Investigate and inhibit microglia support of brain metastases

研究并抑制小胶质细胞对脑转移的支持

基本信息

批准号：
10272360
负责人：
Melanie Hayden Gephart
金额：
$ 35.17万
依托单位：
STANFORD UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-09-21 至 2026-08-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10272360
关键词：
3-Dimensional Affect Air Animal Model Anti-CD47 Antigen Presentation Architecture Biological Assay Blood Circulation Bone Marrow Transplantation Brain Brain Neoplasms Busulfan CD47 gene CRISPR library CSF1 gene Cell Communication Cell Death Cells Cerebrospinal Fluid Clustered Regularly Interspaced Short Palindromic Repeats Coculture Techniques Complex Custom Data Disease Progression Disseminated Malignant Neoplasm Distant Epithelial Evaluation Exhibits Extracellular Matrix Generations Genes Growth Human Immune Immunologic Surveillance Immunological Models Immunotherapy Inflammatory Inflammatory Response Interleukin-6 Interruption Intrinsic factor Knock-out Knowledge Ligands Liquid substance Macrophage Activation Malignant Neoplasms Measures Mediating Messenger RNA Metalloproteases Metastatic malignant neoplasm to brain Methods Microglia Modeling Monitor Mus Myeloid Cells Neoplasm Metastasis Neuroimmune Neurologic Organoids PTPNS1 gene Phagocytosis Phenotype Population Positioning Attribute Primary Neoplasm Production Prognosis Property Protocols documentation Publishing RNA analysis Recurrence Resistance Role Sampling Signal Transduction T-Cell Activation T-Lymphocyte Technology Testing Therapeutic Tumor Cell Invasion Tumor Immunity Tumor-associated macrophages Tumor-infiltrating immune cells aggressive therapy base cancer cell cytokine human disease implantation in vitro Model innovation macrophage migration mortality multidisciplinary neoplastic cell neuropathology novel novel strategies receptor single-cell RNA sequencing small molecule transcriptome tumor tumor microenvironment tumor progression tumor-immune system interactions weapons

项目摘要

1 ABSTRACT – PROJECT 2 2 Brain metastases are highly aggressive, treatment resistant malignancies with debilitating neurological sequelae 3 and a grave prognosis. Efforts to understand and modulate the metastatic brain tumor microenvironment (TME) 4 have great therapeutic potential. Studies have suggested that polarization of macrophages (M1 or M2) may be 5 helpful or harmful in cancer progression (1, 2). Microglia, the brain’s myeloid cells, function in immune 6 surveillance and mediate the tumor-related inflammatory response, including within the context of the metastatic 7 brain tumor microniche (tumor-associated macrophages and tumor-associated microglia). We hypothesize 8 brain metastases are supported in part by native microglia (3), and that these interactions can be 9 manipulated for therapeutic benefit. The role of microglia in brain metastases has been difficult to investigate 10 given the relative inaccessibility of brain metastases samples, inability to isolate human tumor-associated 11 microglia or monitor their interactions within the TME, and lack of models that include or allow manipulation of 12 microglia while still reliably recapitulating the human disease. To overcome these critical hurdles, we employ the 13 following innovative methods: (i) single cell RNA-sequencing (scRNA-seq) on fresh, human brain metastases 14 and normal surrounding brain (Neuropathology Core)(4, 5); (ii) isolation and dynamic array sequencing of cell 15 free messenger RNA (cfRNA) from human cerebrospinal fluid (6, 7); (iii) generation of human and murine-derived 16 brain metastasis 3D air-liquid interface (ALI) organoids that include tumor-associated microglia/macrophages 17 (Toolkit Core); and (iv) repopulation of microglia with circulation derived microglia-like cells (CDMCs) with 18 greater than 90% efficiency throughout the entire brain. These four innovative methods allow us to investigate 19 the role of tumor-associated microglia in human brain metastases, model immune cell interactions, and 20 manipulate their function in an animal model of metastatic malignancy. Our multidisciplinary team is uniquely 21 positioned to test the following three aims: (1) Test the extent to which tumor-associated microglia create an 22 immunosuppressive TME supporting brain metastasis. In addition to relating microglia function to brain 23 metastasis local recurrence and distant progression, we will cross-reference our findings with cancer cell intrinsic 24 factors (Project 1) and systemic macrophage reactivity (Project 3); (2) Model tumor-associated microglia within 25 the brain metastatic TME using ALI organoids. We will model tumor and tumor-associated microglia/macrophage 26 interactions and determine the impact of immunotherapy on microglia/macrophage activation and phagocytosis, 27 T-cell activation states, and cell death. Our findings will be integrated with expanding knowledge of cell-intrinsic 28 factors (Project 1) and brain-periphery immune interactions (Project 3); (3): Determine how high efficiency 29 microglia replacement with CDMCs inhibits brain metastasis. We will replace microglia with “weaponized” 30 CDMCs to inhibit cancer cell implantation and disease progression. 31

1 摘要 – 项目 2 2 脑转移瘤是一种高度侵袭性、难治性的恶性肿瘤，具有使人衰弱的神经系统后遗症 3、努力了解和调节转移性脑肿瘤微环境（TME）。 4 具有巨大的治疗潜力研究表明巨噬细胞（M1 或 M2）的极化可能是。 5 对癌症进展有帮助或有害 (1, 2) 小胶质细胞（大脑的髓样细胞）。 6 监测和介导肿瘤相关的炎症反应，包括在转移性肿瘤的背景下 7.脑肿瘤微生态位（肿瘤相关巨噬细胞和肿瘤相关小胶质细胞）。 8 脑转移瘤部分由天然小胶质细胞支持 (3)，并且这些相互作用可以通过 9 为了有益治疗而操纵小胶质细胞在脑转移中的作用一直难以研究。 10 鉴于脑转移样本相对难以获取，无法分离与人类肿瘤相关的 11 小胶质细胞或监测它们在 TME 内的相互作用，并且缺乏包含或允许操纵的模型 12 小胶质细胞同时仍然可靠地重现人类疾病为了克服这些关键障碍，我们采用了以下 13 种创新方法：(i) 对新鲜人脑转移瘤进行单细胞 RNA 测序 (scRNA-seq) 14 和正常周围大脑（神经病理学核心）(4, 5)；(ii) 细胞分离和动态阵列测序 15 来自人类脑脊液的游离信使 RNA (cfRNA) (6, 7)；(iii) 人类和小鼠来源的产生； 16 个脑转移 3D 气液界面 (ALI) 类器官，包括肿瘤相关小胶质细胞/巨噬细胞 17（工具包核心）；和（iv）用循环衍生的小胶质细胞样细胞（CDMC）重新填充小胶质细胞 18 整个大脑的效率超过 90% 这四种创新方法使我们能够进行研究。 19 肿瘤相关小胶质细胞在人脑转移中的作用、免疫细胞相互作用模型以及 20 在转移性恶性肿瘤动物模型中操纵它们的功能我们的多学科团队是独一无二的。 21 旨在测试以下三个目标：(1) 测试肿瘤相关小胶质细胞产生的程度 22 免疫抑制 TME 除了将小胶质细胞功能与大脑联系起来之外，还支持脑转移。 23 转移局部复发和远处进展，我们将交叉参考我们的发现与癌细胞内在的 24 个因素（项目 1）和全身巨噬细胞反应性（项目 3）；（2）模型内肿瘤相关小胶质细胞 25 使用 ALI 类器官的脑转移性 TME 我们将模拟肿瘤和肿瘤相关的小胶质细胞/巨噬细胞。 26 相互作用并确定免疫疗法对小胶质细胞/巨噬细胞激活和吞噬作用的影响， 27 T 细胞激活状态和细胞死亡将与细胞内在知识的扩展相结合。 28个因素（项目1）和大脑-外周免疫相互作用（项目3）：确定效率有多高； 29 用 CDMC 替代小胶质细胞可抑制脑转移我们将用“武器化”替代小胶质细胞。 30 CDMC 可抑制癌细胞植入和疾病进展。 31