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

基本信息

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
项目类别：
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

项目摘要

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）发生在多达40％的晚期癌症患者中，最常见于非小细胞肺癌（NSCLC）。 BM患者经常患有衰弱的症状，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，导致总体差预后。尽管BM的临床意义得到广泛认可，但我们对潜在分子的理解，但细胞和微环境机制仍然是基本的。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BM的细胞和微环境特征的前所未有的见解。在多模式单细胞RNA，T细胞受体和 NSCLC患者的原发性肿瘤和BM的空间转录组学以及公共分析数据集，我们识别染色体不稳定性（CIN），以及CIN诱导的分子适应为关键驱动力脑部转移性器乐。脑转移生态系统富含蛋白酶髓样和功能失调的T细胞室。在此提案中，我们旨在定义这些的机械基础观察。在AIM 1中，我们将采用一种根本新颖的分析工具，即接触跟踪，以在系统级别绘制基于多模式单细胞测序的BM生态系统中所有细胞细胞相互作用。我们组装了NSCLC标本的大量验证队列，包括原发性肿瘤，脑和颅外转移，该样本进行了全异位测序（WES）和RNA-Seq，将被验证以验证前验证前的前体转移。从单细胞数据得出的命令。此外，我们将额外的NSCLC与已建立的抗体面板组装了多重免疫荧光，以测量CIN，CIN-适应速率及其与髓样和淋巴样免疫浸润的速率。在AIM 2中，通过人类和鼠的功能成像模型我们将CIN视为LKB1缺陷型肿瘤的定义特征。 LKB1损失（或有害突变）是 NSCLC的常见基因组亚型（以及KRAS共同享受），经常向大脑转移并以治疗耐药性和预后不良为特征。我们发现CIN导致滋补促进BM的CGAS刺道途径。我们将剖析CIN诱导的细胞适应的潜在机制，这些机制通过调节脑部移动细胞的调节来赋予脑部转移性器官 CGAMP水解为腺苷，该腺苷渗透到血脑屏障并产生免疫抑制环境。为此，我们将使用体内BM模型的原位生态位标记，并与单细胞/空间转录组学结合来跟踪脑部转移生态系统的演变和细胞相互作用的动力学使用联系跟踪的网络。完成这项工作后，我们将提供机械和临床见解发展为不断发展的脑部转移生态系统的基因组和多细胞特征。这些见解是第一个迈向开发受影响患者的更有效疗法的一步。