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.

项目总结 潜伏感染的具有整合和复制能力的HIV基因组的CD4+T细胞在抗逆转录病毒治疗中持续存在 是根除艾滋病毒的主要障碍。在大多数艾滋病毒携带者(PWH)中，宿主极其稳定 半衰期超过3年。之前所有大幅缩小其尺寸或加速其衰败的尝试都以失败告终。 使用来自梅林B支艾滋病毒初级感染队列(秘鲁利马)参与者的样本，我们观察到 在感染的前3个月开始抗逆转录病毒治疗的个体中，艾滋病毒蓄积物的腐烂速度要快5到10倍 与那些被随机安排在较晚开始抗逆转录病毒治疗的患者相比，这表明早期治疗的人中感染艾滋病毒的细胞 更容易被淘汰。储油层半衰期的差异，至少在 ART的头4年，提供了一个独特的机会来确定可以利用的机制，以减少 水库在所有威尔斯亲王医院的艺术上。在这个项目中，我们建议揭开细胞和病毒特征对 单个艾滋病毒储存细胞的快速清除或长期存留。我们将检验这一假设 储藏细胞长时间存活的能力是由细胞和病毒特征共同驱动的， 这在早期和晚期治疗的个体之间是不同的，并导致不同的储集层衰变。我们会带上 独特的Merlin队列在研究12名启动抗逆转录病毒治疗次数较少的参与者中的艾滋病毒储存细胞方面的优势 感染艾滋病毒后超过3个月(早期抗逆转录病毒治疗：快速衰退)和12名参与者推迟治疗6个月 几个月(晚期艺术：缓慢腐烂)。我们将研究早期、中期和晚期的储集层，使用超低温保存 白细胞(在接受抗逆转录病毒治疗1年和2-3年时采集)和新采集的白细胞连续采集- 他们压抑了7年艺术生涯的参与者。在目标1中，我们将检验这样一个假设，即固有的细胞特征 艾滋病毒储存库细胞的数量是储存库腐烂的不同之处。我们将使用单细胞方法来识别 与储集层稳定性相关的促生存因子(Bcl2、TCF1、FOXO3a等)或者可能会保护受感染的人 来自免疫清除的细胞(免疫检查点分子的配体，Serpin B9，转化生长因子-β)。我们还将评估 假设完整基因组的克隆性扩张将更加普遍 在晚期治疗的参与者中。在目标2中，我们将检验特定病毒特征也起作用的假设。 对艾滋病毒感染细胞的持久性，并将随着时间的推移在ART上逐渐丰富。确定是否特定 病毒在治疗过程中被挑选出来，我们将使用一种将整合位点测序与 HIV转录评估，以确定转录活跃的存储细胞的比例。我们 将使用一种新的方法从潜在的储存库中重建分子病毒克隆，并将测量 这些病毒的适合性，并从功能上评估它们的艾滋病毒蛋白质组。通过这种方式，我们将确定水库是否 逐渐富含编码功能性Nef和VPU的前病毒，这些前病毒导致抗原递呈逃逸 并阻止CTL介导的杀伤以及自身抗体的中和和ADCC。结果来自 这项研究将确定可以有针对性地加速艾滋病毒储存库腐烂的细胞和病毒机制。