Regulation of metastatic progression by an endothelial-derived factor

内皮衍生因子对转移进展的调节

基本信息

批准号：
10155448
负责人：
Sohail F. Tavazoie
金额：
$ 47.67万
依托单位：
ROCKEFELLER UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2019
资助国家：
美国
起止时间：
2019-05-07 至 2024-04-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10155448
关键词：
Affinity Animals Automobile Driving Biology Blocking Antibodies Blood Blood Vessels Blood flow Breast Melanoma Cancer Model Cause of Death Cell Line Cells Clinical Complement Disseminated Malignant Neoplasm Distant Endothelial Cells Endothelium Exhibits Genes Genetic Genetic Models Goals Health Human Image Impairment Investigational Therapies Malignant Neoplasms Mediating Medical Oncology Memorial Sloan-Kettering Cancer Center Methods MicroRNAs Microscopy Modeling Molecular Molecular Genetics Nature Neoplasm Metastasis Nervous system structure Organ Pathogenicity Pathway interactions Patients Pharmacology Positioning Attribute Primary Neoplasm Proteins RNA Regulation Relapse RiboTag Ribosomes Role Series Signal Transduction Signaling Protein Site Skin Stream Testing Therapeutic Therapeutic Uses Tissues Transcript Translations Tumor stage Tumor-Derived Work angiogenesis animal imaging axon guidance cancer cell cancer imaging cancer type cell motility cell type experience human disease in vivo innovation insight malignant breast neoplasm melanoma migration multi-photon multiphoton imaging neoplastic cell next generation sequencing receptor recruit ribosome profiling transcriptomics tumor tumor microenvironment tumor progression

项目摘要

We have previously identified sets of tissue-specific microRNAs that regulate metastatic progression by distinct cancer types. In both breast cancer and melanoma, such metastasis regulating miRNA pathways enable cancer cells to avidly recruit endothelial cells into the primary tumor site (Png et al., Nature, 2011; Pencheva et al., Cell, 2012). A key unknown in the field has been the signal(s) that are provided by such recruited endothelial cells that impact metastatic progression. We have used an innovative unbiased approach wherein ribosomes of tumor endothelial cells are genetically marked with an affinity tag. This enables purification of tumor endothelial ribosomes along with their associated transcripts, which then undergo next-generation sequencing. This allowed us to identify Slit2 as a gene significantly induced in endothelial cells by highly metastatic cells. Slit2 is an axon guidance molecule required for the proper establishment of nervous system connectivity. Our preliminary evidence in syngeneic models reveals that genetic inactivation of Slit2 in the endothelial compartment significantly impairs cancer metastasis from the primary tumor site. We propose a model whereby metastatic cells induce Slit2 in endothelial cells, which serves as a signal that promotes migration of cancer cells within the tumor (low Slit2) towards the vasculature (high Slit2), enabling intravasation and metastasis. This model is supported by preliminary clinical association evidence that reveals that increased Slit2 in endothelial compartment relative to the tumoral compartment associates with higher stage tumors that exhibit higher rates of metastatic relapse. In this application, we propose a series of complementary approaches for rigorously confirming this surprising model and further mechanistically dissecting it. We will modulate Slit2 signal sensing by cancer cells through genetic inactivation of endothelial or tumoral Slit2 using cell-type specific genetic inactivation in a genetically initiated model of cancer progression. We will employ live animal multi-photon microscopy to visualize Slit2-driven tumoral trans-endothelial migration and intravasation. We aim to identify the tumoral receptor that senses Slit2, to use immunohistochemical methods to investigate an association between endothelial Slit2 and human cancer progression and metastatic relapse, and to discover the tumor-derived signal that induces endothelial Slit2. Finally, we will apply these insights by determining if a clinically used therapeutic, which we find induces Slit2 promotes cancer metastasis. This work has the potential for major impact on our understanding of mechanisms of cancer progression by establishing endothelial cells as major orchestrators of metastasis. It could have important impact on human disease given that this pathway governs progression of highly prevalent cancer types and associates with human relapse. Moreover, the cell-type specific ribosomal profiling method we have employed could be applied more broadly to study endless cell-types within the tumor microenvironment.

我们之前已经鉴定出一组组织特异性 microRNA，它们通过不同的方式调节转移进展。癌症类型。在乳腺癌和黑色素瘤中，这种转移调节 miRNA 通路使得癌细胞会贪婪地将内皮细胞募集到原发性肿瘤部位（Png 等人，Nature，2011；Pencheva 等人）等人，细胞，2012）。该领域的一个关键未知数是由此类招募的人员提供的信号影响转移进展的内皮细胞。我们采用了一种创新的公正方法，其中肿瘤内皮细胞的核糖体带有亲和标签进行基因标记。这使得能够纯化肿瘤内皮核糖体及其相关转录本，然后进行下一代测序。这使我们能够将 Slit2 鉴定为通过高度诱导在内皮细胞中显着诱导的基因。转移细胞。 Slit2 是神经系统正常建立所需的轴突引导分子连接性。我们在同基因模型中的初步证据表明，Slit2 的基因失活在内皮区室显着损害原发肿瘤部位的癌症转移。我们提出一个转移细胞在内皮细胞中诱导 Slit2 的模型，该信号作为促进肿瘤内的癌细胞（低 Slit2）向脉管系统（高 Slit2）迁移，从而实现内渗和转移。该模型得到了初步临床关联证据的支持，该证据表明相对于肿瘤区室而言，内皮区室中 Slit2 的增加与更高的分期相关表现出较高转移复发率的肿瘤。在此应用中，我们提出了一系列严格证实这一令人惊讶的模型并进一步机械地补充的方法解剖它。我们将通过内皮细胞或细胞的基因失活来调节癌细胞对 Slit2 信号的感知。肿瘤 Slit2 在基因启动的癌症进展模型中使用细胞类型特异性基因失活。我们将采用活体动物多光子显微镜来观察 Slit2 驱动的肿瘤跨内皮迁移和内渗。我们的目标是识别感知 Slit2 的肿瘤受体，以使用免疫组织化学研究内皮 Slit2 与人类癌症进展和转移之间关联的方法复发，并发现诱导内皮细胞 Slit2 的肿瘤衍生信号。最后，我们将应用这些通过确定临床使用的治疗方法是否会诱导 Slit2 促进癌症转移，我们获得了见解。这项工作有可能对我们对癌症进展机制的理解产生重大影响建立内皮细胞作为转移的主要协调者。它可能对人类产生重要影响鉴于该途径控制高度流行的癌症类型的进展并与人类的复发。此外，我们采用的细胞类型特异性核糖体分析方法可以是更广泛地应用于研究肿瘤微环境中的无限细胞类型。