Loss of Histone H3 K79 Methyltransferase Dot1l Facilitates Kidney Fibrosis by Upregulating Endothelin 1 through Histone Deacetylase 2.

Abstract

BACKGROUND:

The progression rate of CKD varies substantially among patients. The genetic and epigenetic contributions that modify how individual patients respond to kidney injury are largely unknown. Emerging evidence has suggested that histone H3 K79 methyltransferase Dot1l has an antifibrotic effect by repressing Edn1, which encodes endothelin 1 in the connecting tubule/collecting duct.

METHODS:

To determine if deletion of the Dot1l gene is a genetic and epigenetic risk factor through regulating Edn1, we studied four groups of mice: wild-type mice, connecting tubule/collecting duct-specific Dot1l conditional knockout mice (Dot1lAC ), Dot1l and Edn1 double-knockout mice (DEAC ), and Edn1 connecting tubule/collecting duct-specific conditional knockout mice (Edn1AC ), under three experimental conditions (streptozotocin-induced diabetes, during normal aging, and after unilateral ureteral obstruction). We used several approaches (colocalization, glutathione S-transferase pulldown, coimmunoprecipitation, yeast two-hybrid, gel shift, and chromatin immunoprecipitation assays) to identify and confirm interaction of Dot1a (the major Dot1l splicing variant in the mouse kidney) with histone deacetylase 2 (HDAC2), as well as the function of the Dot1a-HDAC2 complex in regulating Edn1 transcription.

RESULTS:

In each case, Dot1lAC mice developed more pronounced kidney fibrosis and kidney malfunction compared with wild-type mice. These Dot1lAC phenotypes were ameliorated in the double-knockout DEAC mice. The interaction between Dot1a and HDAC2 prevents the Dot1a-HDAC2 complex from association with DNA, providing a counterbalancing mechanism governing Edn1 transcription by modulating H3 K79 dimethylation and H3 acetylation at the Edn1 promoter.

CONCLUSIONS:

Our study confirms Dot1l to be a genetic and epigenetic modifier of kidney fibrosis, reveals a new mechanism regulating Edn1 transcription by Dot1a and HDAC2, and reinforces endothelin 1 as a therapeutic target of kidney fibrosis.

• Authors: Long Zhang, Lihe Chen, Chao Gao, Enuo Chen, Andrea R Lightle, Llewellyn Foulke, Bihong Zhao, Paul J Higgins, Wenzheng Zhang
• Journal: Journal of the American Society of Nephrology : JASN (J Am Soc Nephrol) Vol. 31 Issue 2 Pg. 337-349 (02 2020) ISSN: 1533-3450 [Electronic] United States
• PMID: 31843983 (Publication Type: Journal Article, Research Support, N.I.H., Extramural, Research Support, Non-U.S. Gov't)
• Copyright: Copyright © 2020 by the American Society of Nephrology.

Chemical References

• Endothelin-1
• Histones
• DNA
• Dot1l protein, mouse
• Histone-Lysine N-Methyltransferase
• Histone Deacetylase 2

Topics

• Age Factors
• Animals
• Cell Nucleus (metabolism)
• Cells, Cultured
• DNA (metabolism)
• Endothelin-1 (genetics)
• Fibrosis
• Histone Deacetylase 2 (physiology)
• Histone-Lysine N-Methyltransferase (physiology)
• Histones (metabolism)
• Kidney (pathology)
• Mice
• Mice, Inbred C57BL
• Promoter Regions, Genetic
• Up-Regulation