here, we used a radiation chimera in which mice were exposed to densely ionizing 350 MeV/amu Si-particles, γ-radiation, or sham-irradiated and transplanted 3 days later with syngeneic Trp53-null mammary fragments. Trp53-null tumors arising in mice irradiated with Si-particles had a shorter median time to appearance and grew faster once detected compared with those in sham-irradiated or γ-irradiated mice. Tumors were further classified by markers keratin 8/18 (K18, KRT18), keratin 14 (K14, KRT14) and estrogen receptor (ER, ESR1), and expression profiling. Most tumors arising in sham-irradiated hosts were comprised of both K18- and K14-positive cells (K14/18) while those tumors arising in irradiated hosts were mostly K18. Keratin staining was significantly associated with ER status: K14/18 tumors were predominantly ER-positive, whereas K18 tumors were predominantly ER-negative. Genes differentially expressed in K18 tumors compared with K14/18 tumor were associated with ERBB2 and KRAS, metastasis, and loss of E-cadherin. Consistent with this, K18 tumors tended to grow faster and be more metastatic than K14/18 tumors, however, K18 tumors in particle-irradiated mice grew significantly larger and were more metastatic compared with sham-irradiated mice. An expression profile that distinguished K18 tumors arising in particle-irradiated mice compared with sham-irradiated mice was enriched in mammary stem cell, stroma, and Notch signaling genes. These data suggest that carcinogenic effects of densely ionizing radiation are mediated by the microenvironment, which elicits more aggressive tumors compared with similar tumors arising in sham-irradiated hosts.

%B Cancer Res %V 74 %P 7137-48 %8 2014 Dec 1 %G eng %N 23 %1 http://www.ncbi.nlm.nih.gov/pubmed/25304265?dopt=Abstract %R 10.1158/0008-5472.CAN-14-1212 %0 Journal Article %J Cancer Res %D 2014 %T Distinct luminal-type mammary carcinomas arise from orthotopic Trp53-null mammary transplantation of juvenile versus adult mice. %A Nguyen, David H %A Ouyang, Haoxu %A Jiang-Hua Mao %A Hlatky, Lynn %A Barcellos-Hoff, Mary Helen %K Animals %K Breast Neoplasms %K Cell Transformation, Neoplastic %K Cluster Analysis %K Epithelium %K Female %K Gene Expression Regulation, Neoplastic %K Humans %K Mammary Glands, Human %K Mammary Neoplasms, Experimental %K Mice %K Mice, Inbred BALB C %K Phosphorylation %K Receptors, Estrogen %K Receptors, Somatomedin %K Tumor Suppressor Protein p53 %X

Age and physiologic status, such as menopause, are risk factors for breast cancer. Less clear is what factors influence the diversity of breast cancer. In this study, we investigated the effect of host age on the distribution of tumor subtypes in mouse mammary chimera consisting of wild-type hosts and Trp53 nullizygous epithelium, which undergoes a high rate of neoplastic transformation. Wild-type mammary glands cleared of endogenous epithelium at 3 weeks of age were subsequently transplanted during puberty (5 weeks) or at maturation (10 weeks) with syngeneic Trp53-null mammary tissue fragments and monitored for one year. Tumors arose sooner from adult hosts (AH) compared with juvenile hosts (JH). However, compared with AH tumors, JH tumors grew several times faster, were more perfused, exhibited a two-fold higher mitotic index, and were more highly positive for insulin-like growth factor receptor phosphorylation. Most tumors in each setting were estrogen receptor (ER)-positive (80% JH vs. 70% AH), but JH tumors were significantly more ER-immunoreactive (P = 0.0001) than AH tumors. A differential expression signature (JvA) of juvenile versus adult tumors revealed a luminal transcriptional program. Centroids of the human homologs of JvA genes showed that JH tumors were more like luminal A tumors and AH tumors were more like luminal B tumors. Hierarchical clustering with the JvA human ortholog gene list segregated luminal A and luminal B breast cancers across datasets. These data support the notion that age-associated host physiology greatly influences the intrinsic subtype of breast cancer.

%B Cancer Res %V 74 %P 7149-58 %8 2014 Dec 1 %G eng %N 23 %1 http://www.ncbi.nlm.nih.gov/pubmed/25281718?dopt=Abstract %R 10.1158/0008-5472.CAN-14-1440 %0 Journal Article %J Oncotarget %D 2014 %T An interferon signature identified by RNA-sequencing of mammary tissues varies across the estrous cycle and is predictive of metastasis-free survival. %A A Snijders %A Langley, Sasha %A Jiang-Hua Mao %A Bhatnagar, Sandhya %A Bjornstad, Kathleen A %A Rosen, Chris J %A Lo, Alvin %A Huang, Yurong %A Blakely, Eleanor A %A Karpen, Gary H %A Bissell, Mina J %A Wyrobek, Andrew J %K Animals %K Disease-Free Survival %K Estrous Cycle %K Female %K Humans %K Interferons %K Mice %K Mice, Inbred BALB C %K Mice, Inbred C57BL %K Neoplasm Metastasis %K RNA, Messenger %X

The concept that a breast cancer patient's menstrual stage at the time of tumor surgery influences risk of metastases remains controversial. The scarcity of comprehensive molecular studies of menstrual stage-dependent fluctuations in the breast provides little insight in this observation. To gain a deeper understanding of the biological changes in mammary tissue and blood during the menstrual cycle and to determine the influence of environmental exposures, such as low-dose ionizing radiation (LDIR), we used the mouse to characterize estrous-cycle variations in mammary gene transcripts by RNA-sequencing, peripheral white blood cell (WBC) counts and plasma cytokine levels. We identified an estrous-variable and hormone-dependent gene cluster enriched for Type-1 interferon genes. Cox regression identified a 117-gene signature of interferon-associated genes, which correlated with lower frequencies of metastasis in breast cancer patients. LDIR (10cGy) exposure had no detectable effect on mammary transcripts. However, peripheral WBC counts varied across the estrous cycle and LDIR exposure reduced lymphocyte counts and cytokine levels in tumor-susceptible mice. Our finding of variations in mammary Type-1 interferon and immune functions across the estrous cycle provides a mechanism by which timing of breast tumor surgery during the menstrual cycle may have clinical relevance to a patient's risk for distant metastases.

%B Oncotarget %V 5 %P 4011-25 %8 2014 Jun 30 %G eng %N 12 %1 http://www.ncbi.nlm.nih.gov/pubmed/24994117?dopt=Abstract %R 10.18632/oncotarget.2148 %0 Journal Article %J Stem Cells %D 2014 %T Irradiation of juvenile, but not adult, mammary gland increases stem cell self-renewal and estrogen receptor negative tumors. %A Tang, Jonathan %A Fernandez-Garcia, Ignacio %A Vijayakumar, Sangeetha %A Martinez-Ruis, Haydeliz %A Illa-Bochaca, Irineu %A Nguyen, David H %A Jiang-Hua Mao %A Costes, Sylvain V %A Barcellos-Hoff, Mary Helen %K Aging %K Animals %K Biomarkers %K Cell Line %K Cell Lineage %K Cell Proliferation %K Computer Simulation %K Dose-Response Relationship, Radiation %K Epithelial Cells %K Female %K Humans %K Mammary Glands, Animal %K Mammary Neoplasms, Animal %K Mice %K Morphogenesis %K Radiation, Ionizing %K Receptors, Estrogen %K Receptors, Notch %K Stem Cells %K Transforming Growth Factor beta %X

Children exposed to ionizing radiation have a substantially greater breast cancer risk than adults; the mechanism for this strong age dependence is not known. Here we show that pubertal murine mammary glands exposed to sparsely or densely ionizing radiation exhibit enrichment of mammary stem cell and Notch pathways, increased mammary repopulating activity indicative of more stem cells, and propensity to develop estrogen receptor (ER) negative tumors thought to arise from stem cells. We developed a mammary lineage agent-based model (ABM) to evaluate cell inactivation, self-renewal, or dedifferentiation via epithelial-mesenchymal transition (EMT) as mechanisms by which radiation could increase stem cells. ABM rejected cell inactivation and predicted increased self-renewal would only affect juveniles while dedifferentiation could act in both juveniles and adults. To further test self-renewal versus dedifferentiation, we used the MCF10A human mammary epithelial cell line, which recapitulates ductal morphogenesis in humanized fat pads, undergoes EMT in response to radiation and transforming growth factor β (TGFβ) and contains rare stem-like cells that are Let-7c negative or express both basal and luminal cytokeratins. ABM simulation of population dynamics of double cytokeratin cells supported increased self-renewal in irradiated MCF10A treated with TGFβ. Radiation-induced Notch concomitant with TGFβ was necessary for increased self-renewal of Let-7c negative MCF10A cells but not for EMT, indicating that these are independent processes. Consistent with these data, irradiating adult mice did not increase mammary repopulating activity or ER-negative tumors. These studies suggest that irradiation during puberty transiently increases stem cell self-renewal, which increases susceptibility to developing ER-negative breast cancer.

%B Stem Cells %V 32 %P 649-61 %8 2014 Mar %G eng %N 3 %1 http://www.ncbi.nlm.nih.gov/pubmed/24038768?dopt=Abstract %R 10.1002/stem.1533 %0 Journal Article %J Cancer Res %D 2014 %T Stress signaling from human mammary epithelial cells contributes to phenotypes of mammographic density %A DeFilippis, Rosa Anna %A Fordyce, Colleen %A Patten, Kelley %A Hang Chang %A Zhao, Jianxin %A Fontenay, Gerald V %A Kerlikowske, Karla %A Parvin, Bahram %A Tlsty, Thea D %K Antigens, CD36 %K Breast Neoplasms %K DNA Damage %K Epithelial Cells %K Female %K Humans %K Mammary Glands, Human %K Phenotype %K Signal Transduction %X

Telomere malfunction and other types of DNA damage induce an activin A-dependent stress response in mortal nontumorigenic human mammary epithelial cells that subsequently induces desmoplastic-like phenotypes in neighboring fibroblasts. Some characteristics of this fibroblast/stromal response, such as reduced adipocytes and increased extracellular matrix content, are observed not only in tumor tissues but also in disease-free breast tissues at high risk for developing cancer, especially high mammographic density tissues. We found that these phenotypes are induced by repression of the fatty acid translocase CD36, which is seen in desmoplastic and disease-free high mammographic density tissues. In this study, we show that epithelial cells from high mammographic density tissues have more DNA damage signaling, shorter telomeres, increased activin A secretion and an altered DNA damage response compared with epithelial cells from low mammographic density tissues. Strikingly, both telomere malfunction and activin A expression in epithelial cells can repress CD36 expression in adjacent fibroblasts. These results provide new insights into how high mammographic density arises and why it is associated with breast cancer risk, with implications for the definition of novel invention targets (e.g., activin A and CD36) to prevent breast cancer.

%B Cancer Res %V 74 %P 5032-44 %8 2014 Sep 15 %G eng %N 18 %R 10.1158/0008-5472.CAN-13-3390 %0 Journal Article %J Clin Cancer Res %D 2013 %T A 12-gene set predicts survival benefits from adjuvant chemotherapy in non-small cell lung cancer patients. %A Tang, Hao %A Xiao, Guanghua %A Behrens, Carmen %A Schiller, Joan %A Allen, Jeffrey %A Chow, Chi-Wan %A Suraokar, Milind %A Corvalan, Alejandro %A Jiang-Hua Mao %A White, Michael A %A Wistuba, Ignacio I %A Minna, John D %A Xie, Yang %K Adult %K Aged %K Carcinoma, Non-Small-Cell Lung %K Chemotherapy, Adjuvant %K Clinical Trials as Topic %K Female %K Gene Expression Regulation, Neoplastic %K Genome, Human %K Humans %K Kaplan-Meier Estimate %K Lung Neoplasms %K Male %K Middle Aged %K Neoplasm Proteins %K Neoplasm Staging %K Prognosis %K RNA Interference %K Systems Biology %K Treatment Outcome %X

PURPOSE: Prospectively identifying who will benefit from adjuvant chemotherapy (ACT) would improve clinical decisions for non-small cell lung cancer (NSCLC) patients. In this study, we aim to develop and validate a functional gene set that predicts the clinical benefits of ACT in NSCLC.

EXPERIMENTAL DESIGN: An 18-hub-gene prognosis signature was developed through a systems biology approach, and its prognostic value was evaluated in six independent cohorts. The 18-hub-gene set was then integrated with genome-wide functional (RNAi) data and genetic aberration data to derive a 12-gene predictive signature for ACT benefits in NSCLC.

RESULTS: Using a cohort of 442 stage I to III NSCLC patients who underwent surgical resection, we identified an 18-hub-gene set that robustly predicted the prognosis of patients with adenocarcinoma in all validation datasets across four microarray platforms. The hub genes, identified through a purely data-driven approach, have significant biological implications in tumor pathogenesis, including NKX2-1, Aurora Kinase A, PRC1, CDKN3, MBIP, and RRM2. The 12-gene predictive signature was successfully validated in two independent datasets (n = 90 and 176). The predicted benefit group showed significant improvement in survival after ACT (UT Lung SPORE data: HR = 0.34, P = 0.017; JBR.10 clinical trial data: HR = 0.36, P = 0.038), whereas the predicted nonbenefit group showed no survival benefit for 2 datasets (HR = 0.80, P = 0.70; HR = 0.91, P = 0.82).

CONCLUSIONS: This is the first study to integrate genetic aberration, genome-wide RNAi data, and mRNA expression data to identify a functional gene set that predicts which resectable patients with non-small cell lung cancer will have a survival benefit with ACT.

%B Clin Cancer Res %V 19 %P 1577-86 %8 2013 Mar 15 %G eng %N 6 %1 http://www.ncbi.nlm.nih.gov/pubmed/23357979?dopt=Abstract %R 10.1158/1078-0432.CCR-12-2321 %0 Journal Article %J Neoplasia %D 2013 %T Breast fibroblasts modulate early dissemination, tumorigenesis, and metastasis through alteration of extracellular matrix characteristics %A Dumont, Nancy %A Liu, Bob %A DeFilippis, Rosa Anna %A Hang Chang %A Rabban, Joseph T %A Karnezis, Anthony N %A Tjoe, Judy A %A Marx, James %A Parvin, Bahram %A Tlsty, Thea D %K Animals %K Breast %K Cell Line, Tumor %K Cell Transformation, Neoplastic %K Coculture Techniques %K Epithelial Cells %K Extracellular Matrix %K Extracellular Signal-Regulated MAP Kinases %K Female %K Fibroblasts %K Humans %K Lung Neoplasms %K Mammary Neoplasms, Experimental %K Neoplasm Metastasis %K Phenotype %K Proto-Oncogene Proteins c-jun %K rho GTP-Binding Proteins %K Signal Transduction %K Transforming Growth Factor beta %X

A wealth of evidence has now demonstrated that the microenvironment in which a tumorigenic cell evolves is as critical to its evolution as the genetic mutations it accrues. However, there is still relatively little known about how signals from the microenvironment contribute to the early events in the progression to malignancy. To address this question, we used a premalignant mammary model to examine how fibroblasts, and the extracellular matrix (ECM) proteins they secrete, influence progression to malignancy. Their effect on metastatic malignant cells was also assessed for comparison. We found that carcinoma-associated fibroblasts, and the distinct aligned ECM they deposit, can cause both premalignant and malignant mammary epithelial cells to assume a mesenchymal morphology that is associated with increased dissemination and metastasis, while benign reduction mammoplasty fibroblasts favor the maintenance of an epithelial morphology and constrain early dissemination, tumor growth, and metastasis. Our results suggest that normalizing the organization of the ECM could be effective in limiting systemic dissemination and tumor growth.

%B Neoplasia %V 15 %P 249-62 %8 2013 Mar %G eng %N 3 %0 Journal Article %J Epigenetics %D 2013 %T C2ORF40 suppresses breast cancer cell proliferation and invasion through modulating expression of M phase cell cycle genes. %A Lu, Jing %A Wen, Mingxin %A Huang, Yurong %A He, Xiuquan %A Wang, Yunshan %A Wu, Qi %A Li, Zengchun %A Castellanos-Martín, Andrés %A Abad, Mar %A Cruz-Hernandez, Juan J %A Rodriguez, Cesar A %A Perez-Losada, Jesus %A Jiang-Hua Mao %A Wei, Guangwei %K Breast Neoplasms %K Cell Line, Tumor %K Cell Proliferation %K DNA Methylation %K Female %K Gene Expression Regulation, Neoplastic %K Genes, cdc %K Genes, Tumor Suppressor %K Humans %K Mitosis %K Neoplasm Invasiveness %K Neoplasm Proteins %K Prognosis %X

Recently, it has been suggested that C2ORF40 is a candidate tumor suppressor gene in breast cancer. However, the mechanism for reduced expression of C2ORF40 and its functional role in breast cancers remain unclear. Here we show that C2ORF40 is frequently silenced in human primary breast cancers and cell lines through promoter hypermethylation. C2ORF40 mRNA level is significantly associated with patient disease-free survival and distant cancer metastasis. Overexpression of C2ORF4 0 inhibits breast cancer cell proliferation, migration and invasion. By contrast, silencing C2ORF40 expression promotes these biological phenotypes. Bioinformatics and FACS analysis reveal C2ORF40 functions at G2/M phase by downregulation of mitotic genes expression, including UBE2C. Our results suggest that C2ORF40 acts as a tumor suppressor gene in breast cancer pathogenesis and progression and is a candidate prognostic marker for this disease.

%B Epigenetics %V 8 %P 571-83 %8 2013 Jun %G eng %N 6 %1 http://www.ncbi.nlm.nih.gov/pubmed/23770814?dopt=Abstract %R 10.4161/epi.24626 %0 Journal Article %J Int J Oncol %D 2013 %T Hematein, a casein kinase II inhibitor, inhibits lung cancer tumor growth in a murine xenograft model. %A Hung, Ming-Szu %A Xu, Zhidong %A Chen, Yu %A Smith, Emmanuel %A Jiang-Hua Mao %A Hsieh, David %A Lin, Yu-Ching %A Yang, Cheng-Ta %A Jablons, David M %A You, Liang %K Animals %K Apoptosis %K Blotting, Western %K Casein Kinase II %K Enzyme Inhibitors %K Female %K Hematoxylin %K Humans %K Immunoenzyme Techniques %K Lung Neoplasms %K Mice %K Mice, Inbred BALB C %K Phosphorylation %K Proto-Oncogene Proteins c-akt %K T Cell Transcription Factor 1 %K Tumor Cells, Cultured %K Wnt Proteins %K Xenograft Model Antitumor Assays %X

Casein kinase II (CK2) inhibitors suppress cancer cell growth. In this study, we examined the inhibitory effects of a novel CK2 inhibitor, hematein, on tumor growth in a murine xenograft model. We found that in lung cancer cells, hematein inhibited cancer cell growth, Akt/PKB Ser129 phosphorylation, the Wnt/TCF pathway and increased apoptosis. In a murine xenograft model of lung cancer, hematein inhibited tumor growth without significant toxicity to the mice tested. Molecular docking showed that hematein binds to CK2α in durable binding sites. Collectively, our results suggest that hematein is an allosteric inhibitor of protein kinase CK2 and has antitumor activity to lung cancer.

%B Int J Oncol %V 43 %P 1517-22 %8 2013 Nov %G eng %N 5 %1 http://www.ncbi.nlm.nih.gov/pubmed/24008396?dopt=Abstract %R 10.3892/ijo.2013.2087 %0 Journal Article %J Clin Cancer Res %D 2013 %T Murine microenvironment metaprofiles associate with human cancer etiology and intrinsic subtypes. %A Nguyen, David H %A Fredlund, Erik %A Zhao, Wei %A Perou, Charles M %A Balmain, Allan %A Jiang-Hua Mao %A Barcellos-Hoff, Mary Helen %K Animals %K Breast Neoplasms %K Female %K Gene Expression Regulation, Neoplastic %K Humans %K Mammary Neoplasms, Animal %K Mice %K Radiation, Ionizing %K Transcriptome %K Tumor Microenvironment %K Tumor Suppressor Protein p53 %X

PURPOSE: Ionizing radiation is a well-established carcinogen in rodent models and a risk factor associated with human cancer. We developed a mouse model that captures radiation effects on host biology by transplanting unirradiated Trp53-null mammary tissue to sham or irradiated hosts. Gene expression profiles of tumors that arose in irradiated mice are distinct from those that arose in naïve hosts. We asked whether expression metaprofiles could discern radiation-preceded human cancer or be informative in sporadic breast cancers.

EXPERIMENTAL DESIGN: Affymetrix microarray gene expression data from 56 Trp53-null mammary tumors were used to define gene profiles and a centroid that discriminates tumors arising in irradiated hosts. These were applied to publicly available human cancer datasets.

RESULTS: Host irradiation induces a metaprofile consisting of gene modules representing stem cells, cell motility, macrophages, and autophagy. Human orthologs of the host irradiation metaprofile discriminated between radiation-preceded and sporadic human thyroid cancers. An irradiated host centroid was strongly associated with estrogen receptor-negative breast cancer. When applied to sporadic human breast cancers, the irradiated host metaprofile strongly associated with basal-like and claudin-low breast cancer intrinsic subtypes. Comparing host irradiation in the context of TGF-β levels showed that inflammation was robustly associated with claudin-low tumors.

CONCLUSIONS: Detection of radiation-preceded human cancer by the irradiated host metaprofile raises possibilities of assessing human cancer etiology. Moreover, the association of the irradiated host metaprofiles with estrogen receptor-negative status and claudin-low subtype suggests that host processes similar to those induced by radiation underlie sporadic cancers.

%B Clin Cancer Res %V 19 %P 1353-62 %8 2013 Mar 15 %G eng %N 6 %1 http://www.ncbi.nlm.nih.gov/pubmed/23339125?dopt=Abstract %R 10.1158/1078-0432.CCR-12-3554 %0 Journal Article %J Sci Rep %D 2013 %T Nanosensor dosimetry of mouse blood proteins after exposure to ionizing radiation. %A Kim, Dokyoon %A Marchetti, Francesco %A Chen, Zuxiong %A Zaric, Sasa %A Wilson, Robert J %A Hall, Drew A %A Gaster, Richard S %A Lee, Jung-Rok %A Wang, Junyi %A Osterfeld, Sebastian J %A Yu, Heng %A White, Robert M %A Blakely, William F %A Peterson, Leif E %A Bhatnagar, Sandhya %A Mannion, Brandon %A Tseng, Serena %A Roth, Kristen %A Coleman, Matthew %A A Snijders %A Wyrobek, Andrew J %A Wang, Shan X %K Animals %K Biomarkers %K Biosensing Techniques %K Blood Proteins %K Dose-Response Relationship, Radiation %K Female %K Male %K Membrane Proteins %K Mice %K Nanotechnology %K Radiation, Ionizing %K Radiometry %K Reproducibility of Results %K Serum Amyloid A Protein %K Time Factors %X

Giant magnetoresistive (GMR) nanosensors provide a novel approach for measuring protein concentrations in blood for medical diagnosis. Using an in vivo mouse radiation model, we developed protocols for measuring Flt3 ligand (Flt3lg) and serum amyloid A1 (Saa1) in small amounts of blood collected during the first week after X-ray exposures of sham, 0.1, 1, 2, 3, or 6 Gy. Flt3lg concentrations showed excellent dose discrimination at ≥ 1 Gy in the time window of 1 to 7 days after exposure except 1 Gy at day 7. Saa1 dose response was limited to the first two days after exposure. A multiplex assay with both proteins showed improved dose classification accuracy. Our magneto-nanosensor assay demonstrates the dose and time responses, low-dose sensitivity, small volume requirements, and rapid speed that have important advantages in radiation triage biodosimetry.

%B Sci Rep %V 3 %P 2234 %8 2013 %G eng %R 10.1038/srep02234 %0 Journal Article %J Oncotarget %D 2013 %T NFkB disrupts tissue polarity in 3D by preventing integration of microenvironmental signals %A Becker-Weimann, Sabine %A Xiong, Gaofeng %A Furuta, Saori %A Ju Han %A Kuhn, Irene %A Akavia, Uri-David %A Pe'er, Dana %A Bissell, Mina J %A Xu, Ren %K Breast Neoplasms %K Cell Line, Tumor %K Female %K Gene Expression %K Humans %K Imaging, Three-Dimensional %K Microarray Analysis %K NF-kappa B %K Phenotype %K Signal Transduction %K Transcriptional Activation %K Tumor Microenvironment %X

The microenvironment of cells controls their phenotype, and thereby the architecture of the emerging multicellular structure or tissue. We have reported more than a dozen microenvironmental factors whose signaling must be integrated in order to effect an organized, functional tissue morphology. However, the factors that prevent integration of signaling pathways that merge form and function are still largely unknown. We have identified nuclear factor kappa B (NFkB) as a transcriptional regulator that disrupts important microenvironmental cues necessary for tissue organization. We compared the gene expression of organized and disorganized epithelial cells of the HMT-3522 breast cancer progression series: the non-malignant S1 cells that form polarized spheres ('acini'), the malignant T4-2 cells that form large tumor-like clusters, and the 'phenotypically reverted' T4-2 cells that polarize as a result of correction of the microenvironmental signaling. We identified 180 genes that display an increased expression in disorganized compared to polarized structures. Network, GSEA and transcription factor binding site analyses suggested that NFkB is a common activator for the 180 genes. NFkB was found to be activated in disorganized breast cancer cells, and inhibition of microenvironmental signaling via EGFR, beta1 integrin, MMPs, or their downstream signals suppressed its activation. The postulated role of NFkB was experimentally verified: Blocking the NFkB pathway with a specific chemical inhibitor or shRNA induced polarization and inhibited invasion of breast cancer cells in 3D cultures. These results may explain why NFkB holds promise as a target for therapeutic intervention: Its inhibition can reverse the oncogenic signaling involved in breast cancer progression and integrate the essential microenvironmental control of tissue architecture.

%B Oncotarget %V 4 %P 2010-20 %8 2013 Nov %G eng %N 11 %R 10.18632/oncotarget.1451 %0 Journal Article %J Cancer Discov %D 2012 %T CD36 repression activates a multicellular stromal program shared by high mammographic density and tumor tissues %A DeFilippis, Rosa Anna %A Hang Chang %A Dumont, Nancy %A Rabban, Joseph T %A Chen, Yunn-Yi %A Fontenay, Gerald V %A Berman, Hal K %A Gauthier, Mona L %A Zhao, Jianxin %A Hu, Donglei %A Marx, James J %A Tjoe, Judy A %A Ziv, Elad %A Febbraio, Maria %A Kerlikowske, Karla %A Parvin, Bahram %A Tlsty, Thea D %K Adipocytes %K Animals %K Antigens, CD36 %K Breast Neoplasms %K Cell Differentiation %K Female %K Humans %K Mammography %K Mice %K Mice, Knockout %K Risk Factors %K Signal Transduction %K Stromal Cells %X

UNLABELLED: Although high mammographic density is considered one of the strongest risk factors for invasive breast cancer, the genes involved in modulating this clinical feature are unknown. Tissues of high mammographic density share key histologic features with stromal components within malignant lesions of tumor tissues, specifically low adipocyte and high extracellular matrix (ECM) content. We show that CD36, a transmembrane receptor that coordinately modulates multiple protumorigenic phenotypes, including adipocyte differentiation, angiogenesis, cell-ECM interactions, and immune signaling, is greatly repressed in multiple cell types of disease-free stroma associated with high mammographic density and tumor stroma. Using both in vitro and in vivo assays, we show that CD36 repression is necessary and sufficient to recapitulate the above-mentioned phenotypes observed in high mammographic density and tumor tissues. Consistent with a functional role for this coordinated program in tumorigenesis, we observe that clinical outcomes are strongly associated with CD36 expression.

SIGNIFICANCE: CD36 simultaneously controls adipocyte content and matrix accumulation and is coordinately repressed in multiple cell types within tumor and high mammographic density stroma, suggesting that activation of this stromal program is an early event in tumorigenesis. Levels of CD36 and extent of mammographic density are both modifiable factors that provide potential for intervention.

%B Cancer Discov %V 2 %P 826-39 %8 2012 Sep %G eng %N 9 %R 10.1158/2159-8290.CD-12-0107 %0 Journal Article %J Int J Oncol %D 2012 %T Development of a rapid and practical mutation screening assay for human lung adenocarcinoma. %A Choi, Helen %A Kratz, Johannes %A Pham, Patrick %A Lee, Sharon %A Ray, Roshni %A Kwon, Yong-Won %A Jiang-Hua Mao %A Kang, Hio Chung %A Jablons, David %A Kim, Il-Jin %K Adenocarcinoma %K Aged %K Base Sequence %K beta Catenin %K Cell Line, Tumor %K DNA Mutational Analysis %K Female %K Frameshift Mutation %K Genetic Association Studies %K Humans %K Kaplan-Meier Estimate %K Lung Neoplasms %K Male %K Middle Aged %K Molecular Sequence Data %K Mutagenesis, Insertional %K Mutation, Missense %K Prognosis %K Proto-Oncogene Proteins B-raf %K Proto-Oncogene Proteins p21(ras) %K Receptor, Epidermal Growth Factor %K Tumor Suppressor Protein p53 %X

Mortality after initial diagnosis of lung cancer is higher than from any other cancer. Although mutations in several genes, such as EGFR and K-ras, have been associated with clinical outcome, technical complexity, cost and time have rendered routine screening prohibitive for most lung cancer patients prior to treatment. In this study, using both novel and established technologies, we developed a clinically practical assay to survey the status of three frequently mutated genes in lung cancer (EGFR, K-ras and TP53) and two genes (BRAF and β-catenin) with known hotspot mutations in many other cancers. A single 96-well plate was designed targeting a total of 14 fragments (16 exons) from EGFR, K-ras, TP53, BRAF and β-catenin. In 96 lung adenocarcinoma patients, the mutation frequencies of three major genes (EGFR, K-ras and TP53) were between 21-24%. Fifty-six out of 96 (58%) patients had a mutation in at least one of the five genes. K-ras mutations positively correlated with smoking pack-years (p=0.035). EGFR mutations were frequent in never-smokers (p=0.0007), Asians (p=0.0204) and non-stage I lung cancer (p=0.016). There was also a trend towards an association between the presence of any mutation and improved recurrence-free survival (p=0.070). We demonstrate that our novel multigene mutation assay technology can rapidly and cost-effectively screen for mutations in lung adenocarcinoma. This screening assay can be used in the clinical setting for the large-scale validation of prognosis and/or predicting therapeutic response so that the majority of lung cancer patients can benefit from leveraging up-to-date knowledge on how mutation profiles may influence treatment options.

%B Int J Oncol %V 40 %P 1900-6 %8 2012 Jun %G eng %N 6 %1 http://www.ncbi.nlm.nih.gov/pubmed/22407457?dopt=Abstract %R 10.3892/ijo.2012.1396 %0 Journal Article %J PLoS One %D 2012 %T DNA repair and cell cycle biomarkers of radiation exposure and inflammation stress in human blood. %A Budworth, Helen %A A Snijders %A Marchetti, Francesco %A Mannion, Brandon %A Bhatnagar, Sandhya %A Kwoh, Ely %A Tan, Yuande %A Wang, Shan X %A Blakely, William F %A Coleman, Matthew %A Peterson, Leif %A Wyrobek, Andrew J %K Adult %K Biomarkers %K Cell Cycle %K DNA Repair %K Dose-Response Relationship, Radiation %K Female %K Gene Expression Regulation %K Humans %K Inflammation %K Lipopolysaccharides %K Male %K Middle Aged %K Phosphorylation %K Predictive Value of Tests %K Reproducibility of Results %K RNA, Messenger %K Stress, Physiological %K Time Factors %K Transcription, Genetic %K X-Rays %K Young Adult %X

DNA damage and repair are hallmarks of cellular responses to ionizing radiation. We hypothesized that monitoring the expression of DNA repair-associated genes would enhance the detection of individuals exposed to radiation versus other forms of physiological stress. We employed the human blood ex vivo radiation model to investigate the expression responses of DNA repair genes in repeated blood samples from healthy, non-smoking men and women exposed to 2 Gy of X-rays in the context of inflammation stress mimicked by the bacterial endotoxin lipopolysaccharide (LPS). Radiation exposure significantly modulated the transcript expression of 12 genes of 40 tested (2.2E-06

 

%B PLoS One %V 7 %P e48619 %8 2012 %G eng %N 11 %R 10.1371/journal.pone.0048619 %0 Journal Article %J PLoS One %D 2012 %T Genetic differences in transcript responses to low-dose ionizing radiation identify tissue functions associated with breast cancer susceptibility %A A Snijders %A Marchetti, Francesco %A Bhatnagar, Sandhya %A Duru, Nadire %A Ju Han %A Hu, Zhi %A Jiang-Hua Mao %A Gray, Joe W %A Wyrobek, Andrew J %K Animals %K Breast Neoplasms %K Dose-Response Relationship, Radiation %K Female %K Genetic Predisposition to Disease %K Genomic Instability %K Humans %K Mice %K Mice, Inbred BALB C %K Mice, Inbred C57BL %K Radiation, Ionizing %K RNA, Messenger %K Survival Analysis %K Transcription, Genetic %K Tumor Microenvironment %X

High dose ionizing radiation (IR) is a well-known risk factor for breast cancer but the health effects after low-dose (LD, <10 cGy) exposures remain highly uncertain. We explored a systems approach that compared LD-induced chromosome damage and transcriptional responses in strains of mice with genetic differences in their sensitivity to radiation-induced mammary cancer (BALB/c and C57BL/6) for the purpose of identifying mechanisms of mammary cancer susceptibility. Unirradiated mammary and blood tissues of these strains differed significantly in baseline expressions of DNA repair, tumor suppressor, and stress response genes. LD exposures of 7.5 cGy (weekly for 4 weeks) did not induce detectable genomic instability in either strain. However, the mammary glands of the sensitive strain but not the resistant strain showed early transcriptional responses involving: (a) diminished immune response, (b) increased cellular stress, (c) altered TGFβ-signaling, and (d) inappropriate expression of developmental genes. One month after LD exposure, the two strains showed opposing responses in transcriptional signatures linked to proliferation, senescence, and microenvironment functions. We also discovered a pre-exposure expression signature in both blood and mammary tissues that is predictive for poor survival among human cancer patients (p = 0.0001), and a post-LD-exposure signature also predictive for poor patient survival (p<0.0001). There is concordant direction of expression in the LD-exposed sensitive mouse strain, in biomarkers of human DCIS and in biomarkers of human breast tumors. Our findings support the hypothesis that genetic mechanisms that determine susceptibility to LD radiation induced mammary cancer in mice are similar to the tissue mechanisms that determine poor-survival in breast cancer patients. We observed non-linearity of the LD responses providing molecular evidence against the LNT risk model and obtained new evidence that LD responses are strongly influenced by genotype. Our findings suggest that the biological assumptions concerning the mechanisms by which LD radiation is translated into breast cancer risk should be reexamined and suggest a new strategy to identify genetic features that predispose or protect individuals from LD-induced breast cancer.

%B PLoS One %V 7 %P e45394 %8 2012 %G eng %N 10 %R 10.1371/journal.pone.0045394 %0 Journal Article %J PLoS One %D 2012 %T Gremlin is overexpressed in lung adenocarcinoma and increases cell growth and proliferation in normal lung cells. %A Mulvihill, Michael S %A Kwon, Yong-Won %A Lee, Sharon %A Fang, Li Tai %A Choi, Helen %A Ray, Roshni %A Kang, Hio Chung %A Jiang-Hua Mao %A Jablons, David %A Kim, Il-Jin %K Adenocarcinoma %K Cell Line, Tumor %K Cell Proliferation %K Epithelial Cells %K Female %K Fibroblasts %K Gene Expression Regulation, Neoplastic %K Humans %K Immunohistochemistry %K Intercellular Signaling Peptides and Proteins %K Lung Neoplasms %K Male %K Neoplasm Proteins %K Real-Time Polymerase Chain Reaction %K Respiratory Mucosa %K RNA, Messenger %K RNA, Neoplasm %X

BACKGROUND: Gremlin, a member of the Dan family of BMP antagonists, is a glycosylated extracellular protein. Previously Gremlin has been shown to play a role in dorsal-ventral patterning, in tissue remodeling, and recently in angiogenesis. Evidence has previously been presented showing both over- and under-expression of Gremlin in different tumor tissues. Here, we sought to quantify expression of Gremlin in cancers of the lung and performed in vitro experiments to check whether Gremlin promotes cell growth and proliferation.

METHODOLOGY/PRINCIPAL FINDINGS: Expression of Gremlin in 161 matched tumor and normal lung cancer specimens is quantified by quantitative real-time PCR and protein level is measured by immunohistochemistry. GREM1 was transfected into lung fibroblast and epithelial cell lines to assess the impact of overexpression of Gremlin in vitro.

RESULTS: Lung adenocarcinoma but not squamous cell carcinoma shows a significant increase in Gremlin expression by mRNA and protein level. Lung fibroblast and epithelial cell lines transfected with GREM1 show significantly increased cell proliferation.

CONCLUSIONS/SIGNIFICANCE: Our data suggest that Gremlin acts in an oncogenic manner in lung adenocarcinoma and could hold promise as a new diagnostic marker or potential therapeutic target in lung AD or general thoracic malignancies.

%B PLoS One %V 7 %P e42264 %8 2012 %G eng %N 8 %1 http://www.ncbi.nlm.nih.gov/pubmed/22870311?dopt=Abstract %R 10.1371/journal.pone.0042264 %0 Journal Article %J Carcinogenesis %D 2012 %T Independent genetic control of early and late stages of chemically induced skin tumors in a cross of a Japanese wild-derived inbred mouse strain, MSM/Ms. %A Okumura, Kazuhiro %A Sato, Miho %A Saito, Megumi %A Miura, Ikuo %A Wakana, Shigeharu %A Jiang-Hua Mao %A Miyasaka, Yuki %A Kominami, Ryo %A Wakabayashi, Yuichi %K 9,10-Dimethyl-1,2-benzanthracene %K Animals %K Carcinogens %K Crosses, Genetic %K Female %K Genetic Linkage %K Japan %K Male %K Mice %K Mice, Inbred C57BL %K Mice, Inbred Strains %K Mice, Knockout %K Papilloma %K Skin Neoplasms %K Tetradecanoylphorbol Acetate %K Tumor Suppressor Protein p53 %X

MSM/Ms is an inbred mouse strain derived from a Japanese wild mouse, Mus musculus molossinus. In this study, we showed that MSM/Ms mice exhibit dominant resistance when crossed with susceptible FVB/N mice and subjected to the two-stage skin carcinogenesis protocol using 7,12-dimethylbenz(a)anthracene (DMBA)/ 12-O-tetradecanoylphorbol-13-acetate (TPA). A series of F1 backcross mice were generated by crossing p53(+/+) or p53(+/-) F1 (FVB/N × MSM/Ms) males with FVB/N female mice. These generated 228 backcross animals, approximately half of which were p53(+/-), enabling us to search for p53-dependent skin tumor modifier genes. Highly significant linkage for papilloma multiplicity was found on chromosomes 6 and 7 and suggestive linkage was found on chromosomes 3, 5 and 12. Furthermore, in order to identify stage-dependent linkage loci we classified tumors into three categories (<2mm, 2-6mm and >6mm), and did linkage analysis. The same locus on chromosome 7 showed strong linkage in groups with <2mm or 2-6mm papillomas. No linkage was detected on chromosome 7 to papillomas >6mm, but a different locus on chromosome 4 showed strong linkage both to papillomas >6mm and to carcinomas. This locus, which maps near the Cdkn2a/p19(Arf) gene, was entirely p53-dependent, and was not seen in p53 (+/-) backcross animals. Suggestive linkage conferring susceptibility to carcinoma was also found on chromosome 5. These results clearly suggest distinct loci regulate each stage of tumorigenesis, some of which are p53-dependent.

%B Carcinogenesis %V 33 %P 2260-8 %8 2012 Nov %G eng %N 11 %1 http://www.ncbi.nlm.nih.gov/pubmed/22843548?dopt=Abstract %R 10.1093/carcin/bgs250 %0 Journal Article %J Radiat Res %D 2012 %T Low-dose ionizing radiation-induced blood plasma metabolic response in a diverse genetic mouse population. %A Lee, Do Yup %A Bowen, Benjamin P %A Nguyen, David H %A Parsa, Sara %A Huang, Yurong %A Jiang-Hua Mao %A Northen, Trent R %K Animals %K Blood %K Crosses, Genetic %K Dose-Response Relationship, Radiation %K Female %K Genetic Variation %K Male %K Metabolome %K Mice %K Mice, Inbred BALB C %K Species Specificity %K Transcriptome %X

Understanding the biological effects and biochemical mechanisms of low-dose ionizing radiation (LDIR) is important for setting exposure limits for the safe use of nuclear power and medical diagnostic procedures. Although several studies have highlighted the effects of ionizing radiation on metabolism, most studies have focused on uniform genetic mouse populations. Here, we report the metabolic response to LDIR (10 cGy X ray) on a genetically diverse mouse population (142 mice) generated from a cross of radiation-sensitive (BALB/c) and radiation-resistant (SPRET/EiJ) parental strains. GC-TOF profiling of plasma metabolites was used to compare exposed vs. sham animals. From this, 16 metabolites were significantly altered in the LDIR treated vs. sham group. Use of two significantly altered metabolites, thymine and 2-monostearin, was found to effectively segregate the two treatments. Multivariate statistical analysis was used to identify genetic polymorphisms correlated with metabolite abundance (e.g., amino acids, fatty acids, nucleotides and TCA cycle intermediates). Genetic analysis of metabolic phenotypes showed suggestive linkages for fatty acid and amino acid metabolism. However, metabolite abundance was found to be a function of low-dose ionizing radiation exposure, and not of the underlying genetic variation.

%B Radiat Res %V 178 %P 551-5 %8 2012 Dec %G eng %N 6 %1 http://www.ncbi.nlm.nih.gov/pubmed/23051006?dopt=Abstract %R 10.1667/RR2990.1 %0 Journal Article %J Mol Cancer Res %D 2012 %T Pten regulates Aurora-A and cooperates with Fbxw7 in modulating radiation-induced tumor development. %A Kwon, Yong-Won %A Kim, Il-Jin %A Wu, Di %A Lu, Jing %A Stock, William A %A Liu, Yueyong %A Huang, Yurong %A Kang, Hio Chung %A DelRosario, Reyno %A Jen, Kuang-Yu %A Perez-Losada, Jesus %A Wei, Guangwei %A Balmain, Allan %A Jiang-Hua Mao %K Animals %K Aurora Kinase A %K Aurora Kinases %K Binding Sites %K Blotting, Western %K Cell Line %K F-Box Proteins %K Female %K Gamma Rays %K Glycogen Synthase Kinase 3 %K HCT116 Cells %K HEK293 Cells %K Humans %K Male %K Mice %K Mice, Inbred C57BL %K Mice, Knockout %K Mutation %K Neoplasms, Radiation-Induced %K NIH 3T3 Cells %K Protein Binding %K Protein-Serine-Threonine Kinases %K PTEN Phosphohydrolase %K Time Factors %K Ubiquitin %K Ubiquitin-Protein Ligases %X

The Aurora-A kinase gene is frequently amplified and/or overexpressed in a variety of human cancers, leading to major efforts to develop therapeutic agents targeting this pathway. Here, we show that Aurora-A is targeted for ubiquitination and subsequent degradation by the F-box protein FBXW7 in a process that is regulated by GSK3β. Using a series of truncated Aurora-A proteins and site-directed mutagenesis, we identified distinct FBXW7 and GSK3β-binding sites in Aurora-A. Mutation of critical residues in either site substantially disrupts degradation of Aurora-A. Furthermore, we show that loss of Pten results in the stabilization of Aurora-A by attenuating FBXW7-dependent degradation of Aurora-A through the AKT/GSK3β pathway. Moreover, radiation-induced tumor latency is significantly shortened in Fbxw7(+/-)Pten(+/-) mice as compared with either Fbxw7(+/-) or Pten(+/-) mice, indicating that Fbxw7 and Pten appear to cooperate in suppressing tumorigenesis. Our results establish a novel posttranslational regulatory network in which the Pten and Fbxw7 pathways appear to converge on the regulation of Aurora-A level.

%B Mol Cancer Res %V 10 %P 834-44 %8 2012 Jun %G eng %N 6 %1 http://www.ncbi.nlm.nih.gov/pubmed/22513362?dopt=Abstract %R 10.1158/1541-7786.MCR-12-0025 %0 Journal Article %J Blood %D 2012 %T Sequential mutations in Notch1, Fbxw7, and Tp53 in radiation-induced mouse thymic lymphomas. %A Jen, Kuang-Yu %A Song, Ihn Young %A Banta, Karl Luke %A Wu, Di %A Jiang-Hua Mao %A Balmain, Allan %K Animals %K Disease Models, Animal %K F-Box Proteins %K Female %K Loss of Heterozygosity %K Male %K Mice %K Mice, Knockout %K Mutation %K Neoplasms, Radiation-Induced %K Precursor T-Cell Lymphoblastic Leukemia-Lymphoma %K Protein Structure, Tertiary %K Real-Time Polymerase Chain Reaction %K Receptor, Notch1 %K RNA, Messenger %K Thymus Neoplasms %K Tumor Suppressor Protein p53 %K Ubiquitin-Protein Ligases %X

T-cell acute lymphoblastic lymphomas commonly demonstrate activating Notch1 mutations as well as mutations or deletions in Fbxw7. However, because Fbxw7 targets Notch1 for degradation, genetic alterations in these genes are expected to be mutually exclusive events in lymphomagenesis. Previously, by using a radiation-induced Tp53-deficient mouse model for T-cell acute lymphoblastic lymphoma, we reported that loss of heterozygosity at the Fbxw7 locus occurs frequently in a Tp53-dependent manner. In the current study, we show that these thymic lymphomas also commonly exhibit activating Notch1 mutations in the proline-glutamic acid-serine-threonine (PEST) domain. Moreover, concurrent activating Notch1 PEST domain mutations and single-copy deletions at the Fbxw7 locus occur with high frequency in the same individual tumors, indicating that these changes are not mutually exclusive events. We further demonstrate that although Notch1 PEST domain mutations are independent of Tp53 status, they are completely abolished in mice with germline Fbxw7 haploinsufficiency. Therefore, Notch1 PEST domain mutations only occur when Fbxw7 expression levels are intact. These data suggest a temporal sequence of mutational events involving these important cancer-related genes, with Notch1 PEST domain mutations occurring first, followed by Fbxw7 deletion, and eventually by complete loss of Tp53.

%B Blood %V 119 %P 805-9 %8 2012 Jan 19 %G eng %N 3 %1 http://www.ncbi.nlm.nih.gov/pubmed/22117044?dopt=Abstract %R 10.1182/blood-2011-01-327619 %0 Journal Article %J Genes Dev %D 2011 %T Fine-tuning p53 activity through C-terminal modification significantly contributes to HSC homeostasis and mouse radiosensitivity. %A Wang, Yunyuan V %A Leblanc, Mathias %A Fox, Norma %A Jiang-Hua Mao %A Tinkum, Kelsey L %A Krummel, Kurt %A Engle, Dannielle %A Piwnica-Worms, David %A Piwnica-Worms, Helen %A Balmain, Allan %A Kaushansky, Kenneth %A Wahl, Geoffrey M %K Animals %K Cells, Cultured %K Cyclin-Dependent Kinase Inhibitor p21 %K Female %K Gamma Rays %K Gene Dosage %K Gene Expression Regulation %K Gene Knock-In Techniques %K Hematopoietic Stem Cells %K Homeostasis %K Longevity %K Male %K Mice %K Mice, Inbred C57BL %K Mutation %K Proto-Oncogene Proteins c-mdm2 %K Radiation Tolerance %K Tumor Suppressor Protein p53 %X

Cell cycle regulation in hematopoietic stem cells (HSCs) is tightly controlled during homeostasis and in response to extrinsic stress. p53, a well-known tumor suppressor and transducer of diverse stress signals, has been implicated in maintaining HSC quiescence and self-renewal. However, the mechanisms that control its activity in HSCs, and how p53 activity contributes to HSC cell cycle control, are poorly understood. Here, we use a genetically engineered mouse to show that p53 C-terminal modification is critical for controlling HSC abundance during homeostasis and HSC and progenitor proliferation after irradiation. Preventing p53 C-terminal modification renders mice exquisitely radiosensitive due to defects in HSC/progenitor proliferation, a critical determinant for restoring hematopoiesis after irradiation. We show that fine-tuning the expression levels of the cyclin-dependent kinase inhibitor p21, a p53 target gene, contributes significantly to p53-mediated effects on the hematopoietic system. These results have implications for understanding cell competition in response to stresses involved in stem cell transplantation, recovery from adverse hematologic effects of DNA-damaging cancer therapies, and development of radioprotection strategies.

%B Genes Dev %V 25 %P 1426-38 %8 2011 Jul 1 %G eng %N 13 %1 http://www.ncbi.nlm.nih.gov/pubmed/21724834?dopt=Abstract %R 10.1101/gad.2024411 %0 Journal Article %J Integr Biol (Camb) %D 2011 %T Gene transcriptional networks integrate microenvironmental signals in human breast cancer. %A Xu, Ren %A Jiang-Hua Mao %K Breast Neoplasms %K Cadherins %K Cytokines %K Extracellular Matrix %K Extracellular Matrix Proteins %K Female %K Gene Expression Profiling %K Gene Regulatory Networks %K Humans %K Integrins %K Intercellular Signaling Peptides and Proteins %K Mammary Glands, Human %K Oligonucleotide Array Sequence Analysis %K Receptor, Platelet-Derived Growth Factor beta %K Receptors, Cytokine %K Transforming Growth Factor beta %K Tumor Microenvironment %X

A significant amount of evidence shows that microenvironmental signals generated from extracellular matrix (ECM) molecules, soluble factors, and cell-cell adhesion complexes cooperate at the extra- and intracellular level. This synergetic action of microenvironmental cues is crucial for normal mammary gland development and breast malignancy. To explore how the microenvironmental genes coordinate in human breast cancer at the genome level, we have performed gene co-expression network analysis in three independent microarray datasets and identified two microenvironment networks in human breast cancer tissues. Network I represents crosstalk and cooperation of ECM microenvironment and soluble factors during breast malignancy. The correlated expression of cytokines, chemokines, and cell adhesion proteins in Network II implicates the coordinated action of these molecules in modulating the immune response in breast cancer tissues. These results suggest that microenvironmental cues are integrated with gene transcriptional networks to promote breast cancer development.

%B Integr Biol (Camb) %V 3 %P 368-74 %8 2011 Apr %G eng %N 4 %1 http://www.ncbi.nlm.nih.gov/pubmed/21165486?dopt=Abstract %R 10.1039/c0ib00087f %0 Journal Article %J Cancer Cell %D 2011 %T Radiation acts on the microenvironment to affect breast carcinogenesis by distinct mechanisms that decrease cancer latency and affect tumor type. %A Nguyen, David H %A Oketch-Rabah, Hellen A %A Illa-Bochaca, Irineu %A Geyer, Felipe C %A Reis-Filho, Jorge S %A Jiang-Hua Mao %A Ravani, Shraddha A %A Zavadil, Jiri %A Borowsky, Alexander D %A Jerry, D Joseph %A Dunphy, Karen A %A Seo, Jae Hong %A Haslam, Sandra %A Medina, Daniel %A Barcellos-Hoff, Mary Helen %K Animals %K Breast Neoplasms %K Cell Transformation, Neoplastic %K Dose-Response Relationship, Radiation %K Epithelial Cells %K Female %K Gene Expression Profiling %K Gene Expression Regulation, Neoplastic %K Gene Regulatory Networks %K Mammary Glands, Animal %K Mice %K Mice, Inbred BALB C %K Mice, Knockout %K Neoplasms, Radiation-Induced %K Radiation Chimera %K Reaction Time %K Receptors, Estrogen %K Time Factors %K Transforming Growth Factor beta1 %K Tumor Burden %K Tumor Microenvironment %K Tumor Suppressor Protein p53 %K Whole-Body Irradiation %X

Tissue microenvironment is an important determinant of carcinogenesis. We demonstrate that ionizing radiation, a known carcinogen, affects cancer frequency and characteristics by acting on the microenvironment. Using a mammary chimera model in which an irradiated host is transplanted with oncogenic Trp53 null epithelium, we show accelerated development of aggressive tumors whose molecular signatures were distinct from tumors arising in nonirradiated hosts. Molecular and genetic approaches show that TGFβ mediated tumor acceleration. Tumor molecular signatures implicated TGFβ, and genetically reducing TGFβ abrogated the effect on latency. Surprisingly, tumors from irradiated hosts were predominantly estrogen receptor negative. This effect was TGFβ independent and linked to mammary stem cell activity. Thus, the irradiated microenvironment affects latency and clinically relevant features of cancer through distinct and unexpected mechanisms.

%B Cancer Cell %V 19 %P 640-51 %8 2011 May 17 %G eng %N 5 %1 http://www.ncbi.nlm.nih.gov/pubmed/21575864?dopt=Abstract %R 10.1016/j.ccr.2011.03.011 %0 Journal Article %J Clin Cancer Res %D 2011 %T Two distinct routes to oral cancer differing in genome instability and risk for cervical node metastasis. %A Bhattacharya, Aditi %A Roy, Ritu %A A Snijders %A Hamilton, Gregory %A Paquette, Jesse %A Tokuyasu, Taku %A Bengtsson, Henrik %A Jordan, Richard C K %A Olshen, Adam B %A Pinkel, Daniel %A Schmidt, Brian L %A Albertson, Donna G %K Carcinoma, Squamous Cell %K Cohort Studies %K Comparative Genomic Hybridization %K Disease Progression %K DNA Copy Number Variations %K Female %K Genomic Instability %K Head and Neck Neoplasms %K Humans %K Lymphatic Metastasis %K Male %K Middle Aged %K Mouth Neoplasms %K Precancerous Conditions %K Risk %X

PURPOSE: Problems in management of oral cancers or precancers include identification of patients at risk for metastasis, tumor recurrence, and second primary tumors or risk for progression of precancers (dysplasia) to cancer. Thus, the objective of this study was to clarify the role of genomic aberrations in oral cancer progression and metastasis.

EXPERIMENTAL DESIGN: The spectrum of copy number alterations in oral dysplasia and squamous cell carcinomas (SCC) was determined by array comparative genomic hybridization. Associations with clinical characteristics were studied and results confirmed in an independent cohort.

RESULTS: The presence of one or more of the chromosomal aberrations +3q24-qter, -8pter-p23.1, +8q12-q24.2, and +20 distinguishes a major subgroup (70%-80% of lesions, termed 3q8pq20 subtype) from the remainder (20%-30% of lesions, non-3q8pq20). The 3q8pq20 subtype is associated with chromosomal instability and differential methylation in the most chromosomally unstable tumors. The two subtypes differ significantly in clinical outcome with risk for cervical (neck) lymph node metastasis almost exclusively associated with the 3q8pq20 subtype in two independent oral SCC cohorts.

CONCLUSIONS: Two subtypes of oral lesions indicative of at least two pathways for oral cancer development were distinguished that differ in chromosomal instability and risk for metastasis, suggesting that +3q,-8p, +8q, and +20 constitute a biomarker with clinical utility for identifying patients at risk for metastasis. Moreover, although increased numbers of genomic alterations can be harbingers of progression to cancer, dysplastic lesions lacking copy number changes cannot be considered benign as they are potential precursors to non-3q8pq20 locally invasive, yet not metastatic oral SCC.

%B Clin Cancer Res %V 17 %P 7024-34 %8 2011 Nov 15 %G eng %N 22 %R 10.1158/1078-0432.CCR-11-1944 %0 Journal Article %J Histopathology %D 2010 %T Array comparative genomic hybridization analysis indicates that serous carcinomas of the ovary, fallopian tube and endometrium are distinct entities. %A Nowee, Marlies E %A Seeber, Laura Ms %A Horrée, Nicole %A A Snijders %A van Diest, Paul J %A Verheijen, René Hm %A Dorsman, Josephine C %K Biomarkers, Tumor %K Chromosomes, Artificial, Bacterial %K Comparative Genomic Hybridization %K Cystadenocarcinoma, Serous %K Endometrial Neoplasms %K Fallopian Tube Neoplasms %K Female %K Gene Expression Profiling %K Humans %K Microarray Analysis %K Ovarian Neoplasms %B Histopathology %V 57 %P 634-7 %8 2010 Oct %G eng %N 4 %R 10.1111/j.1365-2559.2010.03656.x %0 Journal Article %J J Clin Oncol %D 2010 %T Deletion of the PER3 gene on chromosome 1p36 in recurrent ER-positive breast cancer. %A Climent, Joan %A Perez-Losada, Jesus %A Quigley, David A %A Kim, Il-Jin %A DelRosario, Reyno %A Jen, Kuang-Yu %A Bosch, Ana %A Lluch, Ana %A Jiang-Hua Mao %A Balmain, Allan %K Animals %K Breast Neoplasms %K Chromosomes, Human, Pair 1 %K Disease Models, Animal %K Female %K Gene Dosage %K Gene Expression %K Genetic Predisposition to Disease %K Humans %K Mice %K Neoplasm Recurrence, Local %K Period Circadian Proteins %K Prognosis %K Receptors, Estrogen %K Sequence Deletion %K Survival Analysis %X

PURPOSE: To investigate the role of the PER3 circadian rhythm gene, located within the commonly deleted region of chromosome 1p36, in human breast cancer development.

PATIENTS AND METHODS: The frequency of genetic alterations at 1p36 and PER3 gene copy number status were analyzed in 180 lymph node-negative breast cancers from patients who had received treatment with chemotherapy and/or tamoxifen. The expression levels of PER3 were also analyzed using published microarray profiles from > 400 breast cancer samples. Finally, the effect of loss of Per3 on tumor susceptibility was tested using two mouse models of breast cancer.

RESULTS: Deletion of PER3 is directly related to tumor recurrence in patients with estrogen receptor (ER) - positive breast cancers treated with tamoxifen. Low expression of PER3 mRNA is associated with poor prognosis, particularly in a subset of tumors that are ER positive, and either luminal A or ERBB2-positive tumors. Mice deficient in Per3 showed increased susceptibility to breast cancer induced by carcinogen treatment or by overexpression of Erbb2.

CONCLUSION: Disruption of PER3 function may serve as an indicator of probability of tumor recurrence in patients with ER-positive tumors. Further investigations of this pathway may reveal links between deregulation of sleep homeostasis and breast tumorigenesis.

%B J Clin Oncol %V 28 %P 3770-8 %8 2010 Aug 10 %G eng %N 23 %1 http://www.ncbi.nlm.nih.gov/pubmed/20625127?dopt=Abstract %R 10.1200/JCO.2009.27.0215 %0 Journal Article %J Breast Cancer Res %D 2010 %T The expression level of HJURP has an independent prognostic impact and predicts the sensitivity to radiotherapy in breast cancer. %A Hu, Zhi %A Huang, Ge %A Sadanandam, Anguraj %A Gu, Shenda %A Lenburg, Marc E %A Pai, Melody %A Bayani, Nora %A Blakely, Eleanor A %A Gray, Joe W %A Jiang-Hua Mao %K Biomarkers, Tumor %K Blotting, Western %K Breast Neoplasms %K Cell Line, Tumor %K Disease-Free Survival %K DNA-Binding Proteins %K Female %K Gene Expression Profiling %K Gene Expression Regulation, Neoplastic %K Humans %K Oligonucleotide Array Sequence Analysis %K Predictive Value of Tests %K Prognosis %K RNA Interference %K RNA, Messenger %X

INTRODUCTION: HJURP (Holliday Junction Recognition Protein) is a newly discovered gene reported to function at centromeres and to interact with CENPA. However its role in tumor development remains largely unknown. The goal of this study was to investigate the clinical significance of HJURP in breast cancer and its correlation with radiotherapeutic outcome.

METHODS: We measured HJURP expression level in human breast cancer cell lines and primary breast cancers by Western blot and/or by Affymetrix Microarray; and determined its associations with clinical variables using standard statistical methods. Validation was performed with the use of published microarray data. We assessed cell growth and apoptosis of breast cancer cells after radiation using high-content image analysis.

RESULTS: HJURP was expressed at higher level in breast cancer than in normal breast tissue. HJURP mRNA levels were significantly associated with estrogen receptor (ER), progesterone receptor (PR), Scarff-Bloom-Richardson (SBR) grade, age and Ki67 proliferation indices, but not with pathologic stage, ERBB2, tumor size, or lymph node status. Higher HJURP mRNA levels significantly decreased disease-free and overall survival. HJURP mRNA levels predicted the prognosis better than Ki67 proliferation indices. In a multivariate Cox proportional-hazard regression, including clinical variables as covariates, HJURP mRNA levels remained an independent prognostic factor for disease-free and overall survival. In addition HJURP mRNA levels were an independent prognostic factor over molecular subtypes (normal like, luminal, Erbb2 and basal). Poor clinical outcomes among patients with high HJURP expression were validated in five additional breast cancer cohorts. Furthermore, the patients with high HJURP levels were much more sensitive to radiotherapy. In vitro studies in breast cancer cell lines showed that cells with high HJURP levels were more sensitive to radiation treatment and had a higher rate of apoptosis than those with low levels. Knock down of HJURP in human breast cancer cells using shRNA reduced the sensitivity to radiation treatment. HJURP mRNA levels were significantly correlated with CENPA mRNA levels.

CONCLUSIONS: HJURP mRNA level is a prognostic factor for disease-free and overall survival in patients with breast cancer and is a predictive biomarker for sensitivity to radiotherapy.

%B Breast Cancer Res %V 12 %P R18 %8 2010 %G eng %N 2 %1 http://www.ncbi.nlm.nih.gov/pubmed/20211017?dopt=Abstract %R 10.1186/bcr2487 %0 Journal Article %J PLoS Comput Biol %D 2010 %T Linking changes in epithelial morphogenesis to cancer mutations using computational modeling %A Rejniak, Katarzyna A %A Wang, Shizhen E %A Bryce, Nicole S %A Hang Chang %A Parvin, Bahram %A Jourquin, Jerome %A Estrada, Lourdes %A Gray, Joe W %A Arteaga, Carlos L %A Weaver, Alissa M %A Quaranta, Vito %A Anderson, Alexander R A %K Apoptosis %K Breast Neoplasms %K Cell Proliferation %K Computer Simulation %K Epithelium %K Extracellular Matrix %K Female %K Humans %K Mammary Glands, Human %K Models, Biological %K Morphogenesis %K Mutation %K Receptor, ErbB-2 %X

Most tumors arise from epithelial tissues, such as mammary glands and lobules, and their initiation is associated with the disruption of a finely defined epithelial architecture. Progression from intraductal to invasive tumors is related to genetic mutations that occur at a subcellular level but manifest themselves as functional and morphological changes at the cellular and tissue scales, respectively. Elevated proliferation and loss of epithelial polarization are the two most noticeable changes in cell phenotypes during this process. As a result, many three-dimensional cultures of tumorigenic clones show highly aberrant morphologies when compared to regular epithelial monolayers enclosing the hollow lumen (acini). In order to shed light on phenotypic changes associated with tumor cells, we applied the bio-mechanical IBCell model of normal epithelial morphogenesis quantitatively matched to data acquired from the non-tumorigenic human mammary cell line, MCF10A. We then used a high-throughput simulation study to reveal how modifications in model parameters influence changes in the simulated architecture. Three parameters have been considered in our study, which define cell sensitivity to proliferative, apoptotic and cell-ECM adhesive cues. By mapping experimental morphologies of four MCF10A-derived cell lines carrying different oncogenic mutations onto the model parameter space, we identified changes in cellular processes potentially underlying structural modifications of these mutants. As a case study, we focused on MCF10A cells expressing an oncogenic mutant HER2-YVMA to quantitatively assess changes in cell doubling time, cell apoptotic rate, and cell sensitivity to ECM accumulation when compared to the parental non-tumorigenic cell line. By mapping in vitro mutant morphologies onto in silico ones we have generated a means of linking the morphological and molecular scales via computational modeling. Thus, IBCell in combination with 3D acini cultures can form a computational/experimental platform for suggesting the relationship between the histopathology of neoplastic lesions and their underlying molecular defects.

%B PLoS Comput Biol %V 6 %8 2010 %G eng %N 8 %R 10.1371/journal.pcbi.1000900 %0 Journal Article %J PLoS Comput Biol %D 2010 %T Molecular predictors of 3D morphogenesis by breast cancer cell lines in 3D culture %A Han, Ju %A Chang, Hang %A Giricz, Orsi %A Lee, Genee Y %A Baehner, Frederick L %A Gray, Joe W %A Bissell, Mina J %A Kenny, Paraic A %A Parvin, Bahram %K Biomarkers, Tumor %K Breast Neoplasms %K Cell Culture Techniques %K Cell Line, Tumor %K Female %K Gene Expression Profiling %K Histocytochemistry %K Humans %K Image Processing, Computer-Assisted %K Models, Biological %K Phenotype %K PPAR gamma %K Receptor, ErbB-2 %K Reproducibility of Results %X

Correlative analysis of molecular markers with phenotypic signatures is the simplest model for hypothesis generation. In this paper, a panel of 24 breast cell lines was grown in 3D culture, their morphology was imaged through phase contrast microscopy, and computational methods were developed to segment and represent each colony at multiple dimensions. Subsequently, subpopulations from these morphological responses were identified through consensus clustering to reveal three clusters of round, grape-like, and stellate phenotypes. In some cases, cell lines with particular pathobiological phenotypes clustered together (e.g., ERBB2 amplified cell lines sharing the same morphometric properties as the grape-like phenotype). Next, associations with molecular features were realized through (i) differential analysis within each morphological cluster, and (ii) regression analysis across the entire panel of cell lines. In both cases, the dominant genes that are predictive of the morphological signatures were identified. Specifically, PPARgamma has been associated with the invasive stellate morphological phenotype, which corresponds to triple-negative pathobiology. PPARgamma has been validated through two supporting biological assays.

%B PLoS Comput Biol %V 6 %P e1000684 %8 2010 Feb %G eng %N 2 %R 10.1371/journal.pcbi.1000684 %0 Journal Article %J Nature %D 2009 %T Genetic architecture of mouse skin inflammation and tumour susceptibility. %A Quigley, David A %A To, Minh D %A Perez-Losada, Jesus %A Pelorosso, Facundo G %A Jiang-Hua Mao %A Nagase, Hiroki %A Ginzinger, David G %A Balmain, Allan %K Animals %K Cell Cycle %K Crosses, Genetic %K Female %K Gene Expression Regulation %K Genetic Predisposition to Disease %K Hair Follicle %K Hematopoiesis %K Inflammation %K Male %K Mice %K Quantitative Trait Loci %K Receptors, Calcitriol %K Receptors, G-Protein-Coupled %K Skin %K Skin Neoplasms %X

Germline polymorphisms in model organisms and humans influence susceptibility to complex trait diseases such as inflammation and cancer. Mice of the Mus spretus species are resistant to tumour development, and crosses between M. spretus and susceptible Mus musculus strains have been used to map locations of genetic variants that contribute to skin cancer susceptibility. We have integrated germline polymorphisms with gene expression in normal skin from a M. musculus x M. spretus backcross to generate a network view of the gene expression architecture of mouse skin. Here we demonstrate how this approach identifies expression motifs that contribute to tissue organization and biological functions related to inflammation, haematopoiesis, cell cycle control and tumour susceptibility. Motifs associated with inflammation, epidermal barrier function and proliferation are differentially regulated in backcross mice susceptible or resistant to tumour development. The intestinal stem cell marker Lgr5 is identified as a candidate master regulator of the hair follicle, and the vitamin D receptor (Vdr) is linked to coordinated control of epidermal barrier function, inflammation and tumour susceptibility.

%B Nature %V 458 %P 505-8 %8 2009 Mar 26 %G eng %N 7237 %1 http://www.ncbi.nlm.nih.gov/pubmed/19136944?dopt=Abstract %R 10.1038/nature07683 %0 Journal Article %J Cell Oncol %D 2009 %T Genomic profiling by array comparative genomic hybridization reveals novel DNA copy number changes in breast phyllodes tumours. %A Kuijper, Arno %A A Snijders %A Berns, Els M J J %A Kuenen-Boumeester, Vibeke %A van der Wall, Elsken %A Albertson, Donna G %A van Diest, Paul J %K Breast Neoplasms %K Chromosomes, Human %K Comparative Genomic Hybridization %K Female %K Gene Dosage %K Gene Expression Profiling %K Genome, Human %K Humans %K Oligonucleotide Array Sequence Analysis %K Phyllodes Tumor %X

Breast phyllodes tumour (PT) is a rare fibroepithelial tumour. The genetic alterations contributing to its tumorigenesis are largely unknown. To identify genomic regions involved in pathogenesis and progression of PTs we obtained genome-wide copy number profiles by array comparative genomic hybridization (CGH).DNA was isolated from fresh-frozen tissue samples. 11 PTs and 3 fibroadenomas, a frequently occurring fibroepithelial breast tumour, were analyzed. Arrays composed of 2464 genomic clones were used, providing a resolution of ~1.4 Mb across the genome. Each clone contains at least one STS for linkage to the human genome sequence.No copy number changes were detected in fibroadenomas. On the other hand, 10 of 11 PT (91%) showed DNA copy number alterations. The mean number of chromosomal events in PT was 5.5 (range 0-16) per case. A mean of 2.0 gains (range 0-10) and 3.0 losses (range 0-9) was seen per case of PT. Three cases showed amplifications. DNA copy number change was not related to PT grade. We observed recurrent loss on chromosome 1q, 4p, 10, 13q, 15q, 16, 17p, 19 and X. Recurrent copy number gain was seen on 1q, 2p, 3q, 7p, 8q, 16q, 20.In this study we used array CGH for genomic profiling of fibroepithelial breast tumours. Whereas most PT showed chromosomal instability, fibroadenomas lacked copy number changes. Some copy number aberrations had not previously been associated with PT. Several well-known cancer related genes, such as TP53 and members of the Cadherin, reside within the recurrent regions of copy number alteration. Since copy number change was found in all benign PT, genomic instability may be an early event in PT genesis.

%B Cell Oncol %V 31 %P 31-9 %8 2009 %G eng %N 1 %0 Journal Article %J Clin Cancer Res %D 2007 %T Amplification of PVT1 contributes to the pathophysiology of ovarian and breast cancer. %A Guan, Yinghui %A Kuo, Wen-Lin %A Stilwell, Jackie L %A Takano, Hirokuni %A Lapuk, Anna V %A Fridlyand, Jane %A Jiang-Hua Mao %A Yu, Mamie %A Miller, Melinda A %A Santos, Jennifer L %A Kalloger, Steve E %A Carlson, Joseph W %A Ginzinger, David G %A Celniker, Susan E %A Mills, Gordon B %A Huntsman, David G %A Gray, Joe W %K Apoptosis %K Breast Neoplasms %K Chromosome Aberrations %K Chromosomes, Human, Pair 8 %K Female %K Gene Expression Profiling %K Gene Expression Regulation, Neoplastic %K Genome %K Humans %K In Situ Hybridization, Fluorescence %K Ovarian Neoplasms %K Proteins %K Proto-Oncogene Proteins c-myc %K RNA, Long Noncoding %K Transcription, Genetic %K Treatment Outcome %X

PURPOSE: This study was designed to elucidate the role of amplification at 8q24 in the pathophysiology of ovarian and breast cancer because increased copy number at this locus is one of the most frequent genomic abnormalities in these cancers.

EXPERIMENTAL DESIGN: To accomplish this, we assessed the association of amplification at 8q24 with outcome in ovarian cancers using fluorescence in situ hybridization to tissue microarrays and measured responses of ovarian and breast cancer cell lines to specific small interfering RNAs against the oncogene MYC and a putative noncoding RNA, PVT1, both of which map to 8q24.

RESULTS: Amplification of 8q24 was associated with significantly reduced survival duration. In addition, small interfering RNA-mediated reduction in either PVT1 or MYC expression inhibited proliferation in breast and ovarian cancer cell lines in which they were both amplified and overexpressed but not in lines in which they were not amplified/overexpressed. Inhibition of PVT1 expression also induced a strong apoptotic response in cell lines in which it was overexpressed but not in lines in which it was not amplified/overexpressed. Inhibition of MYC, on the other hand, did not induce an apoptotic response in cell lines in which MYC was amplified and overexpressed.

CONCLUSIONS: These results suggest that MYC and PVT1 contribute independently to ovarian and breast pathogenesis when overexpressed because of genomic abnormalities. They also suggest that PVT1-mediated inhibition of apoptosis may explain why amplification of 8q24 is associated with reduced survival duration in patients treated with agents that act through apoptotic mechanisms.

%B Clin Cancer Res %V 13 %P 5745-55 %8 2007 Oct 1 %G eng %N 19 %1 http://www.ncbi.nlm.nih.gov/pubmed/17908964?dopt=Abstract %R 10.1158/1078-0432.CCR-06-2882 %0 Journal Article %J Biochem Cell Biol %D 2007 %T Characterization of breast cancer by array comparative genomic hybridization. %A Climent, J %A Garcia, J L %A Jiang-Hua Mao %A Arsuaga, J %A Perez-Losada, J %K Breast Neoplasms %K Computational Biology %K Cytogenetic Analysis %K Female %K Gene Expression Profiling %K Genetic Predisposition to Disease %K Humans %K Oligonucleotide Array Sequence Analysis %K Prognosis %X

Cancer progression is due to the accumulation of recurrent genomic alterations that induce growth advantage and clonal expansion. Most of these genomic changes can be detected using the array comparative genomic hybridization (CGH) technique. The accurate classification of these genomic alterations is expected to have an important impact on translational and basic research. Here we review recent advances in CGH technology used in the characterization of different features of breast cancer. First, we present bioinformatics methods that have been developed for the analysis of CGH arrays; next, we discuss the use of array CGH technology to classify tumor stages and to identify and stratify subgroups of patients with different prognoses and clinical behaviors. We finish our review with a discussion of how CGH arrays are being used to identify oncogenes, tumor suppressor genes, and breast cancer susceptibility genes.

%B Biochem Cell Biol %V 85 %P 497-508 %8 2007 Aug %G eng %N 4 %1 http://www.ncbi.nlm.nih.gov/pubmed/17713584?dopt=Abstract %R 10.1139/O07-072 %0 Journal Article %J Cancer Cell %D 2007 %T Crosstalk between Aurora-A and p53: frequent deletion or downregulation of Aurora-A in tumors from p53 null mice. %A Jiang-Hua Mao %A Wu, Di %A Perez-Losada, Jesus %A Jiang, Tao %A Li, Qian %A Neve, Richard M %A Gray, Joe W %A Cai, Wei-Wen %A Balmain, Allan %K Animals %K Apoptosis %K Aurora Kinase A %K Aurora Kinases %K Breast Neoplasms %K Cells, Cultured %K Down-Regulation %K Embryo, Mammalian %K Female %K Fibroblasts %K Gene Deletion %K Gene Dosage %K Gene Expression Profiling %K Genomic Instability %K Heterozygote %K Lymphoma %K Male %K Mice %K Mice, Knockout %K Microarray Analysis %K Neoplasms, Radiation-Induced %K Protein-Serine-Threonine Kinases %K Survival Rate %K Thymus Neoplasms %K Tumor Suppressor Protein p53 %K Whole-Body Irradiation %X

The Aurora-A kinase gene is amplified in a subset of human tumors and in radiation-induced lymphomas from p53 heterozygous mice. Normal tissues from p53-/- mice have increased Aurora-A protein levels, but lymphomas from these mice exhibit heterozygous deletions of Aurora-A and/or reduced protein expression. A similar correlation between low p53 levels and Aurora-A gene deletions and expression is found in human breast cancer cell lines. In vitro studies using mouse embryo fibroblasts demonstrate that inhibition of Aurora-A can have either positive or negative effects on cell growth as a function of p53 status. These data have implications for the design of approaches to targeted cancer therapy involving the crosstalk between Aurora-A kinase and p53 pathways.

%B Cancer Cell %V 11 %P 161-73 %8 2007 Feb %G eng %N 2 %1 http://www.ncbi.nlm.nih.gov/pubmed/17292827?dopt=Abstract %R 10.1016/j.ccr.2006.11.025 %0 Journal Article %J J Pathol %D 2007 %T DNA profiling of primary serous ovarian and fallopian tube carcinomas with array comparative genomic hybridization and multiplex ligation-dependent probe amplification. %A Nowee, M E %A A Snijders %A Rockx, D A P %A de Wit, R M %A Kosma, V M %A Hämäläinen, K %A Schouten, J P %A Verheijen, R H M %A van Diest, P J %A Albertson, D G %A Dorsman, J C %K Base Sequence %K Carcinoma %K Cystadenocarcinoma, Serous %K DNA Fingerprinting %K DNA Probes %K Fallopian Tube Neoplasms %K Female %K Gene Expression Regulation, Neoplastic %K Humans %K Molecular Sequence Data %K Nucleic Acid Amplification Techniques %K Nucleic Acid Hybridization %K Oligonucleotide Array Sequence Analysis %K Ovarian Neoplasms %X

Primary serous ovarian carcinoma (OVCA) and serous Fallopian tube carcinoma (FTC), both belonging to the BRCA-linked tumour spectrum, share many properties and are treated similarly. However, a detailed molecular comparison has been lacking. We hypothesized that comparative genomic studies of serous OVCAs and FTCs should point to gene regions critically involved in their tumorigenesis. Array comparative genomic hybridization (array CGH) analysis indicated that serous OVCAs and serous FTCs displayed common but also more distinctive patterns of recurrent changes. Targeted gene identification using a dedicated multiplex ligation-dependent probe amplification (MLPA) probe set directly identified EIF2C2 on 8q as a potentially important driver gene. Other previously unappreciated gained/amplified genes included PSMB4 on 1q, MTSS1 on 8q, TEAD4 and TSPAN9 on 12p, and BCAS4 on 20q. SPINT2 and ACTN4 on 19q were predominantly found in FTCs. Gains/amplifications of CCNE1 and MYC, often in conjunction with changes in genes of the AKT pathway, EVI1 and PTK2, seemed to be involved at earlier stages, whereas changes of ERBB2 were associated with advanced stages. The only BRCA1-mutated FTC shared common denominators with the sporadic tumours. In conclusion, the data suggest that serous OVCAs and FTCs, although related, exhibit differences in genomic profiles. In addition to known pathways, new genes/pathways are likely to be involved, with changes in an miRNA-associated gene, EIF2C2, as one important new feature. Dedicated MLPA sets constitute potentially important tools for differential diagnosis and may provide footholds for tailored therapy.

%B J Pathol %V 213 %P 46-55 %8 2007 Sep %G eng %N 1 %R 10.1002/path.2217 %0 Journal Article %J Nature %D 2007 %T Promotion of Hras-induced squamous carcinomas by a polymorphic variant of the Patched gene in FVB mice. %A Wakabayashi, Yuichi %A Jiang-Hua Mao %A Brown, Ken %A Girardi, Michael %A Balmain, Allan %K Amino Acid Sequence %K Animals %K Apoptosis %K Carcinoma, Squamous Cell %K Cell Line %K Cell Transformation, Neoplastic %K Crosses, Genetic %K Female %K Gene Expression Regulation, Neoplastic %K Genes, ras %K HSP40 Heat-Shock Proteins %K Humans %K Kruppel-Like Transcription Factors %K Male %K Mice %K Mice, Inbred C57BL %K Mice, Transgenic %K Molecular Sequence Data %K Polymorphism, Genetic %K ras Proteins %K Receptors, Cell Surface %X

Mice of the C57BL/6 strain are resistant to the development of skin squamous carcinomas (SCCs) induced by an activated Ras oncogene, whereas FVB/N mice are highly susceptible. The genetic basis of this difference in phenotype is unknown. Here we show that susceptibility to SCC is under the control of a carboxy-terminal polymorphism in the mouse Ptch gene. F1 hybrids between C57BL/6 and FVB/N strains ((B6FVB)F1) are resistant to Ras-induced SCCs, but resistance can be overcome either by elimination of the C57BL/6 Ptch allele (Ptch(B6)) or by overexpression of the FVB/N Ptch allele (Ptch(FVB)) in the epidermis of K5Hras-transgenic (B6FVB)F1 hybrid mice. The human Patched (PTCH) gene is a classical tumour suppressor gene for basal cell carcinomas and medulloblastomas, the loss of which causes increased signalling through the Sonic Hedgehog (SHH) pathway. SCCs that develop in PtchB6+/- mice do not lose the wild-type Ptch gene or show evidence of increased SHH signalling. Although Ptch(FVB) overexpression can promote SCC formation, continued expression is not required for tumour maintenance, suggesting a role at an early stage of tumour cell lineage commitment. The Ptch polymorphism affects Hras-induced apoptosis, and binding to Tid1, the mouse homologue of the Drosophila l(2)tid tumour suppressor gene. We propose that Ptch occupies a critical niche in determining basal or squamous cell lineage, and that both tumour types can arise from the same target cell depending on carcinogen exposure and host genetic background.

%B Nature %V 445 %P 761-5 %8 2007 Feb 15 %G eng %N 7129 %1 http://www.ncbi.nlm.nih.gov/pubmed/17230190?dopt=Abstract %R 10.1038/nature05489 %0 Journal Article %J BMC Cancer %D 2006 %T Breast tumor copy number aberration phenotypes and genomic instability. %A Fridlyand, Jane %A A Snijders %A Ylstra, Bauke %A Li, Hua %A Olshen, Adam %A Segraves, Richard %A Dairkee, Shanaz %A Tokuyasu, Taku %A Ljung, Britt Marie %A Jain, Ajay N %A McLennan, Jane %A Ziegler, John %A Chin, Koei %A Devries, Sandy %A Feiler, Heidi %A Gray, Joe W %A Waldman, Frederic %A Pinkel, Daniel %A Albertson, Donna G %K Adult %K Aged %K Breast Neoplasms %K Carcinoma, Ductal, Breast %K Chromosome Aberrations %K Chromosomes, Human %K DNA, Neoplasm %K E2F Transcription Factors %K Female %K Gene Dosage %K Gene Expression Regulation, Neoplastic %K Genes, p53 %K Genes, Retinoblastoma %K Genomic Instability %K Humans %K Karyotyping %K Middle Aged %K Neoplasm Proteins %K Nucleic Acid Hybridization %K Oligonucleotide Array Sequence Analysis %K Phenotype %K Retinoblastoma Protein %K Signal Transduction %K Telomere %X

BACKGROUND: Genomic DNA copy number aberrations are frequent in solid tumors, although the underlying causes of chromosomal instability in tumors remain obscure. Genes likely to have genomic instability phenotypes when mutated (e.g. those involved in mitosis, replication, repair, and telomeres) are rarely mutated in chromosomally unstable sporadic tumors, even though such mutations are associated with some heritable cancer prone syndromes.

METHODS: We applied array comparative genomic hybridization (CGH) to the analysis of breast tumors. The variation in the levels of genomic instability amongst tumors prompted us to investigate whether alterations in processes/genes involved in maintenance and/or manipulation of the genome were associated with particular types of genomic instability.

RESULTS: We discriminated three breast tumor subtypes based on genomic DNA copy number alterations. The subtypes varied with respect to level of genomic instability. We find that shorter telomeres and altered telomere related gene expression are associated with amplification, implicating telomere attrition as a promoter of this type of aberration in breast cancer. On the other hand, the numbers of chromosomal alterations, particularly low level changes, are associated with altered expression of genes in other functional classes (mitosis, cell cycle, DNA replication and repair). Further, although loss of function instability phenotypes have been demonstrated for many of the genes in model systems, we observed enhanced expression of most genes in tumors, indicating that over expression, rather than deficiency underlies instability.

CONCLUSION: Many of the genes associated with higher frequency of copy number aberrations are direct targets of E2F, supporting the hypothesis that deregulation of the Rb pathway is a major contributor to chromosomal instability in breast tumors. These observations are consistent with failure to find mutations in sporadic tumors in genes that have roles in maintenance or manipulation of the genome.

%B BMC Cancer %V 6 %P 96 %8 2006 %G eng %R 10.1186/1471-2407-6-96 %0 Journal Article %J Clin Cancer Res %D 2006 %T Chromosomal instability in microsatellite-unstable and stable colon cancer. %A Trautmann, Karolin %A Terdiman, Jonathan P %A French, Amy J %A Roydasgupta, Ritu %A Sein, Nancy %A Kakar, Sanjay %A Fridlyand, Jane %A A Snijders %A Albertson, Donna G %A Thibodeau, Stephen N %A Waldman, Frederic M %K Adenocarcinoma %K Aged %K Aged, 80 and over %K Chromosomal Instability %K Colonic Neoplasms %K Female %K Humans %K In Situ Hybridization, Fluorescence %K Male %K Microsatellite Instability %K Middle Aged %K Nucleic Acid Hybridization %K Oligonucleotide Array Sequence Analysis %X

PURPOSE: The genomic instability in colon cancer can be divided into at least two major types, microsatellite instability (MSI) or chromosomal instability (CIN). Although initially felt to be mutually exclusive, recent evidence suggests that there may be overlap between the two. The aim of this study was to identify chromosomal alterations at high resolution in sporadic colon cancers with high-level microsatellite instability (MSI-H) and to compare them to those present in a set of matched microsatellite stable (MSS) tumors.

EXPERIMENTAL DESIGN: Array-based comparative genomic hybridization was used to analyze a set of 23 sporadic MSI-H and 23 MSS colon cancers matched for location, gender, stage, and age. The arrays consisted of 2,464 bacterial artificial chromosome clones.

RESULTS: MSI and MSS colon cancers differed significantly with respect to frequency and type of chromosomal alterations. The median fraction of genome altered was lower among MSI-H tumors than MSS tumors (2.8% versus 30.7%, P=0.00006). However, the MSI-H tumors displayed a range of genomic alterations, from the absence of detectable alterations to extensive alterations. Frequent alterations in MSI-H tumors included gains of chromosomes 8, 12, and 13, and loss of 15q14. In contrast, the most frequent alterations in MSS tumors were gains of 7, 13, 8q, and 20, and losses of 8p, 17p, and 18. A small, previously uncharacterized, genomic deletion on 16p13.2, found in 35% of MSI-H and 21% of MSS tumors, was confirmed by fluorescence in situ hybridization.

CONCLUSION: MSI and CIN are not mutually exclusive forms of genomic instability in sporadic colon cancer, with MSI tumors also showing varying degrees of CIN.

%B Clin Cancer Res %V 12 %P 6379-85 %8 2006 Nov 1 %G eng %N 21 %R 10.1158/1078-0432.CCR-06-1248 %0 Journal Article %J Nat Genet %D 2006 %T A functional switch from lung cancer resistance to susceptibility at the Pas1 locus in Kras2LA2 mice. %A To, Minh D %A Perez-Losada, Jesus %A Jiang-Hua Mao %A Hsu, Jeff %A Jacks, Tyler %A Balmain, Allan %K Adenoma %K Alleles %K Animals %K Carcinogens %K Female %K Genetic Predisposition to Disease %K Lung Neoplasms %K Male %K Mice %K Models, Genetic %K Mutation %K Oncogenes %K Proto-Oncogene Proteins p21(ras) %K Risk Factors %K Urethane %X

Pulmonary adenoma susceptibility 1 (Pas1) is the major mouse lung cancer susceptibility locus on chromosome 6 (ref. 1). Kras2 is a common target of somatic mutation in chemically induced mouse lung tumors and is a candidate Pas1 gene. M. spretus mice (SPRET/Ei) carry a Pas1 resistance haplotype for chemically induced lung tumors. We demonstrate that the SPRET/Ei Pas1 allele is switched from resistance to susceptibility by fixation of the parental origin of the mutant Kras2 allele. This switch correlates with low expression of endogenous Kras2 in SPRET/Ei. We propose that the Pas1 modifier effect is due to Kras2, and that a sensitive balance between the expression levels of wild-type and mutant alleles determines lung tumor susceptibility. These data demonstrate that cancer predisposition should also be considered in the context of somatic events and could have major implications for the design of human association studies to identify cancer susceptibility genes.

%B Nat Genet %V 38 %P 926-30 %8 2006 Aug %G eng %N 8 %1 http://www.ncbi.nlm.nih.gov/pubmed/16823377?dopt=Abstract %R 10.1038/ng1836 %0 Journal Article %J J Pathol %D 2006 %T Increased gene copy numbers at chromosome 20q are frequent in both squamous cell carcinomas and adenocarcinomas of the cervix. %A Wilting, S M %A Snijders, P J F %A Meijer, G A %A Ylstra, B %A van den Ijssel, P R L A %A A Snijders %A Albertson, D G %A Coffa, J %A Schouten, J P %A van de Wiel, M A %A Meijer, C J L M %A Steenbergen, R D M %K Adenocarcinoma %K Adult %K Aged %K Carcinoma, Squamous Cell %K Cell Line, Tumor %K Chromosome Aberrations %K Chromosome Mapping %K Chromosomes, Human %K Chromosomes, Human, Pair 20 %K DNA (Cytosine-5-)-Methyltransferase %K Female %K Genome, Human %K Humans %K In Situ Hybridization, Fluorescence %K Middle Aged %K Oligonucleotide Array Sequence Analysis %K Papillomaviridae %K RNA, Messenger %K RNA, Neoplasm %K Uterine Cervical Neoplasms %X

Genome-wide microarray-based comparative genomic hybridization (array CGH) was used to identify common chromosomal alterations involved in cervical carcinogenesis as a first step towards the discovery of novel biomarkers. The genomic profiles of nine squamous cell carcinomas (SCCs) and seven adenocarcinomas (AdCAs), as well as four human papillomavirus (HPV)-immortalized keratinocyte cell lines, were assessed. On a genome-wide scale, SCCs showed significantly more gains than AdCAs. More specifically, there was a striking and highly significant difference between the two histological types for gain at 3q12.1-28, which was predominantly observed in SCC. Other frequent alterations included gains of 1q21.1-31.1 and 20q11.21-13.33, and losses of 11q22.3-25 and 13q14.3-21.33. Subsequent FISH analysis for hTR, located at 3q26, confirmed the presence of 3q gain in SCCs and HPV-immortalized cell lines. Fine mapping of chromosome 20q using multiplex ligation-dependent probe amplification (MLPA) showed copy number increases for a number of genes located at 20q11-q12, including DNMT3B and TOP1. For DNMT3B, this correlated with elevated mRNA expression in 79% of cases. In conclusion, the assessment of frequent genomic alterations resulted in the identification of potential novel biomarkers, which may ultimately enable a better risk stratification of high-risk (hr)-HPV-positive women.

%B J Pathol %V 209 %P 220-30 %8 2006 Jun %G eng %N 2 %R 10.1002/path.1966 %0 Journal Article %J Oncogene %D 2005 %T Genomic instability in radiation-induced mouse lymphoma from p53 heterozygous mice. %A Jiang-Hua Mao %A Li, Jiangzhen %A Jiang, Tao %A Li, Qian %A Wu, Di %A Perez-Losada, Jesus %A DelRosario, Reyno %A Peterson, Leif %A Balmain, Allan %A Cai, Wei-Wen %K Animals %K DNA Damage %K DNA Repair %K Female %K Gene Amplification %K Gene Deletion %K Gene Expression Profiling %K Genes, p53 %K Genomic Instability %K Lymphoma %K Male %K Mice %K Mice, Inbred C57BL %K Neoplasms, Radiation-Induced %K Thymus Neoplasms %X

Although radiation can directly induce DNA damage and is a known human and animal carcinogen, the number of genetic changes in radiation-induced tumors, and the pathways responsible for generating them, are unknown. We have used high-density BAC arrays covering >95% of the mouse genome for analysis of genomic patterns of aberrations in spontaneous and radiation-induced mouse lymphomas. The majority of radiation-induced tumors exhibit one of three 'signatures' based on gene copy number changes. Some exhibit extensive scrambling of the genome, with very high numbers of recurrent gains and losses. Two other signatures are characterized by excess gains but relatively few losses, or vice versa. Changes in spontaneous tumors often involve whole chromosomes, whereas radiation-induced tumors exhibit a high frequency of localized deletion/amplification events. The number of copy number abnormalities does not correlate with the latency or pathology of the tumors. We propose that specific early events following radiation exposure induce changes in 'caretaker' genes that control specific downstream pathways involved in DNA damage repair. The nature of these early events may determine the overall genomic signature observed in the resulting tumor.

%B Oncogene %V 24 %P 7924-34 %8 2005 Nov 24 %G eng %N 53 %1 http://www.ncbi.nlm.nih.gov/pubmed/16116484?dopt=Abstract %R 10.1038/sj.onc.1208926 %0 Journal Article %J Nature %D 2004 %T Fbxw7/Cdc4 is a p53-dependent, haploinsufficient tumour suppressor gene. %A Jiang-Hua Mao %A Perez-Losada, Jesus %A Wu, Di %A DelRosario, Reyno %A Tsunematsu, Ryosuke %A Nakayama, Keiichi I %A Brown, Ken %A Bryson, Sheila %A Balmain, Allan %K Animals %K Base Sequence %K Cell Cycle Proteins %K Cell Transformation, Neoplastic %K F-Box Proteins %K Female %K Fibroblasts %K Gene Deletion %K Genes, Tumor Suppressor %K Loss of Heterozygosity %K Male %K Mice %K Mice, Knockout %K Mutation %K Neoplasms %K RNA, Messenger %K RNA, Small Interfering %K Survival Rate %K Tumor Suppressor Protein p53 %K Ubiquitin-Protein Ligases %X

The FBXW7/hCDC4 gene encodes a ubiquitin ligase implicated in the control of chromosome stability. Here we identify the mouse Fbxw7 gene as a p53-dependent tumour suppressor gene by using a mammalian genetic screen for p53-dependent genes involved in tumorigenesis. Radiation-induced lymphomas from p53+/- mice, but not those from p53-/- mice, show frequent loss of heterozygosity and a 10% mutation rate of the Fbxw7 gene. Fbxw7+/- mice have greater susceptibility to radiation-induced tumorigenesis, but most tumours retain and express the wild-type allele, indicating that Fbxw7 is a haploinsufficient tumour suppressor gene. Loss of Fbxw7 alters the spectrum of tumours that develop in p53 deficient mice to include a range of tumours in epithelial tissues such as the lung, liver and ovary. Mouse embryo fibroblasts from Fbxw7-deficient mice, or wild-type mouse cells expressing Fbxw7 small interfering RNA, have higher levels of Aurora-A kinase, c-Jun and Notch4, but not of cyclin E. We propose that p53-dependent loss of Fbxw7 leads to genetic instability by mechanisms that might involve the activation of Aurora-A, providing a rationale for the early occurrence of these mutations in human cancers.

%B Nature %V 432 %P 775-9 %8 2004 Dec 9 %G eng %N 7018 %1 http://www.ncbi.nlm.nih.gov/pubmed/15592418?dopt=Abstract %R 10.1038/nature03155 %0 Journal Article %J Cell Oncol %D 2004 %T Genomic alterations in primary gastric adenocarcinomas correlate with clinicopathological characteristics and survival. %A Weiss, Marjan M %A Kuipers, Ernst J %A Postma, Cindy %A A Snijders %A Pinkel, Daniel %A Meuwissen, Stefan G M %A Albertson, Donna %A Meijer, Gerrit A %K Adenocarcinoma %K Adult %K Aged %K Aged, 80 and over %K Chromosome Aberrations %K Chromosomes %K Female %K Gene Expression Regulation, Neoplastic %K Genetic Linkage %K Genome, Human %K Humans %K Image Processing, Computer-Assisted %K Lymph Nodes %K Lymphatic Metastasis %K Male %K Middle Aged %K Models, Genetic %K Nucleic Acid Hybridization %K Oligonucleotide Array Sequence Analysis %K Sensitivity and Specificity %K Sequence Tagged Sites %K Stomach Neoplasms %K Time Factors %X

BACKGROUND & AIMS: Pathogenesis of gastric cancer is driven by an accumulation of genetic changes that to a large extent occur at the chromosomal level. In order to investigate the patterns of chromosomal aberrations in gastric carcinomas, we performed genome-wide microarray based comparative genomic hybridisation (microarray CGH). With this recently developed technique chromosomal aberrations can be studied with high resolution and sensitivity.

METHODS: Array CGH was applied to a series of 35 gastric adenocarcinomas using a genome-wide scanning array with 2275 BAC and P1 clones spotted in triplicate. Each clone contains at least one STS for linkage to the sequence of the human genome. These arrays provide an average resolution of 1.4 Mb across the genome. DNA copy number changes were correlated with clinicopathological tumour characteristics as well as survival.

RESULTS: All thirty-five cancers showed chromosomal aberrations and 16 of the 35 tumours showed one or more amplifications. The most frequent aberrations are gains of 8q24.2, 8q24.1, 20q13.12, 20q13.2, 7p11.2, 1q32.3, 8p23.1-p23.3, losses of 5q14.1, 18q22.1, 19p13.12-p13.3, 9p21.3-p24.3, 17p13.1-p13.3, 13q31.1, 16q22.1, 21q21.3, and amplifications of 7q21-q22, and 12q14.1-q21.1. These aberrations were correlated to clinicopathological characteristics and survival. Gain of 1q32.3 was significantly correlated with lymph node status (p=0.007). Tumours with loss of 18q22.1, as well as tumours with amplifications were associated with poor survival (p=0.02, both).

CONCLUSIONS: Microarray CGH has revealed several chromosomal regions that have not been described before in gastric cancer at this frequency and resolution, such as amplification of at 7q21-q22 and 12q14.1-q21.1, as well gains at 1q32.3, 7p11.2, and losses at 13q13.1. Interestingly, gain of 1q32.3 and loss of 18q22.1 are associated with a bad prognosis indicating that these regions could harbour gene(s) that may determine aggressive tumour behaviour and poor clinical outcome.

%B Cell Oncol %V 26 %P 307-17 %8 2004 %G eng %N 5-6 %0 Journal Article %J Cancer Res %D 2003 %T Allele-specific Hras mutations and genetic alterations at tumor susceptibility loci in skin carcinomas from interspecific hybrid mice. %A Nagase, Hiroki %A Jiang-Hua Mao %A Balmain, Allan %K Alleles %K Animals %K Chromosome Mapping %K Crosses, Genetic %K Female %K Genes, ras %K Genetic Predisposition to Disease %K Mice %K Mutation %K Skin Neoplasms %X

We have investigated the effects of germ-line variants that influence skin tumor susceptibility loci on the patterns of somatic genetic alterations in mouse skin cancers. Using a two-stage skin carcinogenesis model, we previously identified at least 13 skin tumor susceptibility (Skts) loci in a large interspecific F1 backcross [(NIH/Ola x M. spretus) x NIH/Ola] study. In this report, we describe the analysis of allele-specific alterations at these loci in skin tumors from the same backcross animals. The mouse Hras gene, located close to Skts2 on chromosome 7, had specific activating mutations in the Mus musculus allele in 23 of 26 carcinomas. In all cases, tumors with Hras mutations also showed specific imbalance of chromosome 7 markers that favored the chromosome carrying the mutant allele. Allele-specific quantitative microsatellite analysis was also carried out, using DNA from 62 carcinomas from (NIH/Ola x M. spretus) x NIH/Ola mice. Frequent allelic imbalance was detected at five additional tumor-susceptibility loci on chromosomes 4, 6, 7, 9, and 16 (Skts7, Skts12, Skts1, Skts6, and Skts9, respectively). At all except Skts7, we found loss of the allele inherited from the resistant strain or amplification of the allele from the susceptible strain. We conclude that polymorphisms in some low-penetrance tumor modifier genes are reflected in the pattern of somatic alterations in tumors. Analysis of such allele-specific changes in tumors may facilitate the identification of functional germ-line variants that control tumor susceptibility.

%B Cancer Res %V 63 %P 4849-53 %8 2003 Aug 15 %G eng %N 16 %1 http://www.ncbi.nlm.nih.gov/pubmed/12941805?dopt=Abstract %0 Journal Article %J J Pathol %D 2003 %T Determination of amplicon boundaries at 20q13.2 in tissue samples of human gastric adenocarcinomas by high-resolution microarray comparative genomic hybridization. %A Weiss, Marjan M %A A Snijders %A Kuipers, Ernst J %A Ylstra, Bauke %A Pinkel, Daniel %A Meuwissen, Stefan G M %A van Diest, Paul J %A Albertson, Donna G %A Meijer, Gerrit A %K Adenocarcinoma %K Adult %K Aged %K Aged, 80 and over %K Chromosome Aberrations %K Chromosomes, Artificial, Bacterial %K Chromosomes, Human, Pair 20 %K DNA, Neoplasm %K Female %K Gene Amplification %K Humans %K In Situ Hybridization, Fluorescence %K Male %K Middle Aged %K Oligonucleotide Array Sequence Analysis %K Stomach Neoplasms %X

Comparative genomic hybridization (CGH) of gastric adenocarcinomas frequently shows gains and amplifications of chromosome 20. However, the underlying genetic lesion is unknown and conventional CGH results do not allow specification of the target region. In order to investigate this chromosomal aberration with a higher resolution and sensitivity, microarray-based CGH was performed with both scanning and high-resolution arrays of chromosome 20 in a series of 27 gastric adenocarcinomas. Locus-specific fragments of genomic DNA from bacterial artificial chromosome (BAC) clones were spotted as microarrays. A scanning array contained a set of 27 BAC clones covering chromosome 20q. A high-resolution array contained 27 overlapping BAC clones at 20q13.2. This high-resolution array was used to narrow down the amplicon at 20q13.2 in tumours showing amplification of this chromosomal region with the scanning array. Positive copy number changes on chromosome 20q were detected in 12 of 27 cases (44%). These changes included gain of the whole arm of chromosome 20q in 8 of 27 (30%) cases, amplification restricted to 20q12.1 in one case, and amplifications restricted to 20q13 in three cases (11%). The three tumours showing amplification restricted to 20q13 were analysed further using the high-resolution array. In one tumour, the whole contig was amplified at a constant level. One of the other two tumours had a clear proximal breakpoint, while the other tumour had a clear distal breakpoint within the 20q13.2 region. The proximal and the distal breakpoint were approximately 800 kb apart. In the present study, an amplicon at 20q13.2 has been narrowed down to 800 kb which is likely to harbour one or more putative oncogenes relevant to gastric carcinogenesis, for which ZNF217 and CYP24 are good candidates.

%B J Pathol %V 200 %P 320-6 %8 2003 Jul %G eng %N 3 %R 10.1002/path.1359 %0 Journal Article %J Mol Pathol %D 2003 %T Genome wide array comparative genomic hybridisation analysis of premalignant lesions of the stomach. %A Weiss, M M %A Kuipers, E J %A Postma, C %A A Snijders %A Stolte, M %A Vieth, M %A Pinkel, D %A Meuwissen, S G M %A Albertson, D %A Meijer, G A %K Adenoma %K Carcinoma in Situ %K Chromosome Aberrations %K Disease Progression %K Female %K Genome %K Humans %K Hyperplasia %K Male %K Nucleic Acid Hybridization %K Oligonucleotide Array Sequence Analysis %K Precancerous Conditions %K Stomach %K Stomach Neoplasms %X

BACKGROUND: Gastric cancer is one of the most frequent malignancies in the world, ranking fifth in the Netherlands as a cause of cancer death. Surgery is the only curative treatment for advanced cases, but results of gastrectomy largely depend on the stage of the disease. A better understanding of the mechanisms of progression from a preneoplastic condition through intraepithelial neoplasia to invasive cancer may provide information relevant to designing focused prevention strategies.

METHODS: Because the pattern of chromosomal aberrations in precursors of gastric cancer is unclear, 11 gastric polyps with intraepithelial neoplasia (three hyperplastic polyps and eight adenomas) were analysed by microarray comparative genomic hybridisation to study chromosomal instability in precursors of gastric cancer.

RESULTS: Chromosomal aberrations were detected in all specimens. Adenomas showed no more chromosomal aberrations than did the hyperplastic polyps. The most frequent aberrations were gain of 7q36 and 20q12, and loss of 5q14-q21 in the adenomas, and loss of 15q11-14, 1p21-31, and 21q11-21.2 in the hyperplastic polyps. The most frequent chromosomal aberration in common to both types was loss of 9p21.3.

CONCLUSION: Hyperplastic polyps showed many chromosomal aberrations, confirming that neoplastic transformation can occur in these lesions. These observations are consistent with the existence of two morphologically and genetically distinct pathways to gastric cancer-the hyperplastic polyp pathway and the (intestinal type) adenoma pathway. The relative contribution of each to gastric carcinogenesis in general, and how they compare to patterns of chromosomal aberrations in the more prevalent flat foci of intraepithelial neoplasia remain to be determined.

%B Mol Pathol %V 56 %P 293-8 %8 2003 Oct %G eng %N 5 %0 Journal Article %J Oncogene %D 2003 %T Genome-wide-array-based comparative genomic hybridization reveals genetic homogeneity and frequent copy number increases encompassing CCNE1 in fallopian tube carcinoma. %A A Snijders %A Nowee, Marlies E %A Fridlyand, Jane %A Piek, Jurgen M J %A Dorsman, Josephine C %A Jain, Ajay N %A Pinkel, Daniel %A van Diest, Paul J %A Verheijen, René H M %A Albertson, Donna G %K Carcinoma %K Chromosome Mapping %K Cyclin E %K DNA %K Fallopian Tube Neoplasms %K Female %K Gene Expression Regulation, Neoplastic %K Genome %K Humans %K Models, Genetic %K Mutation %K Nucleic Acid Hybridization %K Oligonucleotide Array Sequence Analysis %K Oncogene Proteins %K Oncogenes %X

Fallopian tube carcinoma (FTC) is a rare, poorly studied and aggressive cancer, associated with poor survival. Since tumorigenesis is related to the acquisition of genetic changes, we used genome-wide array comparative genomic hybridization to analyse copy number aberrations occurring in FTC in order to obtain a better understanding of FTC carcinogenesis and to identify prognostic events and targets for therapy. We used arrays of 2464 genomic clones, providing approximately 1.4 Mb resolution across the genome to map genomic DNA copy number aberrations quantitatively from 14 FTC onto the human genome sequence. All tumors showed a high frequency of copy number aberrations with recurrent gains on 3q, 6p, 7q, 8q, 12p, 17q, 19 and 20q, and losses involving chromosomes 4, 5q, 8p, 16q, 17p, 18q and X. Recurrent regions of amplification included 1p34, 8p11-q11, 8q24, 12p, 17p13, 17q12-q21, 19p13, 19q12-q13 and 19q13. Candidate, known oncogenes mapping to these amplicons included CMYC (8q24), CCNE1 (19q12-q21) and AKT2 (19q13), whereas PIK3CA and KRAS, previously suggested to be candidate driver genes for amplification, mapped outside copy number maxima on 3q and 12p, respectively. The FTC were remarkably homogeneous, with some recurrent aberrations occurring in more than 70% of samples, which suggests a stereotyped pattern of tumor evolution.

%B Oncogene %V 22 %P 4281-6 %8 2003 Jul 3 %G eng %N 27 %R 10.1038/sj.onc.1206621 %0 Journal Article %J Zhongguo Zhong Yao Za Zhi %D 2002 %T [Effects of aqueous extract in herba of Lysimachia christinae on hyperuricemia in mice]. %A Wang, Hai-dong %A Ge, Fei %A Yusong Guo %A Kong, Ling-dong %K Animals %K Drugs, Chinese Herbal %K Female %K Hyperuricemia %K Mice %K Plants, Medicinal %K Primulaceae %K Uric Acid %X

OBJECTIVE: To study hypouricemic effect of aqueous extract of Lysimachia christinae on hyperuricemia in mice.

METHOD: The uricase inhibitor potassium oxonate was used to induce hyperuricemia in mice, and serum uric acid level was determined with the phosphotungstic acid method.

RESULT: The aqueous extract of Lysimachia christinae, when administered orally to the oxonate-induced hyperuricemic mice at the doses of 5.2, 10.4 and 20.8 g.kg-1, was able to elicit dose-dependent hypouricemic effects. At these doses of the extract, the serum urate levels of the oxonate-pretreated mice showed no difference from the normal mice. In normal mice, however, oral administration of the extract at the same doses did not produce any observable hypouricemic effects.

CONCLUSION: The aqueous extract of Lysimachia christinae possesses potent hypuricemic effects on models of hyperuricemia in mice pretreated with oxonate.

%B Zhongguo Zhong Yao Za Zhi %V 27 %P 939-41, 944 %8 2002 Dec %G eng %N 12 %0 Journal Article %J Genome Res %D 2002 %T Fully automatic quantification of microarray image data. %A Jain, Ajay N %A Tokuyasu, Taku A %A A Snijders %A Segraves, Richard %A Albertson, Donna G %A Pinkel, Daniel %K Breast Neoplasms %K Female %K Gene Expression Profiling %K Humans %K Image Processing, Computer-Assisted %K Male %K Nucleic Acid Hybridization %K Oligonucleotide Array Sequence Analysis %K Staining and Labeling %K Tumor Cells, Cultured %X

DNA microarrays are now widely used to measure expression levels and DNA copy number in biological samples. Ratios of relative abundance of nucleic acids are derived from images of regular arrays of spots containing target genetic material to which fluorescently labeled samples are hybridized. Whereas there are a number of methods in use for the quantification of images, many of the software systems in wide use either encourage or require extensive human interaction at the level of individual spots on arrays. We present a fully automatic system for microarray image quantification. The system automatically locates both subarray grids and individual spots, requiring no user identification of any image coordinates. Ratios are computed based on explicit segmentation of each spot. On a typical image of 6000 spots, the entire process takes less than 20 sec. We present a quantitative assessment of performance on multiple replicates of genome-wide array-based comparative genomic hybridization experiments. By explicitly identifying the pixels in each spot, the system yields more accurate estimates of ratios than systems assuming spot circularity. The software, called, runs on Windows platforms and is available free of charge for academic use.

%B Genome Res %V 12 %P 325-32 %8 2002 Feb %G eng %N 2 %R 10.1101/gr.210902 %0 Journal Article %J Nat Biotechnol %D 2002 %T Genome-wide detection of chromosomal imbalances in tumors using BAC microarrays. %A Cai, Wei-Wen %A Jiang-Hua Mao %A Chow, Chi-Wen %A Damani, Shamsha %A Balmain, Allan %A Bradley, Allan %K Animals %K Chromosome Aberrations %K Chromosomes, Artificial, Bacterial %K Female %K Fluorescent Antibody Technique %K Genome %K Genomics %K Genotype %K Loss of Heterozygosity %K Lymphoma %K Male %K Mice %K Microsatellite Repeats %K Oligonucleotide Array Sequence Analysis %X

Chromosomal imbalances such as deletions and amplifications are common rearrangements in most tumors. Specific rearrangements are consistently associated with specific tumor types or stages, implicating the role of the genes in a region of chromosomal imbalance in tumor initiation and progression. The development of comparative genomic hybridization (CGH) has obviated the need to obtain metaphase spreads from tumors, so that the chromosomal imbalances in many solid tumors may be revealed using an extracted genomic DNA sample. However, the resolution of the cytogenetic method remains and the extreme technical difficulty of CGH has restricted its use. Conceptually, DNA microarray-based CGH is an obvious solution to all of the limitations of conventional CGH. Although arrays have been used for CGH studies, their success has been limited by poor specific signal-to-noise ratios. Here we demonstrate a microarray-based CGH method that allows reliable detection of chromosomal deletions and amplifications with high resolution. Our microarray system is fundamentally different from most current microarray technologies in that activated DNA is printed on natural glass surfaces while other systems almost exclusively focus on activating the surfaces, a strategy that invariably introduces hybridization backgrounds. The concept of using pre-modification may be generally applied for making arrays of other biological materials, as modifying the substrates will be more controllable in solution than on surfaces.

%B Nat Biotechnol %V 20 %P 393-6 %8 2002 Apr %G eng %N 4 %1 http://www.ncbi.nlm.nih.gov/pubmed/11923847?dopt=Abstract %R 10.1038/nbt0402-393 %0 Journal Article %J Nat Genet %D 2001 %T Assembly of microarrays for genome-wide measurement of DNA copy number. %A A Snijders %A Nowak, N %A Segraves, R %A Blackwood, S %A Brown, N %A Conroy, J %A Hamilton, G %A Hindle, A K %A Huey, B %A Kimura, K %A Law, S %A Myambo, K %A Palmer, J %A Ylstra, B %A Yue, J P %A Gray, J W %A Jain, A N %A Pinkel, D %A Albertson, D G %K Aneuploidy %K Chromosomes, Artificial, Bacterial %K Cloning, Molecular %K Female %K Gene Dosage %K Genome, Human %K Genomics %K Humans %K Male %K Oligonucleotide Array Sequence Analysis %K Polymerase Chain Reaction %K Polyploidy %K Tumor Cells, Cultured %K X Chromosome %X

We have assembled arrays of approximately 2,400 BAC clones for measurement of DNA copy number across the human genome. The arrays provide precise measurement (s.d. of log2 ratios=0.05-0.10) in cell lines and clinical material, so that we can reliably detect and quantify high-level amplifications and single-copy alterations in diploid, polyploid and heterogeneous backgrounds.

%B Nat Genet %V 29 %P 263-4 %8 2001 Nov %G eng %N 3 %R 10.1038/ng754 %0 Journal Article %J Lab Invest %D 2001 %T Comparative genomic hybridization of microdissected familial ovarian carcinoma: two deleted regions on chromosome 15q not previously identified in sporadic ovarian carcinoma. %A Zweemer, R P %A Ryan, A %A A Snijders %A Hermsen, M A %A Meijer, G A %A Beller, U %A Menko, F H %A Jacobs, I J %A Baak, J P %A Verheijen, R H %A Kenemans, P %A van Diest, P J %K BRCA2 Protein %K Chromosomes, Human, Pair 15 %K Female %K Genes, BRCA1 %K Genetic Markers %K Humans %K Nucleic Acid Hybridization %K Ovarian Neoplasms %K Sequence Deletion %X

The vast majority of familial ovarian cancers harbor a germline mutation in either the breast cancer gene BRCA1 or BRCA2 tumor suppressor genes. However, mutations of these genes in sporadic ovarian cancer are rare. This suggests that in contrast to hereditary disease, BRCA1 and BRCA2 are not commonly involved in sporadic ovarian cancer and may indicate that there are two distinct pathways for the development of ovarian cancer. To characterize further differences between hereditary and sporadic cancers, the comparative genomic hybridization technique was employed to analyze changes in copy number of genetic material in a panel of 36 microdissected hereditary ovarian cancers. Gains at 8q23-qter (18 of 36, 5 cases with high-level amplifications), 3q26.3-qter (18 of 36, 2 cases with high-level amplifications), 11q22 (11 of 36) and 2q31-32 (8 of 36) were most frequent. Losses most frequently occurred (in decreasing order of frequency) on 8p21-pter (23 of 36), 16q22-pter (19 of 36), 22q13 (19 of 36), 9q31-33 (16 of 36), 12q24 (16 of 36), 15q11-15 (16 of 36), 17p12-13 (14 of 36), Xp21-22 (14 of 36), 20q13 (13 of 36), 15q24-25 (12 of 36), and 18q21 (12 of 36). Comparison with the literature revealed that the majority of these genetic alterations are also common in sporadic ovarian cancer. Deletions of 15q11-15, 15q24-25, 8p21-ter, 22q13, 12q24 and gains at 11q22, 13q22, and 17q23-25, however, appear to be specific to hereditary ovarian cancer. Aberrations at 15q11-15 and 15q24-25 have not yet been described in familial ovarian cancer. In these regions, important tumor suppressor genes, including the hRAD51 gene, are located. These and other yet unknown suppressor genes may be involved in a specific carcinogenic pathway for familial ovarian cancer and may explain the distinct clinical presentation and behavior of familial ovarian cancer.

%B Lab Invest %V 81 %P 1363-70 %8 2001 Oct %G eng %N 10 %0 Journal Article %J Cancer Res %D 2001 %T Epistatic interactions between skin tumor modifier loci in interspecific (spretus/musculus) backcross mice. %A Nagase, H %A Jiang-Hua Mao %A de Koning, J P %A Minami, T %A Balmain, A %K Animals %K Epistasis, Genetic %K Female %K Genetic Linkage %K Genetic Predisposition to Disease %K Inbreeding %K Male %K Mice %K Papilloma %K Skin Neoplasms %X

The development of cancer is influenced both by exposure to environmental carcinogens and by the host genetic background. Epistatic interactions between genes are important in determining phenotype in plant and animal systems and are likely to be major contributors to cancer susceptibility in humans. Several tumor modifier loci have been identified from studies of mouse models of human cancer, and genetic interactions between modifier loci have been detected by genome scanning using recombinant congenic strains of mice (R. Fijneman et al., Nat. Genet., 14: 465-467, 1996; T. van Wezel et al., Nat. Genet., 14: 468-470, 1996; W. N. Frankel et al., Nat. Genet., 14, 371-373, 1996). We demonstrate here that strong genetic interactions between skin tumor modifier loci can be detected by hierarchical whole genome scanning of a complete interspecific backcross [outbred Mus spretus X Mus musculus (NIH/Ola)]. A locus on chromosome 7 (Skts1) showed a highly significant interaction with Skts5 on chromosome 12 (P < 10(-16)), whereas additional significant interactions were detected between loci on chromosomes 4 and 5, and 16 and 15. Some of these quantitative trait loci and their interactions, in particular the Skts1-Skts5 interaction, were confirmed in two completely independent backcrosses using inbred spretus strains (SEG/Pas and SPRET/Ei) and NIH/Ola. These results, therefore, illustrate the general use of interspecific crosses between Mus musculus and Mus spretus for the detection of strong genetic interactions between tumor modifier genes.

%B Cancer Res %V 61 %P 1305-8 %8 2001 Feb 15 %G eng %N 4 %1 http://www.ncbi.nlm.nih.gov/pubmed/11245425?dopt=Abstract %0 Journal Article %J Br J Cancer %D 2000 %T Advanced-stage cervix cancer: rapid tumour growth rather than late diagnosis. %A Symonds, P %A Bolger, B %A Hole, D %A Jiang-Hua Mao %A Cooke, T %K Adult %K Aged %K Biopsy %K Bromodeoxyuridine %K DNA %K Female %K Flow Cytometry %K Humans %K Middle Aged %K Ploidies %K Time Factors %K Uterine Cervical Neoplasms %K Vaginal Smears %X

Either diagnostic delay or tumour biology are possible factors governing the degree of spread at diagnosis of cervical cancer. To try to identify the most important parameter contributing to advanced stage, the duration of symptoms were recorded from patients scheduled for radiotherapy (n = 141) or radical hysterectomy (n = 36). In 146 cases tumour proliferation rates were evaluated following in vivo labelling with the DNA precursor BrdUrd. For symptomatic patients there was no association between duration of symptoms and stage at presentation. There was a significant trend for patients with increasing tumour stage to have more rapidly proliferating tumours with higher mean labelling index (LI) measurements (P = 0.001) and a shorter mean potential doubling time (Tpot) (P = 0.023). Socio economic deprivation may be associated with shorter Tpot values. The conclusion from this data is that stage at diagnosis is more dependent on the biological behaviour of the tumour, as expressed by proliferation rates, than delay in presentation.

%B Br J Cancer %V 83 %P 566-8 %8 2000 Sep %G eng %N 5 %1 http://www.ncbi.nlm.nih.gov/pubmed/10944592?dopt=Abstract %R 10.1054/bjoc.2000.1336 %0 Journal Article %J J Clin Pathol %D 2000 %T Apoptosis in cervical squamous carcinoma: predictive value for survival following radiotherapy. %A Paxton, J R %A Bolger, B S %A Armour, A %A Symonds, R P %A Jiang-Hua Mao %A Burnett, R A %K Analysis of Variance %K Apoptosis %K Carcinoma, Squamous Cell %K Female %K Humans %K Predictive Value of Tests %K Survival Rate %K Uterine Cervical Neoplasms %X

BACKGROUND: Apoptosis, or programmed cell death, can be induced by radiotherapy. The extent of apoptosis in a tumour before treatment may have important implications for response to radiotherapy and long term survival.

AIM: To examine the extent of apoptosis in tumour tissue from patients with squamous carcinoma of the cervix before radiotherapy, and to correlate this with response to treatment and prognosis.

METHODS: The percentage of apoptotic cells was assessed in 146 carcinomas of the cervix from patients scheduled to receive radiotherapy. The CAS 200 static image analysis system was used to count the number of tumour nuclei per high power field, while the numbers of apoptotic cells in the same field were visualised simultaneously on the image analyser and recorded manually.

RESULTS: The median apoptotic level was 0.73%. Patients were divided into two groups around the median. There was no statistically significant difference in outcome between the two groups as determined by long term survival following radiotherapy.

CONCLUSIONS: The CAS 200 static image analyser system can be used to assist in the rapid semiautomated assessment of apoptosis in conventionally prepared tissue. The results suggest that the apoptotic state of a tumour before treatment is of no value in predicting response to radiotherapy and subsequent prognosis. Tumour stage, size, and BrdU labelling index, as a measure of proliferation rate, remain the most important prognostic factors in terms of predicting local tumour control.

%B J Clin Pathol %V 53 %P 197-200 %8 2000 Mar %G eng %N 3 %1 http://www.ncbi.nlm.nih.gov/pubmed/10823138?dopt=Abstract %0 Journal Article %J BJOG %D 2000 %T Factors affecting fetal weight distribution in women with type I diabetes. %A Johnstone, F D %A Jiang-Hua Mao %A Steel, J M %A Prescott, R J %A Hume, R %K Birth Weight %K Diabetes Mellitus, Type 1 %K Female %K Fetal Weight %K Hemoglobin A, Glycosylated %K Hemoglobinuria %K Humans %K Hypoglycemia %K Infant, Newborn %K Pregnancy %K Pregnancy in Diabetics %K Prospective Studies %K Risk Factors %K Scotland %K Smoking %X

OBJECTIVE: To identify factors independently affecting fetal weight in women with type I diabetes.

DESIGN: Prospectively recorded data in consecutive women with type I diabetes, between 1975-1992.

SETTING: Simpson Memorial Maternity Hospital, Edinburgh. Population Three hundred and two pregnancies with type I diabetes identified before pregnancy, with antenatal care and delivery in the Simpson Memorial Maternity Hospital, a singleton pregnancy, and the same diabetic physician.

METHODS: Normal ranges for birthweight were established for the total hospital population. All cases and the total population had pregnancy dating by ultrasound. The relation between standardised birthweight and explanatory variables was investigated using correlation analysis, t tests and chi2 tests as appropriate, and subsequently using multiple linear regression.

RESULTS: Standardised birthweight in cases, compared with the reference population, showed a unimodal, approximately normal distribution, markedly shifted to the right (mean + 1.26 SD). The most predictive variable was glycated haemoglobin concentration at 27-33 weeks, which explained 6.3% of the birthweight variance, while smoking explained 2.7% and maternal weight 2.0%. There was a trend towards a negative relationship with glycated haemoglobin concentration at 6-12 weeks. Smoking and glycated haemoglobin concentration were strongly intercorrelated.

CONCLUSIONS: Most of the variance in standardised birthweight remains unexplained, but glycated haemoglobin concentration at 27-33 weeks is the most powerful explanatory variable. Possible reasons why there is not a stronger relationship between markers of maternal glycaemia and birthweight are discussed.

%B BJOG %V 107 %P 1001-6 %8 2000 Aug %G eng %N 8 %1 http://www.ncbi.nlm.nih.gov/pubmed/10955432?dopt=Abstract %0 Journal Article %J Clin Exp Immunol %D 1998 %T Enhanced prostaglandin E2 production by monocytes in atopic dermatitis (AD) is not accompanied by enhanced production of IL-6, IL-10 or IL-12. %A A Snijders %A Van der Pouw Kraan, T C %A Engel, M %A Wormmeester, J %A Widjaja, P %A Zonneveld, I M %A Bos, J D %A Kapsenberg, M L %K Adolescent %K Adult %K Cells, Cultured %K Dermatitis, Atopic %K Dinoprostone %K Female %K Humans %K Interferon-gamma %K Interleukin-10 %K Interleukin-12 %K Interleukin-6 %K Interleukins %K Leukocytes, Mononuclear %K Lipopolysaccharides %K Lymphocyte Activation %K Male %K Middle Aged %K Stimulation, Chemical %X

AD is associated with a bias of the T helper cells to show increased IL-4 and reduced interferon-gamma (IFN-gamma) production. The production of IFN-gamma and IL-4 and the development of Th cells into either high IFN-gamma or high IL-4 producers is strongly influenced by factors produced by antigen-presenting cells (APC), like IL-12 and prostaglandin E2 (PGE2). IL-12 selectively enhances IFN-gamma production and favours the development of IFN-gamma-producing Th cells, whereas PGE2 selectively inhibits IFN-gamma production by Th cells. The aim of this study was to test whether the increased IL-4/IFN-gamma production ratio by Th cells in AD can be explained by an increased PGE2/IL-12 production ratio by the APC. Monocytes were used as APC source. PGE2 and IL-12 production by lipopolysaccharide (LPS)-stimulated monocytes from 12 AD patients and 12 non-atopic controls was determined using two complementary experimental systems, whole blood cultures and purified monocytes. In addition, we determined IL-6 production as a measure of monocyte activation, and IL-10 production because IL-12 production by monocytes is highly influenced by endogenously produced IL-10. The monocytes from AD patients showed normal production levels of IL-6 and IL-10, a two-fold, but non-significant decrease in IL-12 production, and a significantly (three-fold) higher PGE2 production than those from non-atopic controls. Here we show for the first time that enhanced PGE2 production by monocytes in AD is not accompanied by a general rise in cytokine production. We conclude that AD is indeed associated with an increased PGE2/IL-12 production ratio by monocytes.

%B Clin Exp Immunol %V 111 %P 472-6 %8 1998 Mar %G eng %N 3 %0 Journal Article %J Math Biosci %D 1997 %T A two-stage model for childhood acute lymphoblastic leukemia: application to hereditary and nonhereditary leukemogenesis. %A Wheldon, E G %A Lindsay, K A %A Wheldon, T E %A Jiang-Hua Mao %K Aging %K Child %K Child, Preschool %K Embryo, Mammalian %K Female %K Humans %K Incidence %K Male %K Middle Aged %K Models, Genetic %K Models, Theoretical %K Mutation %K Precursor Cell Lymphoblastic Leukemia-Lymphoma %K Sex Factors %K Stochastic Processes %X

A differential equation model is developed to represent a two-stage mutational process leading to childhood acute lymphoblastic leukemia (ALL). Leukemogenesis is modeled as transformation of target stem cells that initially grow rapidly in the embryo but plateau and then decline in postnatal childhood. Inheritance of the first of two leukemogenic mutations is allowed as a possibility in a small minority of leukemic patients who would characteristically develop leukemia at an early age. The model is shown to be capable of providing good fits to incidence data for childhood ALL; these fits allow estimation of some parameters of the model. The analysis shows that individuals inheriting one of the two mutations necessary for ALL would be likely to experience "multiclonal leukemogenesis"; that is, the parallel development of several leukemic clones arising from multiple independent leukemic events. The model suggests that between two and ten such clones would typically have developed in such individuals by the time of diagnosis. The main conclusions of the deterministic investigation were confirmed by stochastic modeling. The existence of multiclonal leukemogenesis is in principle testable by molecular biological methods (clonality analysis) that rely on the random inactivation of one of two X-chromosomes in normal female subjects. It is expected that the mathematical methods developed here will also be useful for more general (N-stage) models of malignant transformation of stem cell populations undergoing growth or decline.

%B Math Biosci %V 139 %P 1-24 %8 1997 Jan 1 %G eng %N 1 %1 http://www.ncbi.nlm.nih.gov/pubmed/9111777?dopt=Abstract %0 Journal Article %J Br J Obstet Gynaecol %D 1996 %T Clinical and ultrasound prediction of macrosomia in diabetic pregnancy. %A Johnstone, F D %A Prescott, R J %A Steel, J M %A Jiang-Hua Mao %A Chambers, S %A Muir, N %K Birth Weight %K Diabetes, Gestational %K Female %K Fetal Macrosomia %K Forecasting %K Gestational Age %K Humans %K Parity %K Pregnancy %K Pregnancy in Diabetics %K Prospective Studies %K Sensitivity and Specificity %K Ultrasonography, Prenatal %X

OBJECTIVE: To study prospectively the prediction power, at different gestations, of clinical and ultrasound measurements for fetal size in diabetic pregnancy.

SETTING: A large combined obstetric diabetic clinic in a teaching hospital.

PARTICIPANTS: One hundred and eighty-one pregnancies in which women had scans at least two of three specific time points and who were delivered of singletons after 34 weeks: 73% were pre-gestational insulin-dependent diabetics, the others were pre-gestational White class A or gestational diabetics.

INTERVENTIONS: Clinical estimates of fundal height and fetal size and ultrasound estimates of abdominal circumference and head circumference were routinely carried out at gestational ages of 28, 34 and 38 weeks or before delivery.

MAIN OUTCOME MEASURES: Standardised birthweight, corrected for gestation and parity. The relation with clinical and ultrasound measurements was investigated using multiple linear regression and the capability of the measurements to predict macrosomic births (> 95th centile of normals) using receiver-operator characteristic curves.

RESULTS: All measurements are poor predictors of eventual standardised birthweight. Prediction improves with closeness to delivery. Prediction is significantly improved by adding ultrasound to clinical information, but at 34 weeks or later this only contributes 8% of the variance. There is no difference in the prediction power for macrosomia between clinical and ultrasound measurements.

CONCLUSIONS: Even regular serial scanning and clinical examination will not always diagnose the macrosomic fetus in diabetic pregnancy. In our hands, clinical examination is as predictive as ultrasound measurements. Ultrasound does add to clinical prediction power but only to a small extent. Ultrasound should be used in a selected way, as defined by clinical findings, and with recognition and understanding of the errors and biases involved.

%B Br J Obstet Gynaecol %V 103 %P 747-54 %8 1996 Aug %G eng %N 8 %1 http://www.ncbi.nlm.nih.gov/pubmed/8760702?dopt=Abstract %0 Journal Article %J Br J Obstet Gynaecol %D 1996 %T Smoking during pregnancy: the dose dependence of birthweight deficits. %A Ellard, G A %A Johnstone, F D %A Prescott, R J %A Ji-Xian, W %A Jian-Hua, M %K Adult %K Birth Weight %K Dose-Response Relationship, Drug %K Female %K Humans %K Logistic Models %K Maternal Age %K Medical History Taking %K Nicotine %K Organ Size %K Parity %K Placenta %K Pregnancy %K Sex Factors %K Smoking %K Truth Disclosure %K Weight Gain %X

OBJECTIVE: To assess whether a simple urine based estimate of relative daily nicotine intake could predict smoking related birthweight deficits more accurately than self-reported cigarette consumption.

DESIGN: Active smokers were identified by a simple qualitative colorimetric urine test procedure and their relative nicotine intakes assessed by determining the ratios of the urinary concentrations of nicotine plus its metabolites to creatinine using automated colorimetric methods.

SETTING: A large teaching hospital.

PARTICIPANTS: Three thousand and thirty-eight mothers from whom smoking histories had been elicited and who gave birth to live singleton babies after 28 weeks of gestation.

MAIN OUTCOME MEASURES: Birthweights (adjusted for maternal weight, maternal age, baby's sex, parity and length of gestation), maternal weight gains during pregnancy and placental weights.

RESULTS: The adjusted birthweight deficits of babies born to proven active smokers averaged 226 g (95% confidence interval 194 g to 258 g), but dose dependent effects were only apparent when nicotine intake was based on urinary nicotine metabolites/creatinine ratios. Among the smokers, adjusted birthweights fell linearly with increasing nicotine intakes but gave a predicted mean birthweight for nonsmokers that was 102 g (95% CI 50 g to 154 g) lighter than that actually found (P < 0.0001). Maternal weight gains during pregnancy were substantially reduced in smokers and correlated more closely with urinary nicotine metabolite excretions than with reported daily cigarette consumptions. Placental weights were unaffected by smoking.

CONCLUSION: There was a closer dose-effect relationship between birthweight deficits and urinary nicotine metabolites/creatinine ratios than with self-reported daily cigarette consumptions. The influence of nicotine exposure on birthweight appears to be biphasic, with one mechanism operating at very low levels of tobacco smoke intake and the other causing seemingly linearly related effects over the whole range of nicotine intakes of active smokers. These findings support recent evidence that passive smoking can cause substantial birthweight deficits.

%B Br J Obstet Gynaecol %V 103 %P 806-13 %8 1996 Aug %G eng %N 8 %1 http://www.ncbi.nlm.nih.gov/pubmed/8760712?dopt=Abstract %0 Journal Article %J Thorax %D 1995 %T An open, prospective comparison of beta 2 agonists given via nebuliser, Nebuhaler, or pressurised inhaler by ambulance crew as emergency treatment. %A Campbell, I A %A Colman, S B %A Jiang-Hua Mao %A Prescott, R J %A Weston, C F %K Acute Disease %K Administration, Inhalation %K Adolescent %K Adult %K Aged %K Albuterol %K Analysis of Variance %K Emergencies %K Emergency Medical Services %K Female %K Humans %K Male %K Middle Aged %K Nebulizers and Vaporizers %K Prospective Studies %K Respiratory Sounds %K Terbutaline %K Wales %X

BACKGROUND: The merits of the use of beta 2 agonists by ambulance crew and best methods of delivery have not been fully explored.

METHODS: A prospective comparison has been made of treatments applied in three districts in South Wales (200 micrograms salbutamol by pressurised inhaler, 5 mg salbutamol via nebuliser, and 5 mg terbutaline via Nebuhaler) by emergency ambulance personnel to acutely wheezy patients en route to hospital. Pulse rate, respiratory rate, peak expiratory flow rate (PEFR), and breathlessness scored on a visual analogue scale were compared before and after treatment. Data were collected on diagnosis, artificial ventilation, cardiorespiratory arrest, and death.

RESULTS: Thirty eight patients received salbutamol inhaler, 51 salbutamol via nebuliser, and 41 terbutaline via Nebuhaler. There were greater reductions in respiratory rate and breathlessness score and more improvement in PEFR in the group receiving nebulised salbutamol than in the other two groups. No patient was ventilated and of the five deaths none was caused by asthma.

CONCLUSIONS: For wheezy, breathless patients treated en route to hospital by emergency ambulance personnel, 5 mg salbutamol given by an oxygen-driven nebuliser was more effective than either 5 mg terbutaline via a Nebuhaler or 200 micrograms salbutamol via a pressurised inhaler.

%B Thorax %V 50 %P 79-80 %8 1995 Jan %G eng %N 1 %1 http://www.ncbi.nlm.nih.gov/pubmed/7886655?dopt=Abstract %0 Journal Article %J Obstet Gynecol %D 1994 %T Insulin requirements during pregnancy in women with type I diabetes. %A Steel, J M %A Johnstone, F D %A Hume, R %A Jiang-Hua Mao %K Diabetes Mellitus, Type 1 %K Female %K Humans %K Insulin %K Pregnancy %K Pregnancy in Diabetics %K Pregnancy Outcome %K Pregnancy Trimester, Third %K Regression Analysis %K Weight Gain %X

OBJECTIVES: To document individual variations in the rise in insulin requirements during type I diabetic pregnancies, to relate the degree of increase to maternal characteristics and fetal outcome, and to examine these factors in a subgroup of patients experiencing a large fall in insulin requirement in the third trimester.

METHODS: Insulin dose was documented in 237 pregnancies in women with type I diabetes. Multiple regression analysis was performed to identify significant associations with maternal and fetal characteristics. Eighteen pregnancies with a fall in insulin requirement of 30% or more in the third trimester were considered in detail.

RESULTS: The mean absolute increase in insulin requirement was 52 units. The degree of rise was significantly related to maternal weight gain between 20-29 weeks and maternal weight at booking, and was inversely related to duration of diabetes. It was not related to the degree of diabetes control, complications of pregnancy, White class, or outcome of pregnancy. In the 18 women experiencing a large fall in insulin requirement, there was no relation with maternal characteristics or fetal outcome.

CONCLUSION: There is a wide individual variation in the change in insulin requirements in type I diabetic pregnancy. The degree of increase is related only to maternal weight gain during weeks 20-29 and maternal weight at booking, and is inversely related to duration of diabetes. Large falls in insulin requirement remain unexplained and may not be associated with placental failure.

%B Obstet Gynecol %V 83 %P 253-8 %8 1994 Feb %G eng %N 2 %1 http://www.ncbi.nlm.nih.gov/pubmed/8290190?dopt=Abstract %0 Journal Article %J Leuk Lymphoma %D 1993 %T Identification of prognostic groups in follicular lymphoma. The Scotland and Newcastle Lymphoma Group Therapy Working Party. %A Cameron, D A %A Leonard, R C %A Jiang-Hua Mao %A Prescott, R J %K Adult %K Age Factors %K Aged %K Female %K Humans %K Lymphoma, Follicular %K Male %K Middle Aged %K Prognosis %K Survival Rate %X

Follicular lymphoma is often seen as an indolent disease with a reasonable medium-term survival. We have used the information in the Scotland and Newcastle Lymphoma Group database to devise an index which is easily calculated and differentiates patients into poor, intermediate and good prognostic groups with 5 years survivals of 24%, 61% and 86% respectively. The key factors at presentation are age, ECOG performance status, stage and the presence or absence of B symptoms or gastro-intestinal tract involvement. The use of such an index permits early identification of patients with a poor prognosis for whom more intensive treatment could be offered.

%B Leuk Lymphoma %V 10 %P 89-99 %8 1993 May %G eng %N 1-2 %1 http://www.ncbi.nlm.nih.gov/pubmed/8374528?dopt=Abstract %R 10.3109/10428199309147361