Cell model for KSHV infection and genetic manipulation

KSHV 感染和基因操作的细胞模型

基本信息

批准号：
10759522
负责人：
Shou-Jiang Gao
金额：
$ 58.49万
依托单位：
UNIVERSITY OF PITTSBURGH AT PITTSBURGH
依托单位国家：
美国
项目类别：
财政年份：
2003
资助国家：
美国
起止时间：
2003-01-13 至 2028-06-30
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10759522
关键词：
3-Dimensional Acceleration Acquired Immunodeficiency Syndrome Amino Acids Anabolism Animals Biological Models Biological Process Carbon Carbon nanoparticle Cell Communication Cell Culture System Cell Proliferation Cell model Cells Citric Acid Cycle Clinic Diphosphates Enzymes Funding Genes Glutamine Goals Growth Herpesviridae Infections Human Human Herpesvirus 8 Kaposi Sarcoma Ligase Malignant Neoplasms Mediating Metabolic Metabolic Pathway Metabolism Modeling Morbidity - disease rate Multicentric Angiofollicular Lymphoid Hyperplasia Mutagenesis Nitrogen Nucleotides Pathogenesis Pathway interactions Patients Prevention Proliferating Proteins Purines Pyrimidine Ribose-Phosphate Pyrophosphokinase Role Small Interfering RNA Societies Spindle Cell Neoplasm System Technology Testing Therapeutic Agents Translating Up-Regulation Viral Viral Genes Warburg Effect aerobic glycolysis cancer cell cancer type carcinogenesis cell transformation effective therapy effectiveness evaluation expectation experimental study genetic manipulation in vivo innovation knock-down locked nucleic acid metabolome metabolomics metaplastic cell transformation mortality multidisciplinary new technology new therapeutic target novel novel therapeutics nucleotide metabolism primary effusion lymphoma reverse genetics therapeutic target three dimensional cell culture tumor tumorigenesis

项目摘要

Cancer cells depend on reprogrammed metabolic pathways for anabolic proliferation. Discovering these cancer metabolic vulnerabilities can reveal novel targets for therapy. Kaposi’s sarcoma-associated herpesvirus (KSHV) is the causal agent of Kaposi’s sarcoma (KS) and several other cancers. Despite intensive studies for several decades, there is currently no effective therapy for KS. Our long-term goal is to delineate the pathogenesis of KSHV-induced cancers, providing a scientific basis for developing novel therapies. Toward this goal, we have previously developed an efficient system of KSHV-induced cellular transformation of primary cells and a reverse genetics system for KSHV mutagenesis. Using these powerful systems, in the current funding period, we have delineated viral and cellular genes that are essential for KSHV-induced cellular transformation and identified novel therapeutic targets and agents that have the potentials for translating into clinics. In particular, we have recently discovered that KSHV-transformed cells are addicted to glutamine. Unlike other types of cancer cells that utilize glutamine to replenish the TCA cycle, glutamine is primarily shunted to nucleotides syntheses by providing the critical g-nitrogen in addition to amino acids. KSHV hijacks numerous rate-limiting enzymes in these pathways, including phosphoribosyl pyrophosphate amidotransferase (PPAT) and phosphoribosyl pyrophosphate synthetases 1 (PRPS1), which is upregulated in KS spindle tumor cells. Significantly, knockdown of these enzymes suppresses the proliferation of KSHV-transformed cells but has no effect on the primary/uninfected cells. Our hypothesis is that KSHV encodes specific gene(s) to hijack the nucleotide synthesis pathways to support the proliferation and survival of KSHV-transformed cells, and hence targeting these pathways is effective for therapy of KSHV-induced tumors. We have developed 3D Culture systems that closely representing in vivo metabolic changes, an innovative nanoparticles carbon-dots (Cdots)- mediated delivery approach for locked nucleic acid (LNA)-siRNAs, and cutting-edge technology of metabolomics for tracing the carbon and nitrogen flows. We will determine the essential roles of the dysregulated nucleotide synthesis pathways for KSHV-induced cellular transformation and tumorigenesis (Aim 1); determine the mechanisms by which KSHV hijacks the nucleotide synthesis pathways for supporting the proliferation and survival of KSHV-transformed cells (Aim 2); and target vulnerable genes in the nucleotide pathways using the Cdots-mediated delivery approach for treating KSHV-induced tumorigenesis (Aim 3). The proposed project is highly significant as it will test a novel hypothesis of KSHV manipulation of key cellular metabolic pathways using multidisciplinary innovative approaches and model systems. It is our expectation that the accomplishment of this project will lead to the identification of novel cancer vulnerabilities of KSHV-induced cancers, which could provide a scientific basis for developing novel therapies.

癌细胞依赖于重编形的代谢途径进行合成代谢增殖。发现这些癌症代谢脆弱性可以揭示治疗的新目标。 Kaposi的肉瘤相关疱疹病毒（KSHV）是卡波西肉瘤（KS）和其他几种癌症的因果特工。尽管对几个几十年来，目前尚无对KS的有效疗法。我们的长期目标是描述 KSHV引起的癌症，为开发新型疗法提供了科学基础。达到这个目标，我们有以前开发了一个有效的KSHV诱导的原代细胞细胞转化系统和反向 KSHV诱变的遗传系统。使用这些功能强大的系统，在当前的资金期间，我们有描述的病毒和细胞基因对于KSHV诱导的细胞转化至关重要，并鉴定出具有转化为诊所的潜力的新型热目标和药物。特别是，我们有最近发现，KSHV转化的细胞沉迷于谷氨酰胺。与其他类型的癌细胞不同利用谷氨酰胺复制TCA循环，谷氨酰胺主要通过除氨基酸外，还提供了关键的G硝基。 KSHV劫持了其中的许多限速酶途径，包括焦磷酸氨基转移酶（PPAT）和磷酸蛋白酶基焦磷酸合酶1（PRPS1），在KS纺锤体肿瘤细胞中上调。值得注意的是，敲低在这些酶中，抑制了KSHV转换细胞的增殖，但对初级/未感染的细胞。我们的假设是KSHV编码特定基因以劫持核苷酸支持KSHV转换细胞的增殖和存活的合成途径，因此靶向这些途径可有效治疗KSHV诱导的肿瘤。我们已经开发了3D文化密切代表体内代谢变化的系统，一种创新的纳米颗粒碳点（CDOTS） - 锁定核酸（LNA）siRNA和代谢组学的尖端技术的介导的输送方法用于追踪碳和氮流。我们将确定失调的核苷酸的基本作用 KSHV诱导的细胞转化和肿瘤发生的合成途径（AIM 1）；确定 KSHV劫持了支持增殖和的机制 KSHV转换细胞的存活（AIM 2）；并使用核苷酸途径中靶向脆弱基因 CDOTS介导的递送方法用于治疗KSHV诱导的肿瘤发生（AIM 3）。拟议的项目是高度显着，因为它将测试关键细胞代谢途径KSHV操纵的新假设使用多学科创新方法和模型系统。我们期望实现该项目将导致识别KSHV引起的新型癌症脆弱性癌症可以为开发新型疗法提供科学基础。