title: Transgene-free disease-specific induced pluripotent stem cells from patients with type 1 and type 2 diabetes.
abstract: The induced pluripotent stem cell (iPSC) technology enables derivation of patient-specific pluripotent stem cells from adult somatic cells without using an embryonic cell source. Redifferentiation of iPSCs from diabetic patients into pancreatic islets will allow patient-specific disease modeling and autologous cell replacement therapy for failing islets. To date, diabetes-specific iPSCs have been generated from patients with type 1 diabetes using integrating retroviral vectors. However, vector integration into the host genome could compromise the biosafety and differentiation propensities of derived iPSCs. Although various integration-free reprogramming systems have been described, their utility to reprogram somatic cells from patients remains largely undetermined. Here, we used nonintegrating Sendai viral vectors to reprogram cells from patients with type 1 and type 2 diabetes (T2D). Sendai vector infection led to reproducible generation of genomic modification-free iPSCs (SV-iPSCs) from patients with diabetes, including an 85-year-old individual with T2D. SV-iPSCs lost the Sendai viral genome and antigens within 8-12 passages while maintaining pluripotency. Genome-wide transcriptome analysis of SV-iPSCs revealed induction of endogenous pluripotency genes and downregulation of genes involved in the oxidative stress response and the INK4/ARF pathways, including p16(INK4a), p15(INK4b), and p21(CIP1). SV-iPSCs and iPSCs made with integrating lentiviral vectors demonstrated remarkable similarities in global gene expression profiles. Thus, the Sendai vector system facilitates reliable reprogramming of patient cells into transgene-free iPSCs, providing a pluripotent platform for personalized diagnostic and therapeutic approaches for diabetes and diabetes-associated complications.
authors.affiliation: Division of Endocrinology, Mayo Clinic, Rochester, MN 55905, USA.
authors.forename: Yogish C
authors.initials: YC
authors.lastname: Kudva
authors.forename: Seiga
authors.initials: S
authors.lastname: Ohmine
authors.forename: Lucas V
authors.initials: LV
authors.lastname: Greder
authors.forename: James R
authors.initials: JR
authors.lastname: Dutton
authors.forename: Adam
authors.initials: A
authors.lastname: Armstrong
authors.forename: Josep Genebriera
authors.initials: JG
authors.lastname: De Lamo
authors.forename: Yulia Krotova
authors.initials: YK
authors.lastname: Khan
authors.forename: Tayaramma
authors.initials: T
authors.lastname: Thatava
authors.forename: Mamoru
authors.initials: M
authors.lastname: Hasegawa
authors.forename: Noemi
authors.initials: N
authors.lastname: Fusaki
authors.forename: Jonathan M W
authors.initials: JM
authors.lastname: Slack
authors.forename: Yasuhiro
authors.initials: Y
authors.lastname: Ikeda
mesh_terms: D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D002478:Cells, Cultured; D002877:Chromosomes, Human / Q000235:genetics / Q000378:metabolism; D050762:Cyclin-Dependent Kinase Inhibitor p15 / Q000235:genetics / Q000378:metabolism; D050759:Cyclin-Dependent Kinase Inhibitor p21 / Q000235:genetics / Q000378:metabolism; D003922:Diabetes Mellitus, Type 1 / Q000473:pathology / Q000628:therapy*; D003924:Diabetes Mellitus, Type 2 / Q000473:pathology / Q000628:therapy*; D005260:Female; D005786:Gene Expression Regulation; D019942:Genes, p16; D005822:Genetic Vectors / Q000235:genetics / Q000378:metabolism; D016679:Genome, Viral; D006801:Humans; D057026:Induced Pluripotent Stem Cells / Q000378:metabolism* / Q000637:transplantation; D015603:Keratinocytes / Q000166:cytology / Q000378:metabolism; D016086:Lentivirus / Q000235:genetics / Q000378:metabolism; D008297:Male; D050814:Octamer Transcription Factor-3 / Q000235:genetics / Q000378:metabolism; D018384:Oxidative Stress; D029082:Sendai virus / Q000235:genetics / Q000378:metabolism; D015398:Signal Transduction; D059467:Transcriptome; D019076:Transgenes
chemical_list: C493956:CDKN1A protein, human; C493966:CDKN2B protein, human; D050762:Cyclin-Dependent Kinase Inhibitor p15; D050759:Cyclin-Dependent Kinase Inhibitor p21; D050814:Octamer Transcription Factor-3; C494173:POU5F1 protein, human