Supplementary Data from A novel hTERT promoter–driven E1A therapeutic for ovarian cancer

Figure S4: ROC overlay demonstrating relationship between FSH values and menopause status. Separate curves were generated for pre (n=333) vs. post (n=317) and vs. post/peri (n=390). Subjects who had undergone hysterectomy were excluded in these analyses.

Supplementary Data from Therapeutic Targeting of ATP7B in Ovarian Carcinoma

Figure S5: Individual ROC curves for Videssa Breast model training and validation. Curves shown correspond to those in Fig. 2.

Figure S1: Breakdown of imaging modalities used to enroll subjects in Provista-002. Some subjects underwent multiple imaging procedures prior to enrollment, these are noted in side boxes for screening mammogram and diagnostic mammogram.

Figure S3: Breakdown of subjects excluded in the Provista-002 trial. Reasons for exclusion shown in legend.

Figure S2: Clinical management workflow with inclusion/exclusion criteria for the PDX-002 clinical trial. *Details on malignancy risk from D'Orsi et al.[3]

Table S1. Enrollment Criteria Table S2. Enrollment Sites Table S3. Serum protein biomarkers (SPB) and tumor-associated autoantibodies (TAAb) evaluated in the current study. Table S4: Clinical conditions and criteria for inclusion. Table S5: Comparison of biomarker measurements between women divided by age and by FSH. Table S6: Clinical performance of Videssa Breast in all samples categorized by breast density (dense: categories a and b, non-dense: categories c and d) and by health exclusion. Table S7: Clinical performance of Videssa Breast in subjects ages 25-49.

The editors are publishing this note to alert readers to a concern about [this article][1] ([1][2]): in Fig. 5A, the representative images for "empty liposome" and "control siRNA-DOPC" were both from the "empty liposome" group. MD Anderson's Office of Research Oversight and Integrity

Introduction Overexpression of the epidermal growth factor receptor (EGFR) found in common subtypes of endometrial cancer has been associated with advanced stage disease and a poor prognosis. The purpose of this phase 2 study was to evaluate the efficacy and safety of cetuximab in patients with recurrent endometrial cancer. Methods The study was an open-label phase 2 clinical trial conducted at two institutions. Patients with recurrent or progressive endometrial cancer of any histologic type with the exception of uterine sarcoma received cetuximab at an initial dose of 400 mg/m2 IV followed by weekly doses of 250 mg/m2. One cycle was considered 4 weeks of treatment. The primary efficacy endpoint was clinical benefit response, defined as a complete or partial response or prolonged stable disease (>8 weeks) by RECIST 1.0 criteria. Results A total of 30 patients were enrolled with a median age of 64 years (range 42–83). Of the 20 evaluable patients, three (15%) had clinical benefit response (one complete response, two stable disease). The patient with a clinical benefit response received a total of 27 cycles and the two patients with stable disease were taken off the study due to progression after four and six cycles, respectively. Of the 10 inevaluable patients, nine received ≤1 cycle due to clinical deterioration and one had an anaphylactic reaction. One patient had a grade 3 rash which resolved after a delay in treatment. No dose reduction was reported. Conclusions In this cohort, single agent therapy with cetuximab was well tolerated and had a 15% clinical benefit response. Further studies are required to better identify patients who may respond to this treatment.

AbstractPurpose: With improvements in breast cancer imaging, there has been a corresponding increase in false-positives and avoidable biopsies. There is a need to better differentiate when a breast biopsy is warranted and determine appropriate follow-up. This study describes the design and clinical performance of a combinatorial proteomic biomarker assay (CPBA), Videssa Breast, in women over age 50 years. Experimental Design: A BI-RADS 3, 4, or 5 assessment was required for clinical trial enrollment. Serum was collected prior to breast biopsy and subjects were followed for 6–12 months and clinically relevant outcomes were recorded. Samples were split into training (70%) and validation (30%) cohorts with an approximate 1:4 case:control ratio in both arms. Results: A CPBA that combines biomarker data with patient clinical data was developed using a training cohort (469 women, cancer incidence: 18.5%), resulting in 94% sensitivity and 97% negative predictive value (NPV). Independent validation of the final algorithm in 194 subjects (breast cancer incidence: 19.6%) demonstrated a sensitivity of 95% and a NPV of 97%. When combined with previously published data for women under age 50, Videssa Breast achieves a comprehensive 93% sensitivity and 98% NPV in a population of women ages 25–75. Had Videssa Breast results been incorporated into the clinical workflow, approximately 45% of biopsies might have been avoided. Conclusions: Videssa Breast combines serum biomarkers with clinical patient characteristics to provide clinicians with additional information for patients with indeterminate breast imaging results, potentially reducing false-positive breast biopsies.

Ovarian cancer is often fatal and incidence in the general population is low, underscoring the necessity (and the challenges) for advancements in screening and early detection. The goal of this study was to design a serum-based biomarker panel and corresponding multivariate algorithm that can be used to accurately detect ovarian cancer. A combinatorial protein biomarker assay (CPBA) that uses CA125, HE4, and 3 tumor-associated autoantibodies resulted in an area under the curve of 0.98. The CPBA Ov algorithm was trained using subjects who were suspected to have gynecological cancer and were scheduled for surgery. As a surgical rule-out test, the clinical performance achieves 100% sensitivity and 83.7% specificity. Although sample size (n = 60) is a limiting factor, the CPBA Ov algorithm performed better than either CA-125 alone or the Risk of Ovarian Malignancy Algorithm.

Abstract This abstract was not presented at the symposium.

Breast density is associated with reduced imaging resolution in the detection of breast cancer. A biochemical approach that is not affected by density would provide an important tool to healthcare professionals who are managing women with dense breasts and suspicious imaging findings. Videssa® Breast is a combinatorial proteomic biomarker assay (CPBA), comprised of Serum Protein Biomarkers (SPB) and Tumor Associated Autoantibodies (TAAb) integrated with patient-specific clinical data to produce a diagnostic score that reliably detects breast cancer (BC) as an adjunctive tool to imaging. The performance of Videssa® Breast was evaluated in the dense (a and b) and non-dense (c and d) groups in a population of n = 545 women under age 50. The sensitivity and specificity in the dense breast group were calculated to be 88.9% and 81.2%, respectively, and 92.3% and 86.6%, respectively, for the non-dense group. No significant differences were observed in the sensitivity (p = 1.0) or specificity (p = 0.18) between these groups. The NPV was 99.3% and 99.1% in non-dense and dense groups, respectively. Unlike imaging, Videssa® Breast does not appear to be impacted by breast density; it can effectively detect breast cancer in women with dense and non-dense breasts alike. Thus, Videssa® Breast provides a powerful tool for healthcare providers when women with dense breasts present with challenging imaging findings. In addition, Videssa® Breast provides assurance to women with dense breasts that they do not have breast cancer, reducing further anxiety in this higher risk patient population.

Objective: Early involvement of specialists is associated with improved survival in ovarian cancer patients. However, only 33-60% of patients are currently being referred. A more effective strategy to get patients with an elevated risk of ovarian cancer to gynecologic oncologists is needed. The objective of this study was to evaluate the clinical utility of a multivariate index assay, OVA1*, by assessing its ability to drive referral of ovarian cancer patients to gynecologic oncologists prior to their first surgical intervention.Research design and methods: Information on patients who received a multivariate index assay was collected retrospectively from 22 obstetricians/gynecologists through a chart review. Physicians were recruited from a variety of practices and hospitals representing major geographic regions within the United States. Clinical utility of the multivariate index assay was assessed by examining the rate at which obstetricians/gynecologists involved a gynecologic oncologist for patients with elevated-risk results prior to first surgical intervention.Results: A total of 136 patients with elevated-risk assay results were assessed, of whom 122 underwent surgery to remove an adnexal mass. Prior to surgery, 98 (80%) of the patients were referred to a gynecologic oncologist with an additional 11 (9%) having a gynecologic oncologist available if required by intra-operative findings. Primary ovarian cancer was found in 65 (53%) patients, and gynecologic oncologists performed 61 (94%) of the initial surgeries these patients. Similar results were found in premenopausal and postmenopausal patients.Conclusions: A high proportion of patients with elevated-risk assay results were referred to a gynecologic oncologist prior to their initial surgery. Nearly all of the patients who had primary ovarian malignancies were appropriately referred to gynecologic oncologists, highlighting the clinical utility of the multivariate index assay. Nearly all patients identified with ovarian cancer received their initial surgery by a gynecologic oncologist - demonstrating a higher rate of gynecologic oncologist involvement in comparison to past studies. The study may be limited by unintentional bias in physician response and recall.

BACKGROUND:Women with adnexal mass suspected of ovarian malignancy are likely to benefit from consultation with a gynecologic oncologist, but imaging and biomarker tools to ensure this referral show low sensitivity and may miss cancer at critical stages.OBJECTIVE:The multivariate index assay (MIA) was designed to improve the detection of ovarian cancer among women undergoing surgery for a pelvic mass. To improve the prediction of benign masses, we undertook the redesign and validation of a second-generation MIA (MIA2G).STUDY DESIGN:MIA2G was developed using banked serum samples from a previously published prospective, multisite registry of patients who underwent surgery to remove an adnexal mass. Clinical validity was then established using banked serum samples from the OVA500 trial, a second prospective cohort of adnexal surgery patients. Based on the final pathology results of the OVA500 trial, this intended-use population for MIA2G testing was high risk, with an observed cancer prevalence of 18.7% (92/493). Coded samples were assayed for MIA2G biomarkers by an external clinical laboratory. Then MIA2G results were calculated and submitted to a clinical statistics contract organization for decoding and comparison to MIA results for each subject. Sensitivity, specificity, positive predictive value (PPV), and negative predictive value (NPV) were calculated, among other measures, and stratified by menopausal status, stage, and histologic subtype.RESULTS:Three MIA markers (cancer antigen 125, transferrin, and apolipoprotein A-1) and 2 new biomarkers (follicle-stimulating hormone and human epididymis protein 4) were included in MIA2G. A single cut-off separated high and low risk of malignancy regardless of patient menopausal status, eliminating potential for confusion or error. MIA2G specificity (69%, 277/401 [n/N]; 95% confidence interval [CI], 64.4-73.4%) and PPV (40%, 84/208; 95% CI, 33.9-47.2%) were significantly improved over MIA (specificity, 54%, 215/401; 95% CI, 48.7-58.4%, and PPV, 31%, 85/271; 95% CI, 26.1-37.1%, respectively) in this cohort. Sensitivity and NPV were not significantly different between the 2 tests. When combined with physician assessment, MIA2G correctly identified 75% of the malignancies missed by physician assessment alone.CONCLUSION:MIA2G specificity and PPV were significantly improved compared with MIA, while sensitivity and NPV were unchanged. The second-generation test significantly improved the predicted efficiency of triage vs MIA without sacrificing high sensitivity and NPV, which are essential for effectiveness.

5556 Background: Many women with ovarian cancer receive non-guideline care. Failure to refer to a gynecologic oncologist is one likely cause. To improve specificity of Multivariate Index Assay (MIA) for ovarian cancer triage, we undertook the re-design and validation of a second-generation test (MIA2G). Methods: A panel of MIA-positive malignant and benign case- controls was used to evaluate candidate biomarkers to improve specificity. These, and the 5 MIA biomarkers were then further ranked and developed into the MIA2G using banked samples from a previously published prospective, multi-site FDA registration trial of MIA (N = 585). Upon verification, the algorithm was encoded into diagnostics-grade software to calculate MIA2G in subsequent validation studies. Clinical validity was established under a double-blinded protocol using banked samples from the OVA500 trial, a second prospective MIA multi-site pivotal trial. Samples were assayed at 3 external sites, results were generated at a commercial laboratory and submitted to an independent statistician for decoding and analysis. MIA2G was compared with physician assessment (PA) or MIA. Results were compared for sensitivity, specificity, positive and negative predictive value (PPV and NPV), positive and negative likelihood ratios, concordance and percent overall accuracy. Results: Three MIA markers (CA125-II, transferrin and Apo A-1) and 2 new biomarkers (FSH and HE4) were selected for inclusion in MIA2G. MIA2G results range from 0-10, with a single cut-off separating hi- and low- malignancy risk of ³5.0. 493 of 519 OVA500 subjects were evaluable (276 pre and 217 post-menopausal). MIA2G specificity (69.1%) and PPV (40.4%) were significantly improved over MIA (53.6% and 31.4%, respectively). Sensitivity and NPV were not significantly different. Overall accuracy was significantly improved from 60.9% (MIA) to 73.2% (MIA2G). When MIA2G was combined with PA sensitivity was 93.5% and specificity was 64.8% Conclusions: The redesigned multivariate index assay, MIA2G, has significantly improved specificity, PPV and accuracy compared with MIA. These changes enable high-sensitivity detection of malignancy while reducing likelihood of over-referral.

The “corded and hyalinized” pattern, described in endometrioid carcinoma, has not been previously reported in association with serous carcinoma. We describe a unique case of serous neoplasm of low malignant potential with low-grade serous carcinoma combined with a distinct pattern of high-grade carcinoma characterized by cords of epithelioid and spindled cells enmeshed in a hyalinized, collagenous stroma. This pattern was the predominant architecture in the patient's recurrence and caused a diagnostic challenge, as the splenic recurrence was initially diagnosed as a second primary high-grade spindle cell neoplasm. Both ovarian and splenic tumors displayed positive immunohistochemical staining for cytokeratin 7, cytokeratin 8/18, estrogen receptor, and paired box gene 8 (PAX-8) in the conventional serous carcinoma and the corded and hyalinized component, confirming the diagnosis of recurrent carcinoma. The behavior in this unique case of serous carcinoma associated with a distinct corded and hyalinized pattern was more aggressive than low-grade serous carcinoma, but more favorable than malignant mixed mullerian tumor.

Objective: This study aims to determine factors that may increase the likelihood of adverse drug events (ADEs) in patients with recurrent endometrial cancer treated with pegylated liposomal doxorubicin (PLD) as well as this agent's impact on clinical outcomes.Methods: The treatment records of patients with endometrial cancer who received PLD at The University of Texas, MD Anderson Cancer Center, from 1996 to 2006 were reviewed. Patient demographics, PLD dose, ADEs, use of supportive care interventions, disease progression, and survival were extracted. Logistical regression analysis was used to identify factors that were associated with higher incidence of ADEs and that influenced survival.Results: A total of 60 patients with recurrent endometrial cancer were identified who experienced 122 ADEs. The most commonly reported ADEs were nausea (18.9%), palmar-plantar erythrodysesthesia (PPE; 16.4%), muscle weakness (12.3%), mucositis (10.7%), and peripheral neuropathy (9.8%). Seventeen patients (28%) required a dose reduction because of ADEs. However, only 5 (8.3%) patients discontinued therapy because of toxicity. Cooling mechanisms were used in 19 patients to prevent PPE, although 9 of these patients still experienced PPE. Treatment with 6 or more cycles of PLD was associated with increased incidence of neutropenia (P = 0.045), peripheral neuropathy (P = 0.004), and PPE (P < 0.001). No differences in progression-free survival or time to progression were found between the doses of PLD; however, there was an assessable trend toward increased survival with doses of 40 mg/m(2).Conclusions: Although there was no association with dose level and ADEs, more cycles received increased the incidence of toxicities, including PPE and neuropathy. There was no association between different doses of PLD and progression-free survival or time to progression.

Objective: To determine prognostic factors, recurrence risk factors, associated malignancies, and outcomes of various treatment methods for Paget's disease of the vulva.