PMID- 34496892 OWN - NLM STAT- MEDLINE DCOM- 20220121 LR - 20220121 IS - 1477-3155 (Electronic) IS - 1477-3155 (Linking) VI - 19 IP - 1 DP - 2021 Sep 8 TI - Extracellular vesicles derived from CD73 modified human umbilical cord mesenchymal stem cells ameliorate inflammation after spinal cord injury. PG - 274 LID - 10.1186/s12951-021-01022-z [doi] LID - 274 AB - BACKGROUND: Spinal cord injury (SCI) is an inflammatory condition, and excessive adenosine triphosphate (ATP) is released into the extracellular space, which can be catabolized into adenosine by CD73. Extracellular vesicles have been designed as nano drug carriers in many diseases. However, their impacts on delivery of CD73 after SCI are not yet known. We aimed to construct CD73 modified extracellular vesicles and explore the anti-inflammatory effects after SCI. METHODS: CD73 engineered extracellular vesicles (CD73+ hucMSC-EVs) were firstly established, which were derived from human umbilical cord mesenchymal stem cells (hucMSCs) transduced by lentiviral vectors to upregulate the expression of CD73. Effects of CD73+ hucMSC-EVs on hydrolyzing ATP into adenosine were detected. The polarization of M2/M1 was verified by immunofluorescence. Furthermore, A2aR and A(2b)R inhibitors and A2bR knockdown cells were used to investigate the activated adenosine receptor. Biomarkers of microglia and levels of cAMP/PKA were also detected. Repetitively in vivo study, morphology staining, flow cytometry, cytokine analysis, and ELISA assay, were also applied for verifications. RESULTS: CD73+ hucMSC-EVs reduced concentration of ATP and promoted the level of adenosine. In vitro experiments, CD73+ hucMSC-EVs increased macrophages/microglia M2:M1 polarization, activated adenosine 2b receptor (A2bR), and then promoted cAMP/PKA signaling pathway. In mice using model of thoracic spinal cord contusion injury, CD73+ hucMSC-EVs improved the functional recovery after SCI through decreasing the content of ATP in cerebrospinal fluid and improving the polarization from M1 to M2 phenotype. Thus, the cascaded pro-inflammatory cytokines were downregulated, such as TNF-alpha, IL-1beta, and IL-6, while the anti-inflammatory cytokines were upregulated, such as IL-10 and IL-4. CONCLUSIONS: CD73+ hucMSC-EVs ameliorated inflammation after spinal cord injury by reducing extracellular ATP, promoting A2bR/cAMP/PKA pathway and M2/M1 polarization. CD73+ hucMSC-EVs might be promising nano drugs for clinical application in SCI therapy. CI - (c) 2021. The Author(s). FAU - Zhai, Xiao AU - Zhai X AD - Department of Orthopedics, Shanghai Changhai Hospital, Naval Medical University, Shanghai, 200433, China. FAU - Chen, Kai AU - Chen K AD - Department of Orthopedics, Shanghai Changhai Hospital, Naval Medical University, Shanghai, 200433, China. FAU - Yang, Huan AU - Yang H AD - Department of Orthopedics, Shanghai Changhai Hospital, Naval Medical University, Shanghai, 200433, China. FAU - Li, Bo AU - Li B AD - Department of Orthopedics, Shanghai Changhai Hospital, Naval Medical University, Shanghai, 200433, China. FAU - Zhou, Tianjunke AU - Zhou T AD - Basic Medicine College, Naval Medical University, Shanghai, 200433, China. FAU - Wang, Haojue AU - Wang H AD - Basic Medicine College, Naval Medical University, Shanghai, 200433, China. FAU - Zhou, Huipeng AU - Zhou H AD - Department of Orthopedics, Shanghai Changhai Hospital, Naval Medical University, Shanghai, 200433, China. FAU - Chen, Shaofeng AU - Chen S AD - Department of Orthopedics, Shanghai Changhai Hospital, Naval Medical University, Shanghai, 200433, China. FAU - Zhou, Xiaoyi AU - Zhou X AD - Department of Orthopedics, Shanghai Changhai Hospital, Naval Medical University, Shanghai, 200433, China. FAU - Wei, Xiaozhao AU - Wei X AD - Department of Orthopedics, Shanghai Changhai Hospital, Naval Medical University, Shanghai, 200433, China. weixianzhao@126.com. FAU - Bai, Yushu AU - Bai Y AD - Department of Orthopedics, Shanghai Changhai Hospital, Naval Medical University, Shanghai, 200433, China. spinebaiys@163.com. FAU - Li, Ming AU - Li M AUID- ORCID: 0000-0002-6245-4468 AD - Department of Orthopedics, Shanghai Changhai Hospital, Naval Medical University, Shanghai, 200433, China. limingchspine@126.com. LA - eng GR - 81701199/National Natural Science Foundation of China/ GR - ZY[2018-2020]-FWTX-2005/State Administration of Traditional Chinese Medicine of the People's Republic of China/ GR - 17441900500/Science and Technology Commission of Shanghai Municipality/ GR - 16DZ0504000/Science and Technology Commission of Shanghai Municipality/ PT - Journal Article DEP - 20210908 PL - England TA - J Nanobiotechnology JT - Journal of nanobiotechnology JID - 101152208 RN - 0 (Cytokines) RN - 0 (Receptor, Adenosine A2B) RN - 8L70Q75FXE (Adenosine Triphosphate) RN - E0399OZS9N (Cyclic AMP) RN - EC 2.7.11.1 (Proto-Oncogene Proteins c-akt) RN - EC 3.1.3.5 (5'-Nucleotidase) SB - IM MH - 5'-Nucleotidase/*metabolism MH - Adenosine Triphosphate/metabolism MH - Animals MH - Cyclic AMP/metabolism MH - Cytokines/metabolism MH - Down-Regulation MH - Extracellular Vesicles/metabolism/*transplantation MH - Humans MH - Inflammation/etiology/*therapy MH - Macrophages/cytology/metabolism MH - Male MH - Mesenchymal Stem Cells/cytology/metabolism MH - Mice MH - Mice, Inbred ICR MH - Proto-Oncogene Proteins c-akt/metabolism MH - RNA Interference MH - Receptor, Adenosine A2B/chemistry/genetics/metabolism MH - Signal Transduction MH - Spinal Cord Injuries/complications/*pathology MH - Umbilical Cord/cytology PMC - PMC8425042 OTO - NOTNLM OT - CD73 OT - Extracellular vesicles OT - Inflammation OT - Mesenchymal stem cell OT - Spinal cord injury COIS- The authors do not have any possible conflicts of interests. EDAT- 2021/09/10 06:00 MHDA- 2022/01/22 06:00 PMCR- 2021/09/08 CRDT- 2021/09/09 05:46 PHST- 2021/05/19 00:00 [received] PHST- 2021/09/01 00:00 [accepted] PHST- 2021/09/09 05:46 [entrez] PHST- 2021/09/10 06:00 [pubmed] PHST- 2022/01/22 06:00 [medline] PHST- 2021/09/08 00:00 [pmc-release] AID - 10.1186/s12951-021-01022-z [pii] AID - 1022 [pii] AID - 10.1186/s12951-021-01022-z [doi] PST - epublish SO - J Nanobiotechnology. 2021 Sep 8;19(1):274. doi: 10.1186/s12951-021-01022-z.