Comprehensive search for novel circulating mirnas and axon guidance pathway proteins associated with risk of eskd in diabetes

Eiichiro Satake, Pierre Jean Saulnier, Hiroki Kobayashi, Manoj K. Gupta, Helen C. Looker, Jonathan M. Wilson, Zaipul I. Md Dom, Katsuhito Ihara, Kristina O'Neil, Bozena Krolewski, Caterina Pipino, Meda E. Pavkov, Viji Nair, Markus Bitzer, Monika A. Niewczas, Matthias Kretzler, Michael Mauer, Alessandro Doria, Behzad Najafian, Rohit N. KulkarniKevin L. Duffin, Marcus G. Pezzolesi, C. Ronald Kahn, Robert G. Nelson, Andrzej S. Krolewski

Research output: Contribution to journalArticlepeer-review

18 Scopus citations


Background Mechanisms underlying the pro gression of diabetic kidney disease to ESKD are not fully understood. Methods We performed global microRNA (miRNA) analysis on plasma from two cohorts consisting of 375 individualswith type 1 andtype 2 diabeteswith latediabetickidney disease, andtargetedproteomics analysis on plasma from four cohorts consisting of 746 individuals with late and early diabetic kidney disease. We examined structural lesions in kidney biopsy specimens from the 105 individuals with early diabetic kidney disease.Human umbilical vein endothelial cellswere used to assess the effects of miRNAmimics or inhibitors on regulation of candidate proteins. Results In the late diabetic kidney disease cohorts, we identified 17 circulating miRNAs, represented by four exemplars (miR-1287-5p, miR-197-5p, miR-339-5p, and miR-328-3p), that were strongly associated with 10-year risk of ESKD. These miRNAs targeted proteins in the axon guidance pathway. Circulating levels of six of these proteins-most notably, EFNA4 and EPHA2-were strongly associated with 10-year risk of ESKD in all cohorts. Furthermore, circulating levels of these proteins correlated with severity of structural lesions in kidney biopsy specimens. In contrast, expression levels of genes encoding these proteins had no apparent effects on the lesions. In in vitro experiments,mimics ofmiR-1287-5p andmiR-197-5p and inhibitors ofmiR-339-5p andmiR-328-3p upregulated concentrations of EPHA2 in either cell lysate, supernatant, or both. Conclusions This study reveals novel mechanisms involved in progression to ESKD and points to the importance of systemic factors in the development of diabetic kidney disease. Some circulatingmiRNAs and axon guidance pathway proteins represent potential targets for newtherapies to prevent and treat this condition.

Original languageEnglish (US)
Pages (from-to)2331-2351
Number of pages21
JournalJournal of the American Society of Nephrology
Issue number9
StatePublished - Sep 2021

Bibliographical note

Funding Information:
We acknowledge support from the NIH Clinical Center grants DK041526 (to A.S. Krolewski), and DK110350 (to M.G. Pezzolesi), and DK100449 (to M. Bitzer); Novo Nordisk Foundation grant NNF14OC0013659 (PROTON; to A.S. Krolewski); JDRF grants 3-SRA-2015-106-Q-R (to A. S. Krolewski) and 5-CDA-2015-89-A-B (to M. A. Niewczas); the Mary K. Iacocca Fellowship; the Sunstar Foundation; the Hiroo Kaneda Scholarship; the Foundation for Growth Science of Japan (to E. Satake); the Uehara Memorial Foundation (postdoctoral fellowship); and the Japan Society for the Promotion of Science (overseas research fellowship; to H. Kobayashi). This research was also supported by the American Diabetes Association Clinical Science Award 1-08-CR-42 (to R. G. Nelson), by the Centers for Disease Control and Prevention Interagency Agreement 16FED1604631 (to R. G. Nelson), by the National Institute of Diabetes and Digestive and Kidney Diseases Intramural Research Program, and by the NIH Clinical Center, Diabetes Endocrinology Research Center grant P30 DK036836 (to Joslin Diabetes Center). This study was also supported by George M. O’Brien Michigan Kidney Translational Core Center, funded by NIH Clinical C enter/NIDDK grant 2P30 DK081943.

Funding Information:
M. Bitzer reports receiving honoraria from the National Institutes of Health (NIH). A. Doria reports receiving honoraria from the American Diabetes Association (honorarium for serving as associate editor), and serving as associate editor for DIABETES. K. L. Duffin reports being employed by, and receiving research funding from, Eli Lilly and Company; and having ownership interest in Eli Lilly and Pfizer. C. R. Kahn reports serving as scientific advisor for, or member of, with Cellarity Biotech, Coh-Bar, ERX Biotech, and Kaleido Biotech; and having consultancy agreements with, ownership interest in, and honoraria from Cellarity Biotech, CohBar, ERX Biotech, Kaleido Biotech, and Sana Biotech. M. Kretzler reports receiving research funding, via a sponsored research project as principal investigator at the University of Michigan, from amfAR, Angion, AstraZe-neca, Boehringer Ingelheim, Certa, Chan Zuckerberg Initiative, Chinook, Elpidera, Gilead, Goldfinch, Ionis, Jansen, JDRF, Lilly, NIH, Novo Nordisk, Regeneron, RenalytixAI, and Travere; having consultancy agreements with Astellas, Boehringer Ingelheim, Certa, Janssen, Novo Nordisk, and Poxel (as an employee of University of Michigan); serving on the editorial boards of JASN, Kidney International,and Kidney Disease;and serving on the advisory board of NephCure Kidney International. R. N. Kulkarni reports receiving research funding from Inversago; having consultancy agreements with Inversago and REDD Pharmaceuticals; and serving as a scientificadvi-sor for, ormemberof, Novo Nordisk. M. Mauerreports having consultancy agreements with Amicus, Avrobio, Bayer, Boeheringer Ingelheim, Freeline Theraputics, Sangano, and Sanofi/Genzyme; receiving honoraria from Amicus, Freeline Therapeutics, and Sanofi/Genzyme; receiving research funding from Amicus and Sanofi/Genzyme; and serving on the North American Fabry Registry Board. B. Najafian reports having consultancy agreements with Amicus, Avrobio, 4D Molecular Therapeutics, Freeline Therapeutics, Sangamo, and Sanofi; serving as a scientific advisor for, or member of, Amicus, Freeline Therapeutics, and Sanofi; and receiving research funding and honoraria from Amicus and Sanofi. M. A. Niewczas reports receiving honoraria from Indiana Diabetes Research Center (honorarium for the pilot and feasibility grant review), and serving as an editorial board member for Journal of Diabetes Research. M. E. Pavkov reports serving as a member of the Kidney Health Initiative Board of Directors. E. Satake repo rts receiving research funding from Novo Nordisk and Sunstar Foundation. J. M. Wilson reports being employed by Lilly. All remaining authors have nothing to disclose.

Publisher Copyright:
© 2021 by the American Society of Nephrology.


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