Regulation of metastatic progression by an endothelial-derived factor

内皮衍生因子对转移进展的调节

• 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
• 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

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.

我们先前已经鉴定出一组组织特异性的microRNA，它们通过不同的方式调节转移性进展 癌症类型。在乳腺癌和黑色素瘤中，这种调节miRNA途径的转移启用 癌细胞狂热地募集内皮细胞进入原发性肿瘤部位（Png等，Nature，2011; Pencheva et Al。，Cell，2012）。该领域未知的关键是该招募的信号 影响转移性进展的内皮细胞。我们使用了一种创新的公正方法 肿瘤内皮细胞的核糖体在遗传上用亲和标签标记。这可以净化 肿瘤内皮核糖体及其相关的成绩单，然后进行下一代 测序。这使我们能够将Slit2识别为高度高度诱导内皮细胞中显着诱导的基因 转移细胞。 slit2是正确建立神经系统所需的轴突引导分子 连接性。我们在合元模型中的初步证据表明，遗传在 内皮室极大地损害了原发性肿瘤部位的癌症转移。我们提出了一个 模型，转移细胞在内皮细胞中诱导裂口2，该模型可作为促进的信号 肿瘤内癌细胞（Low Slit2）迁移到脉管系统（高裂隙2），从而可以侵入 和转移。该模型得到了初步临床协会证据的支持，该证据表明 相对于较高阶段的肿瘤区室相对于内皮室中的SLIT2增加 肿瘤表现出更高的转移性复发率。在此应用中，我们提出了一系列 严格确认这一令人惊讶的模型并进一步机械机制的互补方法 解剖它。我们将通过内皮或内皮遗传失活或 在癌症进展模型中，使用细胞类型特异性遗传失活的肿瘤裂隙。 我们将采用活动物多光子显微镜来可视化狭缝2驱动的肿瘤跨内皮迁移 和侵入。我们旨在鉴定使用免疫组织化学的肿瘤受体 研究内皮裂缝与人类癌进展与转移性之间关联的方法 复发，并发现诱导内皮裂缝的肿瘤衍生信号。最后，我们将应用这些 通过确定临床使用的治疗方法是否诱导SLIT2促进癌症转移来进行见解。 这项工作有可能对我们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编排。它可能会对人类产生重要影响 鉴于该途径控制高度普遍的癌症类型的发展，并与 人类复发。此外，我们使用的细胞类型特异性核糖体分析方法可能是 更广泛地用于研究肿瘤微环境中无尽的细胞类型。