High-grade Serous Ovarian Adenocarcinoma Research Articles

Amplification of Hippo Signaling Pathway Genes Is Governed and Implicated in the Serous Subtype-Specific Ovarian Carcino-Genesis.

Among women, ovarian cancer ranks as the fifth most common cause of cancer-related deaths. This study examined the impact of Hippo signaling pathway on ovarian carcinogenesis. Therefore, the signatures related to Hippo signaling pathway were derived from the molecular signatures database (MSigDB) and were used for further analysis. The Z score-based pathway activation scoring method was employed to investigate the expression patterns of these signatures in the mRNA expression profiles of ovarian cancer cohorts. Compared to other subtype tumors, the results of this study show that the Hippo signaling pathway signatures are dysregulated prominently in serous subtype-specific ovarian carcinogenesis. A receiver operating characteristic (ROC) curve-based results of the Hippo gene set, yes-associated protein 1 (YAP1), and mammalian sterile 20-like kinases 1 (MST1) genes can predict the serous subtype tumors by higher specificity and sensitivity with significant areas under the curve values also further reconfirmed these signaling dysregulations. Moreover, these gene sets were studied further for mutation analysis in the profile of high-grade serous ovarian adenocarcinoma in the cBioPortal database. The OncoPrint results reveal that these Hippo signaling pathway genes are amplified highly during the grade three and stage third or fourth of serous type ovarian tumors. In addition, the results of the Dependency Map (DepMap) plot also clearly show that these genes are amplified significantly across the ovarian cancer cell lines. Finally, overall survival (OS) curve plot investigations also revealed that these gene expressions show poor survival patterns linked to highly expressed conditions in serous subtypes of ovarian cancer patients with significant p-values (p < 0.05). Thus, the current finding would help to develop the targeted therapies treatment for serous subtype ovarian carcinogenesis.

E030 Rheumatologists beware! Digital ischaemia as a presenting feature in metastatic ovarian cancer

Abstract Background/Aims Digital ischaemia is an umbrella term to describe the spectrum of acral vascular phenomena irrespective of aetiology, ranging from temporary (as in Raynaud’s phenomenon) to permanent (necrosis). It can be the presenting feature in multiple cancer types, and can be present with or without other features of connective tissue disease. Methods A 54-year-old woman was referred to Rheumatology with a four-day history of acute pain and ischaemic discolouration of all her fingertips bilaterally. There was no history of Raynaud’s phenomenon and no other clinical features of connective tissue disease. She had never smoked. There was a family history of breast cancer in her maternal aunt. She was admitted under vascular surgery for urgent review. All ten fingertips were ischaemic, with a capillary refill time of greater than five seconds. The history was otherwise significant for abdominal bloating, altered bowel habit and fatigue. Examination revealed normal peripheral pulses with no bruits, normal heart sounds and chest sounds. Her abdomen was soft with a mildly enlarged liver. Results FBC, U&amp;E, and LFTs showed no abnormalities. CRP, ESR, ANA, serum protein electrophoresis and complement levels were all normal. Blood tests for ANCA, rheumatoid factor and anti-CCP were also negative. Lupus anticoagulant, anti-cardiolipin and anti-B2-glycoprotein were all negative. The tumour marker CA125 was markedly elevated at 837kU/L (0.0-35.0kU/L). A CT abdomen &amp; pelvis showed a large right-sided adnexal mass with multiple enhancing peritoneal and omental nodules and moderate ascites. Biopsy of a metastatic deposit in the omentum revealed high grade serous ovarian adenocarcinoma. Partial improvement in digital ischaemia was achieved with iloprost and maintenance apixaban and sildenafil. Four cycles of carboplatin and paclitaxel chemotherapy produced near-complete resolution of digital ischaemia symptoms, corresponding to a fall in her CA125 level and reduction in the size of the ovarian mass and intra-abdominal metastases on CT. Ten cases of digital ischaemia associated with ovarian cancer have been described prior to this. The age of onset ranged from 45 to 71, with a median age of 56. Most patients had digital ischaemia without features of connective tissue disease. Four had no blood test abnormalities other than a raised tumour marker CA125. The time from onset of digital ischaemia to diagnosis of ovarian cancer ranged from less than one week to 2 years, with a median time of four months. Conclusion This case study supports previous findings that patients with digital ischaemia associated with ovarian cancer may have normal markers of inflammatory or rheumatological disease. Marked therapeutic responses can be achieved with systemic anti-cancer therapies. Therefore, underlying cancer should be suspected in patients with new-onset or worsening digital ischaemia over the age of 45. A high index of suspicion is required - rheumatologists beware! Disclosure J.H. Miller: None. P. Merry: None.

StarD13 negatively regulates invadopodia formation and invasion in high-grade serous (HGS) ovarian adenocarcinoma cells by inhibiting Cdc42

Metastasis remains the main challenge to overcome for treating ovarian cancers. In this study, we investigate the potential role of the Cdc42 GAP StarD13 in the modulation of cell motility, invasion in ovarian cancer cells. StarD13 depletion does not affect the 2D motility of ovarian cancer cells. More importantly, StarD13 inhibits matrix degradation, invadopodia formation and cell invasion through the inhibition of Cdc42. StarD13 does not localize to mature TKS4-labeled invadopodia that possess matrix degradation ability, while a Cdc42 FRET biosensor, detects Cdc42 activation in these invadopodia. In fact, StarD13 localization and Cdc42 activation appear mutually exclusive in invadopodial structures. Finally, for the first time we uncover a potential role of Cdc42 in the direct recruitment of TKS4 to invadopodia. This study emphasizes the specific role of StarD13 as a narrow spatial regulator of Cdc42, inhibiting invasion, suggesting the suitability of StarD13 for targeted therapy.

A monoclonal antibody against annexin A2 targets stem and progenitor cell fractions in tumors

The involvement of cancer stem cells (CSCs) in driving tumor dormancy and drug resistance is well established. Most therapeutic regimens however are ineffective in targeting these regenerative populations. We report the development and evaluation of a monoclonal antibody, mAb150, which targets the metastasis associated antigen, Annexin A2 (AnxA2) through recognition of a N-terminal epitope. Treatment with mAb150 potentiated re-entry of CSCs into the cell cycle that perturbed tumor dormancy and facilitated targeting of CSCs as was validated by in vitro and in vivo assays. Epigenetic potentiation further improved mAb150 efficacy in achieving total tumor regression by targeting regenerative populations to achieve tumor regression, specifically in high-grade serous ovarian adenocarcinoma.

Anti-GD1b and anti-GM1 positive, acute inflammatory demyelinating polyneuropathy associated with advanced ovarian carcinoma: A case report

Introduction: Paraneoplastic neurological syndromes (PNSs) are mostly associated with non-epithelial ovarian cancers. We report the case of an advanced high-grade serous ovarian adenocarcinoma, diagnosed during the etiological investigation of a subacute motor-sensory axonal neuropathy variant (AMSAN) of Guillain-Barre syndrome (GBS). According to our literature review, only a single case-control study described a positive association between GBS and cancer. Case Report: A 73-year-old woman was admitted to the emergency department for a progressive symmetrical distal tetraparesis, associated with hypoesthesia and abdominal pain. There was no sign of viral or bacterial infection. Over a period of 10 days, she presented a rapid neurological deterioration with nearly complete paralysis of all extremity, facial, and respiratory muscles. The laboratory testing ruled out active infection but the computed tomography (CT) scanner described diffuse peritoneal carcinomatosis, associated with bilateral ovarian tumors. Laparoscopic biopsies confirmed a high-grade serous ovarian cancer. Cerebrospinal fluid analysis confirmed the presence of elevated protein with a normal white blood cell count and anti-ganglioside autoantibodies anti-GD1b and anti-GM1. The patient received systemic chemotherapy together with plasmapheresis and intravenous immunoglobulins, and she experienced a rapid neurological recovery and long-term tumor remission. Conclusion: Neurological paraneoplastic syndromes are rarely associated with epithelial ovarian cancer. Cerebellar ataxia is most frequently associated with this cancer type. Guillain-Barre syndrome can rarely be associated with high-grade serous ovarian cancer. It can delay the tumor diagnosis and treatment. In the case of our patient, early supportive and etiological treatment was of paramount importance for the complete recovery of our patient.

Robotic Lymph Node Resection Off the Aortic Bifurcation for Presumed Recurrence Management of Advanced Ovarian Cancer

Study Objective To demonstrate a surgical video where-in lymph node resection was performed robotically in a patient with presumed ovarian cancer recurrence. Design Case report and a step-by-step video demonstration of robotic lymph node resection off of the aortic bifurcation (Canadian Task Force classification III). Setting Tertiary referral center. Patients or Participants 42-year-old female with stage IIIA1 high-grade serous ovarian adenocarcinoma. Interventions 42-year-old female was diagnosed with stage IIIA1 high-grade serous ovarian adenocarcinoma in 2018 when she underwent upfront optimal cytoreductive surgery, inclusive of extensive debulking of aortocaval lymphadenopathy. The patient remained disease free until February 2020, when surveillance imaging showed a single hypermetabolic lymph node, anterior to the left common iliac artery. She was taken to the operating room for robotic debulking. Robotic trocars were placed on a horizontal line along the umbilical fold. Exploration of the peritoneal cavity revealed no evidence of disease. There was some adhesive disease over right pelvic sidewall and distal small bowel mesentery was overlying the aortic bifurcation. These adhesions were taken down by paying meticulous attention in order not to injure the small bowel mesentery. The retroperitoneum was explored to the level of the third part of the duodenum. Aortocaval lymph node basins were explored. The IMA was skeletonized and left ureterolysis was performed; allowing safe access to the left para-aortic as well as to the presacral area. The suspicious lymph nodes, including the one which correlated with the hypermetabolic left common iliac lymph node were resected without any complications. Measurements and Main Results Successful robotic secondary debulking was performed. The patient was discharged home on postoperative day 0. Pathology of the suspicious lymph nodes were negative for malignancy. Therefore, the patient was deemed free of disease and has been scheduled for surveillance imaging in three months. Conclusion Robotic secondary debulking of presumed ovarian cancer recurrence is feasible and should be considered in selected patients.

Robotic Interval Cytoreductive Surgery for Stage IV Epithelial Ovarian Cancer

Video Objective To demonstrate a surgical video where-in interval cytoreduction to no gross residual disease was performed robotically in a patient with Stage IV epithelial ovarian cancer. Setting Tertiary referral center. Interventions 43-year-old Caucasian female was diagnosed with stage IV-A high-grade serous ovarian adenocarcinoma after presenting with shortness of breath. Computed tomography showed bilateral pleural effusions, adnexal masses, retroperitoneal lymphadenopathy, omental caking. Thoracentesis confirmed adenocarcinoma of Mullerian primary. She received three cycles of neoadjuvant carboplatinum and paclitaxel with excellent clinical response and was taken to the operating room for robotic-assisted interval cytoreductive surgery. Trocars were placed on a straight horizontal line along the umbilical fold. The rectosigmoid colon was mobilized medially. Pararectal and paravesical spaces were developed. Ureterolysis was completed bilaterally. The uterine vessels were sealed at the hypogastric bifurcation. Infundibulopelvic (IP) ligament was sealed and cut. Bilateral pelvic sidewall peritoneum was resected. Bladder flap was developed. Colpotomy was performed and the hysterectomy specimen was removed. Procedure was then continued with debulking of enlarged lymph nodes, from bilateral pelvic sidewalls and peri-aortic area. The robotic arms were targeted to the upper abdomen for total omentectomy. Access to the lesser sac was gained by resecting short gastic vessels, along the greater curvature of the stomach. The incision was then extended to the splenic flexure and hepatic flexure. Total omentectomy was completed. Remaining subcentimeter tumoral nodules along the peritoneal surfaces were ablated with argon beam coagulator. The patient had an uneventful postoperative course and was discharged home on postoperative day 1. Pathology confirmed high grade serous ovarian carcinoma. She was resumed on chemotherapy two weeks after her cytoreductive surgery. Conclusion Laparoscopic/robotic interval cytoreductive surgery should be considered in advanced ovarian cancer patients, who have an excellent clinical response to NACT. Studies to accurately identify the appropriate patient population for laparoscopic/robotic debulking procedures are urgently encouraged and needed.

Robotic Tumor Debulking off External Iliac Vessels for the Management of Recurrent Ovarian Cancer

To show a surgical video in which an isolated mass was resected off the external iliac vessels for the management of recurrent ovarian cancer. Case report. Tertiary referral center in New Haven, Connecticut. This is a step-by-step demonstration of a robotic tumor debulking in a patient with isolated recurrence of epithelial ovarian cancer [1-3]. The patient is a 70-year-old woman with Lynch syndrome who received a diagnosis for stage IIC high-grade serous ovarian adenocarcinoma and underwent complete debulking in 1996. She had most recently been on pembrolizumab for microsatellite instability-high tumor until February 2019, when she received a diagnosis for isolated hypermetabolic mass in close proximity to the external iliac vessels and right iliac fossa. The patient was placed in dorsal low lithotomy Trendelenburg position, and 15° leftward tilt of the table was obtained to expose the right pelvic sidewall and iliac fossa. To optimally target the surgical field of interest, all robotic trocars were placed in a straight line starting from 5 cm above symphysis pubis on the left side to left subcostal line between the midline vertical and the left midclavicular lines, as per the manufacturer's port placement guidelines (Fig. 1). Robotic resection of the tumor nodule off the external iliac vessels was successfully performed with adequate range of motion provided by the arms and without any complications. Trocar placement should be tailored to the site of surgical interest. Robotic-assisted laparoscopy should be considered as a valid alternative to the traditional open approach, when managing solitary masses in patients with recurrent ovarian cancer.

Functional balance between Tcf21-Slug defines cellular plasticity and migratory modalities in high grade serous ovarian cancer cell lines.

Cellular plasticity and transitional phenotypes add to complexities of cancer metastasis that can be initiated by single cell epithelial to mesenchymal transition (EMT) or cooperative cell migration (CCM). Our study identifies novel regulatory cross-talks between Tcf21 and Slug in mediating phenotypic and migration plasticity in high-grade serous ovarian adenocarcinoma (HGSC). Differential expression and subcellular localization associate Tcf21, Slug with epithelial, mesenchymal phenotypes, respectively; however, gene manipulation approaches identify their association with additional intermediate phenotypic states, implying the existence of a multistep epithelial-mesenchymal transition program. Live imaging further associated distinct migratory modalities with the Tcf21/Slug status of cell systems and discerned proliferative/passive CCM, active CCM and EMT modes of migration. Tcf21-Slug balance identified across a phenotypic spectrum in HGSC cell lines, associated with microenvironment-induced transitions and the emergence of an epithelial phenotype following drug exposure. Phenotypic transitions and associated functionalities following drug exposure were affirmed to ensue from occupancy of Slug promoter E-box sequences by Tcf21. Our study effectively provides a framework for understanding the relevance of ovarian cancer plasticity as a function of two transcription factors.

A novel multiple biomarker panel for the early detection of high-grade serous ovarian carcinoma

IntroductionSince the majority of patients are diagnosed at an advanced stage, ovarian cancer remains the most lethal gynecologic malignancy. There is no single biomarker with the sensitivity and specificity required for effective cancer screening; therefore, we investigated a panel of novel biomarkers for the early detection of high-grade serous ovarian carcinoma. MethodsTwelve serum biomarkers with high differential gene expression and validated antibodies were selected: IL-1Ra, IL-6, Dkk-1, uPA, E-CAD, ErbB2, SLPI, HE4, CA125, LCN2, MSLN, and OPN. They were tested using Simple Plex™, a multi-analyte immunoassay platform, in samples collected from 172 patients who were either healthy, had benign gynecologic pathologies, or had high-grade serous ovarian adenocarcinomas. The receiver operating characteristic (ROC) curve, ROC area under the curve (AUC), and standard error (SE) of the AUC were obtained. Univariate ROC analyses and multivariate ROC analyses with the combination of multiple biomarkers were performed. ResultsThe 4-marker panel consisting of CA125, HE4, E-CAD, and IL-6 had the highest ROC AUC. When evaluated for the ability to distinguish early stage ovarian cancer from a non-cancer control, not only did this 4-marker panel (AUC=0.961) performed better than CA 125 alone (AUC=0.851; P=0.0150) and HE4 alone (AUC=0.870; P=0.0220), but also performed significantly better than the 2- marker combination of CA125+HE4 (AUC=0.922; P=0.0278). The 4-marker panel had the highest average sensitivity under the region of its ROC curve corresponding to specificity ranging from 100% down to ~95%. ConclusionThe four-marker panel, CA125, HE4, E-CAD, and IL-6, shows potential in detecting serous ovarian cancer at earlier stages. Additional validation studies using the biomarker combination in ovarian cancer patients are warranted.

Human cytomegalovirus in high grade serous ovarian cancer possible implications for patients survival.

Patients diagnosed with high grade serous ovarian adenocarcinoma have a poor prognosis. Recently human cytomegalovirus (HCMV) has been detected in several tumors. Here, we evaluated HCMV in ovarian cancer tissue specimens obtained at pre- and postchemotherapy tumor resection.Available paraffin embedded ovarian cancer tissues from matched pre- and postchemotherapy tumor resection specimens (i.e., diagnostic excisional biopsy prechemotherapy; DEBPC) and neoadjuvant chemotherapy followed by interval debulking surgery (NACT + IDS) from 10 patients with stage IIIC-IV high grade serous ovarian carcinoma (HGS) diagnosed between years 2007 and 2008 at Karolinska University Hospital were examined for HCMV immediate-early protein (HCMV-IE), tegument protein pp65, and nucleic acid (β2.7) by immunohistochemistry and in situ hybridization.HCMV-IE and pp65 were detected in 8/10 (80%), 4/9 (44%) and in 4/10 (40%), 5/8 in ovarian cancer tissue specimens from DEBPC and NACT + IDS, respectively. HCMV-β2.7 was detected in all available tissue sections obtained from DEBPC and NACT + IDS. Patients with HCMV-IE or pp65 positive cells in their ovarian tumors at IDS after NACT had a median overall survival of 23.4 and 18.2 months, respectively, compared to 29.6 and 54 months, respectively, in those who did not express HCMV proteins in their tumors.In conclusion, HCMV proteins and nucleic acids are frequently detected at different levels in HGS ovarian carcinoma. Despite the limitation of our study, shorter median overall survival of patients with HCMV-IE and pp65 in their tumor highlights the need to further investigate the role of HCMV in ovarian cancer patients.

The expression profile and prognostic value of APE/Ref-1 and NPM1 in high-grade serous ovarian adenocarcinoma.

To analyze the expression trends and clinical significance of Apurinic/Apyrimidinic Endodeoxyribonuclease 1 (APE1/Ref-1) and Nucleophosmin (NPM1) proteins in high-grade serous ovarian adenocarcinoma (HGSC). The expressions of APE1/Ref-1 and NPM1 proteins in 94 patients with HGSC were determined using the immunohistochemical (IHC) method, and their relationships with clinicopathological features were analyzed by the χ2 test or Fisher's exact test. The follow-up data, Cox proportional hazards univariate and multivariate survival analyses were integrated to evaluate the prognostic factors affecting patients with HGSC. In the normal fallopian tubes, APE1/Ref-1 and NPM1 protein were mainly distributed in the nuclear. The HGSC experienced changes in the cellular localization of APE1/Ref-1 and NPM1 protein expressions, which were abnormally expressed in the cytoplasm. The rates of abnormal cytoplasmic expression of APE1/Ref-1 and NPM1 proteins in 94 patients with HGSC were 69.1% and 73.4%, respectively, which were significantly higher than the normal fallopian tube tissues (p < 0.05). The abnormal cytoplasmic APE1/Ref-1 and NPM1 are significantly correlated with the lymph node metastasis, chemosensitivity, FIGO staging, and prognosis. The COX multivariate survival analysis showed that the abnormal expression of APE1/Ref-1 protein, FIGO staging, and lymph node metastasis are independent prognostic factors. Collectively, the abnormal cytoplasmic APE1/Ref-1 and NPM1 proteins are associated with the oncogenic progression and chemoresistance of HGSC, and predict a poor prognosis.

Influence of a novel histone deacetylase inhibitor panobinostat (LBH589) on the growth of ovarian cancer

BackgroundPre-clinical studies have demonstrated that natural and synthetic histone deacetylase (HDAC) inhibitors can impede the in vitro and in vivo growth of cell lines from a variety of gynecologic and other malignancies. We investigated the anti-tumor activity of panobinostat (LBH589) both in vitro and in vivo as either a single agent or in combination with conventional cytotoxic chemotherapy using patient-derived xenograft (PDX) models of primary serous ovarian tumors.MethodsThe ovarian cancer cell lines OVCAR8, SKOV3 and their paclitaxel-resistant derivatives OVCAR8-TR and SKOV3-TR were treated with increasing doses of LBH589. The effect of LBH589 on cell viability was assessed using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. Serially transplanted primary human high-grade serous ovarian adenocarcinoma tissue was utilized to generate xenografts in 6-week old female NOD/SCID mice. The mice were then randomized into one of 4 treatment groups: (1) vehicle control; (2) paclitaxel and carboplatin (P/C); (3) LBH589; or (4) P/C + LBH589. Mice were treated for 21 days and tumor volumes and mouse weights were obtained every 3 days. These experiments were performed in triplicate with three different patient derived tumors. Wilcoxan rank-sum testing was utilized to assess tumor volume differences.ResultsIn vitro treatment with LBH589 significantly reduced the viability of both taxol-sensitive and taxol-resistant ovarian cancer cell lines (p < 0.01). In vivo treatment with LBH589 alone appeared tumorstatic and reduced tumor growth when compared to vehicle treatment (p < 0.007) after 21 days. This single agent activity was confirmed in two additional experiments with other PDX tumors (p < 0.03, p < 0.05). A potential additive effect of LBH589 and P/C, manifested as enhanced tumor regression with the addition of LBH589 compared to vehicle (p < 0.02), in one of the three analyzed serous PDX models.ConclusionsOur findings suggest that pan-HDAC inhibition with panobinostat precludes the growth of ovarian cancer cell lines in vitro and PDXs in vivo. Added benefit of LBH589 to standard P/C therapy was observed in one of three PDX models suggesting improved response in a subset of serous ovarian cancers.

Association between targeted somatic mutation (TSM) signatures and HGS‐OvCa progression

Evidence already exists that the activation‐induced cytidine deaminase (AID/APOBEC) and the adenosine deaminase (ADAR) families of enzymes are implicated as powerful mutagens in oncogenic processes in many somatic tissues. Each deaminase is identified by the DNA or RNA nucleotide sequence (“motif”) surrounding the nucleotide targeted for deamination. The primary objective of this study is to develop an in silico approach to identify nucleotide sequence changes of the target motifs of key deaminases during oncogenesis. If successful, a secondary objective is to investigate if such changes are associated with disease progression indicators that include disease stage and progression‐free survival time. Using a discovery cohort of 194 high‐grade serous ovarian adenocarcinoma (HGS‐OvCa) exomes, the results confirm the ability of the novel in silico approach used to identify changes in the preferred target motifs for AID, APOBEC3G, APOBEC3B, and ADAR1 during oncogenesis. Using this approach, a set of new cancer‐progression associated signatures (C‐PASs) were identified. Furthermore, it was found that the C‐PAS identified can be used to differentiate between the cohort of patients that remained progression‐free for longer than 60 months, from those in which disease progressed within 60 months (sensitivity 95%, specificity 90%). The spectrum of outcomes observed here could provide a foundation for future clinical assessment of susceptibility variants in ovarian, and several other cancers as disease progresses. The ability of the in silico methodology used to identify changes in deaminase motifs during oncogenesis also suggests new links between immune system function and tumorigenesis.

Second Annual Moffitt Anatomic Pathology Symposium.

Dr Magliocco, chair of the Department of Anatomic Pathology, welcomed a near-capacity audience made up of pathologists, histotechnologists, researchers, and pathology residents and medical students at the 2nd Annual Moffitt Anatomic Pathology Symposium: Practicing Pathology in a Changing World that took place in Clearwater Beach, Florida, on September 26, 2015. Th focus of the symposium was on the updates of contemporary and emerging information and technology that impact practicing pathologists in an ever-changing world.Invited speakers included Drs Odze and Schnitt, both from Harvard Medical School in Boston, who shared their expert knowledge in gastrointestinal (GI) and breast pathology, as well as Theresa B. Burchette, Lawrence Patton Jr, John J. Shelley, and Steven Westra. Experts from the H. Lee Moffitt Cancer Center & Research Institute included Drs Magliocco, Bui, Centeno, Coppola, Hussaini, Khalil, Khazia, and Rosa, as well as Mr Hicks and Ms Balasi, Molina, and Snyder. Drs Minton and Malafa also presented topics in molecular pathology, digital pathology, and GI and breast cancers. The Ask the Expert Digital Pathology session was an interactive discussion of real breast and GI pathology cases among the experts and the audience.A total of 22 scientific abstracts were proffered by students, residents, fellows, and staff. Dr Altiok, chair of the research section of the Department of Anatomic Pathology, presented awards to the 3 best submissions.Dr Ryzhova and colleagues earned the Best Poster award for their work titled “Mutation Profiling of High-Grade Serous Ovarian Adenocarcinomas Suggests STK11 and NOTCH1 as Candidate Biomarkers.” This paper reported identification of STK11 mutations as possible biomarkers for platinum-therapy response and detailed actionable mutations in several patients with ovarian cancer.The Best Research Project prize was awarded to Dr Dettloff and colleagues for their work “Utility of a microRNA-Based Assay for Diagnosing Tumor Origin in Soft Tissue Sarcomas.” The investigators examined the recent experience at Moffitt Cancer Center with noncoding microRNA tumor profiling of tumors of uncertain origin. They listed several situations in which the technique can be particularly useful, such as the identification of germ-cell tumors. They also recognized pitfalls of the technique, such as a bias toward sarcoma, possibly representing a biological shift in recurrent or previously treated tumors. The Best Clinicopathologic Correlation award was won by Dr Liu and colleagues for their poster titled “Spindle Cell/Sclerosing Rhabdomyosarcoma Is a New and Underdiagnosed Entity: A Clinical and Tissue Microarray Immunohistochemical Study With Literature Review.” This work emphasized the utility of using tissue microarray to screen previously encountered cases with antibodies directed against emerging biomarkers.

The clinical characteristics and outcomes of cases with pure ovarian clear cell, mixed type and high-grade serous adenocarcinoma.

This study aimed to compare the clinical characteristics and outcomes of the cases with pure ovarian clear cell adenocarcinoma (OCCC), mixed type (clear plus serous or clear plus endometrioid or serous plus clear plus endometrioid) ovarian adenocarcinoma (MOC) and high-grade serous ovarian adenocarcinoma (HGSOC). A retrospective comparison was conducted in 32 cases with pure OCCC, 23 cases with MOC and 28 cases with HGSOC followed between January 1999 and October 2011. Clinico-pathologic characteristics including histopathologic subtypes, age, stage, presence of ascites and/or peritoneal carcinomatosis, endometriosis and optimal cytoreduction, and also survival rates were compared in these three groups. The cases with pure OCCC were younger than the cases with MOC and HGSOC (p=0.004). The median pre-operative CA-125 level was lower in the pure OCCC than in MOC and HGSOC (p=0.006) (p=0.001, p=<0.001, respectively). Bilaterality and peritoneal carcinomatosis were more frequently seen in the HGSOC group (p=0.001, p<0.001, respectively). The majority of the cases (65%) had stage I or II disease in the pure OCCC group, but most of the cases in the HGSOC group had advanced stage disease (p<0.001). Endometriosis was found in 16 cases (16/55, 29%) in the OCCC group. No significant difference was detected with regard to the median survival rate among three groups (p=0.517), while the stage of disease was found to be the only important factor for survival (p=0.002). Pure OCCC, MOC and HGSOC represent distinct clinical characteristics. Clinical characteristics of MOC are more similar to those of HGSOC.

Dysregulation of X chromosome inactivation in high grade ovarian serous adenocarcinoma.

BackgroundOne of the two copies of the X chromosome is randomly inactivated in females as a means of dosage compensation. Loss of X chromosome inactivation (XCI) is observed in breast and ovarian cancers, and is frequent in basal-like subtype and BRCA1 mutation-associated breast cancers. We investigated the clinical implications of the loss of XCI in ovarian cancer and the association between the loss of XCI and BRCA1 dysfunction.Materials and MethodsWe used open source data generated by The Cancer Genome Atlas (TCGA) Genome Data Analysis Centers. Ward’s hierarchical clustering method was used to classify the methylation status of the X chromosome.ResultsWe grouped 584 high grade serous ovarian adenocarcinomas (HG-SOA) according to methylation status, loss of heterozygosity and deletion or gain of X chromosome into the following five groups: preserved inactivated X chromosome (Xi) group (n = 175), partial reactivation of Xi group (n = 100), p arm deletion of Xi group (n = 35), q arm deletion of Xi group (n = 44), and two copies of active X group (n = 230). We found four genes (XAGE3, ZNF711, MAGEA4, and ZDHHC15) that were up-regulated by loss of XCI. HG-SOA with loss of XCI showed aggressive behavior (overall survival of partial reactivation of Xi group: HR 1.7, 95% CI 1.1–2.5, two copies of active X group: HR 1.4, 95% CI 1.0–1.9). Mutation and hypermethylation of BRCA1 were not frequent in HG-SOA with loss of XCI.ConclusionsLoss of XCI is common in HG-SOA and is associated with poor clinical outcome. The role of BRCA1 in loss of XCI might be limited. XCI induced aberrant expression of cancer-testis antigens, which may have a role in tumor aggressiveness.

High-temperature-required protein A2 as a predictive marker for response to chemotherapy and prognosis in patients with high-grade serous ovarian cancers.

Background:High-temperature-required protein A2 (HtrA2), a protein relating with apoptosis in a caspases-dependent and non-dependent manner, has been reported to be associated with chemosensitivity in several human cancers.Methods:Tissue microarrays made from 142 patients with high-grade serous ovarian adenocarcinoma were evaluated to assess whether HtrA2 expression was related with several clinical parameters.Results:Negative HtrA2 expression was observed in 36 cases (25%) of the patients, and related with significantly lower response rates of primary chemotherapy than those with positive HtrA2 expression (56% vs 83%, P<0.01). In addition, negative HtrA2 expression was identified as an independent worse prognostic factor for progression-free survival and overall survival by multivariate analyses. Furthermore, HtrA2 downregulation modulated sensitivity to platinum in serous ovarian cancer cells in vitro.Conclusions:HtrA2 expression was a predictor for sensitivity to chemotherapy, and could be a candidate of molecular target in the treatment of high-grade serous ovarian cancers.

Abstract 304: Feasibility study of next-generation sequencing on residual formalin-fixed paraffin-embedded tissues

Abstract Next Generation Sequencing (NGS) technologies are being used for detection of somatic mutations in tumors and studies of germline variation. However, most NGS studies used DNA isolated either from whole blood or fresh frozen tissue specimens. Meanwhile, the tissue specimens available from most National Cancer Institute (NCI) funded cohorts and the Surveillance, Epidemiology and End Results (SEER) registries (http://seer.cancer.gov/biospecimen/) are primarily formalin fixed paraffin embedded (FFPE). There are limited data, on a small number of FFPE tissue samples, which suggest NGS is feasible. Much less is known about the feasibility of these technologies for large scale studies or using older FFPE specimens (e.g. 5-30 years old). The main objective of this project was to conduct a pilot study to determine whether the DNA obtained from archival FFPE tissue is of sufficient quality and quantity to conduct NGS. Sixty high-grade serous ovarian adenocarcinomas from FFPE tissues which were between 7 and 31 years old were obtained from three SEER registries. DNA was extracted, quantified, quality assessed, and subjected to whole exome sequencing. DNA extraction (yields and quality) and whole exome sequencing (depths of coverage and exome coverage obtained) results from this study will be presented. Ultimately, data derived from this analysis could serve as the basis for determining the utility of archival FFPE biospecimens for characterization and discovery projects utilizing NGS technologies instead of relying on frozen biospecimens. Citation Format: Danielle Mercatante Carrick, Sean Altekruse, Corrine Camalier, Wendy Cozen, Brenda Hernandez, Charles Lynch, Paul McGregor, Michele G. Mehaffey, Lisa McShane, JoyAnn Phillips Rohan, Mickey Williams, Elizabeth M. Gillanders, Leah E. Mechanic, Sheri Schully. Feasibility study of next-generation sequencing on residual formalin-fixed paraffin-embedded tissues. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 304. doi:10.1158/1538-7445.AM2014-304

Discrete Molecular Classes of Ovarian Cancer Suggestive of Unique Mechanisms of Transformation and Metastases

Tumor heterogeneity and subsistence of high-grade serous ovarian adenocarcinoma (HGSC) classes can be speculated from clinical incidences suggesting passive tumor dissemination versus active invasion and metastases. We explored this theme toward tumor classification through two approaches of gene expression pattern clustering: (i) derivation of a core set of metastases-associated genes and (ii) resolution of independent weighted correlation networks. Further identification of appropriate cell and xenograft models was carried out for resolution of class-specific biologic functions. Both clustering approaches achieved resolution of three distinct tumor classes, two of which validated in other datasets. Networks of enriched gene modules defined biologic functions of quiescence, cell division-differentiation-lineage commitment, immune evasion, and cross-talk with niche factors. Although deviant from normal homeostatic mechanisms, these class-specific profiles are not totally random. Preliminary validation of these suggests that Class 1 tumors survive, metastasize in an epithelial-mesenchymal transition (EMT)-independent manner, and are associated with a p53 signature, aberrant differentiation, DNA damage, and genetic instability. These features supported by association of cell-specific markers, including PAX8, PEG3, and TCF21, led to the speculation of their origin being the fimbrial fallopian tube epithelium. On the other hand, Class 2 tumors activate extracellular matrix-EMT-driven invasion programs (Slug, SPARC, FN1, THBS2 expression), IFN signaling, and immune evasion, which are prospectively suggestive of ovarian surface epithelium associated wound healing mechanisms. Further validation of these etiologies could define a new therapeutic framework for disease management.