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Differential pathway engagement and the biological consequences of KRAS variants in cancer

癌症中 KRAS 变异的差异通路参与和生物学后果

基本信息

批准号：
10469984
负责人：
Yanixa Quinones Aviles
金额：
$ 4.68万
依托单位：
YALE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-09-01 至 2024-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10469984
关键词：
Ablation Affect Affinity Alleles Amino Acids Bar Codes Base Sequence Binding Biochemical Biological Biological Assay Biology CRISPR/Cas technology Cancer Biology Cancer Model Cell Line Cell Proliferation Cell model Cell physiology Cells Clinical Clinical Data Competence Computational Technique Data Dependence Development Evaluation Excision Exhibits Frequencies GTPase-Activating Proteins Gene Expression Gene Expression Profile Genetic Genetically Engineered Mouse Goals Growth Guanosine Triphosphate Human Hydrolysis Immunohistochemistry In Vitro Individual KRAS2 gene Lead Maintenance Malignant Neoplasms Mass Spectrum Analysis Mediating Metabolism Methods Molecular Monitor Monomeric GTP-Binding Proteins Mus Mutate Mutation Oncogenic Oncoproteins Output Pancreatic Ductal Adenocarcinoma Pathway interactions Patient-Focused Outcomes Pharmacology Phenotype Phosphoproteins Point Mutation Population Prevalence Prognosis Property Proteins Proto-Oncogenes Reporting Research Scientist Series Signal Pathway Signal Transduction Signaling Molecule Spottings Supervision System Testing Therapeutic Therapeutic Intervention Tissues Training Transplantation Treatment outcome Tumor Cell Line Variant Western Blotting Work base cancer cell cancer type career cell behavior comparative experimental study fitness improved in vitro Assay in vivo inhibitor insight migration mutant pancreatic ductal adenocarcinoma cell pancreatic ductal adenocarcinoma model patient derived xenograft model patient prognosis patient response phosphoproteomics pre-clinical prevent subcutaneous therapeutic target transcriptome sequencing treatment response tumor tumor initiation tumorigenesis tumorigenic

项目摘要

Project Summary KRAS is the most frequently mutated proto-oncogene in human cancer and encodes a small GTPase that regulates multiple cellular processes such as cell proliferation, metabolism, migration, and survival. Point mutations in amino acids G12, G13 and Q61 prevent KRAS inactivation by regulatory GTPase-activating proteins and facilitate tumorigenesis. Although the frequency of specific KRAS mutant variants differ by cancer type, the mechanistic basis for this observation is unknown. It has been postulated that specific mutants induce a “sweet spot” of signaling alterations to induce a cell state optimized for tumor development and maintenance in specific tissues. In support of this hypothesis, KRAS mutants exhibit different biochemical properties in GTP hydrolysis rates and binding affinity to downstream effectors, supporting divergence in their activation of signaling networks. Furthermore, preclinical and clinical data revealed allele-specific differences in tumor initiation capacity and patient prognosis in pancreatic ductal adenocarcinoma (PDAC), suggesting that divergent signaling output could lead to altered phenotypic properties. A systematic and comprehensive evaluation of allele-specific signaling networks would be valuable to better understand KRAS diversity and reveal variant-specific dependencies. The overarching objective of this proposal is to understand KRAS mutant-specific differences in signaling and how these alter cellular fitness. Our preliminary data revealed differential engagement of global signaling networks and canonical amongst KRAS mutants. Therefore, our central hypothesis is that the biochemical differences between KRAS variants result in differential signaling pathway engagement, affecting cellular behavior, tumorigenic properties, and response to therapy. To test this hypothesis, I will re-express a large panel of KRAS mutants observed in human cancer in our recently generated isogenic KRAS deficient PDAC cell lines to dissect KRAS variant-specific differences in signaling networks using data-independent acquisition mass spectroscopy (DIA-MS). Experiments proposed in Aim 1 will investigate differences in signaling pathway activation and dependency of KRAS mutants and validate them in genetically engineered mouse models, PDX models, and human tumor biospecimens. Aim 2 will explore the cellular fitness of these mutants combining in vitro and in vivo competitive assays. Collectively, these data will provide fundamental insights into the biology of KRAS mutants that could potentially explain differences in clinical prevalence and response to therapy. More broadly, this work could inform new allele-specific therapeutic strategies for PDAC and other KRAS mutant cancers. Finally, this highly interdisciplinary and collaborative effort will train me in the application of cutting-edge molecular, biochemical, and computational techniques to facilitate my career goal of becoming an independent scientist in cancer biology.

项目总结