WDHD1 Leads to Cisplatin Resistance by Promoting MAPRE2 Ubiquitination in Lung Adenocarcinoma

Abstract

Ubiquitin ligases have been shown to regulate drug sensitivity. This study aimed to explore the role of the ubiquitin ligase WD repeat and HMG-box DNA binding protein 1 (WDHD1) in regulating cisplatin sensitivity in lung adenocarcinoma (LUAD). A quantitative analysis of the global proteome identified differential protein expression between LUAD A549 cells and the cisplatin-resistant strain A549/DDP. Public databases revealed the relationship between ubiquitin ligase expression and the prognosis of patients with LUAD. Quantitative real-time polymerase chain reaction and Western blotting were used to estimate the WDHD1 expression levels. Analysis of public databases predicted the substrate of WDHD1. Western blotting detected the effect of WDHD1 on microtubule-associated protein RP/EB family member 2 (MAPRE2) and DSTN. Functional analysis of MAPRE2 verified the interaction between WDHD1 and MAPRE2, as well as the interacting sites by methyl-thiazolyl-tetrazolium assay and flow cytometry, immunoprecipitation, protein stability, and immunofluorescence. Cell and animal experiments confirmed the effect of WDHD1 and MAPRE2 on cisplatin sensitivity in LUAD. Clinical data evaluated the impact of WDHD1 expression level on cisplatin sensitivity. Quantitative analysis of the global proteome revealed ubiquitin-dependent protein catabolism to be more active in A549/DDP cells than in A549 cells. WDHD1 expression was higher in A549/DDP cells than in A549 cells, and knocking out WDHD1 increased the sensitivity of A549/DDP cells to cisplatin. WDHD1 overexpression negatively correlated with the overall survival of LUAD patients. We observed that MAPRE2 was upregulated when WDHD1 was knocked out. A MAPRE2 knockout in A549 cells resulted in increased cell viability while decreasing apoptosis when the A549 cells exposed to cisplatin. WDHD1 and MAPRE2 were found to interact in the nucleus, and WDHD1 promoted the ubiquitination of MAPRE2. Following cisplatin exposure, the WDHD1 and MAPRE2 knockout groups facilitated cell proliferation and migration, inhibited apoptosis in A549/DDP cells, decreased apoptosis, and increased tumor size and growth rate in animal experiments. Gong et al. Chemotherapy Resistance of Lung Adenocarcinoma Immunohistochemistry showed that Ki67 levels increased, and levels of apoptotic indicators significantly decreased in the WDHD1 and MAPRE2 knockout groups. Clinical data confirmed that WDHD1 overexpression negatively correlated with cisplatin sensitivity. Thus, the ubiquitin ligase WDHD1 induces cisplatin resistance in LUAD by promoting MAPRE2 ubiquitination.

Exosomal proteomic signatures correlate with drug resistance and carboplatin treatment outcome in a spontaneous model of canine osteosarcoma

“…The differences in the proteome and potentially miRNA cargoes of these exosomes are most likely the cause for these differences. For example, some proteins with protective effect such as the ribose-phosphate pyrophosphokinase 1 (PRPS1) which is responsible for nucleotide synthesis, and the ubiquitin ligase WD repeat and HMG-box DNA binding protein 1 (WDHD1) were expressed exclusively in the exosomal proteomic cargo of HMPOS-2.5R and may explain their distinct effect on HMPOS-S [ 38 , 39 ]. While it is possible that the delivery and uptake of these proteins were key in promoting drug resistance, it is remained to be determined how other factors contained in the exosomal unit such miRNA might also support this phenomenon.…”

The Involvement of WDHD1 in the Occurrence of Esophageal Cancer as a Downstream Target of PI3K/AKT Pathway

“…It is a kind of DNA binding protein in the nucleus and cytoplasm, which controls mitosis, DNA damage, and repair. Meanwhile, WDHD1 is also a ubiquitin ligase, which is related to tumor development and malignant phenotype [ 17 , 19 , 30 ].WDHD1 is a human homologue of CTF4 (chromosome transmission fidelity factor 4) in Saccharomyces cerevisiae [ 31 ]. WDHD1 is similar to the sequence of CTF4 (722 of 1129 aligned residues have 21% homology and 36% similarity).…”

WDHD1 is essential for the survival of PTEN-inactive triple-negative breast cancer

“…The clinical importance of WDHD1 in TNBC was evaluated in samples obtained from TNBC patients, showing that its levels are increased in TNBC compared to normal breast tissues, and associates with tumour size, stage and proliferation, using Ki67 as a proliferation marker 52 . Moreover, recent reports demonstrated that overexpression of WDHD1 leads to cisplatin resistance in lung adenocarcinoma 53 and metastasis in cholangiocarcinoma 54 . Further studies are required to confirm these findings in TNBC.…”