Derivation of Human Induced Pluripotent Stem Cells from Patients with Maturity Onset Diabetes of the Young*

Abstract

Background: Human induced pluripotent stem cells (hiPSCs) can be harnessed for development of novel therapeutics for metabolic disorders. Results: Karyotypically normal hiPSCs were derived from patients with MODY1, MODY2, MODY3, MODY5, or MODY8. Conclusion: hiPSCs were successfully derived from a variety of MODY patients. Significance: MODY-hiPSCs can be used to explore the role of MODY genes in the development and function of pancreatic islet cells. Maturity onset diabetes of the young (MODY) is an autosomal dominant disease. Despite extensive research, the mechanism by which a mutant MODY gene results in monogenic diabetesis not yet clear due to the inaccessibility of patient samples. Induced pluripotency and directed differentiation toward the pancreatic lineage are now viable and attractive methods to uncover the molecular mechanisms underlying MODY. Here we report, for the first time, the derivation of human induced pluripotent stem cells (hiPSCs) from patients with five types of MODY: MODY1 (HNF4A), MODY2 (GCK), MODY3 (HNF1A), MODY5 (HNF1B), and MODY8 (CEL) with a polycistronic lentiviral vector expressing a Cre-excisable human "stem cell cassette" containing the four reprogramming factors OCT4, KLF4, SOX2, and CMYC. These MODY-hiPSCs morphologically resemble human pluripotent stem cells (hPSCs), express pluripotency markers OCT4, SOX2, NANOG, SSEA-4, and TRA-1-60, give rise to derivatives of the three germ layers in a teratoma assay, and are karyotypically normal. Overall, our MODYhiPSCs serve as invaluable tools to dissect the role of MODY genes in the development of pancreas and islet cells and to evaluate their significance in regulating beta cell function. This knowledge will aid future attempts aimed at deriving functional mature beta cells from hPSCs. Maturity onset diabetes of the young (MODY)2 is an autosomal dominant monogenic diabetic disease typically affecting individuals before the age of 25 years. To date, 11 MODY genes have been identified, comprising both transcription factors and enzymes/hormones: MODY1 (HNF4A), MODY2 (GCK), MODY3 (HNF1A), MODY4 (PDX1), MODY5 (HNF1B), MODY6 (NEUROD1), MODY7 (KLF11), MODY8 (CEL), MODY9 (PAX4), MODY10 (INS), and MODY11 (BLK) (1). Various disease-causing genetic variants have been identified for each of the MODY genes, all of which affect pancreas and beta cell development, ultimately resulting in beta cell dysfunction and diabetes.The advent of induced pluripotency (2) has provided an opportunity to derive human induced pluripotent stem cells (hiPSCs) from MODY patients to model the disease in vitro. Several studies have reported that human somatic cells, commonly skin fibroblasts, can be reprogrammed into hiPSCs. A number of groups have reported the generation of hiPSCs from patients with mitochondrial diabetes, type 1 and type 2 diabetes mellitus (3, 4). However, given the complexity of type 1 and type 2 diabetes mellitus, the use of hiPSCs from these patients requires comparison with multiple controls that are appropriate for the genetic and environmenta...

Endocytosis of Secreted Carboxyl Ester Lipase in a Syndrome of Diabetes and Pancreatic Exocrine Dysfunction

“…Thus, studies of the effect of secreted CEL-MUT on human islets in vitro are now needed. Alternatively, preparation of induced pluripotent stem cells from patients with CEL-MODY mutations could be an interesting option for further studies (49).…”

La thérapie cellulaire du diabète

“…Ces limitations, associées au risque carcinogénique, retardent l'initiation de protocoles de recherche clinique et pourraient constituer un frein à l'enrôlement de candidats volontaires à la transplantation. La valeur ajoutée des CSP induites réside dans la possibilité de produire des dérivés de cellules β autologues, ce qui a été démon-tré chez des patients souffrant de DT1 [26] ou de diabète monogé-nique (autrefois appelé « MODY » pour maturity onset diabetes of the young) [27,60] …”

When is it MODY? Challenges in the Interpretation of Sequence Variants in MODY Genes

“…In the context of MODY, most diseasecausing genes play critical regulatory and developmental roles in more than one tissue type, such as HNF1B in the kidney and GCK, HNF1A, and HNF4A in the hepatocyte. Therefore, patientderived iPS cells provide an opportunity for simultaneous investigations of tissue-specific effects via unique maturation switches [101]. The insulinproducing pancreatic beta-cells, recently developed in vitro from human pluripotent stem cells (hPSCs), also provide an ideal genetic and physiological setting for such interrogations, not to mention their immense potential in cell replacement therapy [102,103].…”