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

• 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
• 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

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开发新的治疗策略。致癌的KRAS突变发生在 大多数患者合并有几个不同的肿瘤抑制基因的变化，包括TP53， CDKN2A、Smad4、ARID1A等。多种不同转录亚型的PDAC也被 观察到的，例如经典的和类似基础的程序，它们定义了不同的疾病子集 预后和治疗反应。新型KRAS小分子抑制剂的研究进展 在PDAC中频繁观察到的靶向KRAS突变有可能改变这种疾病的治疗。 我们和其他人已经证明，原发和获得性耐药机制可以限制 KRAS抑制剂在癌症中的单一治疗；因此，了解对 抑制PDAC中的KRAS对于最大限度地发挥这些治疗的潜力至关重要。这项提议将使用小说 突变选择性KRAS和PAN-RAS抑制剂，PDAC的独特人类器官和小鼠模型，以及 创新的单细胞和功能基因组学方法来定义基因、转录和 影响PDAC对KRAS抑制反应的微环境因素。在目标1中，我们将研究如何 肿瘤抑制基因可以改变CRISPR-Cas12a肿瘤抑制基因对KRAS抑制的反应 在体外和体内系统中进行基因敲除筛选以同时模拟多个PDAC基因型 并系统地定义了对KRAS抑制敏感或抵抗的遗传生物标志物和机制。在……里面 目的2，我们将确定PDAC转录细胞状态在调节对KRAS抑制的反应中的作用 新的同基因小鼠PDAC有机类化合物和人患者来源的PDAC有机类化合物代表基底样， 典型的和神经元样的PDAC亚型。我们将描述特定于亚型的适应机制 对KRAS抑制的反应和评估亚型可塑性，目标是开发联合治疗 针对每个亚型的KRAS抑制策略。在目标3中，我们将在初步的单细胞RNA的基础上 来自人类PDAC活检组织的测序(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。我们将研究对以下问题的回应 新的小鼠基础样亚型、经典型亚型和神经元样亚型PDAC的致癌Kras抑制 并将询问来自TME的旁分泌信号机制在改变恶性细胞状态中的作用 用体外scRNA-seq和药物敏感性分析检测对KRAS抑制的反应。总而言之， 这些研究将为开发新的生物标记物和与KRAS的联合治疗奠定基础 可在未来对PDAC患者的临床试验中评估的抑制作用。