Center for Microbial Interface Biology, Department of Microbial Infection and Immunity, The Ohio State University, Columbus, OH, United States of America.
Texas Biomedical Research Institute, San Antonio, TX, United States of America.
PLoS Pathog. 2018 Jun 21;14(6):e1007100. doi: 10.1371/journal.ppat.1007100. eCollection 2018 Jun.
Peroxisome proliferator-activated receptor (PPAR)γ is a global transcriptional regulator associated with anti-inflammatory actions. It is highly expressed in alveolar macrophages (AMs), which are unable to clear the intracellular pathogen Mycobacterium tuberculosis (M.tb). Although M.tb infection induces PPARγ in human macrophages, which contributes to M.tb growth, the mechanisms underlying this are largely unknown. We undertook NanoString gene expression analysis to identify novel PPARγ effectors that condition macrophages to be more susceptible to M.tb infection. This revealed several genes that are differentially regulated in response to PPARγ silencing during M.tb infection, including the Bcl-2 family members Bax (pro-apoptotic) and Mcl-1 (pro-survival). Apoptosis is an important defense mechanism that prevents the growth of intracellular microbes, including M.tb, but is limited by virulent M.tb. This suggested that M.tb differentially regulates Mcl-1 and Bax expression through PPARγ to limit apoptosis. In support of this, gene and protein expression analysis revealed that Mcl-1 expression is driven by PPARγ during M.tb infection in human macrophages. Further, 15-lipoxygenase (15-LOX) is critical for PPARγ activity and Mcl-1 expression. We also determined that PPARγ and 15-LOX regulate macrophage apoptosis during M.tb infection, and that pre-clinical therapeutics that inhibit Mcl-1 activity significantly limit M.tb intracellular growth in both human macrophages and an in vitro TB granuloma model. In conclusion, identification of the novel PPARγ effector Mcl-1 has determined PPARγ and 15-LOX are critical regulators of apoptosis during M.tb infection and new potential targets for host-directed therapy for M.tb.
过氧化物酶体增殖物激活受体 (PPAR)γ 是一种与抗炎作用相关的全局转录调节剂。它在肺泡巨噬细胞 (AMs) 中高度表达,AMs 无法清除细胞内病原体结核分枝杆菌 (M.tb)。尽管 M.tb 感染会诱导人巨噬细胞中的 PPARγ,这有助于 M.tb 的生长,但其中的机制在很大程度上尚不清楚。我们进行了 NanoString 基因表达分析,以确定新的 PPARγ 效应子,使巨噬细胞更容易感染 M.tb。这揭示了一些在 M.tb 感染期间通过 PPARγ 沉默而差异调节的基因,包括 Bcl-2 家族成员 Bax(促凋亡)和 Mcl-1(促生存)。细胞凋亡是一种重要的防御机制,可以防止包括 M.tb 在内的细胞内微生物的生长,但受到毒力 M.tb 的限制。这表明 M.tb 通过 PPARγ 差异调节 Mcl-1 和 Bax 的表达,以限制细胞凋亡。为此,基因和蛋白质表达分析表明,在人类巨噬细胞中,M.tb 感染期间 Mcl-1 的表达是由 PPARγ 驱动的。此外,15-脂氧合酶 (15-LOX) 对 PPARγ 活性和 Mcl-1 表达至关重要。我们还确定了 PPARγ 和 15-LOX 在 M.tb 感染期间调节巨噬细胞凋亡,并且抑制 Mcl-1 活性的临床前治疗剂可显著限制人巨噬细胞和体外结核肉芽肿模型中的 M.tb 细胞内生长。总之,鉴定新型 PPARγ 效应子 Mcl-1 确定了 PPARγ 和 15-LOX 是 M.tb 感染期间细胞凋亡的关键调节剂,为 M.tb 的宿主导向治疗提供了新的潜在靶点。