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.

脑转移(BM)发生在高达40%的晚期癌症患者，最常见的原因是非小细胞肺癌(NSCLC)。BM患者经常出现虚弱的症状，对现代癌症治疗的反应率较低，并被排除在大多数临床试验之外，导致总体表现不佳。 预后。虽然骨髓的临床意义被广泛认识，但我们对潜在分子的理解， 细胞和微环境机制仍处于初级阶段。在这里，通过几项创新，我们克服了实验、技术和分析障碍，以获得对人类骨髓的细胞和微环境特征的前所未有的洞察。在多模式单细胞RNA综合分析中，T细胞受体和 非小细胞肺癌患者原发肿瘤和骨髓的空间转录分析 数据集，我们发现染色体不稳定(CIN)和CIN诱导的分子适应是 脑转移性器官嗜性。脑转移生态系统富含促肿瘤的髓样细胞和 功能失调的T细胞隔间。在这项提案中，我们的目标是定义这些机制的基础 观察。在目标1中，我们将使用一种全新的分析工具ContactTracing，基于多模式单细胞测序，在系统级别绘制BM生态系统中的所有细胞-细胞相互作用图。我们收集了一大批非小细胞肺癌样本，包括原发肿瘤、脑和颅外转移瘤，进行了完整外显子组测序(WES)和RNA-SEQ，这些样本将被去卷积以验证Pre-Seq 派生自单个单元格数据的词条。此外，我们还组装了额外的NSCLC，用于与已建立的抗体板进行多重免疫荧光，以测量CIN、CIN适应性的比率，以及它们与髓系和淋巴系免疫渗透的空间相关性。在目标2中，通过人和小鼠的功能成像 我们认为CIN是LKB1缺陷肿瘤的一个重要特征。LKB1缺失(或有害突变)是一种 常见的非小细胞肺癌基因组亚型(伴随KRAS共突变)，常转移至脑 且具有耐药、预后差的特点。我们发现，CIN导致紧张性激活 促进骨髓形成的cGAS-STING途径。我们将剖析CIN诱导的细胞适应的潜在机制，通过调节脑转移的生态位来赋予脑转移器质性。 CGAMP水解为腺苷，使血脑屏障通透，创造免疫抑制环境。为此，我们将使用活体骨髓模型的原位生态位标记，结合单细胞/空间转录来跟踪脑转移生态系统的进化，以及细胞相互作用的动力学 使用联系人跟踪的网络。在这项工作完成后，我们将提供机械和临床见解 进化中的脑转移生态系统的基因组和多细胞特征。这些见解是第一次 朝着开发更有效的治疗受影响患者的方法迈进。