Systemic HIV/SIV latency reversal via non-canonical NF-?B signaling in vivo

Molecules mimicking the active N-terminal tetrapeptide of the second mitochondrial-derived activator of caspases (SMACm) potently reverse HIV latency in vitro and ex vivo without the pleotropic cellular effects seen with other LRAs. We verified that SMACm facilitate latency reversal through activation of the non-canonical NF?B pathway as exemplified by rapid degradation of cIAP1, followed by a slower conversion of inactive p100 into active p52. A potent representative of this class, AZD5582, increases cell-associated HIV gag RNA expression in resting CD4+ T cells from ART-suppressed, HIV-infected donors while altering the expression of a restricted number of human genes. These findings represent the first demonstration that SMACm have single agent latency reversal activity in patient-derived cells and support evaluation of SMACm in preclinical animal models. Overall design: Differential Expression Analysis using RNA-seq time-series comparison of treatment vs control vs mock on primary T-cells

本研究中的SMACm，即模拟第二代线粒体衍生半胱天冬酶激活因子（second mitochondrial-derived activator of caspases, SMAC）活性N端四肽的分子，可在体外与离体环境中强效逆转HIV潜伏，且不会产生其他LRAs所引发的多效性细胞效应。本研究证实，SMACm可通过激活非经典NF-κB通路实现HIV潜伏逆转，其典型特征为cIAP1的快速降解，随后将无活性的p100缓慢转化为活性p52。该类分子的强效代表性化合物AZD5582，可在接受抗逆转录病毒治疗（Antiretroviral Therapy, ART）且病毒受抑的HIV感染供者的静息CD4+ T细胞中，提升细胞相关HIV gag RNA的表达水平，同时仅调控有限数量的人类基因。本研究结果首次证实，SMACm在患者来源的细胞中具有单药潜伏逆转活性，为其在临床前动物模型中的评估提供了支持依据。研究设计方案：采用RNA测序（RNA Sequencing, RNA-seq）开展差异表达分析，对比原代T细胞中处理组、对照组与空白对照组的时序转录组数据。