Novel Functional Genes Involved in Transdifferentiation of Canine ADMSCs Into Insulin-Producing Cells, as Determined by Absolute Quantitative Transcriptome Sequencing Analysis.

Pengxiu Dai,Xinke Zhang,Guixiang Qi,Yijing Chen,Yihua Zhang,Jiakai Li,Luwen Zhang,Jinglu Wang

doi:10.3389/fcell.2021.685494

Abstract

The transdifferentiation of adipose-derived mesenchymal stem cells (ADMSCs) into insulin-producing cells (IPCs) is a potential resource for the treatment of diabetes. However, the changes of genes and metabolic pathways on the transdifferentiation of ADMSCs into IPCs are largely unknown. In this study, the transdifferentiation of canine ADMSCs into IPCs was completed using five types of procedures. Absolute Quantitative Transcriptome Sequencing Analysis was performed at different stages of the optimal procedure. A total of 60,151 transcripts were obtained. Differentially expressed genes (DEGs) were divided into five groups: IPC1 vs. ADSC (1169 upregulated genes and 1377 downregulated genes), IPC2 vs. IPC1 (1323 upregulated genes and 803 downregulated genes), IPC3 vs. IPC2 (722 upregulated genes and 680 downregulated genes), IPC4 vs. IPC3 (539 upregulated genes and 1561 downregulated genes), and Beta_cell vs. IPC4 (2816 upregulated genes and 4571 downregulated genes). The gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis of DEGs revealed that many genes and signaling pathways that are essential for transdifferentiation. Hnf1B, Dll1, Pbx1, Rfx3, and Foxa1 were screened out, and the functions of five genes were verified further by overexpression and silence. Foxa1, Pbx1, and Rfx3 exhibited significant effects, can be used as specific key regulatory factors in the transdifferentiation of ADMSCs into IPCs. This study provides a foundation for future work to understand the mechanisms of the transdifferentiation of ADMSCs into IPCs and acquire IPCs with high maturity.

Highlights

MATERIALS AND METHODSDiabetes encompasses a group of lifelong metabolic diseases, and common drug therapies are not able to cure it
These results prove that the isolated cells were canine adipose-derived mesenchymal stem cells (ADMSCs)
The canine ADMSCs were transdifferentiated into insulin-producing cells (IPCs) using five types of procedures

Summary

Introduction

MATERIALS AND METHODSDiabetes encompasses a group of lifelong metabolic diseases, and common drug therapies are not able to cure it. Adipose-derived mesenchymal stem cells (ADMSCs) are abundant in sources, are isolated and cultivated, exhibit pluripotent differentiation potential and show low immunogenicity after transplantation, serving as ideal seed cells for the treatment of diabetes and its complications (Kim et al, 2010; Zhang et al, 2016; Takemura et al, 2019; Wada and Ikemoto, 2019; Tokuda et al, 2020). Growth factors, activators, and inhibitors can transdifferentiate ADMSCs into IPCs and improve IPCs survival and ability to release insulin in vitro (Dayer et al, 2017; Anjum et al, 2018; Ikemoto et al, 2018; Shahjalal et al, 2018; Pavathuparambil Abdul Manaph et al, 2019; Ghoneim et al, 2020). ADMSCs transdifferentiate into functional mature beta cells need more research

Results

Discussion

Conclusion