Mechanisms of KSHV-induced endothelial cell loss of contact inhibition of proliferation

KSHV诱导内皮细胞失去接触抑制增殖的机制

基本信息

批准号：
10762813
负责人：
Eva Henriette Gottwein
金额：
$ 49.45万
依托单位：
NORTHWESTERN UNIVERSITY AT CHICAGO
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：
2023-07-01 至 2028-06-30
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10762813
关键词：
AIDS related cancer Acquired Immunodeficiency Syndrome Actins Address Adherens Junction Adhesions Automobile Driving Binding Blood Vessels Cell Culture Techniques Cell Cycle Cell Cycle Arrest Cell Proliferation Cell Shape Cells Complex Contact Inhibition Cyclin-Dependent Kinase Inhibitor Cytoskeleton Data Endothelial Cells Genome Gentian Violet Herpesviridae Infections Human Human Herpesvirus 8 Individual Infection Kaposi Sarcoma Knowledge Left Lymphatic Endothelial Cells Lytic Malignant Neoplasms Measures Mediating Mentored Clinical Scientist Development Program Messenger RNA MicroRNAs Modeling Normal Cell Oncogenic Pathway interactions Phenotype Process Proliferating Proliferation Marker Proteins Protocols documentation Public Health Repression Role Solid Neoplasm Stains Study models Supporting Cell Testing Therapeutic Therapeutic Intervention Transcript Viral Viral Genes Work cadherin 5 cell type gene product improved inhibitor innovation monolayer mutant neoplastic cell trafficking tumor tumorigenesis

项目摘要

SUMMARY Kaposi’s Sarcoma-associated herpesvirus (KSHV) causes the AIDS-defining cancer Kaposi’s Sarcoma (KS). The KSHV-infected KS tumor cells (KSCs) express proliferation markers, indicating a loss of contact inhibition of proliferation (CIP). CIP is considered a tumor suppressive pathway, and loss of CIP is a crucial feature of oncogenic transformation in solid tumors. How KSHV antagonizes CIP is not known. The KS tumor cells most likely originate from microvascular lymphatic endothelial cells (LECs). While LECs therefore represent a relevant model for studies of KS, KSHV-induced proliferation after de novo infection of primary human LECs has not been demonstrated. We have developed a protocol for KSHV infection of primary human LECs that allows us to measure KSHV-induced loss of CIP. The central hypothesis underlying this application is that KSHV-induced loss of CIP is a critical driving feature of oncogenesis in KS. Our preliminary work shows that the KSHV miR-K10 miRNAs contribute substantially to the KSHV-induced loss of CIP in LECs but are not the only viral determinants of this phenotype. Our results furthermore implicate viral repression of p27, disruption of adherens junctions (AJs), and deregulation of the cytoskeleton and vesicular trafficking in KSHV-mediated loss of CIP. To test our hypothesis and elucidate the mechanisms underlying KSHV-induced loss of CIP, we propose three Specific Aims. In Specific Aim 1, we will determine the expression of the four miR-K10 miRNAs in KSHV-infected LECs and KS. We will also define their individual contributions to the KSHV-induced loss of CIP. In Specific Aim 2, we will identify the mechanisms underlying the miR-K10-induced loss of CIP in KSHV- infected LECs. In Specific Aim 3, we will identify other viral genes that promote the KSHV-induced loss of CIP. The proposed study is innovative because our model provides rigorously defined experimental settings that enable the analysis of KSHV-induced loss of CIP after infection of a primary human cell type with relevance to KS. This work is significant because it will establish the viral determinants of KSHV-induced LEC proliferation, thereby explaining oncogenic mechanisms in KS. Results will be impactful since CIP is a tumor-suppressive mechanism. Understanding how KSHV overcomes CIP will help us to explain how KSHV causes KS and could potentially be exploited for therapeutic intervention in KS.

概括卡波西肉瘤相关疱疹病毒 (KSHV) 会导致艾滋病定义的癌症卡波西肉瘤 (KS)。感染 KSHV 的 KS 肿瘤细胞 (KSC) 表达增殖标记物，表明接触抑制丧失增殖（CIP）被认为是一种肿瘤抑制途径，CIP 的丧失是其一个重要特征。 KSHV 如何拮抗 KS 肿瘤细胞尚不清楚。可能源自微血管淋巴内皮细胞（LEC），因此 LEC 代表了一种细胞。用于研究 KS、KSHV 诱导原代人 LEC 从头感染后增殖的相关模型我们已经开发出一种用于原代人类 LEC 感染的方案。使我们能够测量 KSHV 引起的 CIP 损失。该应用的核心假设是： KSHV 诱导的 CIP 丧失是 KS 肿瘤发生的关键驱动特征。 KSHV miR-K10 miRNA 对 KSHV 诱导的 LEC 中 CIP 损失有很大贡献，但并不是我们的结果还表明，这种表型的唯一病毒决定因素是 p27 的病毒抑制、破坏。 KSHV 介导的粘附连接 (AJ) 以及细胞骨架和囊泡运输的失调为了检验我们的假设并阐明 KSHV 引起的 CIP 丧失的机制，我们提出三个具体目标在具体目标 1 中，我们将确定四种 miR-K10 miRNA 的表达。在 KSHV 感染的 LEC 和 KS 中，我们还将定义它们对 KSHV 引起的损失的个体贡献。 CIP。在具体目标 2 中，我们将确定 KSHV 中 miR-K10 诱导的 CIP 丧失的机制。在特定目标 3 中，我们将鉴定其他促进 KSHV 诱导的 CIP 丧失的病毒基因。拟议的研究具有创新性，因为我们的模型提供了严格定义的实验设置能够分析感染原代人类细胞类型后 KSHV 诱导的 CIP 丢失，并与以下相关 KS。这项工作意义重大，因为它将确定 KSHV 诱导的 LEC 增殖的病毒决定因素，解释 KS 的致癌机制结果将具有影响力，因为 CIP 具有肿瘤抑制作用。了解 KSHV 如何克服 CIP 将有助于我们解释 KSHV 如何导致 KS 以及可能发生的情况。可能被用于 KS 的治疗干预。