PDX tumor-TME acquired resistance

PDX肿瘤-TME获得性耐药

• 批准号: 10517261
• 负责人: Jack Roth
• 金额: $ 35.55万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-09-30 至 2027-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10517261
• 关键词: CRISPR screen; Cells; Clinical Trials; Clustered Regularly Interspaced Short Palindromic Repeats; Combination immunotherapy; Data; Drug Screening; Drug Targeting; Drug resistance; Epidermal Growth Factor Receptor; Future; Gene Expression; Histology; Human; Immune; Immune response; KRAS2 gene; Malignant Neoplasms; Modeling; Molecular; Mutation; Non-Small-Cell Lung Carcinoma; Organoids; PDPK1 gene; Pathway interactions; Patients; Proteins; Proteomics; Residual Tumors; Resistance; Role; Small Interfering RNA; Stromal Cells; Testing; Therapeutic; Tumor-associated macrophages; Tyrosine Kinase Inhibitor; acquired drug resistance; base; drug efficacy; drug testing; humanized mouse; immune checkpoint blockade; inhibitor; innovative technologies; macrophage; molecular subtypes; mutant; novel; novel therapeutics; overexpression; phosphoinositide-dependent kinase 1; pre-clinical; programs; protein expression; resistance mechanism; response; restoration; therapeutic target; transcriptome sequencing; treatment response; treatment strategy; tumor; tumor microenvironment; tumor-immune system interactions

Project hypothesis and Specific Aims: We hypothesize that the molecular subtypes of EGFR and KRAS mutant NSCLC tumors with acquired mutation targeted drug resistance are associated with an immunosuppressive TME including both immune and non-immune stromal cells that can be therapeutically targeted in a way that will enhance the efficacy of drugs targeted to intrinsic resistance pathways. Specific Aim 1: Develop and characterize existing and new models of mutant EGFR and KRAS inhibitor acquired resistance including PDXs and PDX-derived organoids and characterize mechanistic relationships between molecular subtypes and the TME, including immune and non-immune stroma. Characterization will include histology, immunohistochemisry (IHC), RNAseq, and MS proteomics. We will identify specific molecular alterations that can be functionally tested and determine their effect on the TME. Examples being tested currently include 3-phosphoinositide-dependent kinase 1 (PDK1, PDPK1) expression, SETD1B inactivating mutations, and YAP (yes-associated protein) expression. We will test drugs that target specific cells in the TME to enhance the immune response. We are currently targeting M2 tumor associated macrophages (TAM). Osimertinib and G12C KRASi resistant PDXs and organoids will be interrogated to identify acquired drug resistance molecular pathways using MS proteomics, scRNAseq, and CRISPR screens. Specific Aim 2: Define treatment strategies in mutant EGFR and KRAS inhibitor acquired resistance models based on targeting pathways (e.g., PDK1, YAP, NF-B) implicated in current and future studies of acquired resistance caused by tumor-TME cell networks (e.g., CSFR1, CD74). Potential drug targets for overcoming acquired resistance to osimertinib (see preliminary data) include PDK1, YAP, NFkB, and TAM. We will develop therapeutic approaches based on TME characterization. H1975R has a more immune stimulatory TME than PC9 YAP thus providing a rationale for testing checkpoint blockade combined with PDK1 inhibition in H1975R. M2 polarized macrophages are increased in PC9 overexpressing wtYAP thus providing a rationale for testing TAM inhibition in PC9 YAP with a CSFR1 inhibitor. Residual tumors after osimertinib and KRAS G12Ci will be characterized following treatment using MS proteomics, scRNAseq, and CRISPR screens to identify novel acquired resistance pathways. The functional role of specific targets in acquired drug resistance will be validated using CRISPR KO. We will develop new models to include future novel drugs developed to target these and other pathways.

项目假设和具体目的：我们假设EGFR和KRAS的分子亚型 具有获得性突变靶向耐药的突变型NSCLC肿瘤与 免疫抑制性TME，包括免疫和非免疫基质细胞，其可以治疗性地 以增强靶向内在耐药途径的药物的功效的方式靶向。 具体目标1：开发和表征突变型EGFR和KRAS抑制剂的现有模型和新模型 获得性抗性，包括PDX和PDX衍生的类器官，并表征其机制 分子亚型和TME之间的关系，包括免疫和非免疫基质。 表征将包括组织学、免疫组化（IHC）、RNAseq和MS蛋白质组学。我们将 鉴定可以进行功能测试的特定分子改变，并确定其对TME的影响。 目前正在测试的例子包括3-磷酸肌醇依赖性激酶1（PDK 1、PDPK 1）表达， SETD 1B失活突变和雅普（yes相关蛋白）表达。我们将测试针对 TME中的特定细胞以增强免疫应答。我们目前正在针对M2肿瘤相关的 巨噬细胞（TAM）。将询问奥希替尼和G12 C KRASi耐药的PDX和类器官，以确定 使用MS蛋白质组学、scRNAseq和CRISPR筛选获得性耐药分子途径。 具体目标2：确定突变型EGFR和KRAS抑制剂获得性耐药的治疗策略 基于靶向途径的模型（例如，PDK 1、雅普、NF-κ B B）与当前和未来研究有关 由肿瘤-TME细胞网络引起的获得性抗性（例如，CSFR1，CD74）。潜在的药物靶点 克服对奥希替尼的获得性耐药性（参见初步数据）的药物包括PDK 1、雅普、NF κ B和TAM。我们 将开发基于TME特征的治疗方法。H1975 R具有更强的免疫刺激性， TME比PC 9雅普更好，因此为在TME中测试检查点阻断与PDK 1抑制的组合提供了依据。 H1975R。在过表达wtYAP的PC 9中，M2极化的巨噬细胞增加，从而为WtYAP的表达提供了理论基础。 用CSFR 1抑制剂测试PC 9雅普中的TAM抑制。奥希替尼和KRAS G12 Ci治疗后的残留肿瘤 将在治疗后使用MS蛋白质组学、scRNAseq和CRISPR筛选进行表征，以鉴定新的 获得性抗性途径。获得性耐药中特异性靶点的功能作用将得到验证 使用CRISPR KO。我们将开发新的模型，包括未来针对这些和其他目标开发的新药。 途径。