A nonhuman primate model of stem cell transplantation to understand determinants of post-transplant SIV clearance

项目总结 鉴于目前感染艾滋病毒的人数是历史上最多的，遏制艾滋病毒流行仍是当务之急。 联合抗逆转录病毒疗法(CART)可以限制病毒复制，但不能治愈。因此，有一个紧急的 需要设计一种通过消除病毒库来治疗的功能疗法。蒂莫西·布朗，也就是柏林病人， 在白血病相关、MHC匹配的异基因造血后，在没有CART的情况下保持无HIV CCR5缺陷供者的干细胞移植(HSCT)。这种功能性治愈背后的机制 目前尚不清楚，但可能取决于免疫调节方案、移植物抗宿主免疫或 缺乏CCR5的重组免疫系统。非人灵长类动物是感染艾滋病毒的最佳模型，但 由于移植的复杂性，以前的移植研究使用的是自体或MHC不匹配的同种异基因造血干细胞移植 非人灵长类MHC。此外，不存在CCR缺陷的猕猴捐赠者。因此，还没有研究能够 重述蒂莫西·布朗的移植过程。我们已经使用一种新的， 毛里求斯食蟹猴(MCM)。由于最近的瓶颈，MCM极大地简化了遗传学 大约500年前。此外，我们目前正在使用CRISPR技术来生成CCR5 匮乏的捐赠者。因此，我们处于独特的位置来重述蒂莫西·布朗的移植 临床相关动物模型。在具体目标1中，我们将测量清髓性化疗的影响 清除自体造血干细胞移植中的潜伏病毒库。在具体目标2中，我们将充分履行 MHC相合的HSCT，测量移植物抗宿主免疫，并将其与SIV在血浆和 纸巾。在具体目标3中，我们将使用我们的CCR5缺陷捐赠者进行干细胞异基因造血干细胞移植。 缺少CCR5。总体而言，这项研究将使我们能够绘制出HSCT后艾滋病毒清除的决定因素。

PROJECT SUMMARY With the most people ever in history currently living with HIV, stopping the HIV epidemic remains imperative. Combination antiretroviral therapy (cART) limits viral replication, but is not curative. Thus, there is an urgent need to design a functional cure via elimination of the viral reservoir. Timothy Brown, aka the Berlin Patient, remains HIV free in the absence of cART following leukemia-related, MHC-matched, allogeneic hematopoietic stem cell transplantation (HSCT) from a CCR5 deficient donor. The mechanisms underlying this functional cure are not known, but may depend on the immune conditioning regimen, graft-versus-host immunity, or a reconstituted immune system lacking CCR5. Non-human primates are the best model of HIV infection, but previous transplant studies have used autologous or MHC-mismatched allogeneic HSCT due to the complexity of non-human primate MHC. Also, no CCR deficient macaque donors exist. Thus, no studies have been able to recapitulate the transplant of Timothy Brown. We have built an allogeneic HSCT model using a novel, Mauritian cynomolgus macaque (MCM). MCM have extremely simplified genetics due to a recent bottleneck approximately 500 years ago. Furthermore, we are currently using CRISPR technology to generate CCR5 deficient donors. Therefore, we are uniquely positioned to recapitulate Timothy Brown’s transplant in a clinically relevant animal model. In specific aim 1, we will measure the impact of myeloablative chemotherapy on clearing the latent viral reservoir in autologous HSCT transplants. In specific aim 2, we will perform fully MHC-matched HSCT, measure graft-versus host immunity, and correlate it to SIV rebound in the plasma and tissues. In specific aim 3, we will use our CCR5 deficient donors to perform an allogeneic HSCT with stem cells lacking CCR5. Overall, this study will allow us to map the determinants of HIV clearance following HSCT.