Post-TB epigenetic scars' impact on long-term inflammation, immunity and mortality

项目总结： 在明显成功的治疗后，结核病幸存者的死亡率为16.9%，是以前的3倍 高于年龄和性别匹配的对照组。在我们之前的工作中，我们发现在六个月后 成功完成结核病治疗后，幸存者保留了有害的表观遗传疤痕，这些疤痕继续困扰着 宿主免疫和炎症。在这个提案中，我们将使用当前登记的队列来测试我们的提案 这些有害的表观遗传疤痕会导致病理性炎症并降低免疫反应性， 导致结核病后死亡风险增加。 我们以前的工作表明，TB诱导了一千多个DNA甲基化扰动 在与炎症相关的通路中，有肿瘤坏死因子、白介素6和干扰素信号通路。从功能上讲，这些DNA 甲基化紊乱与炎症增加和免疫反应性降低有关。 其他感染(艾滋病毒、巨细胞病毒、血吸虫病)也会导致持久的表观遗传疤痕，但到目前为止，还没有研究 已经确定了这些表观遗传疤痕中的哪些与结核病后死亡有关。因此，在目标1中，我们 将在治疗成功后对结核病患者进行为期30个月的跟踪调查，以确定哪种DNA甲基化 MARK与结核病后死亡率有关。 虽然大多数免疫细胞只能存活几天或几周，但我们之前的工作表明，DNA 甲基化紊乱在成功治疗后至少持续6个月。动物模型和我们的 初步数据支持这样一种假设，即有害的表观遗传疤痕出现在祖细胞和干细胞中。至 为了验证这一假设，我们将使用尖端单细胞表观基因组学(ScATAC)和转录组学(ScRNA) 用测序和伪时间生物信息学技术将祖先耗尽的种群与终末期 精疲力竭的人群。 我们的体外初步数据表明，TCA代谢途径的激活介导了 诱导有害的表观遗传标记和抑制TCA的激活可以减轻诱导的感染 DNA超甲基化和恢复免疫反应。因此，利用我们建立的豚鼠结核病 模型中，我们将评估TCA循环的抑制剂是否可以阻止或逆转有害的表观遗传疤痕和 恢复分枝杆菌免疫力。破坏免疫的特定表观遗传标记的阐明 动态平衡和增加结核病后死亡的风险是发展附属疾病的必要步骤 宿主指导治疗以降低目前成功的结核病治疗后的高死亡率。

PROJECT SUMMARY: After apparently successful treatment, Tuberculosis (TB) survivors have a 16.9% of death, a rate 3-fold higher than age and sex matched controls. In our previous work, we identified that six months after the completion of successful TB therapy, survivors retained detrimental epigenetic scars that continue to perturb host immunity and inflammation. In this proposal, using currently enrolling cohorts, we will test our proposal that these detrimental epigenetic scars induce pathologic inflammation and decrease immune responsiveness, leading to increased risk of post-TB mortality. Our previous work demonstrated that TB induces more than a thousand DNA methylation perturbations in pathways related to inflammation, TNF, IL-6 and IFN signaling pathways. Functionally, these DNA methylation perturbations are associated with increased inflammation and decreased immune responsiveness. Other infections (HIV, CMV, schistosomiasis) also induce long-lasting epigenetic scars, yet, to date, no study has identified which of these epigenetic scars are associated with post-TB mortality. Therefore, in Aim 1, we will follow TB patients for 30 months after completion of successful therapy to identify which DNA methylation marks are associated with post-TB mortality. While most immune cells only live a few days or weeks, our previous work demonstrated that DNA methylation perturbations persist at least 6 months after successful therapy. Animal models and our preliminary data support the hypothesis that detrimental epigenetic scars occur in progenitor and stem cells. To test this hypothesis, we will use cutting-edge single cell epigenomic (scATAC) and transcriptomics (scRNA) sequencing and pseudotime bioinformatic techniques to link progenitor exhausted populations with terminally exhausted populations. Our in vitro preliminary data, demonstrate that activation of the TCA metabolic pathway mediates the induction of detrimental epigenetic marks and that inhibiting the TCA activation can mitigate infection induced DNA hyper-methylation and restore immune responsiveness. Therefore, using our established guinea pig TB model, we will evaluate if inhibitors of the TCA cycle can block or reverse detrimental epigenetic scars and restore mycobacterial immunity. The elucidation of the specific epigenetic marks that subvert immune homeostasis and increase the risk of post-TB mortality is a necessary step in the development of adjunctive host directed therapy to decrease the current high mortality rates that occur after successful TB therapy.