Complex project title: “Multidisciplinary Consortium for Supporting Research Skills in Diagnosing, Treating and Identifying Predictive Factors of Malignant Gynecologic Disorders”
Project acronym: ONCOGIN
Project coordinator: Dr. Suciu Nicolae
Competition: “Complex projects completed in consortia CDI” (PCCI), PN III, Program 1, Subprogram 1.2.
Contracting Authority: Executive Unit for Financing Higher Education, Research, Development and Innovation (UEFISCDI)
Project code: PN-III-P1-1.2-PCCDI2017-0833
Contract number: 68PCCDI/2018
Project duration: 05.04.2018-30.09.2020
Project coordinator: The National Institute for Mother and Child Protection “Prof. Dr. Alessandrescu-Rusescu”
Project Director: Prof. Dr. Nicolae Suciu
P1- Fundeni Clinical Institute
P2-Institute of Oncology Prof. Dr. Alexandru Trestoianu
P3-Carol Davila University of Medicine and Pharmacy, Bucharest
P4-Horia Hulubei National Institute of Physics and Nuclear Engineering (IFIN-HH)
Aim: INSMC, having the role of project coordinator, together with the 4 partners, aims to discover and introduce new methods of diagnosis, treatment and prognosis in the field of oncoginecology by identifying revolutionary biomarkers.
The main objective of the project was to find new communication pathways by identifying new markers of angiogenesis and new immunological treatment targets in order to reduce / inhibit tumor progression.
The two sub-projects have in common research, development of models and innovative methods, creating human resources through the shared use of existing infrastructures. Development of personalized therapy to address new therapeutic targets that will be identified in ovarian cancer using new techniques such as NGS (Next Generation Sequencing) for BRCA 1 and BRCA 2 determination, microarray for miRNA and application of advanced bioanalytical tools. Based on analysis of immunohistochemistry, special imaging and genomics, the following are pursued:
- formation of groups of patients;
- elaboration of a working algorithm that identifies the patients who will develop chemoresistance;
- identification of a back-up test using multigenic analyzes corroborated with imaging methods; drafting informed consent forms; elaboration of the patient evaluation sheet; standardized working procedures described below in the report.
Optimization of the workflow and methods of RNA extraction from paraffin samples, fresh frozen tissue, blood, in order to carry out the molecular biology experiments provided in the project.
Selection criteria for patients with peritoneal carcinomatosis in gynecological malignancies.
Development of conceptual models regarding the optimization of the culture methods of the peritoneum / ascites tumor cells.
Consolidation of the consortium and establishment of synergistic and complementary activities in the HIPEC study in peritoneal carcinomatosis.
Selection and inclusion of patients in the batch. - patients with peritoneal carcinomatosis candidates for cytoreduction and HIPEC. - patients with recurrence after initial surgical and chemotherapy treatment.
Obtaining biospecimens (tumor tissue, peritoneum and liquid biopsy / ascites + blood) from patients in whom cytoreduction and HIPEC were performed. Anatomical-histological characterization of tumor / peritumoral samples ".
Extraction of nucleic acids (RNA / DNA) from tissue samples / ascites / serum obtained
Preparation and preliminary testing of some radiotherapy devices for PET imaging in ovarian and breast cancers.
Identification of molecular predictors of HYPERTHERMIC Intraperitoneal Chemotherapy (HIPEC) response and study of mechanisms involved in resistance to treatment in ovarian cancer
- Patient selection and inclusion in the batch. - patients with peritoneal carcinomatosis who are candidates for cytoreduction and HIPEC. - patients with recurrence after the initial surgical and chemotherapy treatment.
- Evaluation of BRCA1 / 2 mutation in patients selected by targeted sequencing techniques.
- Obtaining biospecimens (tumor tissue, peritoneum and liquid / ascites + blood biopsy) from patients in whom cytoreduction and HIPEC were performed. Anatomical-histological characterization of tumor / peritumoral samples.
- Extraction of nucleic acids (RNA / DNA) from tissue samples / ascites / serum obtained.
- Generation of miRNA of interest by microarray analysis of the samples obtained before and after performing HIPEC.
- Selection, by bioinformatic analysis of genes of interest (mRNA) - targets modeled by miRNA of interest. Highlighting the signaling pathways specific to the response to HIPEC.
- QRT-PCR validation of a miRNA panel obtained after bioinformatics analysis.
- In vitro determination of the expression of HER2 receptors in ovarian cancers with radiotransactors on standardized and cultured cell cultures.
- Corelating miRNA expression with the Peritoneal Carcinomatosis Index; residual disease (R) score
- Identifying new therapeutic targets used for evaluating the in vitro response
- Introducing the patients in the study – Registering the dates of every patient in a database – including the patients in the tumor bank (at the time of the diagnosis, the surgical procedure and after receiving the neoadjuvant treatment)
- Creating a digital platform used for collecting medical data and generating reporting; Analyzing the requirements needed for the digital platform used for collecting medical data and generating reports – Defining the data structure which will be registered in the digital platform – Designing the experimental model for the digital platform used for collecting medical data and generating reports – Digital platform’s validation of function - Populating the digital platform with medical data – Statistic analysis of the data
- Obtaining blood prior and after chemotherapy (including the recurrences of disease while post operatory care); tumoral tissue sampling at the time of the surgical intervention
- Analysis of a patient set with selected markers based on medical literature CA 15.3, CEA, CA27.29
- Analysis of a patient set with selected markers based on medical literature AR,ER, PR, HER2, ki67, CD40 and some of the markers used for differential diagnosis S100, SMA, SMMHC, calponin, CK5, CK14, CK17, CD10 and p63, E-cadherin, CK8 and some of the markers for metastasis GCDFP15, mammaglobin, CEA
- Characterizing patients’ tumors by the treatment response and corelating the therapeutic response with prior described markers and with tissue markers for neoangiogenesis: Angiogenin, Ang2, keratinocyte growth factor (KGF), fibroblast growth factor basic, intercellular adhesion molecule (ICAM)-1, platelet-derived growth factor-BB and VEGF (vascular endothelial growth factor)
- Analysis of the same patient lot for immunologic markers: control point inhibitors for the cellular cycle (CTLA4, PDL1), CD 68, CD163 (macrophage), CD8, CXCL12
- Genomic DNA isolation and integrity analysis using Agilent TapeStation 2200 system
- Analysis of somatic mutations (BRCA1 and BRCA2) in patients with breast cancer using NGS technology (Illumina, Ion Torrent)
- Reviewing the date collected by analyzing the patient set; descriptive analysis of the results and statistic interpretation of the molecular biology data and clinical data. Bioinformatic analysis of the NGS data.
- Imaging tests: contrast MRI and ultrasonography
- Correlating the imaging data with molecular biology data in order to promote personalized treatments