Bioactive extracellular vesicles from a subset of endothelial progenitor cells rescue retinal ischemia and neurodegeneration
Kyle V. Marra, … , Susumu Sakimoto, Martin Friedlander
Kyle V. Marra, … , Susumu Sakimoto, Martin Friedlander
Published May 31, 2022
Citation Information: JCI Insight. 2022;7(12):e155928. https://doi.org/10.1172/jci.insight.155928.
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Research Article Ophthalmology Vascular biology

Bioactive extracellular vesicles from a subset of endothelial progenitor cells rescue retinal ischemia and neurodegeneration

Abstract

Disruption of the neurovascular unit (NVU) underlies the pathophysiology of various CNS diseases. One strategy to repair NVU dysfunction uses stem/progenitor cells to provide trophic support to the NVU’s functionally coupled and interdependent vasculature and surrounding CNS parenchyma. A subset of endothelial progenitor cells, endothelial colony-forming cells (ECFCs) with high expression of the CD44 hyaluronan receptor (CD44hi), provides such neurovasculotrophic support via a paracrine mechanism. Here, we report that bioactive extracellular vesicles from CD44hi ECFCs (EVshi) are paracrine mediators, recapitulating the effects of intact cell therapy in murine models of ischemic/neurodegenerative retinopathy; vesicles from ECFCs with low expression levels of CD44 (EVslo) were ineffective. Small RNA sequencing comparing the microRNA cargo from EVshi and EVslo identified candidate microRNAs that contribute to these effects. EVshi may be used to repair NVU dysfunction through multiple mechanisms to stabilize hypoxic vasculature, promote vascular growth, and support neural cells.

Authors

Kyle V. Marra, Edith Aguilar, Guoqin Wei, Ayumi Usui-Ouchi, Yoichiro Ideguchi, Susumu Sakimoto, Martin Friedlander

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Figure 7

Differentially expressed miRs are neurovasculotrophic and contribute to the in vivo rescue effects of EVshi.

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Differentially expressed miRs are neurovasculotrophic and contribute to ...
(A) Heatmap of differentially expressed (q < 0.05) miRs on small RNA sequencing of EVshi (H, n = 2) and EVslo (L, n = 3). (B) Injection of miR mimics upregulated in EVshi in A rescued NV (miR-7-5p, miR-26a-3p miR-30a-5p, miR-216a-3p, miR-381-3p, miR-503-5p) and VO (miR-30a-5p, miR-216a-3p, miR-503-5p) compared to scrmiR-injected controls. n = 12–16 retinas for miR mimics, n = 72 retinas for scrmiR. (C) Combinatorial injection of miR mimics rescued OIR mice dose-dependently. “Candidate miRs” miR-7-5p, miR-23a-3p, miR-216a-3p, and miR-503-5p were upregulated on small RNA sequencing in A, validated on RT-qPCR, and functional in rescuing OIR mice in B. Combination injection of the 2 most effective miR mimics (miR-216a-3p and miR-503-5p) in B and, separately, all candidate miR mimics rescued OIR mice. n = 9–14 retinas for miR-216a-3p and miR-503-5p, n = 11–16 retinas for all candidate miRs, n = 24 retinas for scrmiR. (D) EVs from CD44hi ECFCs with KD expression of individual or combinatorial miRs no longer rescued OIR mice. Multiple lines of ECFCs were generated with KD expression of both miR-216a-3p and miR-503-5p, all candidate miRs together, and each candidate miR individually. EVs from miR-23a-3p KD CD44hi ECFCs failed to rescue VO; EVs from miR-503-5p KD CD44hi ECFCs failed to rescue NV; and EVs from CD44hi ECFCs with KD expression of all candidate miRs failed to rescue both NV and VO in OIR mice. n = 11–20 retinas for individual miR KD, n = 7 retinas for miR-216a-3p and miR-503-5p KD, n = 12 for all candidate miR KD, n = 18 for scrmiR KD, n = 20 for PBS. One-way ANOVA with Tukey’s analysis. Error bars represent SEM. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001.