In situ and digital spatial profiling of the active HIV reservoir in autopsy-derived tissues

尸检组织中活性 HIV 储存库的原位和数字空间分析

• 批准号: 10614019
• 负责人: Timothy Jensen Henrich
• 金额: $ 49.71万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-05-01 至 2027-04-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10614019
• 关键词: Acute myocardial infarction; Anatomy; Autopsy; B-Lymphocytes; Binding; Biological Assay; Biological Markers; Biopsy; Brain; CD4 Positive T Lymphocytes; Cardiac; Cell Count; Cell Lineage; Cells; Cessation of life; County; Cryopreserved Tissue; DNA; Data; Drug or chemical Tissue Distribution; Environment; Formalin; Freezing; Frequencies; Funding; Gene Expression; Gene Expression Profiling; Genetic Transcription; HIV; Heart; Hemorrhage; Histologic; Human; Immune; Immunohistochemistry; Immunologics; In Situ; In Situ Hybridization; Individual; Investigation; Length; Liver; Lung; Lymphoid Cell; Lymphoid Tissue; Macrophage; Maintenance; Measures; Medical Examiners; Metabolic Clearance Rate; Morbidity - disease rate; Myelogenous; Myeloid Cells; National Heart, Lung, and Blood Institute; Neuroglia; Neurologic; Nucleic Acids; Organ; Overdose; Paraffin Embedding; Participant; Persons; Pharmaceutical Preparations; Phenotype; Proteins; Proteomics; Proviruses; Pulmonary Embolism; RNA; Recrudescences; Residual state; Resolution; Resources; Role; Sampling; San Francisco; Source; Spleen; Time; Tissue Sample; Tissue-Specific Gene Expression; Tissues; Transcript; Viral; Virus; Visualization; Withdrawal; antiretroviral therapy; aspirate; cohort; comorbidity; digital; experience; immune activation; in vivo; innate immune pathways; innovation; interest; lymph nodes; mortality; nano-string; novel; prospective; protein expression; sudden cardiac death; synergism; targeted treatment; transcriptomics; viral RNA; viral rebound

Project Summary/Abstract (Project 2) The study of HIV rebound potential necessitates rigorous viral and immunological characterization directly within tissues in people with HIV (PWH) on ART with minimal comorbidities. As a result, this project leverages the longitudinal San Francisco POstmortemSystematic InvesTigation of Sudden Cardiac Death (POST SCD) Study, a postmortem study to bank samples and autopsy data on PWH and uninfected controls who were victims of sudden cardiac death (SCD). To date we have collected extensive tissue samples, including brain, multiple lymph node chains, liver, spleen, heart, pulmonary vasculature and other tissues of interest from47 HIV-infected and >500 uninfected individuals who experienced SCD. Importantly, ~80% of HIV+ SCD cases were on ART and died suddenly of non-HIV (i.e. cardiac) causes. As a result, the HIV POST SCD cohort is a one-of-kind resource for the study of tissue HIV persistence. This highly innovative project involves in situ hybridization and cutting-edge tissue-based transcriptomic/proteomic nanoString Digital Spatial Profiling (DSP) to clearly define how the reservoirs of intact and HIV-expressing proviruses and biomarker expression differ between tissues and determine how tissue-specific differences in the transcriptionally active HIV reservoir relate to in situ host cell gene and protein expression. Our central hypothesis is that infected CD4 T cells and various myeloid lineage cells within immune privileged histologic environments express full-length, intact HIV RNA transcripts and be capable of rapid viral recrudescence following ART withdrawal. We expect to observe lower expression of host antiviral factors and higher expression of pro-survival factors in cell clusters expressing HIV transcripts. Furthermore, we posit that transcriptional activity and immune states of reservoir cells that are identified as predictors of viral rebound in Project 3 will be visualized within specific lymphoid tissue regions of interest. Our aims are to: 1) measure the total burden of intact and defective proviruses and HIV transcripts across the full spectrum of different organs and tissues in vivo; 2) compare across tissues the cellular burden and phenotypes of cells that spontaneously transcribe HIV transcripts in SCD victims on and off ART at the time of death; and 3) determine the in situ impact of HIV burden and residual transcriptional activity on host cell factors (particularly antiviral restriction and innate immune pathways) in tissue-resident lymphoid and myeloid cells in SCD victims on ART compared to uninfected controls. As a result, this project will have the capacity to identify targets for therapeutic approaches to achieve HIV cure in synergy with Projects 1 & 3.

项目摘要/摘要（项目2） 对HIV反弹潜力的研究需要直接在 艾滋病毒（PWH）的组织中的组织在具有最小合并症的艺术中。结果，该项目利用了 纵向旧金山突变后心脏死亡的术后研究（SCD后）研究， 对银行样本的验尸研究和有关PWH和未感染对照的尸检数据，这些数据是受害者的受害者 猝死（SCD）。迄今为止，我们已经收集了包括大脑在内的大量组织样品 淋巴结链，肝，脾，心脏，肺脉管系统和其他感兴趣的组织47 HIV感染 和> 500名经历过SCD的未感染的人。重要的是，约80％的HIV+ SCD病例是ART 并突然死于非HIV（即心脏）原因。结果，艾滋病毒后SCD队列是一种独一无二的 组织HIV持续性研究的资源。这个高度创新的项目涉及原位杂交和 尖端组织的转录组/蛋白质组织数字空间分析（DSP）以清楚定义 完整和表达艾滋病毒的病毒和生物标志物表达的储层在组织和生物标志物表达之间有何不同 确定转录活性HIV储量中组织特异性差异与原位宿主细胞的关系 基因和蛋白质表达。我们的中心假设是感染的CD4 T细胞和各种髓样谱系 免疫特权的组织学环境中的细胞表达全长，完整的HIV RNA转录本，并且BE 能够在ART撤离后快速病毒复发。我们希望观察到宿主的较低表达 表达HIV转录物的细胞簇中的抗病毒因子和较高的生存因子的表达。 此外，我们认为储层细胞的转录活性和免疫状态被识别为 项目3中病毒反弹的预测因素将在特定的淋巴组织中可视化。 我们的目的是：1）衡量整个完整和有缺陷的原病毒和艾滋病毒成绩单的总负担 体内不同的器官和组织的全光谱； 2）比较跨组织的细胞负担和 在SCD受害者中自发转录HIV转录本在ART和ART时自发转录HIV转录本的细胞表型 死亡; 3）确定HIV负担和残留转录活动对宿主细胞因素的原位影响 （尤其是抗病毒限制和先天免疫途径）在组织居住的淋巴样和髓样细胞中 与未感染的对照相比，SCD受害者的ART受害者。结果，该项目将有能力确定 与项目1和3的协同作用的治疗方法的靶标。