Systemic HIV/SIV latency reversal via non-canonical NF-κB 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 NFκB 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. Differential Expression Analysis using RNA-seq time-series comparison of treatment vs control vs mock on primary T-cells

模拟第二种线粒体源性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（SMACm）可在体外及离体（ex vivo）条件下强效逆转HIV潜伏，且未出现其他潜伏期逆转剂（Latency Reversing Agents，LRAs）所具有的多效性细胞效应。我们验证发现，SMACm通过激活非经典NFκB通路介导潜伏期逆转，具体表现为cIAP1快速降解，随后无活性的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HIV感染的供者的静息CD4+ T细胞中提升细胞相关HIV gag RNA的表达水平，同时仅改变少数人类基因的表达。本研究首次证实SMACm在患者来源的细胞中具备单药潜伏期逆转活性，相关发现支持在临床前动物模型中对SMACm进行评估。本次研究采用的差异表达分析方法为：对原代T细胞的处理组、对照组与空白对照组（mock）开展RNA-seq时间序列对比分析。