Down-regulation of WWP2 aggravates Type 2 diabetes mellitus-induced vascular endothelial injury through modulating ubiquitination and degradation of DDX3X

Abstract

Background Endothelial injury caused by Type 2 diabetes mellitus (T2DM) is considered as a mainstay in the pathophysiology of diabetic vascular complications (DVCs). However, the molecular mechanism of T2DM-induced endothelial injury remains largely unknown. Here, we found that endothelial WW domain-containing E3 ubiquitin protein ligase 2 (WWP2) act as a novel regulator for T2DM-induced vascular endothelial injury through modulating ubiquitination and degradation of DEAD-box helicase 3 X-linked (DDX3X). Methods Single-cell transcriptome analysis was used to evaluate WWP2 expression in vascular endothelial cells of T2DM patients and healthy controls. Endothelial-specific Wwp2 knockout mice were used to investigate the effect of WWP2 on T2DM-induced vascular endothelial injury. In vitro loss- and gain-of-function studies were performed to assess the function of WWP2 on cell proliferation and apoptosis of human umbilical vein endothelial cells. The substrate protein of WWP2 was verified using mass spectrometry, coimmunoprecipitation assays and immunofluorescence assays. The mechanism of WWP2 regulation on substrate protein was investigated by pulse-chase assay and ubiquitination assay. Results The expression of WWP2 was significantly down-regulated in vascular endothelial cells during T2DM. Endothelial-specific Wwp2 knockout in mice significantly aggravated T2DM-induced vascular endothelial injury and vascular remodeling after endothelial injury. Our in vitro experiments showed that WWP2 protected against endothelial injury by promoting cell proliferation and inhibiting apoptosis in ECs. Mechanically, we found that WWP2 is down-regulated in high glucose and palmitic acid (HG/PA)-induced ECs due to c-Jun N-terminal kinase (JNK) activation, and uncovered that WWP2 suppresses HG/PA-induced endothelial injury by catalyzing K63-linked polyubiquitination of DDX3X and targeting it for proteasomal degradation. Conclusion Our studies revealed the key role of endothelial WWP2 and the fundamental importance of the JNK-WWP2-DDX3X regulatory axis in T2DM-induced vascular endothelial injury, suggesting that WWP2 may serve as a new therapeutic target for DVCs.

Identification of key genes and biological pathways in chronic obstructive pulmonary disease using bioinformatics and next generation sequencing data analysis

“…Research has shown that LMNA (lamin A/C) [314], VCAM1 [281], MAPK6 [403], DDX3X [366], SHMT2 [459], PHGDH (phosphoglycerate dehydrogenase) [560], S100A9 [433], SNCA (synuclein alpha) [335], TP63 [313], FOSL2 [1449], FHL2 [321], ATF3 [55], FOSL1 [379], DYSF (dysferlin) [475], CA4 [547], S100A8 [58] and MPO (myeloperoxidase) [521] plays an important role in the pathogenesis of lung cancer. LMNA (lamin A/C) [676], VCAM1 [657], DDX3X [708], S100A9 [745], FKBP5 [747], SNCA (synuclein alpha) [690], FOSL2 [1450], FHL2 [680], ATF3 [63], FOSL1 [718], S100A8 [67] and MPO (myeloperoxidase) [787] altered expression is associated with poor prognosis of diabetes mellitus. LMNA (lamin A/C) [676], VCAM1 [986], PHGDH (phosphoglycerate dehydrogenase) [1100], S100A9 [1045], FKBP5 [1046], FHL2 [1003], ATF3 [85] and MPO (myeloperoxidase) [1084] plays an important role in the development of obesity.…”

“…DDX3X [366], MAPK6 [403], ZFP36 [273], BHLHE40 [1451], TP63 [313], KRT17 [1452], CDC25B [556], SHMT2 [459], TRIB3 [444], PHGDH (phosphoglycerate dehydrogenase) [560], S100A8 [58], hsa-mir-410-3p [1453], hsa-mir-186-5p [1454], hsa-mir-432-5p [1455], BRCA1 [1456], FOXA1 [1457], CREB1 [1458], MAX (MYC associated factor X) [1459], ESR1 [1460], NFIC (nuclear factor I C) [1461] and STAT3 [1462] expression might be regarded as an indicator of susceptibility to lung cancer. DDX3X [708], BHLHE40 [1463], FKBP5 [747], TRIB3 [753], S100A8 [67], FOXA1 [1464], CREB1 [1465], SREBF2 [1466], ESR1 [1467] and STAT3 [1468] are a potential markers for the detection and prognosis of diabetes mellitus. A previous study reported that DDX3X [923], ZFP36 [882], FKBP5 [944], CDC25B [976], TRIB3 [948], hsa-mir-1277-5p [1469], MAX (MYC associated factor X) [1470] and STAT3 [1471] are altered expressed in respiratory viral infections.…”

Endothelial dysfunction in vascular complications of diabetes: a comprehensive review of mechanisms and implications

“…These mice also showed more widespread necrotic cores in plaques and enhanced macrophage infiltration and apoptosis in islets (56). Endothelial WW domain-containing E3 ubiquitin protein ligase 2 (WWP2) can preserve cell survival and prevent apoptosis by targeting DEAD-box helicase 3 X-linked (DDX3X) for K63-linked polyubiquitination and proteasomal degradation, thus preventing endothelial injury (57). In addition, knocking down nuclear factor erythroid 2-related factor 2 (Nrf2) increases apoptosis and hinders tube formation in endothelial progenitor cells (EPCs) sourced from healthy donors and wild-type mice.…”

Degradation meets development: Implications in β‐cell development and diabetes

“…DDX3X has been shown to play a proapoptotic role during DNA damage conditions. Thus WWP2 could be a possible target for diabetes induced vascular endothelial injury(You et al, 2023). Collectively, the UPS and its components such as E3 ligases or deubiquitinases has emerged as a promising therapeutic target for the treatment of diabetes and associated diseases.7 | CONCLUSIONSThe virtue of maintaining the balance between synthesis and degradation of key proteins involved in tissue development indicates the efficiency of embryogenesis.…”