Targeting HNRNPU to overcome cisplatin resistance in bladder cancer

Abstract

Purpose The overall response of cisplatin-based chemotherapy in bladder urothelial carcinoma (BUC) remains unsatisfactory due to the complex pathological subtypes, genomic difference, and drug resistance. The genes that associated with cisplatin resistance remain unclear. Herein, we aimed to identify the cisplatin resistance associated genes in BUC. Experimental design The cytotoxicity of cisplatin was evaluated in six bladder cancer cell lines to compare their responses to cisplatin. The T24 cancer cells exhibited the lowest sensitivity to cisplatin and was therefore selected to explore the mechanisms of drug resistance. We performed genome-wide CRISPR screening in T24 cancer cells in vitro, and identified that the gene heterogeneous nuclear ribonucleoprotein U (HNRNPU) was the top candidate gene related to cisplatin resistance. Epigenetic and transcriptional profiles of HNRNPU-depleted cells after cisplatin treatment were analyzed to investigate the relationship between HNRNPU and cisplatin resistance. In vivo experiments were also performed to demonstrate the function of HNRNPU depletion in cisplatin sensitivity. Results Significant correlation was found between HNRNPU expression level and sensitivity to cisplatin in bladder cancer cell lines. In the high HNRNPU expressing T24 cancer cells, knockout of HNRNPU inhibited cell proliferation, invasion, and migration. In addition, loss of HNRNPU promoted apoptosis and S-phase arrest in the T24 cells treated with cisplatin. Data from The Cancer Genome Atlas (TCGA) demonstrated that HNRNPU expression was significantly higher in tumor tissues than in normal tissues. High HNRNPU level was negatively correlated with patient survival. Transcriptomic profiling analysis showed that knockout of HNRNPU enhanced cisplatin sensitivity by regulating DNA damage repair genes. Furthermore, it was found that HNRNPU regulates chemosensitivity by affecting the expression of neurofibromin 1 (NF1). Conclusions Our study demonstrated that HNRNPU expression is associated with cisplatin sensitivity in bladder urothelial carcinoma cells. Inhibition of HNRNPU could be a potential therapy for cisplatin-resistant bladder cancer.

High expression of RUVBL1 and HNRNPU is associated with poor overall survival in stage I and II non-small cell lung cancer patients

“…Therefore, it cannot be excluded that the inconsistency between studies may be attributed to discrepancies in the methods used. On the other hand, higher levels of mRNA HNRNPU were observed in tumoral vs. adjacent tissue in breast [ 11 ], hepatocellular [ 12 ], and bladder cancers [ 13 ]. Although there were no significant associations between the investigated clinicopathological features and HNRNPU protein expression levels, the survival analyses showed an adverse correlation between a high HNRNPU protein level and clinical outcomes of NSCLC patients.…”

CDC20-Mediated hnRNPU Ubiquitination Regulates Chromatin Condensation and Anti-Cancer Drug Response

“…As previously described, hnRNPU plays a critical role in regulating chromatin dynamics through functional interaction with cohesion complex and chromatin loop formation [ 32 , 33 ]. Dysregulation of hnRNPU results in carcinogenesis and drug resistance [ 34 , 35 , 42 ]. Co-immunoprecipitation assays performed in MDA-MB-468 cells confirmed the interaction between endogenous CDC20 and hnRNPU ( Figure 3 f).…”

“…The impact of hnRNPU in carcinogenesis has been documented previously in different types of tumors [ 34 , 42 ]. To determine the role of hnRNPU in breast cancers, we measured the expression of hnRNPU in normal breast epithelial, DCIS, and different breast cancer cell lines, including TNBC, HER2+, and Luminal subtypes.…”

“…In NB cells, hnRNPU interacts with CTCF resulting in transcriptional alteration of genes associated with tumor progression [ 34 ]. Moreover, hnRNPU expression was associated with cisplatin sensitivity in bladder cancer [ 42 ]. Additionally, hnRNPU has been shown to promote breast cancer cell proliferation, migration, and invasion [ 35 ].…”

Genome Editing Approaches with CRISPR/Cas9 for Cancer Treatment: Critical Appraisal of Preclinical and Clinical Utility, Challenges, and Future Research

“…The following supporting information can be downloaded at: , Table S1: CRISPR/Cas gene targets for the treatment of malignant disorders, and the experimental setup (in vivo/ex vivo and in vivo) used for gene editing (knockout, knockdown, knock-in) with the outcome of the experiment. References [ 104 , 105 , 106 , 107 , 108 , 109 , 110 , 111 , 112 , 113 , 114 , 115 , 116 , 117 , 118 , 119 , 120 , 121 , 122 , 123 , 124 , 125 ] are cited in Supplementary Materials File.…”