Optochemogenetic Stimulation of Transplanted iPS-NPCs Enhances Neuronal Repair and Functional Recovery after Ischemic Stroke

• Type of Material: Article
• Authors: Shan Ping Yu, Emory University; Jack K. Tung, Emory University; Zheng Zachory Wei, Emory University; Dongdong Chen, Emory University; Ken Berglund, Emory University; Weiwei Zhong, Center for Visual and Neurocognitive Rehabilitation, Atlanta Veterans Affairs Medical Center; James Y. Zhang, Emory University; Xiaohuan Gu, Emory University; Mingke Song, Emory University; Robert Gross, Emory University; Shinn Z. Lin, Tzu Chi University; Ling Wei, Emory University
• Language: English
• Date: 2019-08-14
• Publisher: The Society for Neuroscience
• Publication Version: Final Published Version
• Copyright Statement: © 2019 the authors.
• License: Creative Commons Attribution 4.0 International
• Final Published Version (URL): http://dx.doi.org/10.1523/JNEUROSCI.2010-18.2019
• Title of Journal or Parent Work: The Journal of Neuroscience
• Volume: 39
• Issue: 33
• Start Page: 6571
• End Page: 6594
• Grant/Funding Information: Postdoctoral Fellowships POST12080252 to M.S. and POST25710112/CDA34110317 to Z.Z.W.; NS091585 to L.W., NS079268 to R.E.G., NS079757 to R.E.G., NS086433 to J.K.T., VA Merit Award RX000666; This work was supported by National Institutes of Health Grants NS085568 to L.W., S.P.Y., and R.E.G.; The Viral Vector Core of Emory Neuroscience National Institute of Neurological Disorders and Stroke Core Facilities was supported by National Institutes of Health Grant P30NS055077.; RX001473 to S.P.Y., National Science Foundation CBET-1512826 to K.B. and R.E.G., American Heart Association Predoctoral Fellowship PRE31230001 to J.Y.Z
• Keywords: Intranasal delivery; optogenetics; Neurosciences & Neurology; optochemogenetics; Optogenetic control; stroke; Life Sciences & Biomedicine; Neurosciences; Neurothropic factor; Electrical stimulation; Neurite outgrowth; iPS cells; Science & Technology; In-vitro; Mesenchymal stem-cells; Mouse; neuronal repair; functional recovery; Promotes; Brain
• Research Categories: Health Sciences, Medicine and Surgery; Health Sciences, Rehabilitation and Therapy; Biology, Neuroscience

Abstract

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.SIGNIFICANCE STATEMENT Neural network reconnection is critical for repairing damaged brain. Strategies that promote this repair are expected to improve functional outcomes. This study pioneers the generation and application of an optochemogenetics approach in stem cell transplantation therapy after stroke for optimal neural repair and functional recovery. Using induced pluripotent stem cell-derived neural progenitor cells (iPS-NPCs) expressing the novel optochemogenetic probe luminopsin (LMO3), and intranasally delivered luciferase substrate coelenterazine, we show enhanced regenerative properties of LMO3-iPS-NPCs in vitro and after transplantation into the ischemic brain of different genders and ages. The noninvasive repeated coelenterazine stimulation of transplanted cells is feasible for clinical applications. The synergetic effects of the combinatorial cell therapy may have significant impacts on regenerative approach for treatments of CNS injuries.