bluuebunny/crossref_metadata_2025_split huggingface dataset:
Abstract PR05: Selective contribution of the SHOC2 phosphatase complex to ERK pathway dynamics highlights its potential as a therapeutic target
Authors: Boned del Rio Isabel , Young Lucy C. , Sari Sibel , Jones Greg G. , Bhamra Amandeep , Surinova Silvia , Rodriguez-Viciana Pablo 
Published: June 2021

Abstract
The ERK signaling pathway is hyperactivated in the majority of human cancers. However, ERK signaling mediates myriad physiologic responses and on-target toxicity has severely limited the clinical efficacy of current ERK pathway inhibitors. Despite the key role of RAF kinases in physiology and disease, we still lack a complete understanding of their regulation. Here we show that dephosphorylation of the conserved ‘S259’ RAF inhibitory site is a key activating step mediated by the SHOC2-MRAS-PP1 complex. Gain-of-function mutations in SHOC2, MRAS, PP1 and CRAF (clustering around S259) found in Noonan syndrome independently validate the important role of the SHOC2 complex in RAF-ERK pathway regulation. We have used CRISPR/cas9 to knock out SHOC2 and the 3 RAF isoforms in colorectal DLD-1 cancer cells. Our data show that SHOC2 is required for EGF-induced dephosphorylation of S365 BRAF, 14-3-3 and MEK dissociation from BRAF and RAF dimerization. However, whereas SHOC2 function is essential for a rapid transient phase of ERK activation in response to EGF stimulation, it is not required for a slow, sustained phase that is instead driven by palmitoylated H/N-RAS proteins and N-region phosphorylation of CRAF. Redundancy makes SHOC2 dispensable for ERK activation and proliferation in 2D whereas KRAS mutant cells rely on SHOC2 for ERK signaling under anchorage-independent/3D conditions, which thus presents a therapeutic opportunity. This study highlights a context-dependent contribution of SHOC2 to ERK pathway dynamics that is preferentially engaged by KRAS oncogenic signaling and provides a biochemical framework for how targeting the SHOC2 regulatory node would allow for selective ERK pathway inhibition, likely to provide improved therapeutic margins in the clinic.
This abstract is also being presented as Poster A32.
Citation Format: Isabel Boned del Rio, Lucy C. Young, Sibel Sari, Greg G. Jones, Amandeep Bhamra, Silvia Surinova, Pablo Rodriguez-Viciana. Selective contribution of the SHOC2 phosphatase complex to ERK pathway dynamics highlights its potential as a therapeutic target [abstract]. In: Proceedings of the AACR Special Conference on Targeting RAS-Driven Cancers; 2018 Dec 9-12; San Diego, CA. Philadelphia (PA): AACR; Mol Cancer Res 2020;18(5_Suppl):Abstract nr PR05.
Abstract AP02: EXPOSING THE LOCAL FACTORS THAT CONTRIBUTE TO OVARIAN COLONIZATION OF FALLOPIAN TUBE-DERIVED TUMORS
Authors: Burdette Joanna E. , Zink Katherine , Dean Matthew , Sanchez Laura M. 
Published: June 2021

Abstract
High grade serous cancer (HGSC) originates in the fallopian tube and typically invade and colonize the ovary as part of peritoneal metastasis. The purpose of this study was to define the factors that contribute to ovarian specific metastasis of fallopian tube HGSC. While protein factors have been explored that facilitate migration and invasion of fallopian tube cell to the ovary, the small molecule communication between the tissues during metastasis remains largely unexplored. Due to the proximity of ovarian primary metastasis, an experimental design was optimized using imaging mass spectrometry to capture the spatial composition of small molecules uniquely expressed when fallopian tube-derived tumor cells were grown in the microenvironment of the ovary as a model of primary metastasis. The observed mass-to-charge ratios (m/z's) that were induced specifically in co-culture represent small molecules that may contribute to the metastasis of HGSOC selectively to the ovary. Human fallopian tube epithelial HGSOC and tumorigenic murine oviductal epithelial cells, but not normal cell types, repeatedly induced a signal from the ovary at m/z 170. This signal was identified as norepinephrine, which was confirmed to simulate invasion of ovarian cancer cells lacking wildtype p53. Norepinephrine has an established role in ovarian cancer metastasis to secondary sites and the use of beta-blockers in women is associated with increased survival. The use of imaging mass spectrometry also revealed a series of new molecules that are specifically enhanced from the ovary or the fallopian tube during co-culture. These molecules may reveal pathways that contribute to metastasis and biological targets for therapeutic intervention to block ovarian metastasis of fallopian tube-derived HGSOC. The developed mass spectrometry method can be adapted to other mammalian-based model systems for investigation of untargeted metabolomics that facilitate metastasis.
Citation Format: Joanna E. Burdette, Katherine Zink, Matthew Dean, and Laura M. Sanchez. EXPOSING THE LOCAL FACTORS THAT CONTRIBUTE TO OVARIAN COLONIZATION OF FALLOPIAN TUBE-DERIVED TUMORS [abstract]. In: Proceedings of the 12th Biennial Ovarian Cancer Research Symposium; Sep 13-15, 2018; Seattle, WA. Philadelphia (PA): AACR; Clin Cancer Res 2019;25(22 Suppl):Abstract nr AP02.
Abstract AP13: GENOME-SCALE CRISPR KNOCKOUT SCREEN IDENTIFIES TIGAR AS A MODIFIER OF PARP INHIBITOR SENSITIVITY
Authors: Fang Pingping , Minn Kay , Chien Jeremy 
Published: June 2021

Abstract
A key challenge is to understand the mechanisms that contribute to “BRCAness” and hence sensitivity to poly (ADP-ribose) polymerase (PARP) inhibitors. Using the sensitivity to PARP inhibitor olaparib as a surrogate for “BRCAness”, we performed a CRISPR/Cas9-based genome-scale loss-of-function screen and identified TIGAR as a modifier of olaparib response. TIGAR knockdown increases the level of intracellular reactive oxygen species, induces more DNA damage after olaparib treatment, and causes “BRCAness” by downregulating BRCA1 and genes in the Fanconi anemia pathway. Further, TIGAR knockdown induces senescence, reduces spheroid tumor cell growth, and enhances the cytotoxic effect of olaparib. Finally, TIGAR is amplified in ovarian cancer and a higher expression of TIGAR is associated with poor overall survival in high-grade serous ovarian cancer. Our study shows that TIGAR knockdown causes extensive transcriptional reprogramming and enhances sensitivity to PARP inhibitor, demonstrating that TIGAR is a relevant therapeutic target for treating ovarian cancer.
Citation Format: Pingping Fang, Kay Minn, Jeremy Chien. GENOME-SCALE CRISPR KNOCKOUT SCREEN IDENTIFIES TIGAR AS A MODIFIER OF PARP INHIBITOR SENSITIVITY [abstract]. In: Proceedings of the 12th Biennial Ovarian Cancer Research Symposium; Sep 13-15, 2018; Seattle, WA. Philadelphia (PA): AACR; Clin Cancer Res 2019;25(22 Suppl):Abstract nr AP13.
Abstract AP30: BRCA1 INTRON RETENTION GENERATES TRUNCATED PROTEINS THAT AVOID BRCT MUTATION MISFOLDING AND PROMOTE PARP INHIBITOR RESISTANCE
Authors: Wang Yifan , Bernhardy Andrea J. , Nacson Joseph , Krais John J. , Tan Yin-Fei , Slifker Michael , Peri Suraj , Johnson Neil 
Published: June 2021

Abstract
INTRODUCTION: BRCA1 BRCT domain mutations result in protein structural defects, and consequently misfolded proteins are subject to proteasomal degradation. Loss of BRCA1 protein activity results in sensitivity to PARP inhibitor (PARPi) therapy.
EXPERIMENTAL PROCEDURES: In this study, we examined PARPi resistance using the SNU-251 human endometrial ovarian cancer cell line. SNU-251 cells harbor a nonsense mutation c.5444G&gt;A (E1815X) in BRCA1 exon 23; that disrupts BRCT domain folding. Cells were cultured in the presence of increasing concentrations of the PARPi rucaparib until resistant cells emerged. RT-PCR, Western blotting and mass spectrometry were used to measure BRCA1 mRNA and protein levels.
RESULTS: PARPi resistant cell lines all demonstrated elevated BRCA1 protein levels that could be detected with N- but not C-terminal specific antibodies. Notably, the gel migration and molecular weight of BRCA1 was different from the mutation-induced stop codon expected size. Mass spectrometric analyses identified BRCA1 peptides encoded by exons 2-15; however, despite harboring a mutation in exon 23, no peptides encoded by exons 16-24 were present in SNU-251 cells. RT-PCR analyses showed that SNU-251 cells translated protein from exon 15 and into intron 15, terminating at an intron 15 generated stop codon. We overexpressed BRCA1 cDNA that harbored stop codons located in either the BRCT domain or in intron 15. BRCT mutation containing constructs had undetectable protein levels due to protein misfolding, and cells were highly PARPi sensitive. In contrast, cells expressing the BRCA1 intron 15 encoded stop codon had robust protein expression, demonstrated RAD51 foci and chemotherapy resistance in vitro and in vivo.
CONCLUSIONS: In summary, we discovered that BRCA1 intron translation generates new stop codons resulting in loss of the entire BRCT domain. BRCTless proteins avoid mutant protein folding problems and promote residual DNA repair and chemotherapy resistance.
Citation Format: Yifan Wang, Andrea J. Bernhardy, Joseph Nacson, John J. Krais, Yin-Fei Tan, Michael Slifker, Suraj Peri and Neil Johnson. BRCA1 INTRON RETENTION GENERATES TRUNCATED PROTEINS THAT AVOID BRCT MUTATION MISFOLDING AND PROMOTE PARP INHIBITOR RESISTANCE [abstract]. In: Proceedings of the 12th Biennial Ovarian Cancer Research Symposium; Sep 13-15, 2018; Seattle, WA. Philadelphia (PA): AACR; Clin Cancer Res 2019;25(22 Suppl):Abstract nr AP30.
Abstract DP-006: THE EFFECT OF AGE AND OCP USE ON THE INCIDENCE OF PRE-CANCEROUS P53 LESIONS AND THE DEVELOPMENT OF HIGH GRADE SEROUS OVARIAN CARCINOMA
Authors: Greening Kendall , Karnezis Anthony , Cochrane Dawn , Hanley Gillian , Huntsman David 
Published: June 2021

Abstract
BACKGROUND: High grade serous ovarian cancer (HGSOC) accounts for &gt;70% of ovarian cancer related deaths and is the most common ovarian cancer histotype, most originating from pre-cancerous p53 lesions in the fallopian tube (FT) fimbria. Use of oral contraceptive pills (OCPs) for 5 years or more is associated with &gt;40% reduction in risk of HGSOC, but the mechanism is unknown. We hypothesize that OCP use reduces the incidence of p53 lesions. Our preliminary data show higher incidence of p53 lesions in post- compared to pre-menopausal women, therefore we aim quantify p53 lesions in post-menopausal women who previously did or did not use OCPs. This will provide insight into the protective effects of OCPs against HGSOC.
PRELIMINARY RESULTS: We determined the presence of p53 lesions by immunohistochemistry (IHC) in FT of women up to 40 years old (n=27) and &gt;60 years old (n=24) who underwent salpingectomies for non-cancer reasons. p53 lesions were identified in 3/27 cases of the younger cohort (11%) and in 10/24 of the older cohort (42%). Thus, we conclude an increased incidence of p53 lesions in older compared to younger women.
PROPOSED DESIGN: IHC for p53 will be performed on FT fimbria of women &gt;55 years old who received salpingectomy/hysterectomy for non-cancer reasons. Based on an assumed reduction in p53 lesions of 35% in women who used OCPs for 5 years or more compared to non-users (25 vs. 42%), analysis of 190 cases from each group will provide &gt;80% power (p&lt;0.05). Cases will be identified through Population Data BC and blind analysis by tp53 IHC will be performed at the Vancouver General Hospital. Post-menopausal status will be confirmed by endometrium histology and data flowed back to Pop Data BC to compare to OCP data.
CONCLUSION: Our preliminary study found that 42% of post menopausal women had p53 lesions, informing this study design. The study registered through this abstract will be the first to examine the impact of OCPs on the earliest known precursors of HGSOC.
Citation Format: Kendall Greening, Anthony Karnezis, Dawn Cochrane, Gillian Hanley, David Huntsman. THE EFFECT OF AGE AND OCP USE ON THE INCIDENCE OF PRE-CANCEROUS P53 LESIONS AND THE DEVELOPMENT OF HIGH GRADE SEROUS OVARIAN CARCINOMA [abstract]. In: Proceedings of the 12th Biennial Ovarian Cancer Research Symposium; Sep 13-15, 2018; Seattle, WA. Philadelphia (PA): AACR; Clin Cancer Res 2019;25(22 Suppl):Abstract nr DP-006.
Abstract DP-010: INTRAPERITONEAL CHEMOTHERAPY USE AND OUTCOMES: EXPLORATION OF UPMC OVARIAN CANCER REGISTRY DATA
Authors: S Adambekov , S Wang , S Taylor , R Buckanovich , L Coffman , RP Edwards , F Linkov 
Published: June 2021

Abstract
BACKGROUND: Ovarian cancer (OC) is the deadliest gynecologic cancer. Despite randomized clinical trials showing improved survival for patients receiving intraperitoneal chemotherapy (IP), IP has not received a wide use outside of specialty hospitals. The aim of this study was to explore the impact of IP on OC patient survival and to evaluate whether treatment facility type impacted the outcomes.
METHODS: Detailed demographic and clinical information on OC patients (N=2924) who underwent treatment in UPMC facilities between 2000-2016 was obtained from the UPMC Cancer Registry. Duplicate records, rare, grade 1, and non-epithelial tumors were excluded. Kaplan-Meier plots were constructed to compare 10-year survival rates based on the chemotherapy type (IP vs. no-IP), cancer grade, cancer stage, surgery type, neoadjuvant and treatment facility (specialized vs. community). Multivariable Gray's models and logistic regression model were fitted to evaluate the effect of different factors on survival and tumor recurrence respectively. Two tailed P-values &lt;0.05 were considered significant. R software package was used to analyze the data.
RESULTS: The final sample consisted from 1840 patients (250 IP and 1590 no-IP). IP chemotherapy was used only in 14% of OC patients and was associated with improved long-term survival. Similarly, cases reported by specialty treatment facilities (Magee and Passavant) had better survival compared to other hospitals. Multivariable Gray's model showed that IP and younger age were significantly associated with lower hazard of death, whereas higher cancer stage is significantly associated with higher hazard of death. Multivariable logistic regression showed that IP is not significantly associated with recurrence after adjusting for median income in the zip code of patient residence and cancer stage.
CONCLUSIONS: These findings demonstrate enhanced long-term survival of patients treated with IP therapy at specialty centers. Increasing IP therapy use in clinical practice for OC patient treatment may be important strategy to improve OC outcomes.
Citation Format: Adambekov S, Wang S, Taylor S, Buckanovich R, Coffman L, Edwards RP, Linkov F. INTRAPERITONEAL CHEMOTHERAPY USE AND OUTCOMES: EXPLORATION OF UPMC OVARIAN CANCER REGISTRY DATA [abstract]. In: Proceedings of the 12th Biennial Ovarian Cancer Research Symposium; Sep 13-15, 2018; Seattle, WA. Philadelphia (PA): AACR; Clin Cancer Res 2019;25(22 Suppl):Abstract nr DP-010.
Abstract GMM-028: VALIDATION OF RUNX3 AND CAMKIINα HYPERMETHYLATION AS PROGNOSTIC MARKER FOR OVARIAN CARCINOMA
Authors: Heinze Karolin , Jansen Lars , Oliveira-Ferrer Leticia , Schmalfeldt Barbara , Dürst Matthias , Runnebaum Ingo B , Häfner Norman 
Published: June 2021

Abstract
INTRODUCTION: Ovarian cancer is still the most lethal gynecological malignancy with a low five-years-overall survival rate. Except for BRCA1/2 mutations there is no predictive nor prognostic marker available indicating whether a patient is in need and eligible for personalized therapy. In a previous genome-wide array study we identified different methylation patterns in type II epithelial ovarian cancer patients with either good or poor outcome (PFS &lt; vs. &gt; 3 years). Using methylation-specific PCR (MSP) five marker candidates showed high prognostic value whereby the marker combination RUNX3/CaMKIINα was the most promising one [Häfner et al., 2015]. Moreover, in vitro experiments provided evidence that a transcript variant of RUNX3 can mediate Cisplatin sensitivity, further underlining the clinical relevance of this gene [Heinze et al., 2018].
METHODS: In this study we aim to validate the previous described prognostic value of the five candidates, especially of RUNX3 and CaMKIINα via MSP. Therefore, 100 fresh-frozen primary type II EOC samples from two different surgical centers were tested. Furthermore, previously analyzed samples were retested to gain insight on the marker distributions throughout the tumor and possible heterogeneity. The DNA was isolated via phenol/chloroform or QiaAmp Kit, converted to bisulfite DNA and tested via optimized MSP. All samples had similar clinical parameter (histology, FIGO grade, chemotherapy).
RESULTS AND DISCUSSION: By re-testing a subset of EOC samples previous results could been confirmed. Additionally, the methylation pattern was similar after renewed sectioning for DNA extraction pointing towards little tumor heterogeneity in the analyzed samples. Interestingly the methylation rate is not as high as expected from the initial analyses. Statistical analysis of the data is currently ongoing.
REFERENCES
Häfner N, Steinbach D, Jansen L, Diebolder H, Dürst M, Runnebaum IB. RUNX3 and CAMK2N1 hypermethylation as prognostic marker for epithelial ovarian cancer, Int J Cancer. 2016
Heinze K, Kritsch D, Mosig AS, Dürst M, Häfner N, Runnebaum IB. Functional Analyses of RUNX3 and CaMKIINα in Ovarian Cancer Cell Lines Reveal Tumor-Suppressive Functions for CaMKIINα and Dichotomous Roles for RUNX3 Transcript Variants, Int J Mol Sci. 2018
Citation Format: Karolin Heinze, Lars Jansen, Leticia Oliveira-Ferrer, Barbara Schmalfeldt, Matthias Dürst, Ingo B Runnebaum, Norman Häfner. VALIDATION OF RUNX3 AND CAMKIINα HYPERMETHYLATION AS PROGNOSTIC MARKER FOR OVARIAN CARCINOMA [abstract]. In: Proceedings of the 12th Biennial Ovarian Cancer Research Symposium; Sep 13-15, 2018; Seattle, WA. Philadelphia (PA): AACR; Clin Cancer Res 2019;25(22 Suppl):Abstract nr GMM-028.
Abstract GMM-041: OVARIAN CANCER CELLS INDUCE A FEEDBACK RESPONSE TO PLATINUM-BASED CHEMOTHERAPY BY ACTIVATING AN ONCOGENIC RSK-EPHA2-GPRC5A SIGNALING SWITCH
Authors: Moyano-Galceran Lidia , Turunen Siri Pauliina , Pietilä Elina , Alkasalias Twana , Corvigno Sara , Hjerpe Elisabet , Joneborg Ulrika , Bulanova Daria , Eriksson Mina , Dahlstrand Hanna , Carlson Joseph , Lehti Kaisa 
Published: June 2021

Abstract
Despite increased understanding of chemotherapy resistance, the effects of the chemotherapy itself on the oncogenic cancer cell properties and adaptive resistance mechanisms of the therapy-escaping metastatic cancer cells remain largely undefined. In this study, we show that the platinum-based chemotherapeutic drugs cisplatin and carboplatin induce a switch from the canonical EphA2 receptor tyrosine kinase activity to the stemness and invasion-associated, ligand independent signaling via EphA2 S897 phosphorylation. In patient-derived high grade serous ovarian cancer (HGSOC) cells and established ovarian cancer cell lines, the chemotherapy treatment led to a significant increase in the ratio between S897 and Y588 phosphorylation of EphA2 in conjunction with GPRC5A protein upregulation and beta1-integrin activation. Notably, high protein levels of GPRC5A were significantly associated to poor survival in patients with metastatic HGSOC, revealing a previously unknown association of this EphA2 interacting receptor with the clinical HGSOC outcome. Moreover, pharmacological inhibition of RSK serine-threonine kinases blocked both the EphA2 S897 phosphorylation and the GPRC5A induction coincident with markedly increased cisplatin and carboplatin sensitivity of the HGSOC cells ex vivo. These results identify a novel mechanism of chemotherapy-induced adaptive resistance via the RSK-EphA2-GPRC5A oncogenic signaling axis in HGSOC.
Citation Format: Lidia Moyano-Galceran, Siri Pauliina Turunen, Elina Pietilä, Twana Alkasalias, Sara Corvigno, Elisabet Hjerpe, Ulrika Joneborg, Daria Bulanova, Mina Eriksson, Hanna Dahlstrand, Joseph Carlson, Kaisa Lehti. OVARIAN CANCER CELLS INDUCE A FEEDBACK RESPONSE TO PLATINUM-BASED CHEMOTHERAPY BY ACTIVATING AN ONCOGENIC RSK-EPHA2-GPRC5A SIGNALING SWITCH [abstract]. In: Proceedings of the 12th Biennial Ovarian Cancer Research Symposium; Sep 13-15, 2018; Seattle, WA. Philadelphia (PA): AACR; Clin Cancer Res 2019;25(22 Suppl):Abstract nr GMM-041.
Abstract GMM-044: IDENTIFICATION OF A NOVEL CANDIDATE BIOMARKER FOR DIAGNOSIS AND THERAPEUTIC OVARIAN CANCER TARGET
Authors: Park Kyoungsook , Son Chaeyeon , Song Sang Yong 
Published: June 2021

Abstract
Ovarian cancer is the most fatal gynecological cancer with high mortality. It still has no reliable diagnostic marker and effective treatment due to its high drug resistance and high recurrence. To develop a novel biomarker for diagnosis and therapeutic target of intractable ovarian cancer, we selected for an upregulated gene in serous ovarian tumors compared with normal ovary counterparts by microarray analysis and confirmed its elevated mRNA expression in ovarian tumors by RT-PCR. Consistent with its high mRNA level, tumor-specific preferential high expression of candidate protein was detected in the cytoplasm of the ovarian tumor tissues by immunohistochemistry. To our surprise, indirect immunofluorescence staining revealed an unexpected cytoplasmic subcellular localization of a candidate protein. Furthermore, forced expression of a candidate gene in cultured cells with no detectable level showed a marked inhibition of cell proliferation. To address these unexpected findings, we set out to identify a candidate interacting protein by yeast 2-hybrid screening to explore a novel function and its underlying mechanism. Among them, one of cell death associated protein was identified as a novel interacting partner of a candidate protein and their physical interaction in mammalian cells was confirmed by co-immunoprecipitation assay and confocal microscopy. Further domain mapping analysis should reveal critical domains for their interaction and would provide fundamental and pivotal information for a novel function and an underlying molecular mechanism of a candidate gene. This research was supported by Basic Science Research Program through the National Research Foundation of Korea (NRF) funded by the Ministry of Education (2018R1D1A1B07047640).
Citation Format: Kyoungsook Park, Chaeyeon Son, Sang Yong Song. IDENTIFICATION OF A NOVEL CANDIDATE BIOMARKER FOR DIAGNOSIS AND THERAPEUTIC OVARIAN CANCER TARGET [abstract]. In: Proceedings of the 12th Biennial Ovarian Cancer Research Symposium; Sep 13-15, 2018; Seattle, WA. Philadelphia (PA): AACR; Clin Cancer Res 2019;25(22 Suppl):Abstract nr GMM-044.