Mechanisms of response and resistance to KRAS inhibition in pancreatic cancer

胰腺癌中 KRAS 抑制的反应和耐药机制

基本信息

批准号：
10566224
负责人：
Andrew James Aguirre
金额：
$ 56.18万
依托单位：
DANA-FARBER CANCER INST
依托单位国家：
美国
项目类别：
财政年份：
2023
资助国家：
美国
起止时间：
2023-01-01 至 2027-12-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10566224
关键词：
ARID1A gene Biological Assay Biological Markers Biopsy CDKN2A gene Cell Cycle Progression Cells Clinical Clinical Trials Clustered Regularly Interspaced Short Palindromic Repeats Combined Modality Therapy DNA Damage Data Development Disease Engineering Foundations Future Genes Genetic Genetic Determinism Genetic Markers Genetic Transcription Genomic approach Genomics Genotype Goals Human In Vitro KPC model KRAS oncogenesis KRAS2 gene KRASG12D MADH4 gene MAP Kinase Gene Malignant Neoplasms Malignant neoplasm of pancreas Modeling Molecular Mus Mutation Neurons Oncogenic Oncoproteins Organoids Pancreatic Ductal Adenocarcinoma Paracrine Communication Pathway interactions Patients Pharmaceutical Preparations Phenotype Prognosis RAS inhibition Ras Inhibitor Resistance Role Screening Result Shapes Signal Transduction System TP53 gene Therapeutic Therapeutic Intervention Treatment Efficacy Tumor Suppressor Genes Tumor Suppressor Proteins cancer cell cancer type clinically relevant drug sensitivity effective therapy functional genomics genetic approach human RNA sequencing human data human model in vivo inhibitor innovation knockout gene molecular subtypes mouse model mutant neoplastic cell novel novel therapeutic intervention novel therapeutics pancreatic cancer model pancreatic cancer patients pancreatic ductal adenocarcinoma cell pancreatic ductal adenocarcinoma model paracrine predictive marker programs replication stress research clinical testing resistance mechanism response single-cell RNA sequencing small molecule inhibitor targeted treatment transplant model treatment response tumor tumor microenvironment

ARID1A gene Biological Assay Biological Markers Biopsy CDKN2A gene Cell Cycle Progression Cells Clinical Clinical Trials Clustered Regularly Interspaced Short Palindromic Repeats Combined Modality Therapy DNA Damage Data Development Disease Engineering Foundations Future Genes Genetic Genetic Determinism Genetic Markers Genetic Transcription Genomic approach Genomics Genotype Goals Human In Vitro KPC model KRAS oncogenesis KRAS2 gene KRASG12D MADH4 gene MAP Kinase Gene Malignant Neoplasms Malignant neoplasm of pancreas Modeling Molecular Mus Mutation Neurons Oncogenic Oncoproteins Organoids Pancreatic Ductal Adenocarcinoma Paracrine Communication Pathway interactions Patients Pharmaceutical Preparations Phenotype Prognosis RAS inhibition Ras Inhibitor Resistance Role Screening Result Shapes Signal Transduction System TP53 gene Therapeutic Therapeutic Intervention Treatment Efficacy Tumor Suppressor Genes Tumor Suppressor Proteins cancer cell cancer type clinically relevant drug sensitivity effective therapy functional genomics genetic approach human RNA sequencing human data human model in vivo inhibitor innovation knockout gene molecular subtypes mouse model mutant neoplastic cell novel novel therapeutic intervention novel therapeutics pancreatic cancer model pancreatic cancer patients pancreatic ductal adenocarcinoma cell pancreatic ductal adenocarcinoma model paracrine predictive marker programs replication stress research clinical testing resistance mechanism response single-cell RNA sequencing small molecule inhibitor targeted treatment transplant model treatment response tumor tumor microenvironment

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项目摘要

PROJECT SUMMARY/ABSTRACT Pancreatic ductal adenocarcinoma (PDAC) is a devastating cancer, with a five-year survival of only 10%. There is an urgent need to develop new therapeutic strategies for this disease. Oncogenic KRAS mutations occur in most patients and co-occur with alterations in several different tumor suppressor genes, including TP53, CDKN2A, SMAD4, ARID1A and others. Multiple different transcriptional subtypes of PDAC have also been observed, such as the classical and basal-like programs, which define distinct subsets of disease with differing prognosis and therapeutic response. The recent development of new small molecule inhibitors of KRAS that target KRAS mutations frequently observed in PDAC has the potential to transform the treatment of this disease. We and others have shown that primary and acquired resistance mechanisms can limit the clinical benefit of KRAS inhibitor monotherapy in cancer; thus, understanding the mechanisms of response and resistance to KRAS inhibition in PDAC will be critical to maximize the potential of these therapies. This proposal will use novel mutant-selective KRAS and pan-RAS inhibitors, unique human organoid and mouse models of PDAC, and innovative single-cell and functional genomic approaches to define the genetic, transcriptional and microenvironmental factors that impact response to KRAS inhibition in PDAC. In Aim 1 we will investigate how tumor suppressor genotype can modify response to KRAS inhibition using CRISPR-Cas12a tumor suppressor gene knockout screens in both in vitro and in vivo systems to simultaneously model multiple PDAC genotypes and systematically define genetic biomarkers and mechanisms of sensitivity or resistance to KRAS inhibition. In Aim 2, we will define the role of PDAC transcriptional cell state in modifying response to KRAS inhibition using novel isogenic murine PDAC organoids and human patient-derived PDAC organoids representing the basal-like, classical and neuronal-like subtypes of PDAC. We will characterize subtype-specific adaptive mechanisms of response to KRAS inhibition and evaluate subtype plasticity with a goal to develop combination therapy strategies with KRAS inhibition to target each subtype. In Aim 3, we will build on preliminary single-cell RNA sequencing (scRNA-seq) data from human PDAC biopsies showing that the tumor microenvironment (TME) shapes the transcriptional phenotype and therapeutic response of PDAC cells. We will examine response to oncogenic Kras inhibition in new mouse models of the basal-like, classical and neuronal-like subtypes of PDAC and will interrogate the role of paracrine signaling mechanisms from the TME in modifying malignant cell state and response to KRAS inhibition using ex vivo scRNA-seq and drug sensitivity profiling assays. Collectively, these studies will form a foundation for development of new biomarkers and combination therapies with KRAS inhibition that can be evaluated in future clinical trials for PDAC patients.

项目摘要/摘要胰腺导管腺癌（PDAC）是一种毁灭性的癌症，五年生存率仅为10％。那里迫切需要为这种疾病制定新的治疗策略。致癌KRAS突变发生在大多数患者和同时发生的肿瘤抑制基因，包括TP53， CDKN2A，SMAD4，ARID1A等。 PDAC的多种不同的转录亚型也已经观察到的，例如经典和基础样计划，该程序定义了不同的疾病子集预后和治疗反应。最近的新小分子抑制剂的最新发展 PDAC中经常观察到的靶标KRAS突变有可能改变该疾病的治疗方法。我们和其他人表明，主要和获得的抵抗机制可以限制 KRAS抑制剂单一疗法；因此，了解反应的机制和对 PDAC中的KRAS抑制作用对于最大化这些疗法的潜力至关重要。该提议将使用小说突变选择性的KRAS和PAN-RAS抑制剂，独特的人体器官和PDAC的小鼠模型，以及创新的单细胞和功能基因组方法来定义遗传，转录和影响PDAC中KRAS抑制反应的微环境因素。在目标1中，我们将调查如何肿瘤抑制基因型可以使用CRISPR-CAS12A抑制剂来改变对KRAS抑制的反应体外和体内系统中的基因敲除筛选，同时建模多个PDAC基因型并系统地定义了遗传生物标志物以及对KRAS抑制的敏感性或抗性的机制。在 AIM 2，我们将定义PDAC转录细胞状态在修改对KRAS抑制的反应中的作用新型的等基因鼠PDAC类器官和人类衍生的PDAC类器官，代表基底样的， PDAC的经典和神经元状亚型。我们将表征亚型特异性的自适应机制对KRAS抑制作用的反应，并通过建立组合疗法的目标评估亚型可塑性具有KRAS抑制的策略以针对每个亚型。在AIM 3中，我们将基于初步单细胞RNA 来自人类PDAC活检的测序（SCRNA-SEQ）数据表明肿瘤微环境（TME）塑造PDAC细胞的转录表型和治疗反应。我们将检查对 PDAC的基底样，经典和神经元的亚型的新小鼠模型中的致癌KRAS抑制并将询问来自TME的旁分泌信号传导机制在修饰恶性细胞状态中的作用以及使用离体SCRNA-SEQ和药物敏感性分析测定法对KRAS抑制的反应。共同这些研究将构成与KRAS开发新生物标志物和组合疗法的基础可以在PDAC患者的将来的临床试验中评估的抑制作用。