USP11 Promotes Endothelial Apoptosis-Resistance in Pulmonary Arterial Hypertension by Deubiquitinating HINT3

Abstract

Pulmonary arterial hypertension (PAH) is a progressive, lethal, and incurable disease of the pulmonary vasculature. A previous genome-wide association study (GWAS) with Affymetrix microarray analysis data exhibited elevated histidine triad nucleotide-binding protein 3 (HINT3) in the lung samples of PAH compared to control subjects (failed donors, FD) and the positive correlations of HINT3 with deubiquitinase USP11 and B-cell lymphoma 2 (BCL2). In this study, we aim to investigate the roles and interplay of USP11 and HINT3 in the apoptosis resistance of PAH. The levels of USP11 and HINT3 were increased in the lungs of idiopathic PAH (IPAH) patients and Hypoxia/Sugen-treated mice. USP11 and HINT3 interacted physically, as shown by co-immunoprecipitation (co-IP) assay in human pulmonary arterial endothelial cells (HPAECs). HINT3 was degraded by polyubiquitination, which was reversed by USP11. Furthermore, HINT3 interacted with the anti-apoptotic mediator, BCL2. Overexpression of USP11 increased BCL2 content, congruent to elevated lung tissue levels seen in IPAH patients and Hypoxia/Sugen-treated mice. Conversely, the knockdown of HINT3 function led to a depletion of BCL2. Thus, we conclude that USP11 stabilizes HINT3 activation, which contributes to endothelial apoptosis-resistance of pulmonary arterial endothelial cells in PAH. This can potentially be a novel therapeutic target for ubiquitination modulators for PAH.

Keywords

USP11 / Deubiquitination / HINT3 / BCL2 / Pulmonary hypertension / Apoptosis-resistance

Supplementary Materials

The following supporting information can be found at: https://www.sciepublish.com/article/pii/470. Supplementary Figure S1: PAH exhibits upregulated USP11, HINT3, and BCL2 expression.

Acknowledgments

Data/Tissue samples provided by PHBI under the Pulmonary Hypertension Breakthrough Initiative (PHBI). Pharmacogenomics in Pulmonary Arterial Hypertension (5RO1HL478946).

Author Contributions

Conception, hypothesis delineation, and design, B.-Y.K. and A.J.J.; acquisition of data, analysis, and interpretation, B.-Y.K., J.C., V.T., Y.Z., J.Z., R.S.S., W.A.L., V.S., B.T.K., M.J.P., C.P., J.L., R.J.B. and A.J.J.; writing of the article, B.-Y.K., J.C., V.T., Y.Z., J.Z., R.S.S., W.A.L., V.S., B.T.K., M.J.P., C.P., J.L., R.J.B. and A.J.J.

Ethics Statement

In All animal studies were approved by the Institutional Animal Care and Use Committee of Emory University or the Atlanta Veterans Affairs Healthcare System (V004-21, 19 April 2021).

Informed Consent Statement

Not applicable.

Data Availability Statement

The statement is required for all original articles which informs readers about the accessibility of research data linked to a paper and outlines the terms under which the data can be obtained.

Funding

This study was supported by funding from NIH National Heart, Lung, and Blood Institute R01 grants (HL119291 to C.P., HL157164 to Y.Z., HL151513 to J.Z., AA029690-01A1 to V.S. and B.-Y.K., and HL133053 to B.-Y.K.) and the National Center for Advancing Translational Sciences of the National Institutes of Health UL1TR002378 (B.-Y.K.). Data/Tissue samples provided by PHBI under the Pulmonary Hypertension Breakthrough Initiative (PHBI). Funding for the PHBI is provided under an NHLBI R24 grant, #R24HL123767, and by the Cardiovascular Medical Research and Education Fund (CMREF).

Declaration of Competing Interest

The content does not represent the views of the Department of Veterans Affairs or the United States Government.

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