T-cell Transformation by Oncoviruses

肿瘤病毒对 T 细胞的转化

• 批准号: 7732891
• 负责人: Genoveffa Franchini
• 金额: $ 93.08万
• 依托单位: DIVISION OF BASIC SCIENCES - NCI
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/7732891
• 关键词: Adult; Affect; Amino Acid Substitution; Animal Model; Biochemical; Calnexin; Cell model; Cell surface; Cellular Membrane; Chronic; Class; Cleaved cell; Complex; Dendritic Cells; Disease; Endoplasmic Reticulum; Excision; Gene Expression; Generations; Genetic; Goals; Golgi Apparatus; HTLV-I Infections; Human; Human T-Cell Leukemia Viruses; Human T-lymphotropic virus 1; Immune response; In Vitro; Individual; Infection; Interleukin 2 Receptor; Life; Ligation; Localized; Maintenance; Major Histocompatibility Complex; Nuclear; Nucleolar Proteins; Oncornaviruses; Open Reading Frames; Pathway interactions; Patients; Proteins; Recruitment Activity; Retrieval; Retroviridae; Sampling; Signal Transduction; Structural Protein; T-Cell Receptor; T-Cell Transformation; T-Lymphocyte; Taxes; Testing; Trans-Activators; Variant; Viral; Viral Genes; Viral Genome; Viral Load result; Viral load measurement; calreticulin; genetic analysis; human disease; immunological synapse; in vivo; leukemia; leukemia/lymphoma; mutant; positional cloning; prevent; ranpirnase; therapeutic target; trafficking

In this project we have several aims: Aim 1: To integrate information obtained by functional biochemical studies on ectopically expressed non-structural HTLV-I proteins to infectivity and persistence of HTLV-I in relevant cellular models in vitro (dendritic cells and T-cells) Aim 2: To integrate information obtained by functional biochemical studies on ectopically expressed non-structural HTLV-I proteins to infectivity and persistence of HTLV-I in relevant animal models of HTLV-I infection. The viral genome encodes mRNAs for several non-structural proteins that affects cellular pathways and modulate viral replication in vitro. The p12 protein, encoded by orf I, localizes to the ER and Golgi and cellular membranes and different p12I cellular localization and functions are dictated by proteolytic cleavage. The removal of a non-canonical endoplasmic reticulum (ER) retention/retrieval signal within the amino terminus of p12I is necessary for trafficking to the Golgi apparatus and the generation of a completely cleaved 8 kDa protein. The 8 kDa protein in turn traffics to the cell surface, is recruited to the immunological synapse following T-cell receptor (TCR) ligation and down-regulates TCR proximal signaling. The uncleaved form of p12I resides in the ER and interacts with the b and gc chains of the interleukin-2 receptor (IL-2R), the heavy chain of the major histocompatibility complex (MHC) class I, as well as calreticulin and calnexin. Genetic analysis of ORF-I from ex vivo samples of HTLV-1-infected patients reveals predominant amino acid substitutions within ORF-I that inhibits proteolytic cleavage, suggesting that ER associated functions of p12I may be selected in vivo. We plan to use reverse genetic and use animal models to understand the contribution of variants of this viral gene to the maintenance of viral load in the host. The HTLV-I orf II encodes p30II, a nuclear/nucleolar protein that not only regulates viral expression by a post-transcriptional mechanism, but also affects the expression of genes involved in host responses such as TLR-4. In addition Orf II encodes p13, a small protein that we have recently demonstrate to decrease viral replication by targeting and degrading Tax, the viral trans activator. We have generated genetic mutant of HTLV-I that do not express p13 and/or p30 and found that the p30 affects dramatically the ability of HTLV-I to replicate in monocytoid-derived dendritic cells. The effect of p30 and p13 on in vitro infectivity is being tested in animal models. We also created HTLV-I mutants in the HBZ and found that the lack of HBZ result in an increased HTLV-I fusogenic ability.

在这个项目中，我们有几个目标：目标1：整合信息获得的 对异位表达的非结构HTLV-I蛋白的功能生化研究， HTLV-I在体外相关细胞模型（树突状细胞）中的感染性和持久性 目的2：整合功能生化研究获得的信息， 异位表达的非结构性HTLV-I蛋白对HTLV-I的感染性和持久性的影响 在HTLV-I感染的相关动物模型中。病毒基因组编码几种 影响细胞通路和调节病毒复制的非结构蛋白 体外由orf I编码的p12蛋白定位于ER和高尔基体以及细胞膜 并且不同的p12 I细胞定位和功能由蛋白水解切割决定。 去除非典型内质网（ER）内的保留/回收信号， p12 I的氨基末端是运输到高尔基体所必需的， 产生完全裂解的8kDa蛋白。8 kDa蛋白质又转运到 细胞表面，被募集到免疫突触后的T细胞受体（TCR） 连接并下调TCR近端信号传导。p12I的未切割形式存在于 ER并与白细胞介素-2受体（IL-2 R）的B和gc链相互作用， 主要组织相容性复合体（MHC）I类链，以及钙网蛋白和 钙连接蛋白来自HTLV-1感染患者的离体样品的ORF-I的遗传分析揭示了 ORF-I内抑制蛋白水解切割的主要氨基酸取代， 提示p12I的ER相关功能可以在体内选择。我们计划使用 逆转遗传学，并使用动物模型来了解这种病毒的变体的贡献， 基因对维持宿主体内病毒载量的作用。HTLV-I orf II编码p30 II， 核/核仁蛋白，不仅通过转录后调节病毒表达， 它不仅影响免疫应答机制，而且影响参与宿主应答的基因（如TLR-4）的表达。 此外，Orf II编码p13，这是一种小蛋白，我们最近证明， 通过靶向和降解病毒反式激活因子Tax来复制病毒。我们已经生成 HTLV-I的遗传突变体不表达p13和/或p30，并发现p30影响 显着提高HTLV-I在单核细胞衍生的树突状细胞中复制的能力。的 p30和p13对体外感染性的影响正在动物模型中进行测试。我们也 在HBZ中创建HTLV-I突变体，并发现HBZ的缺乏导致HTLV-I增加， 融合能力

项目成果

Tax oncoprotein trans-represses endogenous B-myb promoter activity in human T cells.

Tax 癌蛋白反式抑制人 T 细胞中的内源 B-myb 启动子活性。

• DOI: 10.1089/08892220050193065
• 发表时间: 2000
• 期刊: AIDS research and human retroviruses
• 影响因子: 1.5
• 作者: Nicot,C;Opavsky,R;Mahieux,R;Johnson,JM;Brady,JN;Wolff,L;Franchini,G
• 通讯作者: Franchini,G

Deletion of the p16INK4A gene in ex vivo acute adult T cell lymphoma/leukemia cells and methylation of the p16INK4A promoter in HTLV type I-infected T cell lines.

• DOI: 10.1089/088922200308701
• 发表时间: 2000-05
• 期刊: AIDS research and human retroviruses
• 影响因子: 1.5
• 作者: R. Trovato;A. Cereseto;S. Takemoto;A. Gessain;T. Watanabe;T. Waldmann;G. Franchini

A lysine-to-arginine change found in natural alleles of the human T-cell lymphotropic/leukemia virus type 1 p12(I) protein greatly influences its stability.

在人类 T 细胞嗜淋巴细胞/白血病病毒 1 型 p12(I) 蛋白的天然等位基因中发现的赖氨酸到精氨酸的变化极大地影响了其稳定性。

• DOI: 10.1128/jvi.73.8.6460-6467.1999
• 发表时间: 1999
• 期刊: Journal of virology
• 影响因子: 5.4
• 作者: Trovato,R;Mulloy,JC;Johnson,JM;Takemoto,S;deOliveira,MP;Franchini,G
• 通讯作者: Franchini,G

p53 stabilization and functional impairment in the absence of genetic mutation or the alteration of the p14(ARF)-MDM2 loop in ex vivo and cultured adult T-cell leukemia/lymphoma cells.

• DOI: 10.1182/blood.v95.12.3939
• 发表时间: 2000-06
• 期刊: Blood
• 影响因子: 20.3
• 作者: S. Takemoto;R. Trovato;A. Cereseto;C. Nicot;T. Kislyakova;L. Casareto;T. Waldmann;G. Torelli;G. Franchini

Differential response to genotoxic stress in immortalized or transformed human T-lymphotropic virus type I-infected T-cells.

永生化或转化的人类 T 淋巴细胞病毒 I 型感染的 T 细胞对基因毒性应激的差异反应。

• DOI: 10.1099/0022-1317-80-7-1575
• 发表时间: 1999
• 期刊: The Journal of general virology
• 作者: Cereseto,A;Kislyakova,T;WashingtonParks,R;Nicot,C;Franchini,G
• 通讯作者: Franchini,G