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).

胶质母细胞瘤多形（GBM）是成年人的常见且高度侵略性的脑癌形式 预后和有限的治疗选择。 GBM恶性肿瘤的关键组成部分来自不同的 胶质瘤干细胞的种群（GSC），可通过其能力来促进和维持恶性肿瘤 为了自我更新，细胞适应和多能力。这些样干细胞具有协同关系 周围的微环境以促进肿瘤进展，是肿瘤内的关键驱动因素 异质性，免疫抑制和耐药性。针对驱动GSC的GSC和机制 类似STEM的表型为靶向治疗剂提供了有希望的途径。 该提议的广泛目标是了解驱动维持的细胞内部机制 独特的免疫抑制GSC子集，识别与免疫抑制相关的细胞漏洞 GBM细胞，并开​​发临床前疗法以靶向这些细胞。我们将利用我们的目标来实现这一目标 经过验证的GSC细胞模型，最先进的单细胞测序技术和尖端的空间 已解决的OMICS平台应用于临床GBM标本，并进行晚期分子体外技术 定义免疫抑制性肿瘤细胞群体并确定转录组和代谢变化 与这些细胞群有关，这些细胞群体与治疗靶向相关。 我们的初步发现表明，TGF-βII型受体（TGFBR2）信号传导诱导A GSC的TGFBR2High子集，该子集选择了某些与与之相关的免疫抑制机制 调节性T细胞（Tregs）发挥免疫抑制行为。在F99阶段，我们将研究 通过靶向TGFBR2诱导的该特定子集来增强抗肿瘤免疫反应的潜力 免疫抑制GSC具有类似Treg的能力。为此，我们将利用诱导shrna 构建体和临床上可翻译的小分子药物可抑制TGFBR2在异位肿瘤同种异体移植物中的TGFBR2 免疫能力小鼠，分析对肿瘤生长，免疫细胞浸润和功能的影响，以及 与检查点抑制剂治疗的合作性。在K00阶段，我们将进行空间解决 患者GBM组织标本上的转录组学和代谢组学，以识别潜在的代谢脆弱性 在免疫抑制GBM细胞中。代谢抑制剂将用于利用 尝试减弱这些细胞的转录组和功能性免疫抑制特征。 随后，经过验证的代谢脆弱性将在体内靶向以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TGFBR2抑制方法和 代谢剥削将告知开发新型策略以重新编程GBM微环境 并在与当前出现的免疫治疗剂结合使用时增强抗肿瘤免疫反应（例如 检查点抑制剂，CAR-T细胞技术，疫苗）。