ENSMUST00000147869 regulates proliferation and fibrosis of mesangial cells in diabetic nephropathy by interacting with Hspa9.

Abstract

Our previous study showed that ENSMUST00000147869 was abnormally low expressed in the early stage of diabetic nephropathy (DN). ENSMUST00000147869 could inhibit the fibrosis and proliferation of mouse mesangial cells (MMCs), but the mechanism is still unclear. This study aims to explore the specific mechanism underline ENSMUST00000147869 regulates the proliferation and fibrosis of MMCs in DN. Nucleocytoplasmic fractionation was applied to define the location of ENSMUST00000147869 in MMCs. RNA-protein pulldown, RNA immunoprecipitation and mass spectrometry were used to identify upregulated Hspa9 directly interacting with ENSMUST00000147869. SiRNA and lentivirus packaging were used to clarify the role of Hspa9 downregulated by ENSMUST00000147869 in promoting proliferation and fibrosis in MMCs. CHX and MG132 were used to clarify the regulatory role of ENSMUST00000147869 to Hspa9. Immunoprecipitation confirmed the binding of Hspa9 and HMGB1. HSPA9 was a direct binding protein of ENSMUST00000147869, and ENSMUST00000147869 could inhibit proliferation and fibrosis of MMCs by down-regulating HSPA9 through ubiquitination process. HMGB1 was the downstream binding protein of Hspa9, and ENSMUST00000147869 could inhibit the interaction between Hspa9 and HMGB1. Our data showed that ENSMUST00000147869 regulates Hspa9 through the ubiquitin proteasome pathway and inhibits the binding of Hspa9 and HMGB1. The ENSMUST00000147869/Hspa9/HMGB1 axis may act as a diagnostic molecular marker and an effective therapeutic target for DN.

• Authors: Min Wang, Xin Chen, Henglu Zhang, Lanlan Li, Yang Xu, Weiping Lu, Yibing Lu
• Journal: IUBMB life (IUBMB Life) Vol. 74 Issue 5 Pg. 419-432 (05 2022) ISSN: 1521-6551 [Electronic] England
• PMID: 35103378 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)
• Copyright: © 2022 International Union of Biochemistry and Molecular Biology.

Chemical References

• HMGB1 Protein
• HSP70 Heat-Shock Proteins
• HSPA9 protein, human
• Mitochondrial Proteins
• RNA, Small Interfering

Topics

• Animals
• Cell Proliferation (genetics)
• Diabetes Mellitus (pathology)
• Diabetic Nephropathies (genetics, metabolism)
• Female
• Fibrosis
• HMGB1 Protein (metabolism)
• HSP70 Heat-Shock Proteins (genetics, metabolism)
• Humans
• Male
• Mesangial Cells (metabolism, pathology)
• Mice
• Mitochondrial Proteins (metabolism)
• RNA, Small Interfering (metabolism)