title: Improvement in Mouse iPSC Induction by Rab32 Reveals the Importance of Lipid Metabolism during Reprogramming.
abstract: Induced pluripotent stem cells (iPSCs) have variable expression levels of a series of genes that affect their pluripotent potential, but the regulatory mechanisms controlling reprogramming remain unclear. By testing the efficiency of iPSC generation using Oct4, Sox2, Klf4 (termed OSK) plus one additional gene, we found that Rab32 improved reprogramming efficiency. We established a system for detecting the number and the size of lipid droplets and autophagosomes per cell for tracking their morphological changes during reprogramming. Our results showed that Rab32 increased lipid storage during the early and middle stages, and also increased autophagy during the middle stage of reprogramming. These findings were further confirmed by the up-regulation of lipid biosynthesis and autophagosome formation related genes, of which their expression could improve iPSC induction. The inhibition of lipid biosynthesis and autophagosome formation significantly reduced reprogramming efficiency, and the inhibition of lipid synthesis phenotype could be rescued by the overexpression of Rab32. In addition, the expression of pluripotency genes such as Klf2, Nr5a2 and Tbx3, was up-regulated by Rab32. These results demonstrated that Rab32 could improve the induction of iPSCs through the enhancement of lipid biosynthesis, highlighting the importance of lipid metabolism during reprogramming.
authors.affiliation: State Key Laboratories for Agrobiotechnology, College of Biological Sciences, China Agricultural University, Beijing, 100193, China.
authors.forename: Yangli
authors.initials: Y
authors.lastname: Pei
authors.affiliation: State Key Laboratories for Agrobiotechnology, College of Biological Sciences, China Agricultural University, Beijing, 100193, China.
authors.forename: Liang
authors.initials: L
authors.lastname: Yue
authors.affiliation: State Key Laboratories for Agrobiotechnology, College of Biological Sciences, China Agricultural University, Beijing, 100193, China.
authors.forename: Wei
authors.initials: W
authors.lastname: Zhang
authors.affiliation: State Key Laboratories for Agrobiotechnology, College of Biological Sciences, China Agricultural University, Beijing, 100193, China.
authors.forename: Yanliang
authors.initials: Y
authors.lastname: Wang
authors.affiliation: State Key Laboratories for Agrobiotechnology, College of Biological Sciences, China Agricultural University, Beijing, 100193, China.
authors.forename: Bingqiang
authors.initials: B
authors.lastname: Wen
authors.affiliation: State Key Laboratories for Agrobiotechnology, College of Biological Sciences, China Agricultural University, Beijing, 100193, China.
authors.forename: Liang
authors.initials: L
authors.lastname: Zhong
authors.affiliation: State Key Laboratories for Agrobiotechnology, College of Biological Sciences, China Agricultural University, Beijing, 100193, China.
authors.forename: Jinzhu
authors.initials: J
authors.lastname: Xiang
authors.affiliation: State Key Laboratories for Agrobiotechnology, College of Biological Sciences, China Agricultural University, Beijing, 100193, China.
authors.forename: Junhong
authors.initials: J
authors.lastname: Li
authors.affiliation: State Key Laboratories for Agrobiotechnology, College of Biological Sciences, China Agricultural University, Beijing, 100193, China.
authors.forename: Shaopeng
authors.initials: S
authors.lastname: Zhang
authors.affiliation: State Key Laboratories for Agrobiotechnology, College of Biological Sciences, China Agricultural University, Beijing, 100193, China.
authors.forename: Hanning
authors.initials: H
authors.lastname: Wang
authors.affiliation: State Key Laboratories for Agrobiotechnology, College of Biological Sciences, China Agricultural University, Beijing, 100193, China.
authors.forename: Haiyuan
authors.initials: H
authors.lastname: Mu
authors.affiliation: State Key Laboratories for Agrobiotechnology, College of Biological Sciences, China Agricultural University, Beijing, 100193, China.
authors.forename: Qingqing
authors.initials: Q
authors.lastname: Wei
authors.affiliation: State Key Laboratories for Agrobiotechnology, College of Biological Sciences, China Agricultural University, Beijing, 100193, China.
authors.forename: Jianyong
authors.initials: J
authors.lastname: Han
mesh_terms: D000818:Animals; D001343:Autophagy / Q000235:genetics; D002454:Cell Differentiation; D065150:Cellular Reprogramming; D015870:Gene Expression; D057026:Induced Pluripotent Stem Cells / Q000166:cytology* / Q000378:metabolism*; D000090062:Kruppel-Like Factor 4; D050356:Lipid Metabolism; D051379:Mice; D010588:Phagosomes / Q000378:metabolism; D014157:Transcription Factors / Q000235:genetics / Q000378:metabolism; D020691:rab GTP-Binding Proteins / Q000235:genetics* / Q000378:metabolism*
chemical_list: C000715468:Klf4 protein, mouse; D000090062:Kruppel-Like Factor 4; D014157:Transcription Factors; C478766:Rab32 protein, mouse; D020691:rab GTP-Binding Proteins