A Translational Study to Explore and Overcome Metabolic-driven Resistance Mechanisms to Standard Chemotherapy in High-grade Ovarian Cancer

The study, titled "A Translational Study to Explore and Overcome Metabolic-Driven Resistance Mechanisms to Standard Chemotherapy in High-Grade Ovarian Carcinoma," is designed as a prospective, monocentric, observational cohort study involving the collection and utilization of biological material. The primary scientific objective of this research is to evaluate the predictive capacity of the U-Cup bioreactor system in determining the response to standard chemotherapy (SCT) in patient-derived high-grade ovarian cancer (HGOC) tissues maintained in perfusion culture. By utilizing this advanced bioreactor technology, the study aims to establish whether the ex vivo response of the tumor tissue accurately reflects the clinical outcomes observed in the patients. Complementing this, the secondary objective focuses on the development of a high-throughput platform for the personalized assessment of therapeutic responses in ovarian cancer cases that exhibit resistance to standard chemotherapy, specifically exploring alternative metabolic therapies. The study population consists of women diagnosed with high-grade serous ovarian carcinoma who have not previously undergone chemotherapy and are candidates for surgical intervention at the UOC Ginecologia Oncologica De Iaco - IRCCS AOUBO. To be eligible for enrollment, patients must be at least 25 years of age, provide informed consent, and have a confirmed histological or surgical diagnosis of high-grade serous ovarian carcinoma. Furthermore, inclusion requires that patients have not yet undergone surgical treatment and that there is sufficient primary or metastatic tumor tissue available for sampling. Conversely, patients will be excluded from the study if they present with significant comorbidities or are affected by other concurrent malignancies. The study is projected to span a duration of 36 months, with an anticipated enrollment of 120 patients to ensure a robust dataset. The statistical methodology for this study is rigorous and multifaceted. For all experimental procedures, including the drug response assays outlined in the secondary objective, the values for each measured parameter will initially undergo the Shapiro-Wilk test to assess the normality of their distribution. Continuous variables that follow a normal distribution will be analyzed using the Student's T-test, while those with a non-normal distribution will be evaluated using the Mann-Whitney U test. For categorical variables, the Chi-square test will be applied to normally distributed data, whereas Fisher's exact test will be used for data that does not meet normality assumptions. Regarding the primary objective, clinical data collected from the participants will be used to categorize patients into those with a Chemotherapy Response Score of 3 (CRS3), indicating a good response, versus those with CRS1 or CRS2, indicating a poor or partial response. The T-test or Mann-Whitney test will be employed to identify significant differences in terms of variations in cell proliferation, measured via Ki67-positive nuclei, apoptosis, indicated by cleaved Caspase-3 positivity, and necrosis, as assessed by a pathologist using hematoxylin and eosin (HE) staining. These comparisons will be made between samples treated with SCT in the bioreactor and untreated controls to determine if one or more of these biological parameters can reliably predict the patient's actual clinical response to treatment. Finally, the study will utilize Area Under the Receiver Operating Characteristic (AUROC) curve analysis and logistic regression to define the optimal threshold values for these parameters, aiming to discriminate between CRS3 and CRS1/2 patients with the highest possible levels of specificity and sensitivity, thereby validating the bioreactor as a precision medicine tool.