PMID- 33579361 OWN - NLM STAT- MEDLINE DCOM- 20210705 LR - 20210705 IS - 1757-6512 (Electronic) IS - 1757-6512 (Linking) VI - 12 IP - 1 DP - 2021 Feb 12 TI - Umbilical mesenchymal stem cell-derived exosomes facilitate spinal cord functional recovery through the miR-199a-3p/145-5p-mediated NGF/TrkA signaling pathway in rats. PG - 117 LID - 10.1186/s13287-021-02148-5 [doi] LID - 117 AB - BACKGROUND: Although exosomes, as byproducts of human umbilical cord mesenchymal stem cells (hUC-MSCs), have been demonstrated to be an effective therapy for traumatic spinal cord injury (SCI), their mechanism of action remains unclear. METHODS: We designed and performed this study to determine whether exosomes attenuate the lesion size of SCI by ameliorating neuronal injury induced by a secondary inflammatory storm and promoting neurite outgrowth. We determined the absolute levels of all exosomal miRNAs and investigated the potential mechanisms of action of miR-199a-3p/145-5p in inducing neurite outgrowth in vivo and in vitro. RESULTS: miR-199a-3p/145-5p, which are relatively highly expressed miRNAs in exosomes, promoted PC12 cell differentiation suppressed by lipopolysaccharide (LPS) in vitro through modulation of the NGF/TrkA pathway. We also demonstrated that Cblb was a direct target of miR-199a-3p and that Cbl was a direct target of miR-145-5p. Cblb and Cbl gene knockdown resulted in significantly decreased TrkA ubiquitination levels, subsequently activating the NGF/TrkA downstream pathways Akt and Erk. Conversely, overexpression of Cblb and Cbl was associated with significantly increased TrkA ubiquitination level, subsequently inactivating the NGF/TrkA downstream pathways Akt and Erk. Western blot and coimmunoprecipitation assays confirmed the direct interaction between TrkA and Cblb and TrkA and Cbl. In an in vivo experiment, exosomal miR-199a-3p/145-5p was found to upregulate TrkA expression at the lesion site and also promote locomotor function in SCI rats. CONCLUSIONS: In summary, our study showed that exosomes transferring miR-199a-3p/145-5p into neurons in SCI rats affected TrkA ubiquitination and promoted the NGF/TrkA signaling pathway, indicating that hUC-MSC-derived exosomes may be a promising treatment strategy for SCI. FAU - Wang, Yang AU - Wang Y AD - Department of Spine Surgery, The Third Affiliated Hospital of Sun Yat-sen University, No. 600 Tianhe Road, Tianhe District, Guangzhou, Guangdong Province, China. AD - Guangdong Provincial Center for Engineering and Technology Research of Minimally Invasive Spine Surgery, No. 600 Tianhe Road, Tianhe District, Guangzhou, Guangdong Province, China. AD - Guangdong Provincial Center for Quality Control of Minimally Invasive Spine Surgery, No. 600 Tianhe Road, Tianhe District, Guangzhou, Guangdong Province, China. FAU - Lai, Xunwei AU - Lai X AD - Department of Spine Surgery, The Third Affiliated Hospital of Sun Yat-sen University, No. 600 Tianhe Road, Tianhe District, Guangzhou, Guangdong Province, China. AD - Guangdong Provincial Center for Engineering and Technology Research of Minimally Invasive Spine Surgery, No. 600 Tianhe Road, Tianhe District, Guangzhou, Guangdong Province, China. AD - Guangdong Provincial Center for Quality Control of Minimally Invasive Spine Surgery, No. 600 Tianhe Road, Tianhe District, Guangzhou, Guangdong Province, China. FAU - Wu, Depeng AU - Wu D AD - Department of Spine Surgery, The Third Affiliated Hospital of Sun Yat-sen University, No. 600 Tianhe Road, Tianhe District, Guangzhou, Guangdong Province, China. AD - Guangdong Provincial Center for Engineering and Technology Research of Minimally Invasive Spine Surgery, No. 600 Tianhe Road, Tianhe District, Guangzhou, Guangdong Province, China. AD - Guangdong Provincial Center for Quality Control of Minimally Invasive Spine Surgery, No. 600 Tianhe Road, Tianhe District, Guangzhou, Guangdong Province, China. FAU - Liu, Bin AU - Liu B AD - Department of Spine Surgery, The Third Affiliated Hospital of Sun Yat-sen University, No. 600 Tianhe Road, Tianhe District, Guangzhou, Guangdong Province, China. AD - Guangdong Provincial Center for Engineering and Technology Research of Minimally Invasive Spine Surgery, No. 600 Tianhe Road, Tianhe District, Guangzhou, Guangdong Province, China. AD - Guangdong Provincial Center for Quality Control of Minimally Invasive Spine Surgery, No. 600 Tianhe Road, Tianhe District, Guangzhou, Guangdong Province, China. FAU - Wang, Nanxiang AU - Wang N AD - Department of Spine Surgery, The Third Affiliated Hospital of Sun Yat-sen University, No. 600 Tianhe Road, Tianhe District, Guangzhou, Guangdong Province, China. AD - Guangdong Provincial Center for Engineering and Technology Research of Minimally Invasive Spine Surgery, No. 600 Tianhe Road, Tianhe District, Guangzhou, Guangdong Province, China. AD - Guangdong Provincial Center for Quality Control of Minimally Invasive Spine Surgery, No. 600 Tianhe Road, Tianhe District, Guangzhou, Guangdong Province, China. FAU - Rong, Limin AU - Rong L AUID- ORCID: 0000-0002-4049-870X AD - Department of Spine Surgery, The Third Affiliated Hospital of Sun Yat-sen University, No. 600 Tianhe Road, Tianhe District, Guangzhou, Guangdong Province, China. ronglm@mail.sysu.edu.cn. AD - Guangdong Provincial Center for Engineering and Technology Research of Minimally Invasive Spine Surgery, No. 600 Tianhe Road, Tianhe District, Guangzhou, Guangdong Province, China. ronglm@mail.sysu.edu.cn. AD - Guangdong Provincial Center for Quality Control of Minimally Invasive Spine Surgery, No. 600 Tianhe Road, Tianhe District, Guangzhou, Guangdong Province, China. ronglm@mail.sysu.edu.cn. LA - eng PT - Journal Article PT - Research Support, Non-U.S. Gov't DEP - 20210212 PL - England TA - Stem Cell Res Ther JT - Stem cell research & therapy JID - 101527581 RN - 0 (MIRN145 microRNA, rat) RN - 0 (MicroRNAs) RN - 9061-61-4 (Nerve Growth Factor) SB - IM MH - Animals MH - *Exosomes/genetics MH - *Mesenchymal Stem Cells MH - *MicroRNAs/genetics MH - Nerve Growth Factor/genetics MH - Rats MH - Signal Transduction MH - Spinal Cord MH - *Spinal Cord Injuries/genetics/therapy PMC - PMC7879635 OTO - NOTNLM OT - Exosomes OT - PC12 cells OT - Spinal cord injury OT - TrkA ubiquitination OT - Umbilical cord mesenchymal stem cells OT - microRNAs COIS- No conflicts of interest are declared by all authors. EDAT- 2021/02/14 06:00 MHDA- 2021/07/06 06:00 PMCR- 2021/02/12 CRDT- 2021/02/13 05:35 PHST- 2020/11/24 00:00 [received] PHST- 2021/01/06 00:00 [accepted] PHST- 2021/02/13 05:35 [entrez] PHST- 2021/02/14 06:00 [pubmed] PHST- 2021/07/06 06:00 [medline] PHST- 2021/02/12 00:00 [pmc-release] AID - 10.1186/s13287-021-02148-5 [pii] AID - 2148 [pii] AID - 10.1186/s13287-021-02148-5 [doi] PST - epublish SO - Stem Cell Res Ther. 2021 Feb 12;12(1):117. doi: 10.1186/s13287-021-02148-5.