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Abstract: FR-PO339

Inhibition of the lncRNA 585189 Prevents Podocyte Injury and Mitochondria Dysfunction by Promoting hnRNP A1 and SIRT1 in Diabetic Nephropathy

Session Information

Category: Diabetic Kidney Disease

  • 701 Diabetic Kidney Disease: Basic

Authors

  • Hu, Jinxiu, Shandong Provincial Hospital, Jinan, Shandong, China
  • Liu, Yue, Shandong Provincial Hospital, Jinan, Shandong, China
  • Chen, Huimin, Shandong Provincial Hospital, Jinan, Shandong, China
  • Lv, Zhimei, Shandong Provincial Hospital, Jinan, Shandong, China
  • Wang, Rong, Shandong Provincial Hospital, Jinan, Shandong, China
Background

Dysfunction of podocytes has been identified as a crucial pathologic characteristic of diabetic nephropathy (DN), while the regulatory effect of long noncoding RNAs (lncRNAs) in this process has not been fully elucidated.

Methods

We conducted RNA-seq on renal tissues and identified a significantly upregulated lncRNA ENST00000585189.1 (lncRNA 585189) in patients with DN. Subsequently, we assessed its correlation with clinical indicators and demonstrated its localization with RNA-FISH. In high-glucose-induced human podocytes, the expression of lncRNA 585189 was assessed via real-time PCR and RNA-FISH. Subsequently, gain- and loss-of-function experiments detected the effect of lncRNA 585189. The expression of Desmin, ZO-1, hnRNP A1 and SIRT1 were evaluated using real-time PCR, western blot and immunofluorescence assays. Mitochondrial morphologies were evaluated using MitoTracker, while mitochondrial ROS and membrane potential were assessed using MitoSOX Red and TMRM staining. Mechanistically, bioinformatics analysis predicted the interaction between lncRNA 585189 and hnRNP A1, which was confirmed by RIP, pull-down, and EMSA assays. Moreover, the binding of hnRNP A1 to SIRT1 mRNA was validated through RIP and pull-down assays. Additionally, the stability of hnRNPA1 and SIRT1 was assessed by treatment with Cloheximide, MG-132, and Actinomycin D.

Results

lncRNA 585189 displayed a positive correlation with renal insufficiency and was found to be upregulated in both DN patients and high-glucose-induced human podocytes. Silence of lncRNA 585189 decreased the production of ROS, rescued the aberrant mitochondrial morphology and membrane potential, restored the podocyte damage caused by high glucose. Mechanistically, lncRNA 585189 binds to hnRNP A1, inducing destabilization of hnRNP A1 protein and downregulating its expression. Conversely, hnRNP A1 promotes the expression of lncRNA 585189. Furthermore, the interaction between hnRNP A1 and SIRT1 mRNA promotes the stability of SIRT1 mRNA and enhances its expression. Finally, our findings suggested that lncRNA 585189 inhibited SIRT1 expression through hnRNP A1, hindering the recovery of mitochondrial abnormalities and podocyte damage.

Conclusion

In summary, targeting lncRNA 585189 may represent a promising strategy for reversing mitochondrial dysfunction and treating DN.

Funding

  • Government Support – Non-U.S.