Neural organoid models of the immunological microenvironment of glioblastoma for drug discovery applications

用于药物发现应用的胶质母细胞瘤免疫微环境的神经类器官模型

• 批准号: 10761235
• 负责人: Connie S Lebakken
• 金额: $ 40.65万
• 依托单位: STEM PHARM, INC.
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-07-11 至 2024-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10761235
• 关键词: 3-Dimensional; Address; Adoptive Cell Transfers; Adult; Anti-Inflammatory Agents; Area; Biological Assay; Blood Vessels; Brain; Cell Line; Cell Proliferation; Cells; Coculture Techniques; Combined Modality Therapy; Complex; Data Set; Development; Drug Screening; Effectiveness; Environment; Excision; Extracellular Matrix; Flow Cytometry; Funding; Gene set enrichment analysis; Genes; Genetic Transcription; Glioblastoma; Glioma; Goals; Heterogeneity; Human; Human Biology; Hydrogels; Image Cytometry; Immune; Immune Evasion; Immune response; Immunofluorescence Immunologic; Immunological Models; Immunologics; Immunosuppression; Immunotherapy; In Vitro; Infiltration; Invaded; Life Expectancy; Link; Macrophage; Macrophage Activation; Malignant Neoplasms; Microglia; Modeling; Myelogenous; National Institute of Environmental Health Sciences; Nature; Operative Surgical Procedures; Organoids; Parents; Pathway interactions; Patients; Peripheral; Peripheral Blood Mononuclear Cell; Pharmaceutical Preparations; Phenotype; Population; Pre-Clinical Model; Primary Brain Neoplasms; Proliferating; Radiation; Reproducibility; Research Personnel; Risk; Role; Sampling; Scientist; Shapes; Small Business Innovation Research Grant; Survival Rate; T-Cell Activation; T-Lymphocyte; Technology; Testing; Therapeutic; Tissues; Universities; Validation; Wisconsin; Work; Xenograft Model; cancer cell; cell motility; cell type; chemotherapy; chimeric antigen receptor T cells; cytokine; drug discovery; effectiveness evaluation; experience; human model; human tissue; immune checkpoint blockade; in vitro Model; individual patient; innovation; interest; migration; neural; novel; novel therapeutic intervention; novel therapeutics; personalized medicine; phase 2 study; response; screening; sex; single-cell RNA sequencing; small molecule; standard of care; stem; therapeutic target; tumor; tumor microenvironment; tumor-immune system interactions

Project Summary/Abstract Glioblastoma (GBM) is the most prevalent primary brain tumor in adults with extremely poor survival rates and largely unchanged standard of care. While there are many challenges to developing better GBM treatments, one of the major challenges is the immune-suppressive environment commonly found within GBM tumors. This immune-suppressive nature results in a tumor that is not suitable for mounting an immune response to GBM cells, rendering emerging immunotherapies ineffective. To address this issue, suitable models that can interrogate the complex interactions between GBM cancer cells and microglia and peripherally derived macrophages would be invaluable for target identification, screening of novel therapeutics and for mode of action studies. Tumor-associated microglia and macrophages are of particular interest due to their primary role in shaping the immunological environment of GBM tumors. Human organoid technology is well-suited for modeling complex, multicellular interactions in a human tissue-like environment. Stem Pharm’s hydrogel-enabled neural organoids allow for incorporation of non-neural populations such as microglia and macrophages in a reproducible, 96-well plate format amenable to screening applications. Therefore, work in this proposal will develop and validate a human in vitro glioblastoma organoid model through incorporation of microglia, macrophages, and patient derived GBM cells in our neural organoids. Specific aims will 1) characterize organoids incorporating GBM, evaluate GBM survival, invasion, and proliferation; and characterize cell-type specific transcriptional responses to GBM and compare them to parent tumors and publicly available data sets. 2) demonstrate immunosuppressive activation of microglia and macrophages within the neural organoid in response to infiltrating GBM cells. Multiplex cytokine panels, co-stimulatory and checkpoint molecule expression, and a direct immunosuppression assay with peripheral blood mononuclear cell-derived T-cells will be used to evaluate microglia and macrophage immunosuppression. Finally, treatment with three small molecules known to modulate macrophage activation will be assessed within the organoids to demonstrate the ability to regulate the microglial and macrophage response to GBM cells. Successful completion of these specific aims will result in a robust in vitro organoid model with novel capabilities to interrogate GBM invasion and subsequent microglia and macrophage immunosuppression. This will provide pharma partners with the ability to study therapies that previously failed due to this immunosuppressive environment, and test new therapeutic approaches. Phase II studies will expand the number of available patient-derived samples to better capture the diversity and heterogeneity of GBM tumors, explore sex-linked differences, and evaluate the effectiveness of CAR-Ts and combination therapies within the GBM model with the goal of bringing better treatment options to patients for this devastating condition.

项目概要/摘要 胶质母细胞瘤 (GBM) 是成人中最常见的原发性脑肿瘤，病情极差 尽管存在许多挑战，但生存率和护理标准基本保持不变。 开发更好的 GBM 治疗方法，主要挑战之一是免疫抑制 GBM 肿瘤中常见的环境会导致免疫抑制。 肿瘤不适合对 GBM 细胞产生免疫反应，从而导致新兴肿瘤 为了解决这个问题，可以使用合适的模型来探究免疫疗法的效果。 GBM 癌细胞与小胶质细胞和外周衍生细胞之间复杂的相互作用 巨噬细胞对于靶标识别、新疗法筛选和 肿瘤相关的小胶质细胞和巨噬细胞的作用模式研究特别令人感兴趣。 由于它们在塑造人类 GBM 肿瘤的免疫环境中发挥着主要作用。 类器官技术非常适合模拟人类复杂的多细胞相互作用 Stem Pharm 的水凝胶神经类器官允许合并。 可重复的 96 孔板中的非神经细胞群，例如小胶质细胞和巨噬细胞 因此，本提案中的工作将得到发展和落实。 通过掺入小胶质细胞验证人类体外胶质母细胞瘤类器官模型， 我们的神经类器官中的巨噬细胞和患者来源的 GBM 细胞的具体目标是 1)。 表征包含 GBM 的类器官，评估 GBM 存活、侵袭和增殖； 并表征细胞类型对 GBM 的特异性转录反应，并将其与亲本进行比较 肿瘤和公开数据集2）证明了免疫抑制激活。 神经类器官内的小胶质细胞和巨噬细胞对浸润的 GBM 细胞做出反应。 多重细胞因子组合、共刺激和检查点分子表达以及直接 使用外周血单核细胞衍生的 T 细胞进行免疫抑制测定 最后评价小胶质细胞和巨噬细胞的免疫抑制作用。 将在类器官内评估已知调节巨噬细胞活化的分子，以 证明了调节小胶质细胞和巨噬细胞对 GBM 细胞反应的能力。 成功完成这些具体目标将产生一个强大的体外类器官模型 研究 GBM 侵袭以及随后的小胶质细胞和巨噬细胞的新能力 这将为制药合作伙伴提供研究治疗方法的能力。 以前由于这种免疫抑制环境而失败，并测试新的治疗方法 II 期研究将扩大可用的患者来源样本的数量。 更好地捕捉 GBM 肿瘤的多样性和异质性，探索性别相关差异，以及 评估 GBM 模型中 CAR-T 和联合疗法的有效性 目标是为这种毁灭性疾病的患者提供更好的治疗选择。