iPSCs derived from insulin resistant offspring of type 2 diabetic patients show increased oxidative stress and lactate secretion

Abstract

Background The genetic factors associated with insulin resistance (IR) are not well understood. Clinical studies on first-degree relatives of type 2 diabetic (T2D) patients, which have the highest genetic predisposition to T2D, have given insights into the role of IR in T2D pathogenesis. Induced pluripotent stem cells (iPSCs) are excellent tools for disease modeling as they can retain the genetic imprint of the disease. Therefore, in this study, we aimed to investigate the genetic perturbations associated with insulin resistance (IR) in the offspring of T2D parents using patient-specific iPSCs. Methods We generated iPSCs from IR individuals (IR-iPSCs) that were offspring of T2D parents as well as from insulin-sensitive (IS-iPSCs) individuals. We then performed transcriptomics to identify key dysregulated gene networks in the IR-iPSCs in comparison to IS-iPSCs and functionally validated them. Results Transcriptomics on IR-iPSCs revealed dysregulated gene networks and biological processes indicating that they carry the genetic defects associated with IR that may lead to T2D. The IR-iPSCs had increased lactate secretion and a higher phosphorylation of AKT upon stimulation with insulin. IR-iPSCs have increased cellular oxidative stress indicated by a high production of reactive oxygen species and higher susceptibility to H2O2 -induced apoptosis. Conclusions IR-iPSCs generated from offspring of diabetic patients confirm that oxidative stress and increased lactate secretion, associated with IR, are inherited in this population, and may place them at a high risk of T2D. Overall, our IR-iPSC model can be employed for T2D modeling and drug screening studies that target genetic perturbations associated with IR in individuals with a high risk for T2D.

Deletion of RFX6, a Diabetes-Associated Gene, Impairs iPSC-Derived Islet Organoid Development and Survival, With No Impact on the Generation of PDX1+/NKX6.1+ Progenitors

“…HA-RFX6 tagged H9 hESC line (RFX6 HA/HA H9-hESCs) with its control, H9-hESCs, were obtained from Dr. N. Ray Dunn (A*STAR, Singapore). Wild type iPSCs (WT-iPSCs) generated and fully characterized in our laboratory were used [21, 36]. Cells were cultured and maintained in mTeSR Plus medium (Stem Cell Technologies, Canada) on Matrigel-coated dishes (Corning, USA), following our established approach [37][38].…”

Transcriptome-Powered Pluripotent Stem Cell Differentiation for Regenerative Medicine

“…Transcriptome analysis is beneficial for understanding disease and providing insight into potential regenerative medicines capable of mitigating or treating disease. This commonly encompasses utilizing iPSCs derived from patients' tissues to generate diseased organoids for research (Table 4) [85][86][87][88][89][90][91]. Figure 5 summarizes the key steps in disease modeling through a combination of iPSC technology and transcriptomics for regenerative medicine.…”

“…Technologies for iPSC differentiation into insulin-secreting β-cells have been applied to generate β-cells or endocrine tissue from patients suffering from different subtypes of diabetes, such as type I diabetes (T1D), type II diabetes (T2D), and monogenic diabetes (MD) (Table 4) [88,[98][99][100]. Memon et al focused on utilizing T2D-derived iPSCs to study the mechanism behind insulin resistance in T2D [89]. RNAseq analysis of insulin-resistant iPSCs discovered the downregulation of L1TD1, RIF1, MYSM1, ZNF195, ZNF208, and ZNF770 compared to healthy controls [89].…”

“…Memon et al focused on utilizing T2D-derived iPSCs to study the mechanism behind insulin resistance in T2D [89]. RNAseq analysis of insulin-resistant iPSCs discovered the downregulation of L1TD1, RIF1, MYSM1, ZNF195, ZNF208, and ZNF770 compared to healthy controls [89]. Upregulated genes in the insulin-resistant group compared to healthy controls include AGRN, SLFN13, CD74, SEMA6B, SLC22A17, and TMEM151B [89].…”

“…RNAseq analysis of insulin-resistant iPSCs discovered the downregulation of L1TD1, RIF1, MYSM1, ZNF195, ZNF208, and ZNF770 compared to healthy controls [89]. Upregulated genes in the insulin-resistant group compared to healthy controls include AGRN, SLFN13, CD74, SEMA6B, SLC22A17, and TMEM151B [89]. The T2D iPSC group had increased lactate secretion and increased oxidative stress in the cells, suggesting lowered oxygen availability to the diseased islet cells.…”