bluuebunny/crossref_metadata_2025 huggingface dataset:
Yoga and Emotion Regulation in High School Students: A Randomized Controlled Trial
Authors: Daly Leslie A. , Haden Sara C. , Hagins Marshall , Papouchis Nicholas , Ramirez Paul Michael
Published: August 2015

Middle adolescents (15–17 years old) are prone to increased risk taking and emotional instability. Emotion dysregulation contributes to a variety of psychosocial difficulties in this population. A discipline such as yoga offered during school may increase emotion regulation, but research in this area is lacking. This study was designed to evaluate the impact of a yoga intervention on the emotion regulation of high school students as compared to physical education (PE). In addition, the potential mediating effects of mindful attention, self-compassion, and body awareness on the relationship between yoga and emotion regulation were examined. High school students were randomized to participate in a 16-week yoga intervention (n=19) or regular PE (n=18). Pre-post data analyses revealed that emotion regulation increased significantly in the yoga group as compared to the PE group (F(1,32) = 7.50,p=.01, and eta2= .19). No significant relationship was discovered between the changes in emotion regulation and the proposed mediating variables. Preliminary results suggest that yoga increases emotion regulation capacities of middle adolescents and provides benefits beyond that of PE alone.
Immune Homeostasis in Epithelial Cells: Evidence and Role of Inflammasome Signaling Reviewed
Authors: Peeters Paul M. , Wouters Emiel F. , Reynaert Niki L.
Published: August 2015

The epithelium regulates the interaction between the noxious xenogenous, as well as the microbial environment and the immune system, not only by providing a barrier but also by expressing a number of immunoregulatory membrane receptors, and intracellular danger sensors and their downstream effectors. Amongst these are a number of inflammasome sensor subtypes, which have been initially characterized in myeloid cells and described to be activated upon assembly into multiprotein complexes by microbial and environmental triggers. This review compiles a vast amount of literature that supports a pivotal role for inflammasomes in the various epithelial barriers of the human body as essential factors maintaining immune signaling and homeostasis.
Paracrine Regulation of Steroidogenesis in Theca Cells by Granulosa Cells Derived from Mouse Preantral Follicles
Authors: Liu Xiaoqiang , Qiao Pengyun , Jiang Aifang , Jiang Junyi , Han Haiyan , Wang Li , Ren Chune
Published: August 2015

Interaction partners of follicular cells play a significant role in steroidogenesis, follicular formation, and development. Androgen secreted by theca cells (TCs) can initiate follicle development and ovulation and provide precursor materials for estrogen synthesis. Therefore, studies on ovarian microenvironment will not only lead to better understanding of the steroidogenesis but also have clinical significance for ovarian endocrine abnormalities such as hyperandrogenism in polycystic ovary syndrome (PCOS). This study applied the Transwell coculture model to investigate if the interaction between granulosa and theca cells may affect androgen production in theca cells. Concentrations of testosterone and androstenedione in the spent medium were measured by radioimmunoassay and enzyme linked immunosorbent assay, respectively. The results show that the coculture with granulosa cells (GCs) increases steroidogenesis in TCs. In addition, testosterone and androstenedione productions in response to LH stimulation were also increased in the coculture model. Significantly increased mRNA expressions of steroidogenic enzymes (Star,Cyp11a1,Cyp17a1, andHsd3b2) were observed in the cocultured TCs. Thus, GCs were capable of promoting steroidogenesis and LH responsiveness in TCs. This study provided a basis for further exploration of ovarian endocrine mechanism and pathologies.
Electrostatic Interactions between Complement Regulator CD46(SCR1-2) and Adenovirus Ad11/Ad21 Fiber Protein Knob
Authors: Chen Carl Z. , Gorham Ronald D. , Gaieb Zied , Morikis Dimitrios
Published: August 2015

Adenoviruses bind to a variety of human cells to cause infection. Both the B2 adenovirus 11 and B1 adenovirus 21 use protein knobs to bind to complement regulator CD46(SCR1-2) in order to gain entry into host cells. In each complex, the two proteins are highly negatively charged but bind to each other at an interface with oppositely charged surface patches. We computationally generated single-alanine mutants of charged residues in the complexes CD46(SCR1-2)-Ad11k and CD46(SCR1-2)-Ad21k. We used electrostatic clustering and Poisson-Boltzmann free energy calculations to propose a hypothesis on the role of electrostatics in association. Our results delineate specific interfacial electrostatic interactions that are critical for association in both CD46(SCR1-2)-Ad11k and CD46(SCR1-2)-Ad21k. These results will serve as a predictive tool in the selection of mutants with desired binding affinity in experimental mutagenesis studies. This study will also serve as a foundation for the design of inhibitors to treat adenovirus infections.
Abstract A06: The chemokine receptor CXCR4 is essential for the maintenance of T cell acute lymphoblastic leukemia.
Authors: Pitt Lauren A. , Tikhonova Anastasia N. , Trimarchi Thomas , King Bryan , Hu Hai , Gong Yixiao , Tsirigos Aris , Sanchez-Martin Marta , Littman Dan R. , Ferrando Adolfo , Morrison Sean J. , Fooksman David R. , Aifantis Iannis , Schwab Susan
Published: August 2015

T cell acute lymphoblastic leukemia (T-ALL) is an aggressive hematological malignancy arising in immature T cell progenitors. Despite improved survival rates, relapse remains a major therapeutic obstacle and intensive treatment regimens expose T-ALL patients to serious long-term side-effects. Hence more effective, less toxic treatment options are required. While the localization of leukemia cells determines the course of disease, little is known about T-ALL migration. Upon profiling chemokine receptor expression by T-ALL cells, we observed high surface levels of the CXCL12 receptor, CXCR4. Intravital imaging revealed that T-ALL cells are in direct contact with CXCL12-producing bone marrow stromal cells in vivo. Genetic targeting of CXCR4 in T-ALL cells resulted in a marked reduction of leukemia burden and significantly prolonged survival. Moreover, disruption of CXCR4 signaling with a small molecule inhibitor severely impaired T-ALL progression in mouse and human xenograft models of the disease. We found that CXCR4 promotes c-Myc expression, which is required in leukemia initiating cells. Given the relatively subtle role for CXCR4 in normal developing T cells, CXCR4 inhibition to our knowledge has not been proposed as a therapy for T cell malignancies. Our data strongly suggest that targeting CXCR4 could be a promising strategy for combating T-ALL.

Citation Format: Lauren A. Pitt, Anastasia N. Tikhonova, Thomas Trimarchi, Bryan King, Hai Hu, Yixiao Gong, Aris Tsirigos, Marta Sanchez-Martin, Dan R. Littman, Adolfo Ferrando, Sean J. Morrison, David R. Fooksman, Iannis Aifantis, Susan Schwab. The chemokine receptor CXCR4 is essential for the maintenance of T cell acute lymphoblastic leukemia. [abstract]. In: Proceedings of the AACR Special Conference on Hematologic Malignancies: Translating Discoveries to Novel Therapies; Sep 20-23, 2014; Philadelphia, PA. Philadelphia (PA): AACR; Clin Cancer Res 2015;21(17 Suppl):Abstract nr A06.
Abstract AS12: Integrating genomics, transcriptomics, and proteomics for the discovery of novel biomarkers to complement CA125 in ovarian cancer early detection
Authors: Boldt Clayton , Simmons Archana , Wang Hong , Baggerly Keith , Bast Robert , Hanash Samir
Published: August 2015

Recent work suggests that a 2-stage strategy incorporating serial measurements of the glycoprotein CA125 in patient serum and the Risk of Ovarian Cancer Algorithm (ROCA), followed by transvaginal ultrasound for women with high-risk scores, may have utility as a screening practice in the general population. However, CA125 only has a sensitivity of 50-60% for early stage disease, and is only present in approximately 80% of all ovarian cancers. Multiple marker panels have shown promise at increasing sensitivity and specificity over CA125 alone, but have not improved lead time for early cancer detection.

Therefore, we have taken an unbiased in-depth approach to identify novel plasma biomarkers to complement CA125 in ovarian cancer early detection. Using high-resolution quantitative mass spectrometry, we have profiled the plasma proteome of preclinical patient samples, taken 1 year prior to CA125 elevation and diagnosis, and healthy controls from the Normal Risk Ovarian Screening (NROS) study. With this approach, we have identified over 30 biomarker candidates that are elevated in at least 50% of preclinical samples relative to controls in triplicate experiments, including cancer-specific post-translational modifications.

As a means of preliminary validation of promising preclinical biomarkers, we have profiled the plasma proteome of 24 newly diagnosed, early stage ovarian cancer patients and 24 matched healthy controls. Further, half of these patients were selected with uninformative CA125 levels at diagnosis, allowing for the discovery of additional candidate biomarkers that may complement CA125.

Correlation of preclinical and newly diagnosed patient plasma data with publicly available genomic and transcriptomic data, as well as proteomic data previously collected from ovarian cancer cell lines resulted in a prioritized list of candidate protein biomarkers and autoantibodies in patient plasma that may improve detection and lead time over CA125. For the most promising candidates, monoclonal antibodies and MagPlex assays are being optimized in order to determine their performance as useful biomarkers in additional patient sera. Multimarker panels will be tested in a training set for the development of a screening algorithm that will be evaluated in preclinical sera to determine the optimized set that improves detection and lead time over CA125.

Citation Format: Clayton Boldt, Ph.D., Archana Simmons, Ph.D., Hong Wang, MS, Ph.D., Keith Baggerly, Ph.D., Robert Bast, Jr., M.D., Samir Hanash, M.D., Ph.D.. Integrating genomics, transcriptomics, and proteomics for the discovery of novel biomarkers to complement CA125 in ovarian cancer early detection [abstract]. In: Proceedings of the 10th Biennial Ovarian Cancer Research Symposium; Sep 8-9, 2014; Seattle, WA. Philadelphia (PA): AACR; Clin Cancer Res 2015;21(16 Suppl):Abstract nr AS12.
Abstract AS21: Ovarian carcinoma CDK12 mutations misregulate DNA repair genes via deficient formation and function of the Cdk12/CycK complex
Authors: Ekumi Kingsley M. , Paculova Hana , Pospichalova Vendula , Bosken Christian A. , Bryja Vitezslav , Geyer Matthias , Blazek Dalibor , Barboric Matjaz
Published: August 2015

The Cdk12/CycK complex promotes gene expression by phosphorylating the C-terminal domain of RNA polymerase II. In previous work we found that CDK12/CycK complex maintains genomic stability via regulation of expression of several key DNA damage response genes such as BRCA1, ATR, FANCD2 or FANCI. CDK12 is among only nine genes with recurrent somatic mutations in high-grade serous ovarian carcinoma. However, the influence of these mutations on the Cdk12/CycK complex and their link to cancerogenesis remain ill-defined. Here, we show that most mutations interfere with the Cdk12/CycK complex formation, rendering the kinase inactive. By examining the mutations within the Cdk12/CycK structure, we find that they likely provoke structural rearrangements detrimental to Cdk12 activation. Our mRNA expression analysis of the patient samples containing the CDK12 mutations identifies coordinated down-regulation of key DNA damage response genes. Accordingly, we observed that the mutant Cdk12 proteins fail to promote the repair of DNA double strand breaks via homologous recombination. Together, we provide the molecular basis of how mutated CDK12 ceases to function in ovarian carcinoma. We propose that CDK12 is a tumor suppressor of which the loss-of-function mutations may elicit defects in multiple DNA repair pathways, leading to genomic instability that underlies the genesis of the cancer..

Citation Format: Kingsley M. Ekumi, Hana Paculova,Vendula Pospichalova, Christian A. Bosken,Vitezslav Bryja, Matthias Geyer, Dalibor Blazek, Matjaz Barboric. Ovarian carcinoma CDK12 mutations misregulate DNA repair genes via deficient formation and function of the Cdk12/CycK complex [abstract]. In: Proceedings of the 10th Biennial Ovarian Cancer Research Symposium; Sep 8-9, 2014; Seattle, WA. Philadelphia (PA): AACR; Clin Cancer Res 2015;21(16 Suppl):Abstract nr AS21.
Abstract POSTER-BIOL-1308: Macrophage infiltration in high-grade serous carcinomas of humans and mice
Authors: Pejovic Tanja , Bean Yukie , Kim Jaeyeon , Matzuk Martin M. , Coffelt Seth B.
Published: August 2015

Over the past 10-15 years, focus on the immune system has reinvigorated cancer research and oncology. Both clinical and experimental evidence generated during this time has shown that immune cells and their molecular mediators represent viable targets for therapeutic manipulation. The immune cell composition of high-grade serous carcinoma (HGSC) consists primarily of macrophages and various T cell subsets. However, the functional contribution of each of these cell types is largely unknown. In order to gain insight into whether these cells are pro- or anti-tumorigenic, we generated tissue microarrays containing tumors from 169 patients with pretreated ovarian cancer and quantified the number of CD68+ macrophages in each biopsy. Tumors were divided into high- or low-infiltrated groups based on the median number of macrophages amongst all HGSCs. Interestingly, patients whose tumors were highly infiltrated with macrophages exhibited worsened overall survival, suggesting that macrophages function as pro-tumorigenic cells in these tumors. To investigate the role of macrophages in HGSC progression, we have begun to characterize their abundance and phenotype in tumors of Dicerflox/flox;Ptenflox/flox;Amhr2cre/+ mice, a genetically engineered mouse model of HGSC. Macrophages constituted the predominant stromal and immune cell population in both primary tumors and secondary metastatic lesions. By contrast, macrophages were only found at low frequency in normal oviduct and ovaries. Gene expression analysis of these Dicer/Pten-deficient mouse tumors also revealed the increase in many macrophage-associated genes, as compared with normal oviduct tissue. Taken together, our data from mice and humans strongly indicate a crucial role for macrophages in facilitating ovarian cancer progression. Current efforts are underway to understand how macrophages promote ovarian tumor progression.

Citation Format: Tanja Pejovic, Yukie Bean, Jaeyeon Kim, Martin M. Matzuk, Seth B. Coffelt. Macrophage infiltration in high-grade serous carcinomas of humans and mice [abstract]. In: Proceedings of the 10th Biennial Ovarian Cancer Research Symposium; Sep 8-9, 2014; Seattle, WA. Philadelphia (PA): AACR; Clin Cancer Res 2015;21(16 Suppl):Abstract nr POSTER-BIOL-1308.
Abstract POSTER-BIOL-1317: Releasing the brakes on ovarian cancer cell survival: epigenetic suppression of RGS10 in ovarian cancer chemoresistance
Authors: Hooks Shelley
Published: August 2015

G-protein signaling pathways promote cell survival, which is aberrantly enhanced in chemoresistant ovarian cancer. Regulator of G-protein Signaling (RGS) proteins limit the strength of G-protein signaling, and therefore indirectly control cell survival. We have previously shown that RGS10 suppresses ovarian cancer cell growth and limits survival in the presence of cytotoxic chemotherapeutic drugs. Further, we have shown that RGS10 expression is suppressed in cell models of ovarian cancer chemoresistance, suggesting that ovarian cancer cells silence RGS10 expression to release the brakes on their growth and survival. However, the mechanisms governing RGS10 expression in ovarian cancer cells are poorly understood. In this study, we fully investigate the molecular mechanism of RGS10 suppression in ovarian cancer acquired chemoresistance, revealing key roles for epigenetic regulation by DNA hypermethylation and histone deacetylation. Pharmacological inhibition of DNA methyl-transferases (DNMTs) increases RGS10 expression in chemoresistant ovarian cancer cells, suggesting potential regulation by DNA methylation. Further, bisulfite sequencing analysis identified a region of the RGS10-1 promoter with significantly enhanced DNA methylation in chemoresistant A2780-AD cells relative to parental A2780 cells. More marked differences are observed in histone acetylation of the RGS10-1 promoter. Acetylated histone H3 associates with the RGS10-1 promoter was significantly lower in A2780-AD cells compared to parental cells, with a corresponding increase in histone deacetylase (HDAC) enzyme association. Further, both HDAC1 and DNMT1 exhibit aberrant association with RGS10 promoters in chemoresistant ovarian cancer cells. Knockdown of HDAC1 or DNMT1 expression, and pharmacological inhibition of DNMT or HDAC enzymatic activity, significantly increases RGS10 expression and cisplatin-mediated cell death. Finally, DNMT1 knock down decreases HDAC1 binding to the RGS10 promoter in chemoresistant cells, suggesting HDAC1 recruitment to RGS10 promoters requires DNMT1 activity. Our results suggest that HDAC1 and DNMT1 cooperatively contribute to the suppression of RGS10 during acquired chemoresistance and support inhibition of HDAC1 and DNMT1 as an adjuvant therapeutic approach to overcome ovarian cancer chemoresistance.

Citation Format: Shelley Hooks, PhD. Releasing the brakes on ovarian cancer cell survival: epigenetic suppression of RGS10 in ovarian cancer chemoresistance [abstract]. In: Proceedings of the 10th Biennial Ovarian Cancer Research Symposium; Sep 8-9, 2014; Seattle, WA. Philadelphia (PA): AACR; Clin Cancer Res 2015;21(16 Suppl):Abstract nr POSTER-BIOL-1317.
Abstract POSTER-BIOL-1324A: Loss of DYRK1A promotes transformation of the fallopian tube epithelial cells by oncogenic ras
Authors: Litovchick Larisa , Saini Siddharth , Karst Alison , Hu Lan , Drapkin Ronny , DeCaprio James A.
Published: August 2015

Purpose of the study: High-grade serous ovarian carcinoma (HGSOC) is an aggressive form of ovarian cancer that is characterized by nearly ubiquitous presence of somatic mutations in TP53 gene and widespread gene copy number variations. It is likely that some of these genetic alterations cooperate with TP53 mutations in the pathogenesis of HGSOC; however, the specific roles of individual genes that are frequently gained or lost in HGSOC are only beginning to emerge. We have previously characterised an important role of DYRK1A protein kinase in the ability of human cells to undergo G0/G1 cell cycle arrest and oncogenic-Ras induced senescence. DYRK1A is required for the assembly of the DREAM repressor complex that controls the genes involved in cell cycle progression; this function of DYRK1A could be promoted by LATS kinases of the Hippo tumor suppressor pathway. Analysis of the The Cancer Genome Atlas (TCGA) ovarian dataset revealed loss of heterozygosity (LOH) of DYRK1A gene in over 25% HGSOC tumors. Therefore, we sought to determine whether loss of DYRK1A function can contribute to ovarian carcinogenesis.

Experimental Procedures: A panel of human ovarian cancer cell lines was used to determine the levels of DYRK1A and its ability to suppress cell proliferation. The fallopian tube epithelial cell (FTEC) cell lines with stable shRNA knock-down of DYRK1A were used in transformation assays with ovarian cancer-relevant oncogenes. Bioinformatics analysis of the TGCA ovarian oncogenomic dataset was applied to determine whether DYRK1A loss in HGSOC could lead to perturbation of any specific gene expression programs.

Summary of Data: It was found that depletion of DYRK1A promotes oncogenic transformation of the FTEC cells expressing mutant p53R175H and oncogenic HRASG12V. Rescue of DYRK1A expression in these cells resulted in arrest of proliferation and senescence. DYRK1A protein levels were reduced in a subset of ovarian cancer cell lines; overexpression of active but not the kinase-inactive DYRK1A suppressed the proliferation of these cells. Gene Set Enrichment Analysis (GSEA) of the TCGA HGSOC dataset revealed upregulated expression of the DREAM target genes in a subset of samples with LOH of both DYRK1A and LATS1 genes.

Conclusion: Our findings support the role of DYRK1A as an ovarian tumor supressor and suggest that loss of DYRK1A function in HGSOC could lead to activation of transcription of the genes involved in cell proliferation.

Citation Format: Larisa Litovchick, Siddharth Saini, Alison Karst, Lan Hu, Ronny Drapkin, James A. DeCaprio. Loss of DYRK1A promotes transformation of the fallopian tube epithelial cells by oncogenic ras [abstract]. In: Proceedings of the 10th Biennial Ovarian Cancer Research Symposium; Sep 8-9, 2014; Seattle, WA. Philadelphia (PA): AACR; Clin Cancer Res 2015;21(16 Suppl):Abstract nr POSTER-BIOL-1324A.
Abstract POSTER-BIOL-1327: Small cell carcinoma of the ovary, hypercalcemic type displays frequent inactivating germline and somatic mutations in SMARCA4
Authors: Ramos Pilar , Karnezis Anthony N. , Craig David W. , Sekulic Aleksandar , Russell Megan L. , Hendricks William P.D. , Corneveaux Jason J. , Barrett Michael T. , Shumansky Karey , Yang Yidong , Shah Sohrab P. , Prentice Leah M. , Marra Marco A. , Kiefer Jeffrey , Zismann Victoria L. , McEachron Troy A. , Salhia Bodour , Prat Jaime , Clarke Blaise A. , Pressey Joseph G. , Farley John H. , Anthony Stephen P. , Roden Richard B.S. , Cunliffe Heather E. , Huntsman David G. , Trent Jeffrey M.
Published: August 2015

Introduction: Small cell carcinoma of the ovary of hypercalcemic type (SCCOHT) is arguably the most aggressive ovarian cancer. Most patients are diagnosed at an advanced stage, do not respond to chemotherapy, and die of disease within 1-2 years. It affects children and young women and is reported to occur in families. The cause of the disease is poorly understood. Therefore, we used next generation sequencing technology to identify the genetic basis of the disease.

Method: We performed whole genome, whole exome sequencing or targeted sequencing on tumours and germline samples from 17 SCCOHT patients and on the SCCOHT cell line BIN-67 and SCCOHT-1. Immunohistochemistry (IHC) was performed on formalin-fixed, paraffin-embedded tumors from 23 patients and on a tissue microarray of 485 primary ovarian tumours of other subtypes. BIN-67 cells harbouring biallelic inactivation of SMARCA4 were transduced with a lentivirus expressing wild type SMARCA4.

Result: We identified inactivating germline and somatic mutations in the SWI/SNF chromatin-remodeling gene SMARCA4 in 79% (11/14) of SCCOHT patients, 2 of whom bore germline mutations, and in both cell lines. SMARCA4 protein was lost in 87% (20/23) of SCCOHT tumours but in only 0.4% (2/485) of other primary ovarian tumours. Reintroduction of wild-type SMARCA4 into BIN-67 cells resulted in altered cell morphology and growth arrest.

Conclusions: The mutation spectrum, IHC profile and cell culture phenotype implicate SMARCA4 as a critical tumour suppressor in SCCOHT pathogenesis.

Citation Format: Pilar Ramos, Anthony N. Karnezis, David W. Craig, Aleksandar Sekulic, Megan L. Russell, William P.D. Hendricks, Jason J. Corneveaux, Michael T. Barrett, Karey Shumansky, Yidong Yang, Sohrab P. Shah, Leah M. Prentice, Marco A. Marra, Jeffrey Kiefer, Victoria L. Zismann, Troy A. McEachron, Bodour Salhia, Jaime Prat, Blaise A. Clarke, Joseph G. Pressey, John H. Farley, Stephen P. Anthony, Richard B.S. Roden, Heather E. Cunliffe, David G. Huntsman, Jeffrey M. Trent. Small cell carcinoma of the ovary, hypercalcemic type displays frequent inactivating germline and somatic mutations in SMARCA4 [abstract]. In: Proceedings of the 10th Biennial Ovarian Cancer Research Symposium; Sep 8-9, 2014; Seattle, WA. Philadelphia (PA): AACR; Clin Cancer Res 2015;21(16 Suppl):Abstract nr POSTER-BIOL-1327.
Abstract POSTER-BIOL-1328: Endothelin-1/endothelin a receptor-mediated biased signaling is a new player in modulating human ovarian cancer cell tumorigenesis
Authors: Teoh Jian-peng , Park Kyoung-mi , Wang Yongchao , Hu Qiuping , Kim Sangmi , Wu Guangyu , Huang Shuang , Maihle Nita , Kim Il-man
Published: August 2015

The endothelin-1 (ET-1)/endothelin A receptor (ETAR, a G protein-coupled receptor) axis confers pleiotropic effects on both tumor cells and the tumor microenvironment, modulating chemo-resistance and other tumor-associated processes by activating Gαq- and β-arrestin-mediated pathways. While the precise mechanisms by which these effects occur remain to be elucidated, interference with ETAR signaling has emerged as a promising antitumor strategy in many cancers including ovarian cancer (OC). However, current clinical approaches using ETAR antagonists in the absence of a detailed knowledge of downstream signaling have resulted in multiple adverse side effects and limited therapeutic efficacy. To maximize the safety and efficacy of ETAR-targeted OC therapy, we investigated the role of other G protein subunits such as Gαs in the ETAR-mediated ovarian oncogenic signaling. In HEY (human metastatic OC) cells where the ET-1/ETAR axis is well-characterized, Gαs signaling inhibits ETAR-mediated OC cell migration, wound healing, proliferation and colony formation on soft agar while inducing OC cell apoptosis. Mechanistically, ET-1/ETAR is coupled to Gαs/cAMP signaling in the same ovarian carcinoma-derived cell line. Gαs/cAMP/PKA activation inhibits ETAR-mediated β-arrestin activation of angiogenic/metastatic Calcrl and Icam2 expression. Consistent with our findings, Gαs overexpression is associated with improved survival in OC patients in the analysis of the Cancer Genome Atlas data. In conclusion, our results indicate a novel function for Gαs signaling in ET-1/ETAR-mediated OC oncogenesis and may provide a rationale for a biased signaling mechanism, which selectively activates Gαs-coupled tumor suppressive pathways while blocking Gαq-/β-arrestin-mediated oncogenic pathways, to improve the targeting of the ETAR axis in OC.

Citation Format: Jian-peng Teoh, Kyoung-mi Park, Yongchao Wang, Qiuping Hu, Sangmi Kim, Guangyu Wu, Shuang Huang, Nita Maihle, Il-man Kim. Endothelin-1/endothelin a receptor-mediated biased signaling is a new player in modulating human ovarian cancer cell tumorigenesis [abstract]. In: Proceedings of the 10th Biennial Ovarian Cancer Research Symposium; Sep 8-9, 2014; Seattle, WA. Philadelphia (PA): AACR; Clin Cancer Res 2015;21(16 Suppl):Abstract nr POSTER-BIOL-1328.
Abstract POSTER-BIOL-1333: TRIP6 regulates p27KIP1 to promote ovarian tumorigenesis
Authors: Lin Fang-Tsyr , Lin Victor T. G. , Lin Vivian Y. , Lai Yun-Ju , Chen Chen-Shan , Liu Kang , Lin Weei-Chin
Published: August 2015

Although the nuclear p27KIP1 functions as a negative regulator of G1/S cell cycle progression by binding to and inhibiting cyclin-CDK complexes, the cytosolic p27KIP1 has been shown to promote adhesion disassembly and tumor invasion. Loss of nuclear p27KIP1 and cytosolic mislocalization of p27KIP1 are frequently found during cancer progression, and these events correlate with poor clinical outcome in many types of cancers, including ovarian cancer. However, the mechanisms underlying this dysregulation are not yet fully understood.

TRIP6 is an adaptor protein that is highly expressed in ovarian serous cystadenocarcinoma. It serves as a platform to recruit a number of molecules, such as NF-κB p65, c-Src, p130cas, Crk, LPA2 and Fas/CD95 receptors, to promote cell motility and antiapoptotic signaling. Using an ovarian cancer xenograft mouse model, we find that TRIP6 knockdown significantly reduces ovarian tumor proliferation. We determine that this effect is in part through the regulation of p27KIP1. Our data show that TRIP6 serves as a bridge to promote the recruitment of p27KIP1 to AKT in the cytosol. TRIP6 regulates the membrane translocation and activation of AKT, and facilitates AKT-mediated recognition and phosphorylation of p27KIP1 specifically at T157, thereby promoting the cytosolic mislocalization of p27KIP1. This is required for p27KIP1 to enhance LPA-induced ovarian cancer cell migration. TRIP6 also promotes serum-induced reduction of nuclear p27KIP1 expression through Skp2-dependent and -independent mechanisms. Consequently, knockdown of TRIP6 in ovarian cancer xenografts restores nuclear p27KIP1 expression and impairs tumor proliferation. Since the expression of TRIP6 and the activity of AKT are significantly higher in ovarian cancer, restoring the expression of nuclear p27KIP1 and inhibiting the migratory effect of phospho-T157-p27KIP1 by targeting TRIP6 may prove to be an effective approach to reduce ovarian cancer progression.

Citation Format: Fang-Tsyr Lin, Victor T. G. Lin, Vivian Y. Lin, Yun-Ju Lai, Chen-Shan Chen, Kang Liu, Weei-Chin Lin. TRIP6 regulates p27KIP1 to promote ovarian tumorigenesis [abstract]. In: Proceedings of the 10th Biennial Ovarian Cancer Research Symposium; Sep 8-9, 2014; Seattle, WA. Philadelphia (PA): AACR; Clin Cancer Res 2015;21(16 Suppl):Abstract nr POSTER-BIOL-1333.
Abstract POSTER-BIOL-1335: Threonyl-tRNA synthetase overexpression correlates with angiogenesis and progression of human ovarian cancer
Authors: Wellman Theresa L. , Eckenstein Midori , Wong Cheung , Rincon Mercedes , Ashikaga Takamaru , Mount Sharon L. , Francklyn Christopher S. , Lounsbury Karen M.
Published: August 2015

Ovarian tumors create a dynamic microenvironment that promotes angiogenesis and reduces immune responses. There is growing evidence that tRNA synthetases have a role in cell signaling separate from their function in protein synthesis. Our research has revealed that threonyl-tRNA synthetase (TARS) has an extracellular angiogenic activity. Exploration of protein and mRNA databases further showed a connection between TARS expression and ovarian cancer. The objective of this study was to test the hypothesis that TARS is secreted by ovarian cancer cells and that its expression in clinical samples correlates with angiogenic markers and ovarian cancer progression. TARS secretion by SKOV3 human ovarian cancer cells was induced by hypoxia or tumor necrosis factor-α (TNF-α) as detected by ELISA and Western blot of culture media (p<0.001). For the clinical study, TARS, vascular endothelial growth factor (VEGF) and PECAM were assessed by immunohistochemistry in serial sections of paraffin embedded ovarian tissues from 70 patients diagnosed with epithelial ovarian cancer and 12 control patients. Serum samples from 31 tissue-matched patients were analyzed by ELISA for TARS, CA125, and tumor necrosis factor-α (TNF-α). There was a strong association between the tumor expression of TARS and advancing stage of epithelial ovarian cancer (p<0.001). TARS expression and localization were also correlated with VEGF (p<0.001). A significant proportion of samples included heavy TARS staining of infiltrating leukocytes which also correlated with stage (p=0.017). Patient serum TARS was related to tumor TARS and angiogenic markers, but did not achieve significance with respect to stage. Multivariate Cox proportional hazard models revealed a surprising inverse relationship between TARS expression and mortality risk in late stage disease (p=0.062). Together these results show that TARS is secreted in response to cell stress and that its expression is increased in epithelial ovarian cancer according to stage and angiogenic progression. These findings and the association of TARS with disease survival provide clinical validation that TARS is associated with angiogenesis in ovarian cancer. These results encourage further study of TARS as a regulator of the tumor microenvironment and possible target for diagnosis and/or treatment in ovarian cancer.

Citation Format: Theresa L. Wellman, Midori Eckenstein, Cheung Wong, Mercedes Rincon, Takamaru Ashikaga, Sharon L. Mount, Christopher S. Francklyn, Karen M. Lounsbury. Threonyl-tRNA synthetase overexpression correlates with angiogenesis and progression of human ovarian cancer [abstract]. In: Proceedings of the 10th Biennial Ovarian Cancer Research Symposium; Sep 8-9, 2014; Seattle, WA. Philadelphia (PA): AACR; Clin Cancer Res 2015;21(16 Suppl):Abstract nr POSTER-BIOL-1335.
Abstract POSTER-BIOL-1348: No association between PARK2 and BRCA1-associated gynecologic cancers
Authors: Walsh Christine , Nassanian Hoorig , Agadjanian Hasmik , Miller Carl , Orsulic Sandra , Karlan Beth
Published: August 2015

Purpose of study: We sought to identify genomic modifiers of gynecologic cancer penetrance in BRCA1 185delAG mutation carriers. In a pilot study using the Illumina 610K Quad chip to perform a whole genome analysis of 592,532 single nucleotide polymorphisms (SNP), we found significant SNP genotype differences in the PARK2 gene between BRCA1 185delAG mutation carriers who developed gynecologic cancer (cases) compared to those who did not (controls). These SNPS created a haplotype structure that was present at four-times higher frequency in cases than controls (37% vs. 9%, p=0.0009). We hypothesized that PARK2, which has been shown to be mutated in other cancers and to have a tumor suppressive function, plays a role in the biology of BRCA1-associated gynecologic cancer development.

Experimental Procedures: To validate a PARK2 risk haplotype in an independent dataset, we isolated germline DNA from 85 BRCA1 185delAG mutation carriers and used TaqMan SNP Genotyping to genotype seven SNPs in PARK2. To determine whether a correlation exits between the PARK2 risk haplotype and PARK2 expression levels in tissue, we quantified PARK2 expression in normal ovaries and ovarian cancer tissue using quantitative real-time PCR (qPCR). To determine the rate of PARK2 mutations in 17 BRCA1-associated ovarian cancers, we used a touchdown PCR protocol to amplify and sequence each PARK2 exon. To identify exon deletions and duplications in the PARK2 gene, we utilized the multiplex ligation-dependent probe amplification (MLPA) technique.

Summary of Data: In an independent dataset of 85 BRCA1 185delAG mutation carriers, we found the PARK2 risk haplotype to be present in 35 (17%) of cases. The risk haplotype was present in a higher proportion of cancer cases than benign controls (6/12 or 50% vs. 29/73 or 8%), but this was not significant (p=0.54). We did not find a difference in PARK2 ovarian tissue expression levels in patients with and without the PARK2 risk haplotype. We sequenced the PARK2 gene and performed MPLA analysis from tumor DNA of 17 ovarian cancers from BRCA1 185delAG mutation carriers, but did not identify any deleterious PARK2 mutations or exon deletions and duplications.

Conclusions: We did not validate a PARK2 risk haplotype as a modifier of gynecologic cancer penetrance in BRCA1 185delAG mutation carriers. We did not find evidence for PARK2 inactivation in BRCA1-associated gynecologic tumors.

Citation Format: Christine Walsh, Hoorig Nassanian, Hasmik Agadjanian, Carl Miller, Sandra Orsulic, Beth Karlan. No association between PARK2 and BRCA1-associated gynecologic cancers [abstract]. In: Proceedings of the 10th Biennial Ovarian Cancer Research Symposium; Sep 8-9, 2014; Seattle, WA. Philadelphia (PA): AACR; Clin Cancer Res 2015;21(16 Suppl):Abstract nr POSTER-BIOL-1348.
Abstract POSTER-BIOL-1351: Molecular characterization of cyclin-dependent kinase 1 pathway in newly established epithelial ovarian cancer cell lines
Authors: Yang Wookyeom , Cho Hanbyoul , Sol Kim , Kim Jae-Hoon
Published: August 2015

Background: Epithelial ovarian cancer (EOC) is a heterogeneous gynecologic tumor. To better understand the biological pathways involved in the development of EOC, we performed mRNA expression profiling on newly established EOC cell lines.

Methods: Five newly established EOC cell lines were characterized by transcript profiling. Microarray analyses were validated using quantitative real-time PCR (RT-PCT) and immunoblotting. Furthermore, biological networks of the differentially expressed genes were predicted using the STRING database and literature review.

Results: A total of 239 genes represented by 24,957 probes were identified from microarray analysis, of which 98 and 141 genes are up- (> 2 fold, p < 0.05) or down-regulated (< 3 fold, p < 0.05), respectively, in EOC cell line compare to human ovarian surface epithelial cell lines. We identified a subset of EOC characterized by the numerous genes related to cell cycle and apoptosis, along with cyclin-dependent kinase 1 (CDK1). A total of 24 genes were validated by quantitative RT-PCR, and expressional reliabilities of 4 CDK1 pathway-related genes were confirmed by immunoblotting. Further biological network analysis revealed that the CDK1 pathway might be essential for EOC tumorigenesis.

Conclusions: These data reveal that the CDK1 pathway is important to EOC development and progression, providing new insight into the biology of EOC.

Citation Format: Wookyeom Yang, Hanbyoul Cho, Kim Sol, Jae-Hoon Kim. Molecular characterization of cyclin-dependent kinase 1 pathway in newly established epithelial ovarian cancer cell lines [abstract]. In: Proceedings of the 10th Biennial Ovarian Cancer Research Symposium; Sep 8-9, 2014; Seattle, WA. Philadelphia (PA): AACR; Clin Cancer Res 2015;21(16 Suppl):Abstract nr POSTER-BIOL-1351.
Abstract POSTER-BIOL-1352: Notch3 promotes anoikis resistance in ovarian cancer
Authors: Brown Caitlin W. , Brodsky Alexander S. , Freiman Richard N.
Published: August 2015

Epithelial ovarian cancer (EOC) is the most lethal gynecologic cancer. Recently, a number of genome-wide studies in ovarian cancer patient samples have postulated that the course of disease can influenced by the Notch3 signaling pathway. In one study, 11% of human tumor samples showed mutation or amplification of the Notch3 receptor alone. Subsequent papers have correlated Notch3 overexpression in human samples with shorter progression-free survival as well as overall survival. In vitro studies have shown Notch3 to be involved in apoptosis resistance and chemoresistance, as well as cell cycle alterations. Together, these human studies have implicated Notch3 expression and amplification with decreased outcomes in ovarian cancer patients.

To test the potential mechanism by which Notch3 over-expression promotes ovarian cancer, we examined several molecular and cellular functions of Notch3 in ovarian cancer cells. Decreasing the levels of Notch3 in ovarian cancer cells increases the susceptibility of these cells to apoptosis, which is consistent with previous studies. Using an effective siRNA-mediated knockdown of Notch3 combined with transfer to ultra-low attachment plates revealed a significant decrease in the ability of Notch3-knockdown IGROV-1 cells to remain viable in anchorage-independent conditions over 24 hours. Our data show that not only does Notch3 prevent apoptosis, but specifically promotes anoikis resistance. Further examining the mechanism underlying this novel finding revealed the positive roles of extracellular matrix proteins and FAK in promoting the anchorage-independent growth of IGROV-1 cells. Strikingly, mRNA and protein levels of col4α2 are reduced when Notch3 levels are decreased and exogenous collagen IV supplementation can largely reverse the anoikis sensitivity. Reduction of col4α2 expression by siRNA induces cell death on its own and high Notch3 expression levels correlate with higher col4α2 expression in human ovarian tumor samples. Together, these studies have revealed that Notch3 promotes anoikis resistance through col4α2 expression and may partly explain its positive correlation with poor survival in ovarian cancer patients.

Citation Format: Caitlin W. Brown, Alexander S. Brodsky, Richard N. Freiman. Notch3 promotes anoikis resistance in ovarian cancer [abstract]. In: Proceedings of the 10th Biennial Ovarian Cancer Research Symposium; Sep 8-9, 2014; Seattle, WA. Philadelphia (PA): AACR; Clin Cancer Res 2015;21(16 Suppl):Abstract nr POSTER-BIOL-1352.
Abstract POSTER-CTRL-1209: Ovarian cancer disparities in a high volume academic medical center
Authors: Hunt Hayley , Rodriguez Noah , Oda Keiji , Wagner Robert J. , Swensen Ron E.
Published: August 2015

Objective: To identify disparities in care and survival in ovarian cancer patients treated at a high volume academic center.

Methods: A review of epithelial ovarian cancer cases treated between 01/01/97 and 12/31/07 was conducted. Demographic, clinico-pathologic, insurance carrier, and treatment data were collected. Insurance carrier was categorized into three groups: Commercial, Medicare, and Medicaid. ANOVA or Kruskal-Wallis tests were utilized for continuous variables and chi-square or Fisher’s exact tests for categorical variables. Univariable Cox proportional hazards model was used to estimate hazard ratios for demographic and pathologic variables identified in association with survival. Multivariable Cox regression models were fitted with insurance status as the main exposure. Kaplan-Meier survival curves were generated for race, income, and insurance and compared utilizing log-rank tests.

Results: 279 epithelial ovarian cancer cases were identified out of 460 total ovarian cancer cases. 199 (71%) were White and 80 (29%) were non-White. 159 (57%) had commercial insurance followed by 71 (25%) and 49 (18%) with Medicaid and Medicare, respectively. 166 (60%) of the study group had less than $45,000 as an annual median income. 219 (78%) patients underwent primary debulking surgery. Of the three insurance categories, women with commercial insurance had a survival advantage (p= 0.002). Medicaid was associated with worse survival after adjusting for grade, stage, residual disease, and chemotherapy (p= 0.04). Survival rates in advanced stage disease were similar across the 3 insurance carriers.

Conclusions: Ovarian cancer survival disparities persist at a high volume center but are largely eliminated in advanced stage.

Citation Format: Hayley Hunt, MD, Noah Rodriguez, MD, Keiji Oda, MPH, Robert J. Wagner, MD, Ron E. Swensen, MD. Ovarian cancer disparities in a high volume academic medical center [abstract]. In: Proceedings of the 10th Biennial Ovarian Cancer Research Symposium; Sep 8-9, 2014; Seattle, WA. Philadelphia (PA): AACR; Clin Cancer Res 2015;21(16 Suppl):Abstract nr POSTER-CTRL-1209.
Abstract POSTER-CTRL-1213: Whole exome and targeted resequencing, of population based ovarian cancer cases and controls, identifies susceptibility genes for ovarian cancer
Authors: Ramus Susan J. , Song Honglin , Dicks Ed , Intermaggio Maria , Hayward Jane , Tyrer Jonathan P. , Gronwald Jacek , Kjær Susanne Krüger , Wittemore Alice , Karlan Beth , Odunsi Kunle , Menon Usha , Bowtell David D. , Goode Ellen L. , Pharoah Paul D.P. , Gayther Simon A.
Published: August 2015

The known genetic variance for epithelial ovarian cancer (EOC) accounts for less than half the familial risks of the disease. BRCA1 and BRCA2 are the major high-penetrance EOC susceptibility genes. More recently genes that interact with BRCA1 and BRCA2 in the same biological pathways, but conferring more moderate penetrance (e.g. RAD51C, RAD51D) have also been identified. Mutations in these genes result in defects in homologous recombination and double strand break repair (HR/DSB). We hypothesized that other genes operating in these and other DNA repair pathways may also confer EOC susceptibility. Whole exome sequencing data, available from more than 400 high-grade serous ovarian cancers, were used to identify 24 candidate genes harboring germline truncating mutations in at least 2 cases, followed by replication analysis using targeted resequencing in ~3,000 cases and 3,000 controls. Subjects were also sequenced for the known susceptibility genes listed above. The data confirmed the previously reported contributions of BRCA1, BRCA2, RAD51C and RAD51D to EOC. In addition, we identified BRIP1, and FANCM as candidate EOC susceptibility genes. Surprisingly, for several genes we found truncating mutations were equally prevalent in cases as they were in controls. These mutations may still represent rare low-penetrance susceptibility alleles for EOC, which we are too limited in sample size and power to confirm; or more likely, it indicates that cells can tolerate heterozygous loss-of-function mutations in apparently critical DNA repair genes without any impact on neoplastic development. These findings may have implications for risk prediction and prevention strategies for EOC and possibly for the identification of novel therapeutic targets.

Citation Format: Susan J. Ramus, Honglin Song, Ed Dicks, Maria Intermaggio, Jane Hayward, Jonathan P. Tyrer, Jacek Gronwald, Susanne Krüger Kjær, Alice Wittemore, Beth Karlan, Kunle Odunsi, Usha Menon, David D. Bowtell, Ellen L. Goode, Paul D.P. Pharoah, Simon A. Gayther. Whole exome and targeted resequencing, of population based ovarian cancer cases and controls, identifies susceptibility genes for ovarian cancer [abstract]. In: Proceedings of the 10th Biennial Ovarian Cancer Research Symposium; Sep 8-9, 2014; Seattle, WA. Philadelphia (PA): AACR; Clin Cancer Res 2015;21(16 Suppl):Abstract nr POSTER-CTRL-1213.
Abstract POSTER-TECH-1101: Characterization of genomic landscapes of BRCA1 and BRCA2 implicated ovarian cancer specimens from a founder french canadian population
Authors: AlShehri Eman , Belanger Moria , Arcand Suzanna , Oros Kathleen Klein , Greenwood Celia , Tonin Patricia N.
Published: August 2015

Although molecular genetic profiling of ovarian cancers harboring germline BRCA1/BRCA2 mutations suggest that pathways in common with sporadic cases are involved in the pathogenesis of the disease, overall survival differs between BRCA1, BRCA2 and sporadic disease. To further dissect molecular pathways involved, which could account for differences in pathogenesis of the disease, we have investigated genomic landscapes in the BRCA1 and BRCA2 mutated ovarian cancers. Chromosomal anomalies were assessed in 28 specimens with BRCA1 (n=15), BRCA2 (n=12) or BRCA1 and BRCA2 (n=1) mutations using high-density Illumina SNP arrays. The majority (22/28) are serous adenocarcinomas while the remaining samples were either endometroid (2/28) or mixed adenocarcinomas (4/28). The cases harbor BRCA1/BRCA2 mutations from a French Canadian population, which exhibit strong founder effects. Allelic imbalance, copy number differences, intrachromosomal breaks and homozygous deletions were inferred visually using the Genome Viewer module of the BeadStudio software followed by ASCAT analysis. A statistical analysis was used to directly compare genotypes of BRCA1 versus BRCA2 positive samples. TP53 gene mutation status was also assessed. All samples were found to harbor a somatic TP53 mutation comprised of missense (19/28), nonsense (2/28), frame-shift (4/28) and (3/28) splice mutations. The results were compared to independently derived data generated previously from our group, which was largely comprised of sporadic cases (Wojnarowicz et al 2012), and to the genomic data from the Cancer Genome Atlas project (TCGA). The genomic landscapes of BRCA1 and BRCA2 mutated cancers overlap those reported in previous studies. However, there were significant differences in the genomic landscapes involving chromosome 6q between BRCA1 and BRCA2 mutation-positive cancers. This genomic distinction between BRCA1 and BRCA2 ovarian cancer samples may point to a region containing genes important in the etiology or progression of hereditary cancer.

Citation Format: Eman AlShehri, Moria Belanger, Suzanna Arcand, Kathleen Klein Oros, Celia Greenwood, Patricia N. Tonin. Characterization of genomic landscapes of BRCA1 and BRCA2 implicated ovarian cancer specimens from a founder french canadian population [abstract]. In: Proceedings of the 10th Biennial Ovarian Cancer Research Symposium; Sep 8-9, 2014; Seattle, WA. Philadelphia (PA): AACR; Clin Cancer Res 2015;21(16 Suppl):Abstract nr POSTER-TECH-1101.