NCT02158039.).

For patients with primary lung cancer, accurate determination of the tumor type significantly influences treatment decisions. However, techniques and methods for lung cancer typing lack standardization. In particular, owing to limited tumor sample amounts and the poor quality of some samples, the classification of primary lung cancers using small preoperative biopsy specimens presents a diagnostic challenge using current tools. We previously described a microRNA-based assay (miRview squamous; Rosetta Genomics Ltd., Rehovot, Israel) that accurately differentiates between squamous and nonsquamous non-small cell lung cancer. Herein, we describe the development and validation of an assay that differentiates between the four main types of lung cancer: squamous cell carcinoma, nonsquamous non-small cell lung cancer, carcinoid, and small cell carcinoma. The assay, miRview lung (Rosetta Genomics Ltd.), is based on the expression levels of eight microRNAs, measured using a sensitive quantitative RT-PCR platform. It was validated on an independent set of 451 samples, more than half of which were preoperative cytologic samples (fine-needle aspiration and bronchial brushing and washing). The assay returned a result for more than 90% of the samples with overall accuracy of 94% (95% CI, 91% to 96%), with similar performance observed in pathologic and cytologic samples. Thus, miRview lung is a simple and reliable diagnostic assay that offers an accurate and standardized classification tool for primary lung cancer using pathologic and cytologic samples.

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This study is to examine our hypothesis that CD4+CD25 high Foxp3+ regulatory T cells (Tregs) have an interleukin-2 (IL-2) withdrawal-triggered apoptosis pathway, and modulation of Treg apoptosis pathway affects development of vascular inflammation. We found that pro-apoptotic protein Bax upregulation in Tregs is induced by IL-2 withdrawal. Treg apoptosis induced by IL-2 withdrawal is inhibited by a Bax inhibitor, suggesting that highly expressed Bax is functional. To define the role of upregulated Bax in Treg apoptosis, we established a Tregs-specific Bax transgenic mouse model. Enforced expression of Bax in Tregs promotes Treg apoptosis triggered by IL-2 withdrawal and other apoptosis stimuli, suggesting pro-apoptotic role of highly expressed Bax in wild-type Tregs. Finally, higher expression of Bax in Tregs decreases the striking threshold of vascular inflammation due to the failure of suppression of inflammatory cells resulting from Treg apoptosis. These results have demonstrated the proof of principle that the modulation of Tregs apoptosis/survival could be used as a new therapeutic approach for inflammatory cardiovascular diseases. KeywordsRegulatory T Cells; Apoptosis; Bax; Inflammation; Vasculitis INTRODUCTIONCD4+CD25 high Foxp3+ regulatory T cells (Tregs), comprising 5-10% of CD4+ T cells(1), exhibit potent immunosuppressive functions(2) in the regulation of autoimmunity and inflammatory atherosclerosis (3,4). Naturally occurring Treg cells (thymus-generated, nTreg cells), as an independent subset, are engaged in the maintenance of immunological selftolerance and inhibition of various immune responses (5) and inflammatory atherogenesis. nTregs (Tregs in the rest of paper) appear to have specific apoptosis pathways since Treg cells have higher susceptibility to apoptosis (6), especially to IL-2 withdrawal-induced apoptosis. Tregs are poor IL-2 producers(7), implying that insufficient paracrine IL-2 supply to Tregs in pathological conditions could be responsible for higher susceptibility of Tregs to apoptosis. However, intracellular regulation of IL-2 insufficiency-triggered Treg apoptosis remains poorly defined. The Bcl-2/Bcl-xL protein family members play a central role in the Send correspondence to: Xiao-Feng Yang, Department of Pharmacology, Temple University School of Medicine, 3420 North Broad Street, MRB, Rm. 325, Philadelphia, PA 19140, Tel: 215-707-5985, Fax: 215-707-7068 MATERIALS AND METHODS Construction of CD25+ T cell-targeting mouse IL-2Rα promoter-Bax transgenic miceMouse IL-2 receptor α chain (CD25) promoter −2539 to +93 (GenBank Accession Number: M16398) vector pmIL2Rα-CAT1 (6.9 kb) was generously provided by P. Reichenbach and M. Nabholz(14). The construction of the CD25+ T cell targeting vector pCD25-Tg was described previously (10). The transgenic vector pCD25-Bax-Tg was verified by DNA sequencing by SeqWright Company (Houston, TX). The 3.818 kb transgenic DNA fragment "Nru I-CD25 promoter-C-Myc-Bax-Sex AI" was prepared by digesting the pCD25-Bax-Tg vector with three restrict...

2. Introduction. 3. Human Breast Epithelial Cells (HBEC) in Culture 4. Factors Influencing Susceptibility of HBEC to Cell Transformation 4.1. Lobular differentiation 4.2. Genetic predisposition 4.3. Cell immortalization 5. Molecular Mechanisms of Cell Immortalization 5.1. Activation of telomerase 5.2. Abrogation of cell cycle control 5.3. Genes preferentially expressed during cell immortalization 6. Molecular Mechanisms of Cell Transformation 6.1. Epigenetic mechanisms 6.2. Genetic mechanisms 7. Genomic changes in Immortalization and Transformation of HBEC 7.1. Genomic changes in cell immortalization 7.2. Genomic changes in cell transformation 8. Genomic Changes in Human Breast Lesions 9. Functional Roles of Chromosomes 11 and 17 in Transformed Phenotype Expression of HBEC 10. Summary and Perspectives 11. Acknowledgments 12. References

Gadd45a plays a pivotal role as a stress sensor that modulates cellular responses to various stress stimuli including oncogenic stress. We reported that the stress sensor Gadd45a gene functions as a tumor suppressor in Ras-driven breast tumorigenesis via increasing JNK-mediated apoptosis and p38-mediated senescence. In contrast, here, we show that Gadd45a promotes Myc-driven breast cancer by negatively regulating MMP10 via GSK3 b/b-catenin signaling, resulting in increased tumor vascularization and growth. These novel findings indicate that Gadd45a functions as either tumor promoter or suppressor, is dependent on the oncogenic stress, and is mediated via distinct signaling pathways. Collectively, these novel findings highlight the significance of the type of oncogenic alteration on how stress response genes function during initiation and progression of tumorigenesis. Because Gadd45a is a target for BRCA1 and p53, these findings have implications regarding BRCA1/p53 tumor suppressor functions. Cancer Res; 70(23); 9671-81. Ó2010 AACR.

Preeclampsia, which affects approximately 5-8% of all pregnancies and is one of the leading causes of maternal and fetal morbidity and mortality, is a pregnancy induced complex of multiple pathological changes, including elevated blood pressure, proteinuria and edema manifested after 20 weeks gestation. There is growing evidence that placental stresses during pregnancy, notably hypoxia, and an increase in circulating soluble Flt-1 (sFlt-1) are important in the etiopathogenesis of preeclampsia. How placental stress results in elevated sFlt-1 expression is currently unknown. Here we provide novel data implicating the Gadd45a stress sensor protein as an upstream modulator of pathophysiological changes observed in preeclampsia. It is shown that Gadd45a expression and activation of its downstream effector p38 kinase are elevated in preeclamptic placentas compared to non-preeclamptic controls, and correlate with elevated sFlt-1. Furthermore, a regulatory loop is demonstrated where stress, including hypoxia, IL-6 or hypertonic stress, caused induction of Gadd45a, leading to p38 activation and ultimately increasing sFlt-1 secretion in endothelial cells. These data provide a compelling working frame to further test the role of Gadd45 stress sensors in the etiology of preeclampsia, and set the stage for considering novel therapeutic regimens, including p38 inhibitors, for treatment of preeclampsia.

Purpose: Clear cell ovarian carcinoma (CCOC) is an aggressive disease that often demonstrates resistance to standard chemotherapies. Approximately 25% of CCOC show a strong APOBEC mutation signature. Here, we determine which APOBEC3 enzymes are expressed in CCOC, establish clinical correlates, and identify a new biomarker for detection and intervention.Experimental Designs: APOBEC3 expression was analyzed by immunohistochemistry and RT-qPCR in a pilot set of CCOC specimens (n=9 tumors). The immunohistochemistry analysis of APOBEC3B was extended to a larger cohort to identify clinical correlates (n=48). Dose response experiments with platinum-based drugs in CCOC cell lines and carboplatin treatment of patient-derived xenografts (PDX) were done to address mechanistic linkages.Results: One DNA deaminase, APOBEC3B, is overexpressed in a formidable subset of CCOC tumors and is low or absent in normal ovarian and fallopian tube epithelial tissues. High APOBEC3B expression associates with improved progression-free survival (p=0.026) and moderately with overall survival (p=0.057). Cell-based studies link APOBEC3B activity and subsequent uracil processing to sensitivity to cisplatin and carboplatin. PDX studies extend this mechanistic relationship to CCOC tissues. Conclusions:These studies demonstrate that APOBEC3B is overexpressed in a subset of CCOC and, contrary to initial expectations, associated with improved (not worse) clinical outcomes. A likely molecular explanation is that DNA damage caused APOBEC3B sensitizes cells to additional genotoxic stress by cisplatin. Thus, APOBEC3B is a molecular determinant and a candidate predictive biomarker of the therapeutic response to platinum-based chemotherapy. These findings may have broader translational relevance, as APOBEC3B is overexpressed in many different cancer types.