Dissecting FAK-regulated oncogenic signaling programs in ovarian cancer

剖析卵巢癌中 FAK 调节的致癌信号传导程序

• 批准号: 10403441
• 负责人: David D Schlaepfer
• 金额: $ 46.06万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN DIEGO
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-05-01 至 2025-04-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10403441
• 关键词: 3-Dimensional; ATAC-seq; Adhesions; Automobile Driving; Back; Bioinformatics; Biological Assay; Biological Markers; Biopsy; Cancer Etiology; Cancer Model; Cancer Patient; Cancer Relapse; Cause of Death; Cell Adhesion; Cell Culture Techniques; Cells; Cellular biology; Cessation of life; Chemoresistance; Chromosomes; Clinical Trials; Clinical trial protocol document; Cluster Analysis; Clustered Regularly Interspaced Short Palindromic Repeats; Collection; Complement; Complex; DDR1 gene; DNA Repair; DNA Repair Gene; Databases; Defect; Dependence; Disease; Drug Targeting; Environment; Exhibits; FOXM1 gene; Focal Adhesion Kinase 1; Fostering; Genes; Genetic; Genomics; Goals; Growth; Human; Hypoxia; In Vitro; Integrins; KRAS2 gene; Knock-out; Knowledge; Link; Malignant Female Reproductive System Neoplasm; Malignant Neoplasms; Malignant neoplasm of ovary; Modeling; Molecular; Molecular Conformation; Mus; Neoadjuvant Therapy; Nuclear; Oncogenes; Oncogenic; Organoids; Ovarian; PTK2 gene; Paclitaxel; Patient-Focused Outcomes; Patients; Peritoneal; Pharmacology; Phenotype; Phosphotransferases; Platinum; Pre-Clinical Model; Protein Tyrosine Kinase; RNA; Recurrence; Resistance; Role; Sampling; Serous; Signal Pathway; Signal Transduction; Signaling Molecule; Sleeping Beauty; Stress; Supporting Cell; System; TP53 gene; Testing; The Cancer Genome Atlas; Time; Transplantation; Tyrosine Phosphorylation; Validation; Woman; base; bench to bedside; beta catenin; cell motility; chemotherapy; clinical trial analysis; combinatorial; epigenome; genetic signature; in vivo; insight; mechanotransduction; model development; mutant; neoplastic cell; novel; ovarian neoplasm; prognostic; programs; reconstitution; research clinical testing; response; single cell sequencing; single-cell RNA sequencing; standard of care; stem; stemness; three dimensional cell culture; transcription factor; transcriptome sequencing; transcriptomics; treatment response; tumor; tumor growth; tumor heterogeneity; tumor microenvironment

Title: Dissecting FAK-regulated oncogenic signaling programs in ovarian cancer High-Grade Serous Ovarian Cancer (HGSOC) kills four of five women within sixty months. HGSOC is genetically complex, which has slowed past preclinical model development. To this end, we have molecularly characterized a new and in vivo evolved, aggressive, implantable, and syngeneic murine ovarian cancer model. These cells display many spontaneous acquired copy number changes, including K-Ras, Myc, and FAK/PTK2 genes (herein termed KMF cells) among other striking similarities to HGSOC. FAK (focal adhesion kinase) is a tyrosine kinase canonically supporting integrin signaling, motility and mechano-sensing. Our collective approaches in HGSOC and KMF cells, including pharmacological inhibition, FAK knockout, FAK re-expression, complementation, and bioinformatic analyses reveal that non-canonical adhesion-independent FAK signaling sustains intrinsic resistance to platinum chemotherapy in part via b-catenin activation and the elevation of transcription factors supporting stemness and DNA repair genes. FAK is activated in patient tumors surviving chemotherapy and acquired platinum resistance can facilitate ovarian tumorsphere dependence on FAK for growth. We identified a gene set associated with FAK expression and a subset linked to intrinsic FAK activity in 3D organoid cell culture. Exogenous activated b-catenin expression was sufficient to rescue FAK loss or inactivation phenotypes in 3D culture, but b-catenin did not promote FAK-null tumor growth in mice. Thus, FAK selectively promotes oncogenic signaling in vivo and FAK senses the tumor microenvironment. Our proposal will test the hypothesis that stress-induced FAK activation in tumorspheres surviving within a mouse peritoneal environment triggers specific cellular reprogramming, fostering stem-like state of heightened oncogenicity. In Aim-1, our unique gene-edited human and murine ovarian tumor systems will be used together with an inducible FAK expression system to characterize FAK localization- and kinase-dependent signals driving malignancy. In Aim-2, total and single cell RNA-seq will be performed on cells isolated from tumor-bearing mice to determine FAK regulated targets in vivo. Combined single cell RNA-seq and ATAC-seq will determine how subpopulations of cells are derived in response to time-dependent FAK activation and the interrelationship of gene markers in cell subpopulations. In Aim-3, we will use molecular and immunohistochemical analyses of patient clinical trial samples to identify and biomarkers associated with FAK inhibition and patient outcome. Our proposal encompasses cell biology, advanced RNA sequencing, epigenome mapping as well as single cell sequencing with bioinformatic clustering analysis. These approaches, together with the evaluation of clinical trial patient samples, will identify a “FAK-dependent” cell biomarker gene signature that can be re-tested for significance within KMF and HGSOOC tumor models. These studies will provide important insights into a targetable signaling pathway sustaining HGSOC malignancy.

标题：剖析卵巢癌中fak调控的致癌信号程序