Journal Article

Young Sca-1+ bone marrow stem cell-derived exosomes preserve visual function via the miR-150-5p/MEKK3/JNK/c-Jun pathway to reduce M1 microglial polarization

Ye, Xin-qi, Bai, Ying, Zhou, Yu-hang, Liu, Xin-na, Yuan, Hui-ping, Wang, Yuan, Wang, Xin-lin, Li, Xiang-hui, Shao, Zheng-bo, Wang, Qi, Zhang, Yan, Liu, Pan, Qin, Wan-yun

2023

Overview

Background Polarization of microglia, the resident retinal immune cells, plays important roles in mediating both injury and repair responses post-retinal ischemia–reperfusion (I/R) injury, which is one of the main pathological mechanisms behind ganglion cell apoptosis. Aging could perturb microglial balances, resulting in lowered post-I/R retinal repair. Young bone marrow (BM) stem cell antigen 1-positive (Sca-1 + ) cells have been demonstrated to have higher reparative capabilities post-I/R retinal injury when transplanted into old mice, where they were able to home and differentiate into retinal microglia. Methods Exosomes were enriched from young Sca-1 + or Sca-1 − cells, and injected into the vitreous humor of old mice post-retinal I/R. Bioinformatics analyses, including miRNA sequencing, was used to analyze exosome contents, which was confirmed by RT-qPCR. Western blot was then performed to examine expression levels of inflammatory factors and underlying signaling pathway proteins, while immunofluorescence staining was used to examine the extent of pro-inflammatory M1 microglial polarization. Fluoro-Gold labelling was then utilized to identify viable ganglion cells, while H&E staining was used to examine retinal morphology post-I/R and exosome treatment. Results Sca-1 + exosome-injected mice yielded better visual functional preservation and lowered inflammatory factors, compared to Sca-1 − , at days 1, 3, and 7 days post-I/R. miRNA sequencing found that Sca-1 + exosomes had higher miR-150-5p levels, compared to Sca-1 − exosomes, which was confirmed by RT-qPCR. Mechanistic analysis found that miR-150-5p from Sca-1 + exosomes repressed the mitogen-activated protein kinase kinase kinase 3 (MEKK3)/JNK/c-Jun axis, leading to IL-6 and TNF-α downregulation, and subsequently reduced microglial polarization, all of which contributes to reduced ganglion cell apoptosis and preservation of proper retinal morphology. Conclusion This study elucidates a potential new therapeutic approach for neuroprotection against I/R injury, via delivering miR-150-5p-enriched Sca-1 + exosomes, which targets the miR-150-5p/MEKK3/JNK/c-Jun axis, thereby serving as a cell-free remedy for treating retinal I/R injury and preserving visual functioning.

• Publisher: BioMed Central,BioMed Central Ltd,Springer Nature B.V,BMC
• Subject: Analysis / Animals / Antigens / Apoptosis / Bioinformatics / Biotechnology / Bone marrow / Bone Marrow Cells - metabolism / Bone marrow Sca-1+ cell / c-Jun protein / Chemistry / Chemistry and Materials Science / Complications and side effects / Cytokines / Cytology / Exosomes / Exosomes - metabolism / Ganglion cells / Health aspects / Immune system / Immunofluorescence / Inflammation / Injuries / Interleukin 6 / Ischemia / Ischemia/reperfusion injury / Kinases / Labeling / Laboratory animals / Light / MAP kinase / MEKK3 / Mice / Microbalances / Microglia / Microglia - metabolism / MicroRNA / MicroRNAs / MicroRNAs - metabolism / miR-150-5p / miRNA / Molecular Medicine / Morphology / Nanotechnology / Neuroprotection / Polarization / Protein kinases / Proteins / Reperfusion / Reperfusion Injury - metabolism / Retina / Signal transduction / Staining / Stem cell antigen 1 / Stem cells / Transcription factors / Tumor necrosis factor-TNF / Visual perception / Vitreous humour