Optochemogenetic Stimulation of Transplanted iPS-NPCs Enhances Neuronal Repair and Functional Recovery after Ischemic Stroke
Optochemogenetic Stimulation of Transplanted iPS-NPCs Enhances Neuronal Repair and Functional Recovery after Ischemic Stroke
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DOI：
10.1523/jneurosci.2010-18.2019
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发表时间：
2019-08-14
影响因子：
5.3
通讯作者：
Wei, Ling
中科院分区：
文献类型：
--
作者：
Yu, Shan Ping;Tung, Jack K.;Wei, Ling
细胞移植治疗为神经修复提供了再生策略。我们测试了一个假设，即移植的诱导多能干细胞衍生的神经祖细胞（iPS-NPC）的选择性激发可以重现一个富含活性的微环境，为中风的治疗提供再生益处。用新型光化学遗传学融合蛋白，发光蛋白3（LMO 3）转导小鼠iPS-NPC，所述发光蛋白3由生物发光荧光素酶Gaussia荧光素酶和视蛋白团藻视紫红质1组成。这些LMO 3-iPS-NPC可以通过使用光的光刺激或通过荧光素酶底物腔肠素（CTZ）来激活。LMO 3-iPS-NPC的体外刺激增加了突触蛋白-1、突触后密度95、脑源性神经营养因子（BDNF）和基质细胞衍生因子1的表达，并促进了神经突生长。移植到小鼠缺血皮质后，LMO 3-iPS-NPC分化为成熟神经元。用免疫金电子显微镜和膜片钳记录鉴定植入和宿主神经元之间的突触形成。每天鼻内给予CTZ刺激移植细胞增强轴突髓鞘形成、突触传递、改善丘脑皮质连接和功能恢复。脑片膜片钳和多电极阵列记录表明，CTZ或光刺激促进突触传递和诱导神经可塑性模仿的EPSPs的LTP。中风小鼠接受组合的LM 0 3-iPS-NPC/CTZ治疗，而不是单独的细胞或CTZ，在梗塞周围区域显示增强的神经网络连接，促进雄性和雌性、年轻和老年小鼠中风后的最佳功能恢复。因此，通过非侵入性光化学遗传学治疗激发移植细胞提供了一种新的综合细胞疗法，具有中风后的全面再生益处。
Cell transplantation therapy provides a regenerative strategy for neural repair. We tested the hypothesis that selective excitation of transplanted induced pluripotent stem cell-derived neural progenitor cells (iPS-NPCs) could recapitulate an activity-enriched microenvironment that confers regenerative benefits for the treatment of stroke. Mouse iPS-NPCs were transduced with a novel optochemogenetics fusion protein, luminopsin 3 (LMO3), which consisted of a bioluminescent luciferase, Gaussia luciferase, and an opsin, Volvox Channelrhodopsin 1. These LMO3-iPS-NPCs can be activated by either photostimulation using light or by the luciferase substrate coelenterazine (CTZ). In vitro stimulations of LMO3-iPS-NPCs increased expression of synapsin-1, postsynaptic density 95, brain derived neurotrophic factor (BDNF), and stromal cell-derived factor 1 and promoted neurite outgrowth. After transplantation into the ischemic cortex of mice, LMO3-iPS-NPCs differentiated into mature neurons. Synapse formation between implanted and host neurons was identified using immunogold electron microscopy and patch-clamp recordings. Stimulation of transplanted cells with daily intranasal administration of CTZ enhanced axonal myelination, synaptic transmission, improved thalamocortical connectivity, and functional recovery. Patch-clamp and multielectrode array recordings in brain slices showed that CTZ or light stimulation facilitated synaptic transmission and induced neuroplasticity mimicking the LTP of EPSPs. Stroke mice received the combined LMO3-iPS-NPC/CTZ treatment, but not cell or CTZ alone, showed enhanced neural network connections in the peri-infarct region, promoted optimal functional recoveries after stroke in male and female, young and aged mice. Thus, excitation of transplanted cells via the noninvasive optochemogenetics treatment provides a novel integrative cell therapy with comprehensive regenerative benefits after stroke.