Tiffany M. Hebert

Insulin-like Growth Factor 2 Expression Modulates Taxol Resistance and Is a Candidate Biomarker for Reduced Disease-Free Survival in Ovarian Cancer

Purpose: This study was undertaken to examine the role of the insulin-like growth factor (IGF) signaling pathway in the response of ovarian cancer cells to Taxol and to evaluate the significance of this pathway in human epithelial ovarian tumors. Experimental Design: The effect of Taxol treatment on AKT activation in A2780 ovarian carcinoma cells was evaluated using antibodies specific for phospho-AKT. To study the drug-resistant phenotype, we developed a Taxol-resistant cell line, HEY-T30, derived from HEY ovarian carcinoma cells. IGF2 expression was measured by real-time PCR. A type 1 IGF receptor (IGF1R) inhibitor, NVP-AEW541, and IGF2 small interfering RNA were used to evaluate the effect of IGF pathway inhibition on proliferation and Taxol sensitivity. IGF2 protein expression was evaluated by immunohistochemistry in 115 epithelial ovarian tumors and analyzed in relation to clinical/pathologic factors using the χ2 or Fishers exact tests. The influence of IGF2 expression on survival was studied with Cox regression. Results: Taxol-induced AKT phosphorylation required IGF1R tyrosine kinase activity and was associated with upregulation of IGF2. Resistant cells had higher IGF2 expression compared with sensitive cells, and IGF pathway inhibition restored sensitivity to Taxol. High IGF2 tumor expression correlated with advanced stage (P < 0.001) and tumor grade (P < 0.01) and reduced disease-free survival (P < 0.05). Conclusions: IGF2 modulates Taxol resistance, and tumor IGF2 expression is a candidate prognostic biomarker in epithelial ovarian tumors. IGF pathway inhibition sensitizes drug-resistant ovarian carcinoma cells to Taxol. Such novel findings suggest that IGF2 represents a therapeutic target in ovarian cancer, particularly in the setting of Taxol resistance. Clin Cancer Res; 16(11); 2999–3010. ©2010 AACR.

Extragenital adenosarcoma: A case report, review of the literature, and management discussion

BACKGROUND Müllerian adenosarcoma is a rare mixed epithelial-mesenchymal tumor. An extragenital site of origin and sarcomatous overgrowth are associated with aggressive clinical behavior. CASE We present a rare case of extragenital adenosarcoma with sarcomatous overgrowth and coexistent endometriosis. She was treated with initial cytoreductive surgery and chemotherapy. She underwent a second surgery for management of a high-grade bowel obstruction, due to pathologically confirmed recurrent intraperitoneal adenosarcoma. A complete clinical response was achieved with liposomal doxorubicin, and the patient remains disease-free eighteen months after completion of chemotherapy. CONCLUSION Liposomal doxorubicin appears to be an active agent for the treatment of adenosarcoma with sarcomatous overgrowth. In addition, we conclude from our review of all reported cases of extragenital adenosarcoma that concurrent endometriosis may represent a favorable prognostic factor.

RNA-seq Identification of RACGAP1 as a Metastatic Driver in Uterine Carcinosarcoma.

Purpose: Uterine carcinosarcoma is a rare aggressive malignancy frequently presenting at advanced stage of disease with extrauterine metastases. Median survival is less than 2 years due to high relapse rates after surgery and poor response to chemotherapy or radiotherapy. The goal of this study was to identify novel therapeutic targets. Experimental Design: We applied RNA-seq analysis to prospectively collected uterine carcinosarcoma tumor samples from patients undergoing primary surgical resection and for comparison, normal endometrial tissues from postmenopausal women undergoing hysterectomy for benign indications. Functional assays were done in primary carcinosarcoma cell lines developed from patients and in established cell lines, as well as a cell line–derived xenograft model. Validation was done by analysis of an independent cohort of patients with uterine carcinosarcoma from The Cancer Genome Atlas (TCGA). Results: Rac GTPase–activating protein 1 (RACGAP1) was identified to be highly upregulated in uterine carcinosarcoma. Functional assays showed that RACGAP1 mediates motility and invasion via regulation of STAT3 phosphorylation and survivin expression. RACGAP1 depletion or survivin inhibition abrogated motility and invasiveness of carcinosarcoma cells, while RACGAP1 overexpression conferred invasiveness to endometrial adenocarcinoma cells. In the TCGA cohort, RACGAP1 expression correlated with survivin expression and extrauterine spread of disease. Conclusions: The RACGAP1–STAT3–survivin signaling pathway is required for the invasive phenotype of uterine carcinosarcoma and is a newly identified therapeutic target in this lethal disease. Clin Cancer Res; 22(18); 4676–86. ©2016 AACR.

Hemoglobin parameters from diffuse reflectance data

Abstract. Tissue vasculature is altered when cancer develops. Consequently, noninvasive methods of monitoring blood vessel size, density, and oxygenation would be valuable. Simple spectroscopy employing fiber optic probes to measure backscattering can potentially determine hemoglobin parameters. However, heterogeneity of blood distribution, the dependence of the tissue-volume-sampled on scattering and absorption, and the potential compression of tissue all hinder the accurate determination of hemoglobin parameters. We address each of these issues. A simple derivation of a correction factor for the absorption coefficient, μa, is presented. This correction factor depends not only on the vessel size, as others have shown, but also on the density of blood vessels. Monte Carlo simulations were used to determine the dependence of an effective pathlength of light through tissue which is parameterized as a ninth-order polynomial function of μa. The hemoglobin bands of backscattering spectra of cervical tissue are fit using these expressions to obtain effective blood vessel size and density, tissue hemoglobin concentration, and oxygenation. Hemoglobin concentration and vessel density were found to depend on the pressure applied during in vivo acquisition of the spectra. It is also shown that determined vessel size depends on the blood hemoglobin concentration used.

A rare case of chondroma of the parotid gland.

Patient: A 39-year-old Hispanic woman. History of Present Illness: The patient had swelling of the left side of her neck, which she had first noticed 3 to 4 months before consultation and which did not subside after 2 courses of antibiotics. She reported no tenderness, dysphagia, odynophagia, dysphonia, otalgia, fevers, chills, or weight changes. Past medical history: The patient had a past history of gastroesophageal reflux disease, arthritis (knee and cervical disease), and a prior abnormal Pap smear result (high grade squamous intraepithelial lesion). The cervical lesion was treated with a loop electrosurgical excision procedure (LEEP). Her past surgical history is remarkable for cholecystectomy and a left breast biopsy with benign results. Social history: Noncontributory. Family history: Noncontributory. Physical exam: The patient harbored a firm, nontender, fully mobile 2- to 3-cm left parotid tail mass without other abnormalities; her facial nerve function was intact in all branches. Principle Laboratory Findings: See [Image 1][1], [Image 2][2], [Image 3][3], [Image 4][4], [Image 5][5], and [Image 6][6]. * FNA : fine needle aspiration LEEP : loop electrosurgical excision procedure CT : computed tomography [1]: #F1 [2]: #F2 [3]: #F3 [4]: #F4 [5]: #F5 [6]: #F6

Protocol for the study of cervical cancer screening technologies in HIV-infected women living in Rwanda

Introduction The optimal method(s) for screening HIV-infected women, especially for those living in sub-Saharan Africa, for cervical precancer and early cancer has yet to be established. Methods and analysis A convenience sample of >5000 Rwandan women, ages 30–54 years and living with HIV infection, is being consented and enroled into a cross-sectional study of cervical cancer screening strategies. Participants are completing an administered short risk factor questionnaire and being screened for high-risk human papillomavirus (hrHPV) using the Xpert HPV assay (Cepheid, Sunnyvale, California, USA), unaided visual inspection after acetic acid (VIA) and aided VIA using the Enhanced Visual Assessment (EVA) system (Mobile ODT, Tel Aviv, Israel). Women positive for hrHPV and/or by unaided VIA undergo colposcopy, which includes the collection of two cervical specimens prior to undergoing a four-quadrant microbiopsy protocol. The colposcopy-collected specimens are being tested by dual immunocytochemical staining for p16INK4a and Ki-67 (CINtec PLUS Cytology, Ventana, Tucson, Arizona, USA) and for E6 or E7 oncoprotein for 8 hrHPV genotypes (HPV16, 18, 31, 33, 35, 45, 52 and 58) using the next-generation AV Avantage hrHPV E6/E7 test (Arbor Vita Corporation, Freemont, California, USA). Women with a local pathology diagnosis of cervical intraepithelial neoplasia grade 2 (CIN2) or more severe (CIN2+) or pathology review diagnosis of CIN grade three or more severe (CIN3+) will receive treatment. Clinical performance and cost-effectiveness (eg, sensitivity, specificity and predictive values) of different screening strategies and algorithms will be evaluated. Ethics and dissemination The protocol was approved by local and institutional review boards for human subjects research. At the completion of the study, results will be disseminated to the scientific community through peer-reviewed publication and to the Rwandan stakeholders through an external advisory panel.

Abstract 5131: Establishment of uterine carcinosarcoma primary cell lines for chemosensitivity testing and evaluation of targeted therapy

To establish primary cell lines from uterine carcinosarcoma (CS) patient samples and to evaluate combination treatment with standard chemotherapy with or without the addition of an anti-IGF1R antibody. CS tumor tissue was obtained under IRB approval at the time of primary surgery. We reported the establishment of patient-derived xenografts elsewhere. Tissue pieces were minced extensively and digested by collagenase treatment. The single cell solution was then seeded in F-media containing EGF, insulin, hydrocortisone, adenine, cholera toxin and the Rock inhibitor Y-27632. A previously established carcinosarcoma cell line CS99 (Schulten et al, 2008) was used for comparison. To verify origin, Short Tandem Repeat (STR) profiles of the primary cell lines CS13, CS18, CS19, CS21 and CS22 were compared with the patient9s tumor samples. Drug sensitivity to Taxol, cisplatin and carboplatin was determined individually or in combination using the sulforhodamine B (SRB) proliferation assay. The effect of IMC-A12 (anti-IGF1R antibody, Eli Lily / Imclone) alone or with chemotherapy was also determined with the SRB assay. Total and promoter-specific IGF2 mRNA levels were determined by reverse transcriptase quantitative PCR. All STR profiles of the primary cell lines matched their patient sample counterparts. IGF2 mRNA expression levels of the cell lines were similar to the patient samples, and over 10,000-fold higher than in CS99. IGF2 promoter-specific primers showed that the IGF2 mRNA transcripts of the primary cell lines were initiated at the oncofetal IGF2 promoters P3 and P4. For Taxol, the IC50 (inhibitory concentration of 50%) ranged from 4.7 nM for CS21 to 9.8 nM for CS22. For cisplatin, the IC50 ranged from 1.0 μM for CS99 to 4.8 μM for CS19. For carboplatin, CS99 cells had the lowest IC50 of 12.5 μM while CS19 had the highest IC50 of 58.1 μM. In the presence of 10 μg/ml IMC-A12, the primary cell lines CS19, CS21 and CS22 showed significant sensitization to Taxol (∼50% decrease in IC50), to carboplatin (∼65% decrease in IC50), and to combination Taxol/carboplatin treatment (∼50% decrease in IC50). We conclude that establishing primary cell lines of this rare cancer is a promising approach for testing sensitivity to current and novel chemotherapies and combination treatment strategies. We also found that high oncofetal promoter-driven IGF2 mRNA expression was observed in all CS samples tested and that IGF1R blockade sensitized the primary cell lines to standard chemotherapy. Citation Format: Jurriaan Brouwer-Visser, Eirwen Scott, Shijun Mi, Maria J. Cossio, Tiffany Hebert, Gloria S. Huang. Establishment of uterine carcinosarcoma primary cell lines for chemosensitivity testing and evaluation of targeted therapy. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 5131. doi:10.1158/1538-7445.AM2015-5131