Insulin-producing cells derived from 'induced pluripotent stem cells' of patients with fulminant type 1 diabetes: Vulnerability to cytokine insults and increased expression of apoptosis-related genes

Yoshiya Hosokawa; Taro Toyoda; Kenji Fukui; Megu Yamaguchi Baden; Michinori Funato; Yasushi Kondo; Tomomi Sudo; Hiromi Iwahashi; Marina Kishida; Chihiro Okada; Akira Watanabe; Isao Asaka; Kenji Osafune; Akihisa Imagawa; Iichiro Shimomura

doi:10.1111/jdi.12727

Insulin-producing cells derived from 'induced pluripotent stem cells' of patients with fulminant type 1 diabetes: Vulnerability to cytokine insults and increased expression of apoptosis-related genes

J Diabetes Investig. 2017 Aug 10;9(3):481-493. doi: 10.1111/jdi.12727. Online ahead of print.

Authors

Yoshiya Hosokawa¹, Taro Toyoda², Kenji Fukui¹, Megu Yamaguchi Baden¹, Michinori Funato^{2

3}, Yasushi Kondo^{2

4}, Tomomi Sudo², Hiromi Iwahashi^{1

5}, Marina Kishida^{2

6}, Chihiro Okada^{2

7}, Akira Watanabe^{2

6}, Isao Asaka², Kenji Osafune², Akihisa Imagawa^{1

8}, Iichiro Shimomura¹

Affiliations

¹ Department of Metabolic Medicine, Osaka University Graduate School of Medicine, Osaka, Japan.
² Center for iPS Cell Research and Application (CiRA), Kyoto University, Kyoto, Japan.
³ Department of Clinical Research, National Hospital Organization, Nagara Medical Center, Gifu, Japan.
⁴ Department of Diabetes, Endocrinology and Nutrition, Kyoto University, Kyoto, Japan.
⁵ Department of Diabetes Care Medicine, Osaka University Graduate School of Medicine, Osaka, Japan.
⁶ Japan Agency for Medical Research and Development (AMED)-CREST, Japan.
⁷ Mitsubishi Space Software, Tokyo, Japan.
⁸ Department of Internal Medicine (I), Osaka Medical College, Osaka, Japan.

Abstract

Aims/introduction: The present study was carried out to generate induced pluripotent stem cells (iPSCs) from patients with fulminant type 1 diabetes, and evaluate the cytokine-induced apoptotic reactions of β-like insulin-producing cells differentiated from the iPSCs.

Materials and methods: iPSCs were generated from fibroblasts of patients with fulminant type 1 diabetes by inducing six reprogramming factors. Insulin-producing cells were differentiated from the iPSCs in vitro. The proportion of cleaved caspase-3-positive or terminal deoxynucleotidyl transferase 2'-deoxyuridine, 5'-triphosphate nick end labeling-positive cells among insulin (INS)-positive cells derived from fulminant type 1 diabetes iPSC and control human iPSC lines was evaluated under treatment with tumor necrosis factor-α, interleukin-1β and interferon-γ. Ribonucleic acid sequencing was carried out to compare gene expressions in INS-positive cells derived from fulminant type 1 diabetes iPSC and control human iPSC lines.

Results: Two iPSC clones were established from each of three patients with fulminant type 1 diabetes. The differentiation of insulin-producing cells from fulminant type 1 diabetes iPSC was confirmed by immunofluorescence analysis and KCl-induced C-peptide secretion. After treatment with pro-inflammatory cytokines, these INS-positive cells showed higher expression of cleaved caspase-3 than those derived from control human iPSCs. Altered expression levels of several apoptosis-related genes were observed in INS-positive cells derived from the fulminant type 1 diabetes iPSCs by ribonucleic acid sequencing.

Conclusions: We generated iPSCs from patients with fulminant type 1 diabetes and differentiated them into insulin-producing cells. This in vitro disease model can be used to elucidate the disease mechanisms of fulminant type 1 diabetes.

Keywords: Fulminant type 1 diabetes; Induced pluripotent stem cell; β-Cell.