Rolland Maude, Li Xiaojun, Sellier Yann, Martin Hélène, Perez-Berezo Teresa, Rauwel Benjamin, Benchoua Alexandra, Bessières Bettina, Aziza Jacqueline, Cenac Nicolas, Luo Minhua, Casper Charlotte, Peschanski Marc, Gonzalez-Dunia Daniel, Leruez-Ville Marianne, Davrinche Christian, Chavanas Stéphane
Centre de Physiopathologie Toulouse Purpan, INSERM UMR 1043, CNRS UMR 5282, Université Paul Sabatier, Toulouse, France.
Laboratory of Virology, Wuhan Institute of Virology, Chinese Academy of Sciences, Wuhan, China.
PLoS Pathog. 2016 Apr 14;12(4):e1005547. doi: 10.1371/journal.ppat.1005547. eCollection 2016 Apr.
Congenital infection by human cytomegalovirus (HCMV) is a leading cause of permanent sequelae of the central nervous system, including sensorineural deafness, cerebral palsies or devastating neurodevelopmental abnormalities (0.1% of all births). To gain insight on the impact of HCMV on neuronal development, we used both neural stem cells from human embryonic stem cells (NSC) and brain sections from infected fetuses and investigated the outcomes of infection on Peroxisome Proliferator-Activated Receptor gamma (PPARγ), a transcription factor critical in the developing brain. We observed that HCMV infection dramatically impaired the rate of neuronogenesis and strongly increased PPARγ levels and activity. Consistent with these findings, levels of 9-hydroxyoctadecadienoic acid (9-HODE), a known PPARγ agonist, were significantly increased in infected NSCs. Likewise, exposure of uninfected NSCs to 9-HODE recapitulated the effect of infection on PPARγ activity. It also increased the rate of cells expressing the IE antigen in HCMV-infected NSCs. Further, we demonstrated that (1) pharmacological activation of ectopically expressed PPARγ was sufficient to induce impaired neuronogenesis of uninfected NSCs, (2) treatment of uninfected NSCs with 9-HODE impaired NSC differentiation and (3) treatment of HCMV-infected NSCs with the PPARγ inhibitor T0070907 restored a normal rate of differentiation. The role of PPARγ in the disease phenotype was strongly supported by the immunodetection of nuclear PPARγ in brain germinative zones of congenitally infected fetuses (N = 20), but not in control samples. Altogether, our findings reveal a key role for PPARγ in neurogenesis and in the pathophysiology of HCMV congenital infection. They also pave the way to the identification of PPARγ gene targets in the infected brain.
人巨细胞病毒(HCMV)的先天性感染是中枢神经系统永久性后遗症的主要原因,包括感音神经性耳聋、脑瘫或严重的神经发育异常(占所有出生病例的0.1%)。为深入了解HCMV对神经元发育的影响,我们使用了来自人类胚胎干细胞的神经干细胞(NSC)以及受感染胎儿的脑切片,并研究了感染对过氧化物酶体增殖物激活受体γ(PPARγ)的影响,PPARγ是发育中的大脑中一种关键的转录因子。我们观察到,HCMV感染显著损害了神经发生的速率,并强烈增加了PPARγ的水平和活性。与这些发现一致,在受感染的神经干细胞中,已知的PPARγ激动剂9-羟基十八碳二烯酸(9-HODE)的水平显著增加。同样,将未感染的神经干细胞暴露于9-HODE可重现感染对PPARγ活性的影响。它还增加了HCMV感染的神经干细胞中表达IE抗原的细胞比例。此外,我们证明:(1)异位表达的PPARγ的药理学激活足以诱导未感染神经干细胞的神经发生受损;(2)用9-HODE处理未感染的神经干细胞会损害神经干细胞的分化;(3)用PPARγ抑制剂T0070907处理HCMV感染的神经干细胞可恢复正常的分化速率。在先天性感染胎儿(N = 20)的脑生发区中检测到核PPARγ的免疫反应,但在对照样本中未检测到,这有力地支持了PPARγ在疾病表型中的作用。总之,我们的发现揭示了PPARγ在神经发生以及HCMV先天性感染的病理生理学中的关键作用。它们还为识别受感染大脑中的PPARγ基因靶点铺平了道路。