Human sensory neurons derived from induced pluripotent stem cells support varicella-zoster virus infection.
Human sensory neurons derived from induced pluripotent stem cells support varicella-zoster virus infection.
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DOI：
10.1371/journal.pone.0053010
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发表时间：
2012
期刊：
影响因子：
3.7
通讯作者：
Levin MJ
中科院分区：
文献类型：
--
作者：
Lee KS;Zhou W;Scott-McKean JJ;Emmerling KL;Cai GY;Krah DL;Costa AC;Freed CR;Levin MJ
初次感染后，水痘-带状疱疹病毒（VZV）在背根和三叉神经节的神经元中建立潜伏期。关于VZV发病机制的许多问题仍然没有答案，部分原因是严格的宿主嗜性和从尸检和流产中获得的人体组织的不一致性。近年来诱导多能干细胞（iPS）的发展为许多疾病的研究提供了巨大的潜力。我们先前通过用非整合腺病毒引入四个重编程基因从皮肤成纤维细胞产生人iPS细胞。在这项研究中，我们开发了一种新的协议，从iPS细胞产生感觉神经元。将人iPS细胞暴露于小分子抑制剂10天，其有效地将多能细胞转化为神经祖细胞（NPC）。然后将NPC暴露于转化为感觉神经元所需的生长因子中两周。通过免疫细胞化学、流式细胞术、RT-qPCR和电生理学表征iPS细胞衍生的感觉神经元。分化后，约80%的总细胞群表达神经元特异性蛋白β III-微管蛋白。重要的是，总细胞群的15%共表达标记物Brn 3a和外周蛋白，表明这些细胞是感觉神经元。这些感觉神经元可以被VZV和一种相关的α疱疹病毒单纯疱疹病毒（HSV）感染。由于有限的神经元群体能够支持整个VZV和HSV生命周期，我们的iPS衍生的感觉神经元模型可能被证明对研究α疱疹病毒潜伏期和再激活有用。
After primary infection, varicella-zoster virus (VZV) establishes latency in neurons of the dorsal root and trigeminal ganglia. Many questions concerning the mechanism of VZV pathogenesis remain unanswered, due in part to the strict host tropism and inconsistent availability of human tissue obtained from autopsies and abortions. The recent development of induced pluripotent stem (iPS) cells provides great potential for the study of many diseases. We previously generated human iPS cells from skin fibroblasts by introducing four reprogramming genes with non-integrating adenovirus. In this study, we developed a novel protocol to generate sensory neurons from iPS cells. Human iPS cells were exposed to small molecule inhibitors for 10 days, which efficiently converted pluripotent cells into neural progenitor cells (NPCs). The NPCs were then exposed for two weeks to growth factors required for their conversion to sensory neurons. The iPS cell-derived sensory neurons were characterized by immunocytochemistry, flow cytometry, RT-qPCR, and electrophysiology. After differentiation, approximately 80% of the total cell population expressed the neuron-specific protein, βIII-tubulin. Importantly, 15% of the total cell population co-expressed the markers Brn3a and peripherin, indicating that these cells are sensory neurons. These sensory neurons could be infected by both VZV and herpes simplex virus (HSV), a related alphaherpesvirus. Since limited neuronal populations are capable of supporting the entire VZV and HSV life cycles, our iPS-derived sensory neuron model may prove useful for studying alphaherpesvirus latency and reactivation.