RNA binding protein HuD mediates the crosstalk between β cells and islet endothelial cells by the regulation of Endostatin and Serpin E1 expression

RNA binding protein HuD plays essential roles in gene expression by regulating RNA metabolism, and its dysregulation is involved in the pathogenesis of several diseases, including tumors, neurodegenerative diseases, and diabetes. Here, we explored HuD-mediated differential expression of secretory proteins in mouse insulinoma beta TC6 cells using a cytokine array. Endostatin and Serpin E1 that play anti-angiogenic roles were identified as differentially expressed proteins by HuD. HuD knockdown increased the expression of alpha chain of collagen XVIII (Col18a1), a precursor form of endostatin, and Serpin E1 by associating with the 3 & PRIME;-untranslated regions (UTRs) of Col18a1 and Serpin E1 mRNAs. Reporter analysis revealed that HuD knockdown increased the translation of EGFP reporters containing 3 & PRIME;UTRs of Col18a1 and Serpin E1 mRNAs, which suggests the role of HuD as a translational repressor. Co-cultures of beta TC6 cells and pancreatic islet endothelial MS1 cells were used to assess the crosstalk between beta cells and islet endothelial cells, and the results showed that HuD downregulation in beta TC6 cells inhibited the growth and migration of MS1 cells. Ectopic expression of HuD decreased Col18a1 and Serpin E1 expression, while increasing the markers of islet vascular cells in the pancreas of db/db mice. Taken together, these results suggest that HuD has the potential to regulate the crosstalk between beta cells and islet endothelial cells by regulating Endostatin and Serpin E1 expression, thereby contributing to the maintenance of homeostasis in the islet microenvironment.

Automated summary

The RNA binding protein HuD plays a crucial role in gene expression and its dysregulation is associated with various diseases. This study investigated the differential expression of secretory proteins mediated by HuD in mouse insulinoma beta TC6 cells. The results identified endostatin and Serpin E1 as differentially expressed proteins regulated by HuD. HuD knockdown increased the expression of Col18a1, a precursor form of endostatin, and Serpin E1 by interacting with their 3’ untranslated regions. The study also revealed the role of HuD as a translational repressor. HuD downregulation in beta TC6 cells inhibited the growth and migration of islet endothelial cells, and the ectopic expression of HuD in mice affected the expression of Col18a1, Serpin E1, and markers of islet vascular cells.