Identifying vulnerabilities in the long-lived HIV reservoir to accelerate its decay

识别长期艾滋病毒储存库的脆弱性以加速其腐烂

• 批准号: 10673309
• 负责人: Nicolas Chomont
• 金额: $ 108.86万
• 依托单位: FRED HUTCHINSON CANCER CENTER
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-04-01 至 2028-03-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10673309
• 关键词: Acceleration; Acute; Affect; Anti-Retroviral Agents; Antibodies; Antigen Presentation; Antigens; Autologous; BCL2 gene; BCL2L1 gene; Biological Assay; CD4 Positive T Lymphocytes; Cells; Characteristics; Chromatin; Clonal Expansion; Clonality; Coupled; Couples; Cryopreservation; Data; Early Diagnosis; Ensure; FOXO3A gene; Future; Genetic Transcription; Genome; Goals; HIV; HIV Genome; HIV Infections; HIV envelope protein; Half-Life; Immunologics; Individual; Infection; Leukapheresis; Ligands; Longterm Follow-up; Maintenance; Measures; Mediating; Memory; Molecular; NCOA6 gene; Nature; Participant; Persons; Pharmaceutical Preparations; Phenotype; Predisposition; Process; Production; Proliferating; Proteome; Proviruses; Randomized; Reporting; Resistance; Sampling; Serpins; Source; Testing; Time; Transforming Growth Factor beta; Viral; Viral Genome; Virus; cohort; decay acceleration; experimental study; fitness; genome sequencing; immune checkpoint; immune clearance; integration site; novel strategies; response; tool; whole genome

PROJECT SUMMARY Latently infected CD4+ T cells harboring integrated and replication-competent HIV genomes persist during ART and are the main obstacle to HIV eradication. In most people with HIV (PWH), the reservoir is extremely stable with a half-life of over 3 years. All prior attempts to significantly reduce its size or accelerate its decay have failed. Using samples from participants in the MERLIN clade B primary HIV infection cohort (Lima, Peru), we observed 5 to 10-times faster decay of the HIV reservoir in individuals initiating ART during the first 3 months of infection compared to those randomized to start ART later, suggesting that HIV-infected cells in people treated early are more susceptible to elimination. Differences in the half-life of the reservoir, which are maintained during at least the first 4 years of ART, offer a unique opportunity to identify mechanisms that could be harnessed to reduce the reservoir in all PWH on ART. In this project, we propose to unravel the cellular and viral features responsible for the rapid clearance or long-term persistence of individual HIV reservoir cells. We will test the hypothesis that the capacity of reservoir cells to persist for prolonged periods is driven by a combination of cellular and viral features, which differ between early and late treated individuals and result in differential reservoir decay. We will take advantage of the unique MERLIN cohort to study HIV reservoir cells in 12 participants who initiated ART less than 3 months after HIV acquisition (early ART: rapid decay) and 12 participants who deferred treatment for 6 months (late ART: slow decay). We will study the early, intermediate and late reservoirs, using cryopreserved leukaphereses (collected at 1 & 2-3 years of ART) and newly collected leukapheresis from the same continually- suppressed participants at >7 years of ART. In Aim 1, we will test the hypothesis that intrinsic cellular features of HIV reservoir cells underlie differences in reservoir decay. We will employ a single cell approach to identify pro-survival factors associated with reservoir stability (Bcl-2, TCF-1, FOXO3A etc.) or that may protect infected cells from immune clearance (ligands of immune checkpoint molecules, Serpin B9, TGF-β). We will also evaluate the clonality of the reservoir with the hypothesis that clonal expansions of intact genomes will be more common in late treated participants. In Aim 2, we will test the hypothesis that specific viral characteristics also contribute to the persistence of HIV-infected cells and will be gradually enriched over time on ART. To determine if specific viruses are selected against during therapy, we will use an assay that couples integration site sequencing with HIV transcription assessment, to determine the proportion of reservoir cells that are transcriptionally active. We will use a novel approach to reconstruct molecular viral clones from the latent reservoir, and will measure the fitness of these viruses and functionally assess their HIV proteomes. In this way, we will determine if the reservoir is gradually enriched in proviruses encoding functional Nef and Vpu, which cause escape of antigen presentation and impede CTL-mediated killing as well as neutralization and ADCC by autologous antibodies. Results from this study will identify cellular and viral mechanisms that can be targeted to accelerate decay of the HIV reservoir.

项目摘要 在ART期间，携带综合和复制能力的HIV基因组的潜在感染的CD4+ T细胞持续存在 这是消除艾滋病毒的主要障碍。在大多数艾滋病毒（PWH）的人中，水库非常稳定 半衰期超过3年。所有先前的尝试显着降低其大小或加速其衰减的尝试都失败了。 使用Merlin进化枝B原发性HIV感染队列（Lima，Peru）的样本，我们观察到 在感染的前三个月，艾滋病毒水库的艾滋病毒水库的衰减速度更快 与后来随机开始艺术的那些相比，表明早期治疗的人的艾滋病毒感染细胞是 更容易消除。储层的半衰期差异，至少在 艺术的前4年，提供了一个独特的机会来确定可以利用的机制来减少 所有PWH的水库。在这个项目中，我们建议阐明负责的细胞和病毒特征 单个HIV储层细胞的快速清除或长期持久性。我们将检验以下假设 储层细胞长期持续的能力是由细胞和病毒特征的组合驱动的 在早期和晚期治疗的个体之间有所不同，并导致储层衰减。我们会接受的 在12名参与者中研究HIV储层细胞的独特Merlin队列的优势 艾滋病毒收购后的3个月（早期艺术：快速衰减）和12名推迟治疗的参与者6 月份（后期艺术：缓慢的衰减）。我们将使用冷冻保存研究早期，中间和晚期储量 Leukapheres（在1和2 - 3年的艺术品中收集），并从同一连续地收集了白细胞术 在> 7年的艺术品上被压制的参与者。在AIM 1中，我们将测试固有细胞特征的假设 HIV储层细胞的含量差异是储层衰减的差异。我们将采用单元方法来识别 与储层稳定性（BCL-2，TCF-1，FOXO3A等）相关的阳离子因素，或者可能保护感染 免疫清除的细胞（免疫检查点分子的配体，Serpin B9，TGF-β）。我们还将评估 储层的克隆性具有以下假设：完整基因组的克隆膨胀将更为普遍 在AIM 2中，我们将检验以下假设：特定的病毒特征也有助于 为了持续的HIV感染细胞，随着时间的流逝，将逐渐富集。确定是否具体 在治疗期间选择病毒，我们将使用一项评估，以使整合位点测序与 HIV转录评估，以确定转录活性的储层细胞的比例。我们 将使用一种新颖的方法从潜在储层重建分子病毒克隆，并将测量 这些病毒的适应性并在功能上评估其HIV蛋白质组。这样，我们将确定水库是否 逐渐富集在编码功能性NEF和VPU的Provirus中，这会导致抗原表现的逃脱 并阻碍CTL介导的杀戮以及自体抗体的谈判和ADCC。结果 这项研究将确定可靶向加速HIV储量衰减的细胞和病毒机制。