Dissecting the tumor cell-immune TME axis to identify therapeutically actionable vulnerabilities that potentiate immunotherapy in GBM

剖析肿瘤细胞免疫 TME 轴，以确定可增强 GBM 免疫治疗的治疗上可操作的漏洞

• 批准号: 10743534
• 负责人: Amanda Johnson
• 金额: $ 4.92万
• 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: 2023-09-01 至 2025-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10743534
• 关键词: Adult; Allografting; Attenuated; Automobile Driving; Behavior; Bioinformatics; Biological Assay; Biological Models; Brain Neoplasms; Cell Survival; Cell model; Cell physiology; Cells; Characteristics; Clinical; Clonal Evolution; Coculture Techniques; Data; Dedications; Dependence; Development; Endowment; Environment; Event; Fatal Outcome; Gene Expression Profile; Genetic; Glioblastoma; Glioma; Goals; Grant; Growth; Heterogeneity; Hypoxia; Immune; Immune Evasion; Immune Targeting; Immune checkpoint inhibitor; Immune response; Immunocompetent; Immunosuppression; Immunotherapeutic agent; Immunotherapy; In Vitro; Interdisciplinary Study; Maintenance; Malignant Neoplasms; Malignant neoplasm of brain; Mesenchymal; Metabolic; Methods; Modeling; Molecular; Mus; Nutrient; Oncogenic; Patients; Pharmaceutical Preparations; Phase; Phenotype; Population; Prognosis; Receptor Inhibition; Receptor Signaling; Regulatory T-Lymphocyte; Research; Resistance; Signal Induction; Signal Transduction; Specimen; T-Lymphocyte; TGFBR2 gene; Techniques; Technology; Therapeutic; Tissues; Training; Transgenic Organisms; Tumor Immunity; Tumor Promotion; Tumor-infiltrating immune cells; Vaccines; anti-tumor immune response; cancer stem cell; cell dedifferentiation; checkpoint therapy; chimeric antigen receptor T cells; clinical translation; effective therapy; immune cell infiltrate; improved; in vivo; inhibitor; interdisciplinary approach; knock-down; metabolic profile; metabolomics; molecular subtypes; neoplastic cell; novel; novel strategies; novel therapeutics; pharmacologic; pre-clinical; pressure; programs; response; self-renewal; single cell sequencing; single-cell RNA sequencing; small hairpin RNA; small molecule; small molecule inhibitor; stem; stem cell biology; stem cell function; stem cells; stem-like cell; targeted treatment; therapeutic target; therapy resistant; trait; transcription factor; transcriptional reprogramming; transcriptome sequencing; transcriptomics; translatable strategy; tumor; tumor growth; tumor heterogeneity; tumor microenvironment; tumor progression

Glioblastoma multiforme (GBM) is a common and highly aggressive form of brain cancer in adults with a dismal prognosis and limited therapeutic options. A critical component of GBM malignancy derives from the distinct population of glioma stem cells (GSCs) that function to promote and maintain malignancy through their capacity for self-renewal, cellular adaptation and multipotency. These stem-like cells engage in a synergistic relationship with the surrounding microenvironment to promote tumor progression and are key drivers of intratumoral heterogeneity, immune-suppression, and therapy resistance. Targeting GSCs and mechanisms that drive the stem-like phenotype presents a promising avenue for targeted therapeutics. The broad goal of this proposal is to understand the cell-intrinsic mechanisms driving maintenance of a unique, immunosuppressive GSC subset, identify cellular vulnerabilities associated with immunosuppressive GBM cells, and develop preclinical therapeutics to target these cells. We will achieve this goal by utilizing our validated GSC cell models, state-of-the art single-cell sequencing technology, and cutting-edge spatially resolved omics platforms applied to clinical GBM specimens, and advanced molecular in vitro techniques to define immunosuppressive tumor cell populations and determine the transcriptomic and metabolic changes associated with these cell populations that are amenable to therapeutic targeting. Our preliminary findings demonstrate that TGF-beta type II receptor (TGFBR2) signaling induces a TGFBR2high subset of GSCs that co-opt certain immunosuppressive mechanisms associated with and utilized by regulatory T cells (Tregs) to exert immunosuppressive behavior. In the F99 phase, we will investigate the potential for boosting the anti-tumor immune response by targeting this specific subset of TGFBR2-induced immunosuppressive GSCs endowed with Treg-like capabilities. To do so, we will utilize inducible shRNA constructs and a clinically translatable small-molecule drug to inhibit TGFBR2 in orthotopic tumor allografts in immune-competent mice and analyze the effects on tumor growth, immune cell infiltration and function, and cooperativity with check point inhibitor therapy. In the K00 phase, we will conduct spatially resolved transcriptomics and metabolomics on patient GBM tissue specimens to identify potential metabolic vulnerabilities in immunosuppressive GBM cells. Metabolic inhibitors will be utilized to exploit candidate vulnerabilities in an attempt to attenuate the transcriptomic and functional immune-suppressive characteristics of these cells. Subsequently, validated metabolic vulnerabilities will be targeted in vivo to assess the effects on tumor growth and the anti-tumor immune response. Our proposed methods of pharmacological TGFBR2 inhibition and metabolic exploitation will inform the development of novel strategies to reprogram the GBM microenvironment and enhance anti-tumor immune responses when combined with current emerging immunotherapeutics (e.g. checkpoint inhibitors, CAR-T cell technology, vaccines).

多形性胶质母细胞瘤（GBM）是一种常见且高度侵袭性的成人脑癌， 预后和有限的治疗选择。GBM恶性肿瘤的一个关键组成部分来自于不同的 神经胶质瘤干细胞（GSC）群体，通过其能力促进和维持恶性肿瘤 自我更新、细胞适应和多能性。这些类干细胞参与了一种协同关系 与周围微环境结合促进肿瘤进展，是肿瘤内 异质性、免疫抑制和治疗抗性。以全球供应链为目标， 干细胞样表型为靶向治疗提供了有希望的途径。 这项提议的主要目标是了解细胞内在机制驱动维持一个细胞的功能。 独特的免疫抑制性GSC亚群，识别与免疫抑制性GSC相关的细胞脆弱性， GBM细胞，并开发针对这些细胞的临床前疗法。我们将通过利用我们的 经过验证的GSC细胞模型，最先进的单细胞测序技术， 适用于临床GBM标本的解析组学平台，以及先进的体外分子技术， 确定免疫抑制肿瘤细胞群并确定转录组学和代谢变化 与这些细胞群相关的基因，这些基因适合于治疗靶向。 我们的初步研究结果表明，TGF-β II型受体（TGFBR 2）信号转导诱导了一种新的免疫应答。 GSC的TGFBR 2高亚群，其选择某些免疫抑制机制，这些机制与以下相关并被以下利用： 调节性T细胞（Tcells）发挥免疫抑制行为。在F99阶段，我们将研究 通过靶向TGFBR 2诱导的这种特异性亚群来增强抗肿瘤免疫应答的潜力 具有Treg样能力的免疫抑制性GSC。为了做到这一点，我们将利用可诱导的shRNA 构建体和临床上可转化的小分子药物在原位肿瘤同种异体移植物中抑制TGFBR 2， 免疫活性小鼠，并分析对肿瘤生长、免疫细胞浸润和功能的影响， 与检查点抑制剂疗法的协同性。在K 00阶段，我们将进行空间分辨 对患者GBM组织标本进行转录组学和代谢组学研究，以确定潜在的代谢脆弱性 在免疫抑制性GBM细胞中。代谢抑制剂将被用来利用候选漏洞在一个 试图减弱这些细胞的转录组学和功能性免疫抑制特征。 随后，将在体内针对经验证的代谢脆弱性来评估对肿瘤生长的影响 和抗肿瘤免疫反应。我们提出的药理学TGFBR 2抑制和 代谢利用将为开发重新编程GBM微环境的新策略提供信息 并且当与当前出现的免疫治疗剂（例如， 检查点抑制剂、CAR-T细胞技术、疫苗）。