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Circulating miRNAs in diabetic kidney disease: case–control study and in silico analyses

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Abstract

Aims

The aim of this study was to investigate a miRNA expression profile in type 1 diabetes mellitus (T1DM) patients with DKD (cases) or without this complication (controls).

Methods

Expression of 48 miRNAs was screened in plasma of 58 T1DM patients (23 controls, 18 with moderate DKD, and 17 with severe DKD) using TaqMan Low Density Array cards (Thermo Fisher Scientific). Then, five of the dysregulated miRNAs were selected for validation in an independent sample of 10 T1DM controls and 19 patients with DKD (10 with moderate DKD and 9 with severe DKD), using RT-qPCR. Bioinformatic analyses were performed to explore the putative _target genes and biological pathways regulated by the validated miRNAs.

Results

Among the 48 miRNAs investigated in the screening analysis, 9 miRNAs were differentially expressed between DKD cases and T1DM controls. Among them, the five most dysregulated miRNAs were chosen for validation in an independent sample. In the validation sample, miR-21-3p and miR-378-3p were confirmed to be upregulated in patients with severe DKD, while miR-16-5p and miR-29a-3p were downregulated in this group compared to T1DM controls and patients with moderate DKD. MiR-503-3p expression was not validated. Bioinformatic analyses indicate that the four validated miRNAs regulate genes from PI3K/Akt, fluid shear stress and atherosclerosis, AGE-RAGE, TGF-β1, and relaxin signaling pathways.

Conclusions

Our study found four miRNAs differentially expressed in patients with severe DKD, providing significant information about the biological pathways in which they are involved.

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Funding

This study was supported by Grants from the Conselho Nacional de Desenvolvimento Científico e Tecnológico (Grant number: 482525/2013-4), Fundação de Amparo à Pesquisa do Estado do Rio Grande do Sul (Edital FAPERGS/CNPq PRONEX 12/2014: 16-2551-0000476-5), Coordenação de Aperfeiçoamento de Pessoal de Nível Superior, and Fundo de Incentivo à Pesquisa e Eventos at Hospital de Clínicas de Porto Alegre (Grant number: 14–0213). D. Crispim, L. H. Canani and T. S. Assmann are recipients of scholarships from CNPq.

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Authors and Affiliations

  1. Endocrine Division, Hospital de Clínicas de Porto Alegre, Ramiro Barcelos Street 2350, Building 12; 4th floor, Porto Alegre, Rio Grande Do Sul, 90035-003, Brazil

    Taís S. Assmann, Aline R. Costa, Luís H. Canani, Andrea C. Bauer & Daisy Crispim

  2. Postgraduation Program in Endocrinology, Faculdade de Medicina, Universidade Federal do Rio Grande do Sul, Porto Alegre, Rio Grande Do Sul, Brazil

    Taís S. Assmann, Luís H. Canani, Andrea C. Bauer & Daisy Crispim

  3. Institute of Informatics, Universidade Federal do Rio Grande do Sul, Porto Alegre, Rio Grande Do Sul, Brazil

    Mariana Recamonde-Mendoza

  4. Experimental Research Center, Hospital de Clínicas de Porto Alegre, Porto Alegre, Rio Grande Do Sul, Brazil

    Mariana Recamonde-Mendoza

  5. Instituto da Criança com Diabetes, Hospital Nossa Senhora da Conceição, Porto Alegre, Rio Grande Do Sul, Brazil

    Márcia Puñales & Balduíno Tschiedel

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  1. Taís S. Assmann
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  6. Luís H. Canani
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  7. Andrea C. Bauer
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  8. Daisy Crispim
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Contributions

TSA designed the study, acquired and analyzed the data, and drafted the manuscript. MRM performed the bioinformatics analysis and reviewed the manuscript. ACB, MP, BT, and LHC interpreted the data and reviewed the manuscript. DC supervised the study, analyzed the data, and drafted the manuscript. All authors approved the final version.

Corresponding author

Correspondence to Daisy Crispim.

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All authors declare no conflict of interest.

Ethical approval

All procedures performed in studies involving human participants were in accordance with the ethical standards of the Hospital de Clínicas de Porto Alegre research committee (number of approval 14–0213) and with the 1964 Helsinki Declaration and its amendments or comparable ethical standards.

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Informed consent was obtained from all individual participants included in the study.

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Assmann, T.S., Recamonde-Mendoza, M., Costa, A.R. et al. Circulating miRNAs in diabetic kidney disease: case–control study and in silico analyses. Acta Diabetol 56, 55–65 (2019). https://doi.org/10.1007/s00592-018-1216-x

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