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Satellite cell-derived exosome-mediated delivery of microRNA-23a/27a/26a cluster ameliorates the renal tubulointerstitial fibrosis in mouse diabetic nephropathy

Acta Pharmacologica Sinica volume 44, pages 2455–2468 (2023)Cite this article

Abstract

Renal tubulointerstitial fibrosis (TIF) is considered as the final convergent pathway of diabetic nephropathy (DN) without effective therapies currently. MiRNAs play a key role in fibrotic diseases and become promising therapeutic targets for kidney diseases, while miRNA clusters, formed by the cluster arrangement of miRNAs on chromosomes, can regulate diverse biological functions alone or synergistically. In this study, we developed clustered miR-23a/27a/26a-loaded skeletal muscle satellite cells–derived exosomes (Exos) engineered with RVG peptide, and investigated their therapeutic efficacy in a murine model of DN. Firstly, we showed that miR-23a-3p, miR-26a-5p and miR-27a-3p were markedly decreased in serum samples of DN patients using miRNA sequencing. Meanwhile, we confirmed that miR-23a-3p, miR-26a-5p and miR-27a-3p were primarily located in proximal renal tubules and highly negatively correlated with TIF in db/db mice at 20 weeks of age. We then engineered RVG-miR-23a/27a/26a cluster loaded Exos derived from muscle satellite cells, which not only enhanced the stability of miR-23a/27a/26a cluster, but also efficiently delivered more miR-23a/27a/26a cluster homing to the injured kidney. More importantly, administration of RVG-miR-23a/27a/26a-Exos (100 μg, i.v., once a week for 8 weeks) significantly ameliorated tubular injury and TIF in db/db mice at 20 weeks of age. We revealed that miR-23a/27a/26a-Exos enhanced antifibrotic effects by repressing miRNA cluster-targeting Lpp simultaneously, as well as miR-27a-3p-targeting Zbtb20 and miR-26a-5p-targeting Klhl42, respectively. Knockdown of Lpp by injection of AAV-Lpp-RNAi effectively ameliorated the progression of TIF in DN mice. Taken together, we established a novel kidney-targeting Exo-based delivery system by manipulating the miRNA-23a/27a/26a cluster to ameliorate TIF in DN, thus providing a promising therapeutic strategy for DN.

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Fig. 1: Downregulated miR-23a-3p, miR-26a-5p, and miR-27a-3p were associated with TIF in DN.
Fig. 2: Engineered RVG-modified Exos homing to injured kidneys.
Fig. 3: RVG-miR-23a/27a/26a-Exos attenuate TIF in the mice with DN.
Fig. 4: RVG-miR-23a/27a/26a-Exos inhibit ECM accumulation in HG-treated TECs.
Fig. 5: Synergistic antifibrotic activity of RVG-miR-23a/27a/26a-Exos in mice with DN.
Fig. 6: RVG-cluster 3-Exos exert antifibrotic activity by targeting multiple genes.
Fig. 7: Inhibition of Lpp contributes to the recovery of TIF in the mice with DN.
Fig. 8: Schematic illustration of RVG-miR-23a/27a/26a-Exos for the treatment of DN.

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Acknowledgements

This work was supported by the National Natural Science Foundation of China (No. 81970664, 82000648, 82070735); the Natural Science Foundation of Jiangsu Province (No. BK20211385, BK20200363); the 789 Outstanding Talent Program of the Second Affiliated Hospital of Nanjing Medical University (No. 789ZYRC202080119, 789ZYRC202090251); the Science and Technology Development Foundation of Nanjing Medical University (No. NMUB2020049) and the Fundamental Research Funds for the Central Universities (No. 2242023K40046). We are grateful for the help provided by Lab Center, the Second Affiliated Hospital of Nanjing Medical University and Institute of Nephrology, Southeast University.

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  1. These authors contributed equally: Jia-ling Ji, Hui-min Shi, Zuo-lin Li

Authors and Affiliations

  1. Department of Pediatric Nephrology, the Second Affiliated Hospital of Nanjing Medical University, Nanjing, 210003, China

    Jia-ling Ji, Hui-min Shi, Ran Jin, Gao-ting Qu, Hui Zheng, E. Wang, Yun-yang Qiao, Xing-yue Li & Wei-hua Gan

  2. Institute of Nephrology, Zhong Da Hospital, Southeast University School of Medicine, Nanjing, 210009, China

    Zuo-lin Li & Bin Wang

  3. Department of Pediatrics, the Fourth Affiliated Hospital of Nanjing Medical University, Nanjing, 210031, China

    Ling Ding & Ai-qing Zhang

  4. Department of Endocrinology, the Second Affiliated Hospital of Nanjing Medical University, Nanjing, 210003, China

    Da-fa Ding

  5. Department of Urology, the Second Affiliated Hospital of Nanjing Medical University, Nanjing, 210003, China

    Liu-cheng Ding

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Contributions

AQZ and BW designed the experiments. JLJ analyzed and interpreted the data and wrote the paper. JLJ, HMS, and ZLL conducted the experiments and analyzed the data. RJ drew the graphical abstract. GTQ, HZ, EW, YYQ, XYL, LD, LCD, and WHG contributed to the data analysis. DFD provided serum samples of patients.

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Correspondence to Bin Wang or Ai-qing Zhang.

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Ji, Jl., Shi, Hm., Li, Zl. et al. Satellite cell-derived exosome-mediated delivery of microRNA-23a/27a/26a cluster ameliorates the renal tubulointerstitial fibrosis in mouse diabetic nephropathy. Acta Pharmacol Sin 44, 2455–2468 (2023). https://doi.org/10.1038/s41401-023-01140-4

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