RNA binding protein YWHAZ mediates specific mRNA translation and regulates cell proliferation and apoptosis in diabetic foot ulcer

<h4>Background</h4>Diabetic foot ulcer (DFU), a condition marked by high rates of recurrence, amputation, and mortality, represents one of the major challenges in diabetes management. RNA-binding proteins (RBPs) are pivotal for post-transcriptional regulation in diabetic complications. However, the aberrantly expressed RBP genes and their regulatory mechanisms in DFU remain unclear. This study aimed to investigate the potential functions and molecular interactions of YWHAZ, a dysregulated RBP identified in DFU tissues.<h4>Methods</h4>YWHAZ was selected for further investigation based on its dysregulation in DFU tissues as identified through an independent public RNA-seq dataset. Reverse transcription quantitative polymerase chain reaction (RT-qPCR) and immunohistochemistry were used to validate YWHAZ expression. The biological behavior of HaCaT cells with YWHAZ knockdown (Si_YWHAZ) was compared with that of control cells. Differentially expressed genes (DEGs) were identified by RNA-seq. Additionally, improved RNA immunoprecipitation (iRIP)-seq was employed to investigate potential binding interactions of YWHAZ in DFU tissues.<h4>Results</h4>YWHAZ was significantly upregulated and validated as an RBP in DFU by RT-qPCR and immunohistochemistry. Cellular experiments revealed that Si_YWHAZ facilitated proliferation and migration while inhibiting apoptosis, consistent with its upregulation in DFU. RNA-seq identified 1,072 DEGs in Si_YWHAZ cells. Upregulated genes were significantly enriched for cell proliferation-related processes and included <i>AREG</i>, <i>FOSL1</i>, <i>HAS2,</i> and <i>IL7R</i>, whereas downregulated genes were associated with cell adhesion, including <i>LAMB3</i>, <i>SLAMF7</i>, <i>COL12A1,</i> and <i>ITGA5</i>. iRIP-seq results demonstrated that YWHAZ interacts with a large number of mRNAs and is located in the CD and intron region of the genome. Furthermore, the ABLIFE algorithm indicated that YWHAZ binds to GC-rich motifs. Integrated iRIP-seq and RNA-seq analyses identified 57 DEGs that were selectively bound by YWHAZ, most of which were downregulated. Notably, <i>SREBF1</i>, which is positively associated with type 2 diabetes risk. KEGG pathway analysis revealed that <i>SREBF1</i> is enriched in both the insulin resistance and insulin signaling pathways.<h4>Conclusion</h4>The study results suggest the cellular functions and molecular targets of YWHAZ, indicating its potential regulatory role in DFU development. This study provides valuable insights into the regulatory mechanisms of YWHAZ in DFU for future investigations.