title: Reprogramming adipose tissue-derived mesenchymal stem cells into pluripotent stem cells by a mutant adeno-associated viral vector.
abstract: Induced pluripotent stem (iPS) cells have great potential for personalized regenerative medicine. Although several different methods for generating iPS cells have been reported, improvement of safety and efficiency is imperative. In this study, we tested the feasibility of using a triple tyrosine mutant AAV2 (Y444+500+730F) vector, designated AAV2.3m, to generate iPS cells. We developed a polycistronic rAAV2.3m vector expressing three reprogramming factors, Klf4, Oct4, and Sox2, and then used this vector to infect mouse adipose-derived mesenchymal stem cells (AT-MSCs) to induce the generation of iPS cells. We demonstrated that (1) the triple tyrosine mutant AAV2 vector is able to reprogram mouse adult adipose tissue-derived stem cells into the pluripotent state. Those rAAV2.3m-derived iPS (rAAV2.3m-iPS) cells express endogenous pluripotency-associated genes including Oct4, Sox2, and SSEA-1, and form teratomas containing multiple tissues in vivo; (2) c-myc, an oncogene, is dispensable in rAAV2.3m-mediated cellular reprogramming; and (3) transgene expression is undetectable after reprogramming, whereas vector DNA is detectable, indicating that transgenes are silenced. These results indicated the rAAV vector may have some advantages in generating iPS cells.
authors.affiliation: 1 Department of Pharmaceutics, University of Florida , Gainesville, FL 32610.
authors.forename: Mong-Jen
authors.initials: MJ
authors.lastname: Chen
authors.forename: Yuanqing
authors.initials: Y
authors.lastname: Lu
authors.forename: Takashi
authors.initials: T
authors.lastname: Hamazaki
authors.forename: Hsin-Yin
authors.initials: HY
authors.lastname: Tsai
authors.forename: Kirsten
authors.initials: K
authors.lastname: Erger
authors.forename: Thomas
authors.initials: T
authors.lastname: Conlon
authors.forename: Ahmed S
authors.initials: AS
authors.lastname: Elshikha
authors.forename: Hong
authors.initials: H
authors.lastname: Li
authors.forename: Arun
authors.initials: A
authors.lastname: Srivastava
authors.forename: Chunli
authors.initials: C
authors.lastname: Yao
authors.forename: Mark
authors.initials: M
authors.lastname: Brantly
authors.forename: Vince
authors.initials: V
authors.lastname: Chiodo
authors.forename: William
authors.initials: W
authors.lastname: Hauswirth
authors.forename: Naohiro
authors.initials: N
authors.lastname: Terada
authors.forename: Sihong
authors.initials: S
authors.lastname: Song
mesh_terms: D000273:Adipose Tissue / Q000166:cytology*; D000818:Animals; D065150:Cellular Reprogramming; D000229:Dependovirus / Q000235:genetics*; D005822:Genetic Vectors / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D000090062:Kruppel-Like Factor 4; D051741:Kruppel-Like Transcription Factors / Q000235:genetics / Q000378:metabolism; D016256:Lewis X Antigen / Q000235:genetics / Q000378:metabolism; D059630:Mesenchymal Stem Cells / Q000166:cytology* / Q000378:metabolism; D051379:Mice; D008810:Mice, Inbred C57BL; D008822:Mice, Transgenic; D050814:Octamer Transcription Factor-3 / Q000235:genetics / Q000378:metabolism; D039904:Pluripotent Stem Cells / Q000166:cytology*; D016271:Proto-Oncogene Proteins c-myc / Q000235:genetics / Q000378:metabolism; D055748:SOXB1 Transcription Factors / Q000235:genetics / Q000378:metabolism; D013724:Teratoma / Q000473:pathology; D014161:Transduction, Genetic
chemical_list: C000715467:KLF4 protein, human; C000715468:Klf4 protein, mouse; D000090062:Kruppel-Like Factor 4; D051741:Kruppel-Like Transcription Factors; D016256:Lewis X Antigen; D050814:Octamer Transcription Factor-3; D016271:Proto-Oncogene Proteins c-myc; D055748:SOXB1 Transcription Factors