PMID- 35547766 OWN - NLM STAT- MEDLINE DCOM- 20220517 LR - 20220716 IS - 1838-7640 (Electronic) IS - 1838-7640 (Linking) VI - 12 IP - 7 DP - 2022 TI - hUC-MSC-mediated recovery of subacute spinal cord injury through enhancing the pivotal subunits beta3 and gamma2 of the GABA(A) receptor. PG - 3057-3078 LID - 10.7150/thno.72015 [doi] AB - Rationale: Spinal cord injury (SCI) remains an incurable neurological disorder leading to permanent and profound neurologic deficits and disabilities. Human umbilical cord-derived mesenchymal stem cells (hUC-MSCs) are particularly appealing in SCI treatment to curtail damage, restore homeostasis and possible neural relay. However, the detailed mechanisms underlying hUC-MSC-mediated functional recovery of SCI have not been fully elucidated. The purpose of our current study is to identify novel therapeutic targets and depict the molecular mechanisms underlying the hUC-MSC-mediated recovery of subacute SCI. Methods: Adult female rats suffering from subacute incomplete thoracic SCI were treated with intrathecal transplantation of hUC-MSCs. The beneficial effects of hUC-MSCs on SCI repair were evaluated by a series of behavioral analyses, motor evoked potentials (MEPs) recording of hindlimb and immunohistochemistry. We carried out extensive transcriptome comparative analyses of spinal cord tissues at the lesion site from the subacute phase of SCI (sub-SCI) either treated without (+PBS) or with hUC-MSCs (+MSC) at 0 (sub-SCI), 1, 2, and 4 weeks post-transplantation (wpt), as well as normal spinal cord segments of intact/sham rats (Intact). Adeno-associated virus (AAV)-mediated neuron-specific expression system was employed to functionally screen specific gamma-aminobutyric acid type A receptor (GABA(A)R) subunits promoting the functional recovery of SCI in vivo. The mature cortical axon scrape assay and transplantation of genetically modified MSCs with either overexpression or knockdown of brain-derived neurotrophic factor (BDNF) were employed to demonstrate that hUC-MSCs ameliorated the reduction of GABA(A)R subunits in the injured spinal cord via BDNF secretion in vitro and in vivo, respectively. Results: Comparative transcriptome analysis revealed the GABAergic synapse pathway is significantly enriched as a main target of hUC-MSC-activated genes in the injured spinal cord. Functional screening of the primary GABA(A)R subunits uncovered that Gabrb3 and Garbg2 harbored the motor and electrophysiological recovery-promoting competence. Moreover, targeting either of the two pivotal subunits beta3 or gamma2 in combination with/without the K(+)/Cl(-) cotransporter 2 (KCC2) reinforced the therapeutic effects. Mechanistically, BDNF secreted by hUC-MSCs contributed to the upregulation of GABA(A)R subunits (beta3 & gamma2) and KCC2 in the injured neurons. Conclusions: Our study identifies a novel mode for hUC-MSC-mediated locomotor recovery of SCI through synergistic upregulation of GABA(A)R beta3 and gamma2 along with KCC2 by BDNF secretion, indicating the significance of restoring the excitation/inhibition balance in the injured neurons for the reestablishment of neuronal circuits. This study also provides a potential combinatorial approach by targeting the pivotal subunit beta3 or gamma2 and KCC2, opening up possibilities for efficacious drug design. CI - (c) The author(s). FAU - Cao, Tingting AU - Cao T AD - Department of Cell Biology, School of Basic Medical Sciences, Southern Medical University, Guangzhou, 510515, China. FAU - Chen, Huan AU - Chen H AD - Department of Cell Biology, School of Basic Medical Sciences, Southern Medical University, Guangzhou, 510515, China. FAU - Huang, Weiping AU - Huang W AD - Department of Cell Biology, School of Basic Medical Sciences, Southern Medical University, Guangzhou, 510515, China. FAU - Xu, Sisi AU - Xu S AD - Department of Cell Biology, School of Basic Medical Sciences, Southern Medical University, Guangzhou, 510515, China. FAU - Liu, Peilin AU - Liu P AD - Department of Cell Biology, School of Basic Medical Sciences, Southern Medical University, Guangzhou, 510515, China. FAU - Zou, Weiwei AU - Zou W AD - Department of Cell Biology, School of Basic Medical Sciences, Southern Medical University, Guangzhou, 510515, China. FAU - Pang, Mao AU - Pang M AD - Department of Spine Surgery, The Third Affiliated Hospital of Sun Yat-sen University, Guangzhou, 510515, China. AD - Guangdong Provincial Center for Engineering and Technology Research of Minimally Invasive Spine Surgery, 510630, China. AD - Guangdong Provincial Center for Quality Control of Minimally Invasive Spine Surgery, 510630, China. FAU - Xu, Ying AU - Xu Y AD - Department of Spine Surgery, The Third Affiliated Hospital of Sun Yat-sen University, Guangzhou, 510515, China. AD - Guangdong Provincial Center for Engineering and Technology Research of Minimally Invasive Spine Surgery, 510630, China. AD - Guangdong Provincial Center for Quality Control of Minimally Invasive Spine Surgery, 510630, China. FAU - Bai, Xiaochun AU - Bai X AD - Department of Cell Biology, School of Basic Medical Sciences, Southern Medical University, Guangzhou, 510515, China. FAU - Liu, Bin AU - Liu B AD - Department of Spine Surgery, The Third Affiliated Hospital of Sun Yat-sen University, Guangzhou, 510515, China. AD - Guangdong Provincial Center for Engineering and Technology Research of Minimally Invasive Spine Surgery, 510630, China. AD - Guangdong Provincial Center for Quality Control of Minimally Invasive Spine Surgery, 510630, China. FAU - Rong, Limin AU - Rong L AD - Department of Spine Surgery, The Third Affiliated Hospital of Sun Yat-sen University, Guangzhou, 510515, China. AD - Guangdong Provincial Center for Engineering and Technology Research of Minimally Invasive Spine Surgery, 510630, China. AD - Guangdong Provincial Center for Quality Control of Minimally Invasive Spine Surgery, 510630, China. FAU - Cui, Zhong-Kai AU - Cui ZK AD - Department of Cell Biology, School of Basic Medical Sciences, Southern Medical University, Guangzhou, 510515, China. FAU - Li, Mangmang AU - Li M AD - Department of Cell Biology, School of Basic Medical Sciences, Southern Medical University, Guangzhou, 510515, China. LA - eng PT - Journal Article PT - Research Support, Non-U.S. Gov't DEP - 20220328 PL - Australia TA - Theranostics JT - Theranostics JID - 101552395 RN - 0 (Brain-Derived Neurotrophic Factor) RN - 0 (Receptors, GABA-A) RN - 0 (Symporters) RN - 56-12-2 (gamma-Aminobutyric Acid) SB - IM MH - Animals MH - Brain-Derived Neurotrophic Factor/metabolism MH - Female MH - *Mesenchymal Stem Cell Transplantation MH - Rats MH - Receptors, GABA-A MH - *Spinal Cord Injuries/pathology MH - *Symporters MH - Umbilical Cord/metabolism MH - gamma-Aminobutyric Acid PMC - PMC9065192 OTO - NOTNLM OT - K+-Cl(-) cotransporter 2 OT - human umbilical cord mesenchymal stem cells OT - spinal cord injury OT - transcriptome OT - gamma-aminobutyric acid type A (GABAA) receptor COIS- Competing Interests: The authors have declared that no competing interest exists. EDAT- 2022/05/14 06:00 MHDA- 2022/05/18 06:00 PMCR- 2022/01/01 CRDT- 2022/05/13 11:17 PHST- 2022/02/14 00:00 [received] PHST- 2022/03/12 00:00 [accepted] PHST- 2022/05/13 11:17 [entrez] PHST- 2022/05/14 06:00 [pubmed] PHST- 2022/05/18 06:00 [medline] PHST- 2022/01/01 00:00 [pmc-release] AID - thnov12p3057 [pii] AID - 10.7150/thno.72015 [doi] PST - epublish SO - Theranostics. 2022 Mar 28;12(7):3057-3078. doi: 10.7150/thno.72015. eCollection 2022.