Evaluating the role of the novel apoptosis inhibitor TRAILshort, in maintaining HIV persistence

评估新型细胞凋亡抑制剂 TRAILshort 在维持 HIV 持续存在方面的作用

• 批准号: 10427482
• 负责人: ANDREW D BADLEY
• 金额: $ 60.69万
• 依托单位: MAYO CLINIC ROCHESTER
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-06-15 至 2023-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10427482
• 关键词: Acute; Affinity; Antibodies; Antigens; Apoptosis; Apoptosis Inhibitor; Apoptotic; Autologous; Autologous Tumor Cell; B lymphoid malignancy; Binding; Biochemical; Biochemistry; Biological Models; CD4 Positive T Lymphocytes; Cancer Model; Cancerous; Cell Death; Cell Proliferation; Cell physiology; Cells; Cessation of life; Communicable Diseases; Complex; Cryopreservation; Cysteine; Cytotoxic T-Lymphocytes; Data; Data Set; Defect; Dominant-Negative Mutation; Event; Exposure to; Genetic; HIV; HIV Infections; Homeostasis; Human; IL2RA gene; Immune response; Immune system; Immunohistochemistry; Immunologic Surveillance; Immunosuppression; Impairment; In Situ Hybridization; In Vitro; Infection; Interferons; Length; Ligand Binding; Ligands; Ligation; Lysosomes; MAPK8 gene; Maintenance; Malignant Neoplasms; Maps; Mediating; Natural Killer Cells; Pathway interactions; Patients; Phosphorylation; Production; Progressive Disease; Proliferating; Proteins; Proteomics; Publishing; RNA Splicing; Receptor Signaling; Research; Resistance; Role; Sampling; Signal Pathway; Signal Transduction; T cell response; T-Cell Activation; T-Cell Proliferation; T-Cell Receptor; T-Lymphocyte; TLR7 gene; TNF gene; TNF-related apoptosis-inducing ligand; TNFRSF10B gene; Techniques; Testing; Variant; Viral; Virus; Virus Replication; Western Blotting; ZAP-70 Gene; acute infection; antigen-specific T cells; biobank; cancer cell; cell killing; cytokine; exhaustion; extracellular vesicles; humanized antibody; knock-down; novel; p38 Mitogen Activated Protein Kinase; phosphoproteomics; prevent; receptor; small molecule; transcriptome sequencing

PROJECT SUMMARY HIV infected cells escape normal host immune responses. One pathway that the immune system uses to kill virally infected cells is TRAIL, which is expressed on activated T cells or NK cells. We have studied TRAIL involvement in HIV infection for ~15 years and discovered that (i) despite expressing TRAIL receptors, HIV infected cells are paradoxically resistant to the pro-death effects of TRAIL, and (ii) we discovered a splice variant of TRAIL that is produced by HIV+ cells which we have called TRAILshort. TRAILshort binds to TRAIL receptors and prevents normal (full length) TRAIL from killing these cells. We therefore created fully-humanized anti-TRAILshort-specific antibodies that sequester TRAILshort, and tested it in acute in vitro HIV infection. Anti TRAILshort antibody (or genetic inhibition of TRAILshort production) causes more HIV infected cells to die during acute infection, resulting in reduced HIV viral replication. We next analyzed publicly available RNAseq datasets from 253,200 human samples and identified TRAILshort exclusively in samples from donors with active infectious diseases, and/or human malignancy. We have since published that ~40% of human cancers express TRAILshort (by immunohistochemistry and in situ hybridization), and that primary B cell malignancies are inefficiently killed by autologous T cells, yet in the presence of TRAILshort antibody, that killing is significantly enhanced. We also observed that T cells exposed to cognate antigen proliferated more in the presence of anti-TRAILshort antibody, than in the absence, suggesting that TRAILshort might also directly impact T cell function and proliferation. Proteomic data presented herein show that TRAILshort treatment of primary T cells results in intracellular T signaling, changes in the cellular phosphorome, alterations in regulators of T cell activation and function (e.g. p38, ERK, JNK and Akt). In primary T cells treated with TRAILshort protein, we observe p38 phosphorylation at residues 180/182 by western blot, and impaired T cell activation induced by T-cell receptor (TCR) ligation (reduced CD25 and 69, less CFSE dilution and reduced Zap70 Lat phosphorylation by western blot), altogether indicating that TRAILshort is immunosuppressive to T cells. We will advance our understanding of the effect of TRAILshort on HIV specific T cell function by (i) testing the effect of TRAILshort antagonism on restoring HIV-specific T cell killing of productively HIV infected cells and latently HIV infected CD4 T cells induced to reactivate from latency, (ii) using advanced phospho-proteomic and biochemical techniques to understand how TRAILshort binding to TRAIL receptor 2 alters T cell homeostasis and (iii) study TCR-induced cell activation in the presence or absence of TRAILshort, to define defects in TCR signaling, reversing the defect using genetic and small molecule approaches, as well as TRAILshort antibodies.

项目摘要 HIV感染的细胞逃脱正常的宿主免疫反应。免疫系统用来杀死的一条途径 病毒感染的细胞是Trail，该细胞在活化的T细胞或NK细胞上表达。我们研究了踪迹 参与艾滋病毒感染约15年，发现（i）尽管表达了越野赛受体，艾滋病毒 感染的细胞矛盾地抵抗了小径的亲死亡影响，（ii）我们发现了一个剪接 艾滋病毒+细胞产生的小径变体，我们称之为Trailshort。步伐与步道结合 受体并防止正常（全长）踪迹杀死这些细胞。 因此，我们创建了完全人性化的抗trailshort特异性抗体，这些抗体隔离了轨迹，并且 在急性体外HIV感染中对其进行了测试。抗TRAILSHORT抗体（或遗传抑制径向生产） 导致更多的HIV感染细胞在急性感染期间死亡，从而减少HIV病毒复制。 下一 仅在患有活跃感染疾病的捐助者和/或人类恶性肿瘤的样本中。从那以后我们就有了 出版了约40％的人类癌症表达径流（通过免疫组织化学和原位 杂交），而原发性B细胞恶性肿则被自体T细胞杀死，但在 径向抗体的存在，杀戮显着增强。 我们还观察到暴露于同源抗原的T细胞在存在抗Trailshort的情况下增加了更多的增殖 抗体比不存在的抗体表明，径向也可能直接影响T细胞功能和 增殖。本文介绍的蛋白质组学数据表明，对原代T细胞的尾巴处理导致 细胞内T信号传导，细胞磷酸组的变化，T细胞活化调节剂的改变和 函数（例如p38，ERK，JNK和AKT）。在用Trailshort蛋白处理的原代T细胞中，我们观察到p38 通过蛋白质印迹在180/182的残基上的磷酸化，T细胞受体诱导的T细胞激活受损 （TCR）连接（减少CD25和69，CFSE稀释率较小，ZAP70 LAT磷酸化降低了。 斑点），完全表明Trailshort对T细胞进行免疫抑制。 通过（i）测试 跟踪拮抗作用对恢复HIV特异性的T细胞杀死感染的HIV感染细胞和 潜在的HIV感染的CD4 T细胞诱导了从潜伏期中重新激活的（ii），使用晚期磷酸蛋白酶 和生化技术，以了解跟踪与跟踪受体的结合如何改变T细胞 稳态和（iii）研究TCR诱导的细胞在存在或不存在的情况下定义 TCR信号的缺陷，使用遗传和小分子方法逆转缺陷，以及 径向抗体。

项目成果

Correction: Both HIV-Infected and Uninfected Cells Express TRAILshort, Which Confers TRAIL Resistance upon Bystander Cells within the Microenvironment.

更正：感染 HIV 的细胞和未感染的细胞都表达 TRAILshort，从而赋予微环境中旁观者细胞 TRAIL 抗性。

• DOI: 10.4049/jimmunol.1800867
• 发表时间: 2018
• 期刊: Journal of immunology (Baltimore, Md. : 1950)
• 作者: Nie,Zilin;Aboulnasr,Fatma;Natesampillai,Sekar;Burke,StephenP;Krogman,Ashton;Bren,GaryD;Chung,ThomasDY;Anderson,JeffR;Smart,MicheleK;Katzmann,DavidJ;Rajagopalan,Govindarajan;Cummins,NathanW;Badley,AndrewD
• 通讯作者: Badley,AndrewD

The long road to TRAIL therapy: a TRAILshort detour.

• DOI: 10.18632/oncotarget.27902
• 发表时间: 2021-03-30
• 期刊: Oncotarget
• 作者: Chandrasekar AP;Maynes M;Badley AD
• 通讯作者: Badley AD

The TRAIL: TRAILshort Axis in HIV Immunopathology.

• DOI: 10.1615/critrevimmunol.2019029632
• 发表时间: 2018
• 期刊: Critical reviews in immunology
• 影响因子: 1.3
• 作者: Aboulnasr F;Paranjape G;Badley AD
• 通讯作者: Badley AD

Plasma IL-6 levels following corticosteroid therapy as an indicator of ICU length of stay in critically ill COVID-19 patients.

• DOI: 10.1038/s41420-021-00429-9
• 发表时间: 2021-03-15
• 期刊: Cell death discovery
• 影响因子: 7
• 作者: Awasthi S;Wagner T;Venkatakrishnan AJ;Puranik A;Hurchik M;Agarwal V;Conrad I;Kirkup C;Arunachalam R;O'Horo J;Kremers W;Kashyap R;Morice W 2nd;Halamka J;Williams AW;Faubion WA Jr;Badley AD;Gores GJ;Soundararajan V
• 通讯作者: Soundararajan V