Multi-cellular interactions defining the human brain metastatic niche

多细胞相互作用定义人脑转移生态位

• 批准号: 10651257
• 负责人: Benjamin Izar
• 金额: $ 70.86万
• 依托单位: COLUMBIA UNIVERSITY HEALTH SCIENCES
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-05-01 至 2028-04-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10651257
• 关键词: Address; Adenosine; Advanced Malignant Neoplasm; Affect; Aneuploidy; Antibodies; Biological Assay; Biology; Blood - brain barrier anatomy; Brain; Cancer Etiology; Cancer Model; Cancer Patient; Cell Communication; Cell Compartmentation; Cell physiology; Cells; Cephalic; Cessation of life; Chromosomal Instability; Chronic; Clinical; Clinical Trials; Communities; Coupled; Cytosol; Data; Data Set; Dependence; Development; Disease; Ecosystem; Environment; Evolution; Exclusion; Exhibits; Functional Imaging; Genes; Genetic; Genomics; Genotype; Goals; Heterogeneity; Human; Hydrolysis; Immune; Immune System Diseases; Immune response; Immunocompetent; Immunocompromised Host; Immunofluorescence Immunologic; In Situ; Innate Immune Response; KRAS2 gene; Knock-out; Knowledge; Label; Ligands; Link; Lymphoid; Malignant Neoplasms; Maps; Measures; Metabolic; Metastatic malignant neoplasm to brain; Methods; Modeling; Modernization; Molecular; Mus; Mutation; Myelogenous; Nature; Neoplasm Metastasis; Non-Small-Cell Lung Carcinoma; Pathway interactions; Patients; Phenotype; Primary Neoplasm; Prognosis; RNA; Research; Role; Rupture; STK11 gene; Sampling; Shapes; Signal Transduction; Specimen; Stains; Stimulator of Interferon Genes; System; T-Cell Receptor; T-Lymphocyte; Testing; Tissues; Tumor-Derived; Validation; Work; analytical tool; blood-brain barrier permeabilization; cancer cell; cancer therapy; case control; cell type; clinically significant; cohort; disabling symptom; ds-DNA; ecto-nucleotidase; effective therapy; exome sequencing; human data; human model; immune cell infiltrate; improved; in vivo; in vivo Model; innovation; insight; lung Carcinoma; melanoma; micronucleus; mouse model; multimodality; neoplastic cell; novel; novel therapeutics; programs; response; single cell sequencing; single-cell RNA sequencing; spatiotemporal; therapeutic development; therapeutic target; therapy resistant; transcriptome sequencing; transcriptomics; tumor; tumor microenvironment; tumorigenic

Brain metastasis (BM) occurs in up to 40% of patients with advanced cancers, most frequently arising from non-small cell lung cancer (NSCLC). Patients with BM frequently suffer from debilitating symptoms, have worse response rates to modern cancer therapies and are excluded from most clinical trials, resulting in an overall poor prognosis. While the clinical significance of BM is broadly recognized, our understanding of underlying molecular, cellular and microenvironmental mechanisms remains rudimentary. Here, through several innovations, we overcome experimental, technical, and analytical barriers to gain unprecedented insight into cellular and microenvironmental features of human BM. In an integrative analysis of multi-modal single-cell RNA, T cell receptor and spatial transcriptomics of primary tumors and BM from patients with NSCLC, coupled with analyses of public data sets, we identify chromosomal instability (CIN), and CIN-induced molecular adaptations as key driver of brain-metastatic organotropism. The brain metastatic ecosystem is enriched with a pro-tumorigenic myeloid and dysfunctional T cell compartment. In this proposal, we aim to define the mechanistic underpinnings of these observations. In Aim 1, we will employ a fundamentally novel analytical tool, ContactTracing, to map at a systems level all cell-cell interactions in the BM ecosystem based on multi-modal single-cell sequencing. We assembled a large validation cohort of NSCLC specimen, including primary tumors, brain and extracranial metastases, that underwent whole-exome sequencing (WES) and RNA-seq which will be deconvolved to validate pre- dictions derived from single-cell data. Furthermore, we assembled additional NSCLC for multiplexed immunofluorescence with established antibody panels to measure the rate of CIN, CIN-adaption, and their spatial association with myeloid and lymphoid immune infiltrates. In Aim 2, through functional imaging of human and murine models we identify CIN as a defining feature of LKB1-deficient tumors. LKB1 loss (or deleterious mutation) is a common genomic subtype of NSCLC (along with KRAS co-mutation) that frequently metastasizes to the brain and is characterized by treatment resistance and poor prognosis. We find that CIN results in tonic activation of the cGAS-STING pathway to promote BM. We will dissect underlying mechanisms of CIN-induced cellular adaptations that confer brain-metastatic organotropism through modulation of the brain-metastatic niche through cGAMP hydrolysis to adenosine, which permeabilizes the blood-brain-barrier and creates an immunosuppressive environment. For this purpose, we will use in situ niche-labeling of in vivo BM models, coupled with single-cell/spatial transcriptomics to track the evolution of brain-metastatic ecosystem, and dynamics of cell interaction networks using ContactTracing. Upon completion of this work, we will provide mechanistic and clinical insights into genomic and multi-cellular features of the evolving brain-metastatic ecosystem. These insights are the first step towards development of more effective therapies of affected patients.

高达 40% 的晚期癌症患者会发生脑转移 (BM)，其中最常见的是非小细胞肺癌 (NSCLC)。 BM 患者经常出现衰弱症状，对现代癌症疗法的反应率较差，并且被排除在大多数临床试验之外，导致整体效果不佳 预后。虽然 BM 的临床意义已得到广泛认可，但我们对潜在分子、 细胞和微环境机制仍然处于初级阶段。在这里，通过多项创新，我们克服了实验、技术和分析障碍，获得了对人类骨髓细胞和微环境特征前所未有的洞察。在多模式单细胞 RNA、T 细胞受体和 NSCLC 患者原发肿瘤和 BM 的空间转录组学，以及公众的分析 数据集，我们确定染色体不稳定性（CIN）和 CIN 诱导的分子适应是关键驱动因素 脑转移性器官倾向。脑转移生态系统富含促肿瘤骨髓和 T 细胞区室功能失调。在本提案中，我们的目标是定义这些的机制基础 观察。在目标 1 中，我们将采用一种全新的分析工具 ContactTracing，基于多模式单细胞测序在系统层面绘制 BM 生态系统中所有细胞间相互作用的图谱。我们组建了一个大型 NSCLC 样本验证队列，包括原发性肿瘤、脑部和颅外转移瘤，这些样本经过全外显子组测序 (WES) 和 RNA-seq，这些样本将进行去卷积以验证预检测结果。 来自单细胞数据的词典。此外，我们用已建立的抗体组组装了额外的 NSCLC 进行多重免疫荧光，以测量 CIN、CIN 适应率及其与骨髓和淋巴免疫浸润的空间关联。在目标 2 中，通过人类和小鼠的功能成像 在模型中，我们将 CIN 确定为 LKB1 缺陷肿瘤的一个定义特征。 LKB1 丢失（或有害突变）是 经常转移至大脑的 NSCLC 常见基因组亚型（以及 KRAS 共突变） 其特点是治疗耐药、预后差。我们发现 CIN 会导致强直激活 cGAS-STING 途径促进 BM。我们将剖析 CIN 诱导的细胞适应的潜在机制，这些适应通过调节脑转移生态位来赋予脑转移器官向性 cGAMP 水解为腺苷，使血脑屏障通透并创造免疫抑制环境。为此，我们将使用体内 BM 模型的原位生态位标记，结合单细胞/空间转录组学来追踪脑转移生态系统的演变以及细胞相互作用的动态 使用 ContactTracing 的网络。完成这项工作后，我们将提供机制和临床见解 研究不断发展的脑转移生态系统的基因组和多细胞特征。这些见解是第一 为受影响的患者开发更有效的疗法迈出了一步。