title: Autophagy governs erythroid differentiation both in vitro and in vivo.
abstract: OBJECTIVE
  Although the importance of autophagy in determination of cell fate has been much explored in recent years, its definite role in regulating erythroid differentiation remains unclear.
  
  
  METHODS
  In this study, human erythroleukemic cell line K562 was employed as a cell model for studying erythroid differentiation in vitro. Starvation and rapamycin were used to induce autophagy, whereas Baf-A1 and CRISPR/Cas9/Atg7 were used to inhibit late and early phase of autophagy, respectively. The mice model of autophagy activation and autophagy deletion were established, and red blood cell counts and flow cytometry were used to analyze erythroid differentiation in vivo.
  
  
  RESULTS
  The results showed that the transcriptional levels of α-globin and γ-globin, and the ratio of hemoglobin-positive cells all significantly increased in K562 cells with starvation and rapamycin treatment, which were blocked by autophagy inhibitor, Baf-A1 or Atg7 gene knockout. In the autophagy-enhanced mouse model, the number of mature erythrocytes significantly increased, while in Atg7-deleted mouse model, erythroid differentiation was severely blocked.
  
  
  CONCLUSIONS
  It is concluded that autophagy is indispensible in the development, maturation, and homeostasis of erythroid cells both in vitro and in vivo. Our findings support the potential strategy for erythroleukemia treatment and production of erythroblasts in vitro by modulating autophagy.
authors.affiliation: a Hematology Center of Cyrus Tang Medical Institute , Soochow University School of Medicine , Suzhou 215123 , Jiangsu , China.
authors.forename: Yan
authors.initials: Y
authors.lastname: Cao
authors.affiliation: a Hematology Center of Cyrus Tang Medical Institute , Soochow University School of Medicine , Suzhou 215123 , Jiangsu , China.|b Current address: State key Laboratory of Reproductive Medicine , Nanjing Medical University , Nanjing 210029 , Jiangsu , China.
authors.forename: Jinyang
authors.initials: J
authors.lastname: Cai
authors.affiliation: a Hematology Center of Cyrus Tang Medical Institute , Soochow University School of Medicine , Suzhou 215123 , Jiangsu , China.
authors.forename: Xin
authors.initials: X
authors.lastname: Li
authors.affiliation: a Hematology Center of Cyrus Tang Medical Institute , Soochow University School of Medicine , Suzhou 215123 , Jiangsu , China.
authors.forename: Na
authors.initials: N
authors.lastname: Yuan
authors.affiliation: a Hematology Center of Cyrus Tang Medical Institute , Soochow University School of Medicine , Suzhou 215123 , Jiangsu , China.
authors.forename: Suping
authors.initials: S
authors.lastname: Zhang
mesh_terms: D000818:Animals; D001343:Autophagy; D002454:Cell Differentiation; D004900:Erythroblasts / Q000378:metabolism*; D006801:Humans; D020014:K562 Cells; D051379:Mice; D008822:Mice, Transgenic
keywords: Atg7; K562 cells; autophagy; erythroid differentiation; erythroleukemia