AAV-delivered HIV inhibitors for SHIV therapy

AAV 递送的 HIV 抑制剂用于 SHIV 治疗

• 批准号: 10683342
• 负责人: Matthew Ryan Gardner
• 金额: $ 80.95万
• 依托单位: EMORY UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-08-15 至 2027-05-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10683342
• 关键词: Anti-Retroviral Agents; Antibodies; Antibody Response; Binding Sites; Clinical Trials; Data; Dependovirus; Dose; Engineering; Evaluation; HIV; HIV Entry Inhibitors; HIV-1; HIV-2; Human; Immune Targeting; Immune response; Immune system; In Vitro; Individual; Infection; Infusion procedures; Injecting drug user; Intramuscular; Lifting; Macaca; Macaca mulatta; Mediating; Monkeys; Mus; Muscle; Muscle Cells; Needle Sharing; Pathway interactions; Persons; Pharmaceutical Preparations; Pharmacotherapy; Pilot Projects; Proteins; Reporting; Resistance; SIV; Safety; Serum; Skeletal Muscle; Technology; Testing; Therapeutic; Time; Tissues; Transgenes; Treatment Efficacy; United States; Viral; Viral Load result; Viral reservoir; Viremia; Virus; Virus Replication; Virus-like particle; Woman; adeno-associated viral vector; antiretroviral therapy; cohort; design; gene therapy; humanized mouse; immune checkpoint; improved; in vivo; inhibitor; men; neutralizing antibody; novel; programmed cell death ligand 1; receptor binding; simian human immunodeficiency virus; small molecule; success; therapy development; transgene expression; transmission process; viral DNA; viral RNA; viral rebound

PROJECT SUMMARY More 10% of the new infections in the United States were from contaminated needle sharing among people who inject drugs (PWID). Importantly, only about 50% of men and 57% of women of the PWID group had suppressed viremia. Despite the lack of a sterilizing HIV-1 cure, antiretroviral drug therapies (ART) effectively suppress viral replication in people living with HIV-1. One limitation of ART is that these small molecule drugs cannot eliminate the viral reservoir. Broadly neutralizing antibodies (bNAbs) could supplement ART and be used to reduce the viral reservoir. Several studies have shown that a single dose of a bNAb can decrease viremia in HIV-1 infected individuals, with viral rebound occurring as the bNAb concentration decreases. Adeno-associated virus (AAV) vectors may provide the means for long-term expression of bNAbs at concentrations capable of maintaining viral suppression via intramuscular inoculations. However, we and others have shown that the emergence of anti-drug antibodies (ADA) to bNAbs limits their overall expression. Here, we show that we have made significant strides overcome this host immune response. First is through utilizing immune checkpoints that regulate immune system pathways. We observed a 21-fold increase in concentrations of the HIV-1 bNAb 10-1074 in rhesus macaques when macaques were co-inoculated with an AAV vector encoding rhesus macaque PD-L1. Second, we have developed a novel, HIV-1 entry inhibitor, eCD4-Ig, which tends to be more tolerated in rhesus macaques when expressed from AAV vectors. Unlike bNAbs, we have shown that AAV vectors encoding eCD4-Ig can express the inhibitor in macaques for over a year and the ADA response against eCD4-Ig decreases over time. Because eCD4-Ig neutralizes all HIV-1, HIV-2, and SIV isolates and is difficult to escape, it may be useful when included in a therapy strategy. Our pilot studies show that low concentrations AAV-expressed eCD4-Ig can suppress SHIV infection in rhesus macaques for two years, yet viremia is still detectable. This proposal combines our AAV advancements into a single strategy to determine whether AAV-expressed inhibitors can suppress a SHIV infection and reduce the viral reservoir. In Aim 1, we will assess the therapeutic efficacy of the combination of AAV-delivered eCD4-Ig and 10-1074 after ART is lifted in SHIV-infected rhesus macaques. In Aim 2, we will determine whether suppressing an established SHIV infection with AAV- delivered eCD4-Ig and 10-1074 results in a quantitatively different viral reservoir compared to ART. In Aim 3, we will improve the safety of AAV gene therapy by developing an irreversible “kill-switch” to turn off transgene expression.

项目摘要 在美国，超过10%的新感染病例来自受污染的针头共用， 注射毒品的人（PWID）重要的是，只有大约50%的男性和57%的女性的PWID 组抑制了病毒血症。尽管缺乏一种能消除HIV-1病毒的治疗方法， (ART)有效抑制HIV-1感染者的病毒复制。ART的一个局限性是， 这些小分子药物无法消除病毒储存库。广泛中和抗体 （bNAb）可以补充ART并用于减少病毒库。几项研究表明 单剂量的bNAb可以减少HIV-1感染者的病毒血症， 随着bNAb浓度的降低。腺相关病毒（AAV）载体可以提供 在能够维持病毒感染的浓度下长期表达bNAb的方法 通过肌肉注射抑制。然而，我们和其他人已经表明， 针对bNAb的抗药抗体（ADA）限制了它们的总体表达。在这里，我们表明，我们已经取得了 克服这种宿主免疫反应的重大进展。首先是利用免疫检查点 调节免疫系统的通路。我们观察到HIV-1的浓度增加了21倍 bNAb 10-1074在恒河猴中的表达 恒河猴PD-L1。其次，我们开发了一种新型HIV-1进入抑制剂eCD 4-IG， 当从AAV载体表达时，在恒河猴中趋于更耐受。与bNAb不同，我们 已经显示编码eCD 4-IG的AAV载体可以在猕猴中表达抑制剂超过一年 并且针对eCD 4-IG的ADA应答随时间降低。因为eCD 4-IG中和所有HIV-1， HIV-2和SIV分离株，并且难以逃脱，当包括在治疗策略中时可能有用。 我们的初步研究表明，低浓度的AAV表达的eCD 4-IG可以抑制SHIV感染。 恒河猴两年了，但病毒血症仍然可以检测到。该提案结合了我们的AAV 研究进展到一个单一的策略，以确定是否AAV表达的抑制剂可以抑制 SHIV感染和减少病毒库。在目标1中，我们将评估 在SHIV感染的恒河猴中ART解除后AAV递送的eCD 4-IG和10-1074的组合 猕猴在目标2中，我们将确定是否用AAV抑制已建立的SHIV感染。 与ART相比，递送的eCD 4-IG和10-1074导致定量上不同的病毒储库。 目的3、通过研制一种不可逆的“开关”， 关闭转基因表达。