bluuebunny/crossref_metadata_2025 huggingface dataset:
Notch Represses Transcription by PRC2 Recruitment to the Ternary Complex
Authors: Han Xiaoqing , Ranganathan Prathibha , Tzimas Christos , Weaver Kelly L. , Jin Ke , Astudillo Luisana , Zhou Wen , Zhu Xiaoxia , Li Bin , Robbins David J. , Capobianco Anthony J.
Published: June 2017

It is well established that Notch functions as a transcriptional activator through the formation of a ternary complex that comprises Notch, Maml, and CSL. This ternary complex then serves to recruit additional transcriptional cofactors that link to higher order transcriptional complexes. The mechanistic details of these events remain unclear. This report reveals that the Notch ternary complex can direct the formation of a repressor complex to terminate gene expression of select target genes. Herein, it is demonstrated that p19Arf and Klf4 are transcriptionally repressed in a Notch-dependent manner. Furthermore, results indicate that Notch recruits Polycomb Repressor Complex 2 (PRC2) and Lysine Demethylase 1 (KDM1A/LSD1) to these promoters, which leads to changes in the epigenetic landscape and repression of transcription. The demethylase activity of LSD1 is a prerequisite for Notch-mediated transcriptional repression. In addition, a stable Notch transcriptional repressor complex was identified containing LSD1, PRC2, and the Notch ternary complex. These findings demonstrate a novel function of Notch and provide further insight into the mechanisms of Notch-mediated tumorigenesis.

Implications: This study provides rationale for the targeting of epigenetic enzymes to inhibit Notch activity or use in combinatorial therapy to provide a more profound therapeutic response. Mol Cancer Res; 15(9); 1173–83. ©2017 AACR.
Abstract AP06: SUBCLINICAL SIGNIFICANCE OF CIRCULATING TUMOR CELLS IN EPITHELIAL OVARIAN CANCER AS AN EARLY INDICATEOR OF RECURRENCE
Authors: Ruiz Carmen , Zheng Serena , Duggan Bridgette , Hicks James , Kuhn Peter
Published: June 2017

BACKGROUND: After diagnosis of cancer and subsequent treatment, a fraction of epithelial ovarian cancer (EOC) patients relapse with distant metastatic disease. While the fraction of the treated population that will relapse is predictable, there is not yet a way to determine the specific cases that will become metastatic. This transition between localized and disseminated cancer is clinically identified by the appearance of the patient's original cancer in a new anatomic location by various imaging methodologies or the increase of surrogate serum biomarkers. The actual biologic transition point, however, clearly occurs at some point prior to the time when metastasis becomes clinically evident. This becomes relevant during periods when disease is not clinically evident, where more sensitive identification methods of relapsing cancer may offer opportunities for therapy before tumor burden rises to an incurable level.

PURPOSE: With the observation that circulating tumor cells (CTCs) are found in cancer patients with no clinical evidence of disseminated disease prior to clinically evident relapse, we sought to investigate whether the presence of CTCs in the circulatory compartment could possibly indicate tumor dissemination at its earliest time point and whether that could be leveraged to develop predictive models and markers that identify patients prior to clinical evidence of recurrent disease.

METHODS: Peripheral blood samples collected after chemotherapy from 18 patients diagnosed with serous EOC were evaluated using the High Definition– Single Cell Assay (HD–SCA) for HD–CTCs enumeration, cytomorphological analysis and DNA copy number variation (CNV) profiling.

RESULTS: HD–CTCs (≥1 CTC/mL) were detected in 9 (50%) of these patients, 3 of whom relapsed. Despite the genomic heterogeneity of the CTCs analyzed, single cell CNV profiles from one of these recurrent patients showed clonal cyclin E1 amplification on chromosome 19 and ARID1A tumor suppressor gene deletion on chromosome 1, genomic alterations consistent with EOC.

CONCLUSION: The data suggests malignancy of multiple, single CTCs in recurrent patients, and further analysis of CTCs from non–recurrent patients could provide the means to determine the subclinical significance of CTCs in ovarian cancer. Earlier diagnosis of relapse, and subsequent personalized treatment, could eventually lead to more favorable prognoses in EOC patients.

Citation Format: Carmen Ruiz, Serena Zheng, Bridgette Duggan, James Hicks and Peter Kuhn. SUBCLINICAL SIGNIFICANCE OF CIRCULATING TUMOR CELLS IN EPITHELIAL OVARIAN CANCER AS AN EARLY INDICATEOR OF RECURRENCE [abstract]. In: Proceedings of the 11th Biennial Ovarian Cancer Research Symposium; Sep 12-13, 2016; Seattle, WA. Philadelphia (PA): AACR; Clin Cancer Res 2017;23(11 Suppl):Abstract nr AP06.
Abstract AP10: A NOVEL PAN–ALDH1A INHIBITOR INDUCES NECROPTOSIS IN OVARIAN CANCER STEM–LIKE CELLS AND SYNERGIZES WITH CHEMOTHERAPY TO IMPROVE OUTCOMES
Authors: Chefetz Ilana , Hong Linda , Yang Kun , Huddle Brandt , Landen Charles , Hurley Thomas , Larsen Scott D. , Buckanovich Ronald J.
Published: June 2017

BACKGROUND: We recently demonstrated that Aldehyde dehydrogenase (ALDH) activity and the stem cell marker CD133 can be used to define an ovarian cancer cell differentiation hierarchy. We demonstrated that even small increases in multipotent ALDHbrightCD133+ cancer stem–like cells (CSC) significantly increase chemotherapy resistance and tumor initiation capacity. ALDH1A family members (ALDH1A1, 1A2, and 1A3) have been implicated as the primary enzymes identifying CSC and have been strongly linked with chemotherapy resistance. Given the expression of ALDH1A family members in CSC and their role in chemotherapy resistance we hypothesized they could be important therapeutic targets.

METHODS AND RESULTS: We developed a panel of novel ALDH inhibitors (ALDHi). We identified a DEAB analog 673540 (673A) which inhibits all three ALDH1A1 enzymes (IC50 ~220nM). Pan–ALDH1A inhibition, induced necroptotic cell death specifically in ALDHbright and CD133+ ovarian CSC. 673A induced necroptosis was driven by the induction of the mitochondrial uncoupling proteins, UCP1 and UCP3, and reduction in OXPHOS capacity. Necroptosis was caspase and RIP1 independent, but is associated with MLKL translocation to membrane, and DRP1 activation and translocation to mitochondria. 673A therapy was highly synergistic with chemotherapy, reducing tumor initiation capacity, and resulting in a 60% tumor cure rate in multiple tumor models in–vivo.

CONCLUSIONS: ALDHi are highly active in experimental models of ovarian cancer. As ALDHi induce metabolic necroptosis, ALDHi are active in apoptosis resistant ovarian cancer cell lines. These studies support the development of ALDHi as novel therapeutics for ovarian cancer.

Citation Format: Ilana Chefetz, Linda Hong, Kun Yang, Brandt Huddle, Charles Landen, Thomas Hurley, Scott D. Larsen, and Ronald J. Buckanovich. A NOVEL PAN–ALDH1A INHIBITOR INDUCES NECROPTOSIS IN OVARIAN CANCER STEM–LIKE CELLS AND SYNERGIZES WITH CHEMOTHERAPY TO IMPROVE OUTCOMES [abstract]. In: Proceedings of the 11th Biennial Ovarian Cancer Research Symposium; Sep 12-13, 2016; Seattle, WA. Philadelphia (PA): AACR; Clin Cancer Res 2017;23(11 Suppl):Abstract nr AP10.
Abstract AP14: THERAPEUTIC TARGETING USING TUMOR SPECIFIC PEPTIDES INHIBITS LONG NON–CODING RNA HOTAIR ONCOGENIC ACTIVITY IN OVARIAN CANCER
Authors: Özeş Ali R. , Wang Yinu , Fang Fang , Nephew Kenneth P.
Published: June 2017

PURPOSE: Most women diagnosed with advanced-stage ovarian cancer develop chemo-resistant tumor recurrence, which is uniformly fatal. Long non-coding RNAs (lncRNAs) play key roles in human diseases, including cancer. Functional studies of the lncRNA HOTAIR (HOX transcript antisense RNA) provide compelling evidence for therapeutic targeting of HOTAIR in ovarian cancer to impact chemoresistance, but targeting lncRNAs in vivo has proven to be difficult.

EXPERIMENTAL DESIGN: In the current study, we describe a peptide nucleic acids (PNA)-based approach to effectively block the ability of HOTAIR to interact with EZH2 and subsequently inhibit oncogenic HOTAIR-EZH2 activity. In order to deliver the anti-lncRNA to the acidic (pH approximately 6) microenvironment of solid tumors in vivo, PNAs were conjugated to pH-low insertion peptide (pHLIP).

RESULTS: Treatment of HOTAIR-overexpressing ovarian cancer cell lines with PNAs decreased cancer cell invasion and increased chemotherapy sensitivity. The mechanism of action of the anti-lncRNA agent correlated with reduced nuclear factor-kappaB (NF-κB) activation and decreased expression of key NF-κB target genes including matrix metalloprotease 9 and interleukin 6. PNA treatment of ALDH1A1 positive/ovarian tumor initiating cells overexpressing HOTAIR markedly decreased ALDH1A1 activity and reduced the ovarian cancer stem cell population. In mice harboring ovarian tumor xerographs and treated with pHLIP-PNA constructs, HOTAIR oncogenic activity was suppressed, ovarian tumors were resensitized to platinum, and a significant (64%) survival improvement was observed.

CONCLUSIONS: Toward developing new therapeutic approaches against chemoresistant ovarian cancer, we designed a PNA-targeting strategy for HOTAIR that effectively blocks the ability of HOTAIR to interact with EZH2 and inhibits oncogenic HOTAIR-EZH2. This first report on pHLIP-PNA targeting HOTAIR in vivo offer pre-clinical proof-of-concept and represents a possible treatment alternative for chemotherapy-resistant ovarian cancer.

Citation Format: Ali R. Özeş, Yinu Wang, Fang Fang, and Kenneth P. Nephew. THERAPEUTIC TARGETING USING TUMOR SPECIFIC PEPTIDES INHIBITS LONG NON–CODING RNA HOTAIR ONCOGENIC ACTIVITY IN OVARIAN CANCER [abstract]. In: Proceedings of the 11th Biennial Ovarian Cancer Research Symposium; Sep 12-13, 2016; Seattle, WA. Philadelphia (PA): AACR; Clin Cancer Res 2017;23(11 Suppl):Abstract nr AP14.
Abstract AP21: LIPID DESATURATION IS A METABOLIC MARKER AND THERAPEUTIC TARGET OF OVARIAN CANCER STEM CELLS
Authors: Li Junjie , Condello Salvatore , Thomes-Pepin Jessica , Ma Xiaoxiao , Xia Yu , Hurley Thomas D. , Cheng Ji-Xin , Matei Daniela
Published: June 2017

OBJECTIVES: Lack of sensitive single-cell analysis tools has limited the characterization of the metabolic activity of cancer stem cells (CSCs). The objectives of this study were to identify and target metabolic pathways specifically deregulated in ovarian CSCs.

METHODS: We developed a new technique based on hyperspectral stimulated Raman scattering imaging of single living cells coupled with electrospray ionization mass spectrometry (ESI-MS) analysis of extracted lipids to identify unique metabolic spectra of ovarian CSCs. SiRNA knockdown and pharmacological inhibitors were used to block fatty acid desaturases and to measure effects on the ovarian CSCs charcateristics.

RESULTS: We report significantly increased levels of unsaturated lipids in flow-sorted ovarian CSCs (ALDH+CD133+) compared to non-CSCs (ALDH-CD133-). Higher lipid unsaturation levels were also detected in CSC-enriched spheroids compared to monolayer cultures of ovarian cancer cell lines or primary cells derived from human ovarian tumors. ESI-MS identified that the elevated unsaturation in lipid droplets involved an enhanced conversion from saturated fatty acids (18:0) to unsaturated fatty acids (18:1). Expression of fatty acid desaturases was increased in ovarian CSCs compared to non-CSCs. Pharmacological inhibition or siRNA knockdown of lipid desaturases Δ9 (SCD1) or Δ6 effectively eliminated CSCs, suppressed sphere formation in vitro, and blocked tumor initiation capacity in vivo, as measured by a tumor initiation dilution assay. Mechanistically, the lipid desaturase inhibitors suppressed ovarian cancer cell stemness by blocking the NF-κB survival pathway. Chromatin immunoprecipitation and overexpression of RelA demonstrated that NF-κB transcriptionally regulates SCD1 expression. Collectively, our data suggest that a positive feedback loop involving lipid desaturases, NF-κB, and the stem cell marker ALDH1 can be effectively blocked by SCD1 inhibitors, suppressing ovarian CSCs.

CONCLUSIONS: These results demonstrate that increased lipid unsaturation is a novel metabolic marker for ovarian CSCs and a target for CSC-specific therapy.

Citation Format: Junjie Li, Salvatore Condello, Jessica Thomes-Pepin, Xiaoxiao Ma, Yu Xia, Thomas D. Hurley, Ji-Xin Cheng, Daniela Matei. LIPID DESATURATION IS A METABOLIC MARKER AND THERAPEUTIC TARGET OF OVARIAN CANCER STEM CELLS [abstract]. In: Proceedings of the 11th Biennial Ovarian Cancer Research Symposium; Sep 12-13, 2016; Seattle, WA. Philadelphia (PA): AACR; Clin Cancer Res 2017;23(11 Suppl):Abstract nr AP21.
Abstract KP01: UNITED KINGDOM COLLABORATIVE TRIAL OF OVARIAN CANCER SCREENING – (UKCTOCS) – EXPLORING THE RESULTS
Authors: Menon Usha
Published: June 2017

The continued high case fatality ratio in ovarian cancer has underpinned efforts to develop a screening strategy that could impact on mortality. In the general population randomised control trial - United Kingdom Collaborative Trial of Ovarian Cancer Screening (UKCTOCS) two screening strategies (multimodal MMS and ultrasound USS) were compared to no screening (control C). Of 202,638 women randomised between April 2001-October 2005, 50,624 MMS; 50,623 USS; and 101,299 C were eligible for analysis. By the end of screening (December 2011) 345,570 MMS and 327,775 USS annual screens were performed. At censorship for the initial mortality analysis on 31st December 2014, there was an overall significant stage shift in invasive epithelial ovarian/tubal/peritoneal cancer in the MMS group compared to C in an ‘intention to screen’ analysis. More detailed analysis has shown this stage shift seen in the MMS group was limited to Type II (high grade serous) ovarian cancers. This was associated with a delayed but not yet statistically significant mortality reduction. Disease specific mortality rates in the C arm were continuing to rise while they appeared to have plateaued in the screen arms. Further follow-up is now underway to confirm the mortality reduction and determine its full extent. Meanwhile a focus on detecting low volume disease using cancer specific markers, novel biospecimens such as endocervical samples, targeted imaging and time series algorithms for interpreting marker profile suggest that a new era in ovarian cancer screening is underway.

Citation Format: Usha Menon. UNITED KINGDOM COLLABORATIVE TRIAL OF OVARIAN CANCER SCREENING – (UKCTOCS) – EXPLORING THE RESULTS [abstract]. In: Proceedings of the 11th Biennial Ovarian Cancer Research Symposium; Sep 12-13, 2016; Seattle, WA. Philadelphia (PA): AACR; Clin Cancer Res 2017;23(11 Suppl):Abstract nr KP01.
Abstract KP03: NOVEL IMMUNOTHERAPEUTICS FOR OVARIAN CANCER
Authors: Disis Mary L.
Published: June 2017

In the last decade we have seen significance advances in the understanding of how the immune system modulates the ovarian tumor microenvironment and cancer growth. A number of studies have shown that high levels of tumor infiltrating adaptive immune system cells (B-cells and T-cells) found in ovarian cancer are associated with a favorable prognosis, especially if there is evidence of reduced regulatory T-cells. Robust T-cell tumor infiltration (TIL) is associated with PD-L1 upregulation. Almost half of ovarian cancers demonstrate enhanced PD-L1 expression on tumor cells and/or tumor infiltrating immune system cells. Immune checkpoint inhibitor monoclonal antibodies block receptors such as PD-1 and PD-L1 which suppress T-cell function. The largest study of immune checkpoint inhibition in ovarian cancer evaluated the blockade of PD-L1 using avelumab. In this phase I/II study 124 patients with recurrent disease, considered chemotherapy refractory, were treated with 10mg/kg avelumab every 2 weeks (65% of patients with 3 or greater salvage regimens). The overall response rate was 10% with a stable disease rate of 44% resulting in a disease control rate of 54%. Overall survival at 12 months was 44%. Clinical response did not correlate with either PDL-1 expression on tumor or immune infiltrating cells or BRCA mutation status. A randomized trial of avelumab in combination with pegylated liposomal doxorubicin in cisplatin refractory ovarian cancer has been initiated. Pre-clinical studies have shown that clinical responses to immune checkpoint inhibitor therapy in ovarian cancer may be enhanced by increasing Type I T-cell infiltration concurrent with therapy. Clinical strategies aimed at increasing TIL include the use of vaccines immunizing against ovarian cancer antigens to stimulate T-cells trafficking to tumor. Specific chemotherapy may result in increased TIL as some cytotoxic agents have been shown to modulate both the level and phenotype of TIL in ovarian cancer. An alternate approach is to suppress alternate pathways of tumor immune escape such as depletion of macrophage or inhibition of the IDO pathway to enhance immune activation. Several promising combination immunotherapy strategies are currently being evaluated in ovarian cancer.

Citation Format: Mary L. Disis, MD. NOVEL IMMUNOTHERAPEUTICS FOR OVARIAN CANCER [abstract]. In: Proceedings of the 11th Biennial Ovarian Cancer Research Symposium; Sep 12-13, 2016; Seattle, WA. Philadelphia (PA): AACR; Clin Cancer Res 2017;23(11 Suppl):Abstract nr KP03.
Abstract MIP-047: INHIBITION OF POLY ADP–RIBOSE POLYMERASE (PARP) IN OVARIAN CANCER CELLS ENRICHES FOR CD133 AND CD117 POSITIVE POPULATIONS
Authors: Bellio Chiara , Foster Rosemary , Growdon Whitfield B. , Konstantinopoulos Panagiotis , Rueda Bo R.
Published: June 2017

Keywords: chemoresistance, cancer stem cells (CSC), DNA repair pathways, PARP inhibition

INTRODUCTION: Ovarian cancer (OvCa) is the most lethal gynecologic malignancy as women commonly present with metastatic disease that ultimately develops chemotherapy resistance. The development of OvCa has been linked to alterations in DNA repair pathways and the poly (adenosine diphosphate [ADP]–ribose) polymerase (PARP) inhibitor olaparib has demonstrated antitumor activity and is an FDA approved monotherapy for the treatment of recurrent OvCa in patients with a germline BRCA mutations. Despite promising response rates, not all patients derive benefit, and the majority develop resistance to this therapy that is poorly understood. We hypothesized that ultimate resistance to olaparib is in part mediated by small populations of cancer stem cells (CSCs) that are unaffected by olaparib therapy and may promote tumor resurgence.

METHODS: UWB1.289 (BRCA1 mutated), UWB1.289 (WT), OVCAR4 (WT), and OVCAR3 (WT) cell lines and primary cell lines derived from serous OvCa obtained with IRB approval were treated in vitro with 0.1 μM, 1 μM and 10 μM olaparib for 7 days. We assessed cell viability and CSC marker expression (CD133 and CD117) via flow cytometric analysis, and sphere forming capacity via extreme limiting dilution assay (ELDA).

RESULTS: Consecutive treatment over 7 days with olaparib decreased the viability of the non–sorted tumor cell population relative to the vehicle control treated cells. Despite the decrease in total cell number, the relative frequency of CD133+ and CD117+ cells increased (all lines p <0.05) in response to treatment with olaparib. This shift was not observed in vehicle treated cells. The increase in CD133+ and CD117+ cell frequency in response to olaparib was not different in the BRCA1 mutant line when compared to the BRCA1 wild type lines. ELDA revealed an increase in the frequency of sphere formation following olaparib treatment relative to that observed with vehicle treated cells, though mutation status did not impact this frequency.

CONCLUSIONS: These results suggest that olaparib may enrich for CSC populations that could contribute to resistance and recurrence. These data expose a potential resistance mechanism to olaparib therapy that is in part independent of BRCA mutation status.

Citation Format: Chiara Bellio, Rosemary Foster, Whitfield B. Growdon Panagiotis Konstantinopoulos, and Bo R. Rueda. INHIBITION OF POLY ADP–RIBOSE POLYMERASE (PARP) IN OVARIAN CANCER CELLS ENRICHES FOR CD133 AND CD117 POSITIVE POPULATIONS [abstract]. In: Proceedings of the 11th Biennial Ovarian Cancer Research Symposium; Sep 12-13, 2016; Seattle, WA. Philadelphia (PA): AACR; Clin Cancer Res 2017;23(11 Suppl):Abstract nr MIP-047.
Abstract NTOC-101: INTRAPERITONEAL (IP) THERAPY FOR ADVANCED STAGE EPITHELIAL OVARIAN CANCER (EOC): THE CURRENT SPOTLIGHT IS ON CARBOPLATIN AND QUALITY OF LIFE
Authors: Muggia Franco , Wenzel Lari
Published: June 2017

PURPOSE: First-line treatments for (EOC) have been platinum-based for more than three decades; 16 years ago the landmark Gynecologic Oncology Group (GOG158) non-inferiority trial for optimal stage III patients led to the universal adoption of IV carboplatin/paclitaxel as the standard systemic regimen. However, IV cisplatin-based doublets were the comparators in the first 3 GOG studies assessing IP cisplatin-based regimens. Our purpose is to describe recent randomized trials that have assessed IP carboplatin and highlight quality of life (QOL) findings.

METHODS: Gynecology Oncology Group (GOG) studies comparing IP to IV cisplatin regimens formed the basis of the National Cancer Institute (NCI) 2005 Clinical Announcement that considered IP cisplatin-based therapy as a new standard for optimally debulked EOC. Recently, randomized trials with IV carboplatin/paclitaxel-based regimens as comparators have been part of preliminary reports: 1) GOG252 on March 2016 at the Society of Gynecologic Oncology (SGO), completing accrual in 2015, and 2) OV21/PETROC neoadjuvant randomized ‘pick the winner’ study at the June 2016 American Society of Clinical Oncology (ASCO). Beyond 2016, additional data is expected from the ongoing randomized study by the Japan GOG of IP versus IV carboplatin, --both with IV paclitaxel.

RESULTS: GOG252, the largest Phase III study of IV versus IP platinum-based therapy for the first line treatment of ovarian cancer, is the first IP GOG trial with a IV carboplatin-based control arm. None of the 3 arms differed in median progression-free survival. However, the companion QOL study, --contrary to prior GOG studies utilizing IV cisplatin-based controls—showed a clear advantage for the two non-cisplatin arms (not different whether IV or IP). In the OV21/PETROC trial, adverse toxicities coupled with no obvious therapeutic advantage led to dropping the IP cisplatin arm. With additional accrual in the two remaining arms, IP carboplatin was superior to IV carboplatin in time to progression.

CONCLUSIONS: Cisplatin has an unfavorable impact on QOL in women with ovarian cancer whether given IP or IV relative to carboplatin. With IV carboplatin as part of the comparator, unfavorable effects of cisplatin-based chemotherapy are further highlighted. IP carboplatin randomized studies and their final publications are awaited. On the other hand, revisiting the role of IP cisplatin at this time is not warranted.

Citation Format: Franco Muggia MD, Lari Wenzel, PhD. INTRAPERITONEAL (IP) THERAPY FOR ADVANCED STAGE EPITHELIAL OVARIAN CANCER (EOC): THE CURRENT SPOTLIGHT IS ON CARBOPLATIN AND QUALITY OF LIFE [abstract]. In: Proceedings of the 11th Biennial Ovarian Cancer Research Symposium; Sep 12-13, 2016; Seattle, WA. Philadelphia (PA): AACR; Clin Cancer Res 2017;23(11 Suppl):Abstract nr NTOC-101.
Abstract NTOC-105: RUXOLITINIB SENSITIZES OVARIAN CANCER CELLS TO LOW–DOSE PACLITAXEL THERAPY
Authors: Reeves Patrick , Abbaslou Mojgan , Poznansky Mark C.
Published: June 2017

In ovarian cancer chemotherapy initially reduces tumor burden; however, >70% of patients experience tumor recurrence, creating a need for additional therapies. We evaluated the ability of Ruxolitinib, a FDA approved JAK 1/2 kinase inhibitor, as a potential adjunctive therapy for paclitaxel treatment of ovarian cancer. Using murine ID8 and human TOV-112D ovarian cancer cells we tested the ability of Ruxolitinib alone and in combination with low-dose paclitaxel to limit cell proliferation and colony formation in vitro. Additionally, using flow cytometry we assessed the capacity of paclitaxel to induce the expression of cell surface markers associated with stemness and the cellular stress response. We next determined if Ruxolitinib could reduce paclitaxel-induced expression of stemness and stress markers. Finally daily administration of Ruxolitinib, both alone and in combination with a single low-dose administration of paclitaxel, was tested for the capacity to reduce tumor growth and extend survival using the syngeneic ID8 luciferase-expressing model of ovarian cancer in C57Bl/6 mice.

Our in vitro data demonstrate that Ruxolitinib can sensitize both murine ID8 and human TOV-112D cells to low concentrations of paclitaxel, as evidenced by a significant reduction of cell proliferation and colony formation. We find that pre-incubation with Ruxolitinib is sufficient to sensitize cells to subsequent incubation with paclitaxel. Paclitaxel-induced expression of stemness and stress markers (SCA-1, GRP78 and CD133) is reduced by co-incubation with Ruxolitinib. In vivo, Ruxolitinib plus administration of a single low-dose paclitaxel treatment is sufficient to limit tumor growth and significantly extend median survival from in mice bearing ID8 ovarian cancer tumors (Control 59days vs Ruxolitinib-paclitaxel 77 days).

Together, these data indicate that Ruxolitinib can increase the susceptibility of both murine and human ovarian cancer cells to reduced concentrations paclitaxel chemotherapy in vitro and that combination therapy with Ruxolitinib and a single low-dose paclitaxel administration extends survival in mice. Our data provide important pre-clinical data to support the use of Ruxolitinib to enhance the efficacy of paclitaxel therapy for patients with ovarian cancer.

Citation Format: Patrick Reeves PhD, Mojgan Abbaslou MD, Mark C. Poznansky MD PHD. RUXOLITINIB SENSITIZES OVARIAN CANCER CELLS TO LOW–DOSE PACLITAXEL THERAPY [abstract]. In: Proceedings of the 11th Biennial Ovarian Cancer Research Symposium; Sep 12-13, 2016; Seattle, WA. Philadelphia (PA): AACR; Clin Cancer Res 2017;23(11 Suppl):Abstract nr NTOC-105.
Abstract NTOC-113: TGF–β SIGNALING INHIBITION AS A POTENTIAL APPROACH TO TARGET SUBOPTIMALLY DEBULKED OVARIAN TUMORS
Authors: Wei Wei , Birrer Michael J.
Published: June 2017

The current standard of care for all patients with high-grade, advanced-stage serous ovarian cancer is surgery followed by adjuvant chemotherapy. Resection of all visible residual disease to less than 1 cm is associated with a favorable prognosis, but this optimal debulking can only be achieved in ~70% of patients. Patients with suboptimal cytoreduction are presumed to have lower response rates to subsequent chemotherapy and are known to have a poor prognosis.

Recently, we performed a meta-analysis of 1,525 publicly available expression profiling assays and identified a 200-gene expression signature of primary ovarian tumors to predict the outcome of debulking surgery. The signature was validated using two independent sets of tumors. This debulking signature consists of transcripts positively regulated by TGF-β, indicating that hyperactivation of TGF-β signaling may be responsible of the aggressive biological and clinical behavior of suboptimally debulked tumors.

In this study, we evaluated the potential anti-tumor effect of TGF-β signaling blockade in ovarian cancer cells in vitro and in vivo. Integrated TGF-β signaling was observed in 8 of 10 ovarian cancer cell lines tested by a SMAD-binding element based luciferase assay. The potential anti-tumor effect of a small molecular TGFBR1 kinase inhibitor LY2157299 was further investigated in SKOV3 cell line, which presents both endogenous and inducible TGF-β signaling and is highly disseminative when inoculated into immunodeficient mice. Although imposing minimal effect on in vitro cellular proliferation, LY2157299 significantly inhibited tumor-dissemination related biological processes in SKOV3 cells including migration, invasion and mesothelial cell adhesion. In an interperitoneal SKOV3 xenograft model, administration of LY2157299 significantly increased cisplatin related cytotoxicity and tumor burden reduction. Taken together, our study has provided evidence for the potential clinical use of LY2157299 to improve the treatment and survival of women with suboptimal surgery.

Citation Format: Wei Wei and Michael J. Birrer. TGF–β SIGNALING INHIBITION AS A POTENTIAL APPROACH TO TARGET SUBOPTIMALLY DEBULKED OVARIAN TUMORS [abstract]. In: Proceedings of the 11th Biennial Ovarian Cancer Research Symposium; Sep 12-13, 2016; Seattle, WA. Philadelphia (PA): AACR; Clin Cancer Res 2017;23(11 Suppl):Abstract nr NTOC-113.
Abstract TMEM-027: TARGETING RGNEF (P190RHOGEF/ARHGEF28) IMPAIRS OVARIAN TUMOR INITIATION AND PROGRESSION
Authors: Kleinschmidt Elizabeth G. , Miller Nichol L.G. , Tancioni Isabelle , Connolly Denise C. , Schlaepfer David D.
Published: June 2017

Rho GTPase regulatory and integrin signaling pathways are associated with poor high-grade serous ovarian carcinoma (HGSOC) patient survival1. Focal adhesion kinase (FAK) transmits signals from integrins and promotes HGSOC tumor progression through mechanisms involving tumor-stromal cell interactions2. We previously found that the Rho guanine nucleotide exchange factor Rgnef (also named p190RhoGEF or Arhgef28) activates FAK as well as RhoA GTPase activity in mouse fibroblasts3. Here we find that Rgnef protein expression is elevated in Stage 3-4 HGSOC and that Rgnef knockout prevents spontaneous ovarian tumor formation in the MISIIR-T-antigen (TAg) induced C57BL/6 mouse model. At 17 weeks, Rgnef-/-;TAg ovarian tumors were significantly smaller (p<0.001) than Rgnef+/+;TAg controls. Primary Rgnef-/-;TAg and Rgnef+/+;TAg tumor cells were isolated and expanded ex vivo. Although loss of Rgnef did not alter growth as 3D spheroids, Rgnef-/-;TAg orthotopic (ovarian bursa) and intraperitoneal tumors were significantly smaller (p<0.01) than Rgnef+/+;TAg tumors in MISIIR-TAg-Low C57BL/6 syngeneic mice. Notably, FAK activation (as measured by FAK Y397 phosphorylation) was decreased in Rgnef-/-;TAg ascites-associated tumor cells. Together, these studies show that Rgnef loss impedes tumor growth independently of stromal Rgnef status and supports the notion that an Rgnef-FAK signaling linkage facilitates ovarian tumor initiation and progression.

1.Zhang, H., et al. Integrated Proteogenomic Characterization of Human High-Grade Serous Ovarian Cancer. Cell 166 (2016).

2.Sulzmaier, F.J., Jean, C. & Schlaepfer, D.D. FAK in cancer: mechanistic findings and clinical applications. Nat. Rev. Cancer 14, 598-610 (2014).

3.Miller, N.L., et al. A non-canonical role for Rgnef in promoting integrin-stimulated focal adhesion kinase activation. J. Cell Sci. 126, 5074-5085 (2013).

Citation Format: Elizabeth G. Kleinschmidt, Nichol L.G. Miller, Isabelle Tancioni, Denise C. Connolly, David D. Schlaepfer. TARGETING RGNEF (P190RHOGEF/ARHGEF28) IMPAIRS OVARIAN TUMOR INITIATION AND PROGRESSION [abstract]. In: Proceedings of the 11th Biennial Ovarian Cancer Research Symposium; Sep 12-13, 2016; Seattle, WA. Philadelphia (PA): AACR; Clin Cancer Res 2017;23(11 Suppl):Abstract nr TMEM-027.
Abstract TMEM-039: THE ROLE OF SNAIL IN CELL DIFFERENTIATION STATUS: EPITHELIAL-MESENCHYMAL TRANSITION AND CANCER STEM CELLS
Authors: Wang Hanmin , Huisken-Hill Alyse , Chirshev Evgeny , Unternaehrer-Hamm Juli
Published: June 2017

PURPOSE: The epithelial-mesenchymal transition (EMT) is a process by which epithelial cells lose their cell polarity and cell-cell adhesion, and become mesenchymal cells with migratory and invasive properties. EMT is essential for numerous developmental processes such as gastrulation, and is involved in the initiation of metastasis for cancer progression.

Cancer cells that undergo EMT are enabled to metastasize to form secondary tumors. One characteristic of advanced tumors is a low expression level of let-7, a miRNA known to be vital in both maintenances of cellular differentiation and tumor suppression. There is a gap in the literature concerning the mechanism of let-7 downregulation. Our purpose in this study is to present data that shows Snail, an EMT inducer, directly represses let-7, hence inducing cancer stem cells.

EXPERIMENTAL PROCEDURES: Ovarian, pancreatic and breast cancer cell lines treated with TGFβ or EGF, inducers of EMT, were studied in our experiments. Expression of several markers at protein level was visualized with immunofluorescence and quantified with flow cytometry. Gene expression of mesenchymal, epithelial and pluripotency factors and miRNAs was detected by q-RT-PCR. Chromatin immunoprecipitation of Snail was performed in ovarian cancer lines. Let-7i promoter luciferase with and without Snail were co-transfected into OVSAHO and 293T cell lines and followed by luciferase assays for detection of bioluminescence. Snail was knocked down by lentiviral small hairpin RNA. Snail was overexpressed by estrogen receptor fusion protein. Orthotopic xenografts of patient-derived ovarian cancer cells were used to test the effect of Snail knockdown on tumor burden.

RESULTS: In selected ovarian cancer lines, levels of Snail expression correlated directly with Nanog, and indirectly with let-7 expression. Snail bound promoters of let-7 in cell lines tested. Luciferase assays demonstrated direct repression of let-7 transcription by Snail. Metastatic burden was significantly reduced in cell line xenografts in which Snail was knocked down.

CONCLUSIONS: Let-7 is confirmed to be negatively related to Snail. Our preliminary data also shows Snail directly represses let-7, therefore induces properties of stemness in cancer cells. Cell line and patient-derived data support the relationship between Snail expression, let-7 downregulation, and the induction of cancer stem cells. Snail knockdown appears to disrupt the cancer stem cell state and reduces disease progression.

Citation Format: Hanmin Wang, Alyse Huisken-Hill, Evgeny Chirshev, Juli Unternaehrer-Hamm. THE ROLE OF SNAIL IN CELL DIFFERENTIATION STATUS: EPITHELIAL-MESENCHYMAL TRANSITION AND CANCER STEM CELLS [abstract]. In: Proceedings of the 11th Biennial Ovarian Cancer Research Symposium; Sep 12-13, 2016; Seattle, WA. Philadelphia (PA): AACR; Clin Cancer Res 2017;23(11 Suppl):Abstract nr TMEM-039.
Abstract TMEM-042: MALIGNANT EXTRACELLULAR VESICLES CAUSE MESOTHELIAL CELL DAMAGE TOWARD PERITONEAL DISSEMINATION IN OVARIAN CANCER
Authors: Yokoi Akira , Ochiya Takahiro
Published: June 2017

PURPOSE: To elucidate novel mechanisms of peritoneal dissemination by focusing on extracellular vesicles

BACKGROUNDS AND SUMMARY: Metastasis to the abdominal cavity is frequently observed in ovarian cancer patients and is one of the reasons for the unfavorable outcomes. Extracellular vesicles (EVs), including exosomes and microvesicles, are small membrane vesicles that contain various bioactive molecules, such as microRNAs, mRNAs and proteins, and they are released from all cell types and play key physiological roles in intercellular communication. Recent evidence has demonstrated that cancer cells secrete EVs to both proximal surrounding cells and distal sites, thereby enabling the development of a cancer microenvironment that in turn promotes cancer metastasis.

We established orthotopic mouse models by using 4 ovarian carcinoma cell lines with different metastatic traits and investigated the functions of cancer-derived EVs. The EVs from highly metastatic cells strongly induced metastatic behavior in moderately metastatic tumors. Notably, the metastatic cancer EVs efficiently induced apoptotic cell death in mesothelial cells in vitro and in vivo, thus resulting in the destruction of the peritoneal mesothelium barrier. Whole transcriptome analysis showed that MMP1 was significantly elevated in mesothelial cells treated with highly metastatic cancer EVs, and intact MMP1 mRNAs were selectively packaged in the EVs. Importantly, MMP1 expression in ovarian cancer is tightly correlated with a poor prognosis, particularly in stage I patients. Moreover, we found MMP1 mRNA-carrying EVs in the ascites of cancer patients, and these EVs also induced apoptosis in mesothelial cells.

CONCLUSIONS: Our findings provide a promise for the understanding of a previously unknown mechanism for peritoneal dissemination that is associated with cancer-derived EVs, and which may be novel biomarkers for prognosis and can provide a new therapeutic strategy for inhibiting metastasis by disrupting the EVs.

Citation Format: Akira Yokoi and Takahiro Ochiya. MALIGNANT EXTRACELLULAR VESICLES CAUSE MESOTHELIAL CELL DAMAGE TOWARD PERITONEAL DISSEMINATION IN OVARIAN CANCER [abstract]. In: Proceedings of the 11th Biennial Ovarian Cancer Research Symposium; Sep 12-13, 2016; Seattle, WA. Philadelphia (PA): AACR; Clin Cancer Res 2017;23(11 Suppl):Abstract nr TMEM-042.
House Dust Mites Induce Production of Endothelin-1 and Matrix Metalloproteinase-9 in Keratinocytes via Proteinase-Activated Receptor-2 Activation
Authors: Yamada Yoshihito , Matsumoto Tatsumi
Published: June 2017

Background: Atopic dermatitis (AD) is a chronic inflammatory skin disease characterized by skin barrier dysfunction and abnormal immune response. House dust mites (HDM) are a major source of allergens, some of which have cysteine and serine protease activities. Keratinocytes stimulated by HDM-derived proteases have been suggested to contribute to the pathogenesis of AD by producing various cytokines. However, whether keratinocytes contribute to the induction of pruritus in AD, especially by producing pruritus-related mediators upon stimulation with HDM-derived proteases, has not been fully elucidated. Methods: We examined whether the production of endothelin-1 (ET-1), matrix metalloproteinase (MMP)-2, and MMP-9 in keratinocytes can be induced by stimulation with Dermatophagoides farinae extracts, and if so, whether pretreatment with a protease inhibitor or proteinase-activated receptor-2 (PAR-2) antagonist affects the production of these mediators in keratinocytes. Results: Although MMP-2 levels were undetectable in the culture supernatants, the production of ET-1 and MMP-9 was increased upon stimulation with HDM extracts in a concentration- and time-dependent manner and suppressed by pretreatment of HDM extracts with serine protease inhibitor, but not with cysteine protease inhibitor. Mite-derived serine proteases also induced ET-1 and MMP-9 production in a concentration- and time-dependent manner. Moreover, pretreatment with a PAR-2 antagonist inhibited the production of ET-1 and MMP-9 in keratinocytes. Conclusion: These results suggest that the activation of PAR-2 on keratinocytes by HDM-derived serine proteases induces the production of ET-1 and MMP-9, and may contribute to the induction of pruritus in AD.
Clinical Features and Recurrence Pattern of Perianal Abscess in Patients with Acute Myeloid Leukemia
Authors: Chang Hung , Kuo Ming-Chung , Tang Tzung-Chih , Lin Tung-Liang , Wu Jin-Hou , Hung Yu-Shin , Wang Po-Nan
Published: June 2017

Introduction: Perianal abscess may develop during neutropenia periods in patients with acute myeloid leukemia (AML). The standard of care for perianal abscess in AML is unclear. Methods: We retrospectively collected patient data in our institute from 2009 to 2012. Results: Two hundred ninety-two patients with AML were analyzed. In total, 1,051 chemotherapy sessions were administered. Twenty-three patients experienced perianal abscess. Patients with perianal abscess were younger than those without (44 vs. 60 years, p < 0.0001). Perianal abscess developed in various phases of treatment and in the stem cell transplantation period. Twelve recurrences developed in 6 patients. Patients with a prior perianal abscess have a 10-fold risk of developing a subsequent abscess following further chemotherapy. The microbiology profile revealed that most pathogens were derived from the intestinal tracts, which was similar to the findings of previous studies. The 28-day mortality was 14.3% and the direct cause of death was not perianal abscess in any case. Surgical interventions had no impact on recurrence or survival. Conclusion: In patients with AML, perianal abscess results from gastrointestinal tract pathogens. Many patients do not require surgical interventions. The mortality is low but recurrence is common following subsequent chemotherapies. Therefore, awareness of recurrence is important for the timely management of perianal abscess in AML.
Validation of Surrogates of Urine Osmolality in Population Studies
Authors: Youhanna Sonia , Bankir Lise , Jungers Paul , Porteous David , Polasek Ozren , Bochud Murielle , Hayward Caroline , Devuyst Olivier
Published: June 2017

Background: The importance of vasopressin and/or urine concentration in various kidney, cardiovascular, and metabolic diseases has been emphasized recently. Due to technical constraints, urine osmolality (Uosm), a direct reflect of urinary concentrating activity, is rarely measured in epidemiologic studies. Methods: We analyzed 2 possible surrogates of Uosm in 4 large population-based cohorts (total n = 4,247) and in patients with chronic kidney disease (CKD, n = 146). An estimated Uosm (eUosm) based on the concentrations of sodium, potassium, and urea, and a urine concentrating index (UCI) based on the ratio of creatinine concentrations in urine and plasma were compared to the measured Uosm (mUosm). Results: eUosm is an excellent surrogate of mUosm, with a highly significant linear relationship and values within 5% of mUosm (r = 0.99 or 0.98 in each population cohort). Bland-Altman plots show a good agreement between eUosm and mUosm with mean differences between the 2 variables within ±24 mmol/L. This was verified in men and women, in day and night urine samples, and in CKD patients. The relationship of UCI with mUosm is also significant but is not linear and exhibits more dispersed values. Moreover, the latter index is no longer representative of mUosm in patients with CKD as it declines much more quickly with declining glomerular filtration rate than mUosm. Conclusion: The eUosm is a valid marker of urine concentration in population-based and CKD cohorts. The UCI can provide an estimate of urine concentration when no other measurement is available, but should be used only in subjects with normal renal function.
Serum Levels of MicroRNA371a-3p: A Highly Sensitive Tool for Diagnosing and Staging Testicular Germ Cell Tumours: A Clinical Case Series
Authors: Anheuser Petra , Radtke Arlo , Wülfing Christian , Kranz Jennifer , Belge Gazanfer , Dieckmann Klaus-Peter
Published: June 2017

Introduction: MicroRNA (miR)371a-3p was suggested to be a sensitive and specific new serum biomarker of germ cell tumours (GCTs); however, its clinical usefulness remains unproven. Patients, Methods: In 312 consecutive cases with various testicular diseases, serum levels of miR371a-3p were measured. Measurement results became available only after completion of treatment. Five patients with testicular seminoma were selected for review because of unanticipated clinical courses. Results: In each two patients, elevated miR levels heralded undetected primary testicular GCT and metastases despite inconclusive radiological findings. In one case, a normal miR371a-3p level correctly pointed to the absence of metastases contrary to clinical assessment. In all cases, knowledge about the miR371a-3p levels would have altered the clinical management. Conclusions: These cases highlight the exceptional usefulness of the new GCT biomarker. In contrast to classical markers, miR371a-3p can identify primary testicular GCT. The marker can aid in clinical decision making in cases with ambiguous clinical findings.
Comparison of the Clinical Effectiveness of Ultrasound-Guided Corticosteroid Injection with and without Needle Release of the Transverse Carpal Ligament in Carpal Tunnel Syndrome
Authors: Guo Xuan-Yan , Xiong Mao-xiang , Zhao Yu , He Fan-Ding , Cheng Xue-Qing , Wu Yan-Yan , Chen Kai , Lu Man
Published: June 2017

Background: To compare the clinical effectiveness of ultrasound-guided corticosteroid injection with and without needle release of the transverse carpal ligament (TCL) in carpal tunnel syndrome (CTS). Methods: Between May 2014 and June 2016, 52 patients (56 wrists) with CTS were included in this study. Among these patients, 28 wrists were treated with ultrasound-guided corticosteroid injection plus needle release of the TCL (group A) and 28 wrists were treated with a single ultrasound-guided corticosteroids injection (group B). The following parameters were assessed and compared including postoperative results of procedure based on relief of symptoms, electrophysiological parameters (distal motor latency, sensory conduction velocity, and sensory nerve action potential of median nerve), and ultrasound parameters (anteroposterior diameter and cross-sectional area of the median nerve at the levels of pisiform and hamate bone, and the thicknesses of TCL on the cross-section at the level of hamate bone). Results: The overall excellent and good rate regarding the postoperative results of procedure based on the relief of symptoms at 1 month postoperatively was 82.1% in group A and 46.4% in group B (p = 0.004). There was significant difference in the above electrophysiological and ultrasound parameters between the preoperative and postoperative values in both groups (all p < 0.05). Furthermore, a significant difference was also observed in the postoperative values of the above-mentioned electrophysiological and ultrasound parameters in the 2 groups (all p < 0.05). Conclusions: Both approaches had treatment benefit in CTS. Ultrasound-guided corticosteroid injection in combination with needle release of the TCL is superior to the single ultrasound-guided corticosteroids injection.
Um Estudo de Equações Diferenciais
Authors: Noce Eduardo Della
Published: June 2017

O objetivo desse texto é apresentar conceitos básicos e fundamentais para iniciantes sobre equações diferenciais, seus tipos e algumas soluções. Destina-se ao público no geral que deseja começar os estudos no tema ou deseja rever algo importante.
Conto Odontotrônico: Prótese Personalizada
Authors: Dos Santos Gabriel Emídio , Pereira Macario Ferreira Maria Cristina
Published: June 2017

Pairava sobre o consultório o aroma de café sendo feito, como habitualmente ocorria. A paciente Irene estava ansiosa para poder visualizar de maneira tangível as próteses que iriam compor, por um longo período de tempo, seu sorriso. [...]