Advancing RAS pathway targeted therapy in NF1-MPNST: effects of SHP2 and CDK4/6 inhibitors on the tumor and the tumor immune microenvironment

推进NF1-MPNST的RAS通路靶向治疗：SHP2和CDK4/6抑制剂对肿瘤和肿瘤免疫微环境的影响

• 批准号: 10660326
• 负责人: Christine Anne Pratilas
• 金额: $ 55.42万
• 依托单位: JOHNS HOPKINS UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-06-12 至 2028-05-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10660326
• 关键词: Affect; Animal Model; Biochemical; Biological; Biological Assay; Biological Markers; CDK4 gene; Cell Culture Techniques; Cells; Chemotherapy and/or radiation; Clinic; Clinical; Clinical Trials; Combined Modality Therapy; Conditioned Reflex; Data; Development; Dimensions; Disease; Elements; Feedback; Flow Cytometry; Future; GTPase-Activating Proteins; Genetic Transcription; Genomics; Human; Immune; Immune response; Immunocompetent; Immunohistochemistry; Immunotherapeutic agent; Immunotherapy; In Vitro; Infiltration; Investigation; Knowledge; Location; MEK inhibition; MEKs; Mediator; Modeling; Molecular Target; Morbidity - disease rate; Mus; Myelogenous; Myeloid Cells; Myeloid-derived suppressor cells; NF1 gene; Neoplasm Metastasis; Neurofibrosarcoma; Oncogenic; Operative Surgical Procedures; PTPN11 gene; Pathologic; Pathway interactions; Patient Selection; Patients; Persons; Pharmaceutical Preparations; Pre-Clinical Model; Predisposition; Primary Neoplasm; Proteomics; Ras Inhibitor; Receptor Protein-Tyrosine Kinases; Reporting; Research; Resistance; Role; Safety; Schwann Cells; Signal Pathway; Signal Transduction; Technology; Testing; Therapeutic; Time; Toxic effect; Translating; Tumor Immunity; Tumor Suppressor Genes; Tumor-infiltrating immune cells; Up-Regulation; Validation; cell type; checkpoint inhibition; chemotherapy; combinatorial; design; early phase clinical trial; efficacy evaluation; genetic manipulation; hyperactive Ras; immune cell infiltrate; immunoregulation; improved; in vivo evaluation; inhibitor; loss of function; mouse model; neoplastic cell; new therapeutic target; novel; novel drug class; novel therapeutic intervention; novel therapeutics; patient derived xenograft model; patient response; pre-clinical; programs; protein function; rare cancer; ras GTPase-Activating Proteins; rational design; resistance mechanism; response; sarcoma; small molecule inhibitor; small molecule therapeutics; targeted agent; targeted treatment; therapy design; tool; transcriptomics; tumor; tumor microenvironment; tumor-immune system interactions

PROJECT SUMMARY/ ABSTRACT There has been little advancement in overall survival for patients with malignant peripheral nerve sheath tumor (MPNST), despite decades of research and many clinical trials, and thus novel therapies are needed. While loss of NF1 GTPase-activating protein function suggests that targeting RAS may be a logical therapeutic approach, there is currently no approved drug that effectively and directly targets wild-type RAS. The design of novel therapeutic combinations requires a deep understanding of the signaling pathways regulated by hyperactive RAS and feedback that conditions the response to their inhibition. MEK, SHP2, and CDK4/6 are critical nodes in RAS effector signaling in MPNST tumors, and combinations of small molecule inhibitors that target them may have synergistic anti-tumor activity. Further, adaptive signaling changes in response to RAS effector pathway inhibition occurs not only in tumor cells but also in cells that comprise the tumor immune micro- environment (TIME). Thus, the characterization of the TIME in our murine model of MPNST, and its modulation via small-molecule inhibitors of the RAS pathway, will identify key immune pathways leading to the reprogramming of intratumoral myeloid cells, enhancing endogenous immune responses. We propose three Aims: 1. Identify and functionally validate mechanisms of acquired resistance to SHP2 inhibitors in MPNST in order to develop novel therapeutic combinations. We will determine how tumors become resistant to SHP2i, as well as to SHP2i +CDK4/6i in combination, and will perform genetic manipulation studies in order to functionally validate priority hits. 2. Determine the efficacy and tolerability of combination small- molecule inhibitors of RAS signaling, particularly SHP2i + CDK4/6i, in in vitro, patient-derived xenografts (PDX), and immune-competent syngeneic Nf1 -/-/Ink4a/Arf -/- mouse models. We will test the effects of SHP2i and CDK4/6i single agents or combinations as tools to probe the biochemical and biological changes that occur upon pathway perturbation, and will determine the mechanism and anti-tumor activity of these combinations, with a priority focus on the SHP2i+CDK4/6i combination, based on our advanced preliminary data. 3. Reprogram the intratumoral pathological myeloid cells via treatment with the combination of SHP2i and CDK4/6i in murine MPNST. Our preliminary data suggest that the landscape of MPNST is densely populated by tumor infiltrating myeloid cells. To evaluate the effects of these agents and their combinations on the TIME, we will utilize single cell transcriptional analysis, multiparameter flow cytometry and multiplex immunohistochemistry to decipher the specific effects of these drugs on cell types that comprise the tumor, including both immune-infiltrating and primary tumor cells. By determining the efficacy and mechanism of these rationally-designed therapeutic strategies in both PDX and immune-competent mouse models, our studies will provide mechanism-based, promising combinatorial approaches that may be rapidly and successfully translated to the clinic, and will inform effective patient selection strategies and the development of novel clinical trials for patients with MPNST.

项目摘要/摘要 对于恶性周围神经鞘肿瘤的患者，总体存活率几乎没有进步 （MPNST），尽管进行了数十年的研究和许多临床试验，因此仍需要新的疗法。而损失 NF1 GTPase激活蛋白功能的功能表明，靶向RA可能是一种逻辑治疗方法， 目前尚无有效的批准药物，并直接针对野生型RA。小说的设计 治疗组合需要深入了解由过度活跃调节的信号通路 RAS和反馈条件，该反应对其抑制作用。 MEK，SHP2和CDK4/6是关键节点 在MPNST肿瘤中的RAS效应子信号中，以及针对它们的小分子抑制剂的组合可能 具有协同的抗肿瘤活性。此外，自适应信号传导响应于RAS效应器途径 抑制不仅发生在肿瘤细胞中 环境（时间）。因此，我们的MPNST鼠模型中时间的表征及其调制 通过RAS途径的小分子抑制剂，将识别导致的关键免疫途径 重编程肿瘤内髓样细胞，增强内源性免疫反应。 我们提出了三个目标：1。识别并在功能上验证获得对SHP2的抗性的机制 MPNST中的抑制剂是为了开发新的治疗组合。我们将确定肿瘤的变化 对SHP2I的耐药性以及对SHP2I +CDK4/6i的抗性，并将进行遗传操纵研究 为了在功能上验证优先级命中。 2。确定组合小的功效和耐受性 在体外，患者衍生的异种移植物（PDX）中，RAS信号传导的分子抑制剂，尤其是SHP2I + CDK4/6i 和免疫功能的合成性NF1 - / - /INK4A/ARF - / - 鼠标模型。我们将测试SHP2I和 CDK4/6i单位代理或组合作为探测生化和生物学变化的工具 途径扰动，并将确定这些组合的机制和抗肿瘤活性 基于我们的先进初步数据，优先级关注SHP2I+CDK4/6i组合。 3。重新编程 肿瘤内病理髓样细胞通过鼠类在鼠中的SHP2I和CDK4/6i的结合进行处理 mpnst。我们的初步数据表明，MPNST的景观密集被肿瘤浸润 髓样细胞。为了评估这些代理的效果及其在时间上的组合，我们将利用单个 细胞转录分析，多参数流式细胞术和多重免疫组织化学，以破译 这些药物对构成肿瘤的细胞类型的特定作用，包括免疫浸润和 原发性肿瘤细胞。通过确定这些理性设计的治疗的功效和机制 PDX和免疫能力的小鼠模型中的策略，我们的研究将提供基于机制的， 有希望的组合方法可能会迅速并成功地转化为诊所，并将告知 有效的患者选择策略和MPNST患者的新型临床试验的发展。