Comprehensive Characterization and Global Transcriptome Analysis of Human Fetal Liver Terminal Erythropoiesis

Yongshuai Han; Shihui Wang; Yaomei Wang; Yumin Huang; Chengjie Gao; Xinhua Guo; Lixiang Chen; Huizhi Zhao; Xiuli An

doi:10.1016/j.gpb.2023.07.001

Comprehensive Characterization and Global Transcriptome Analysis of Human Fetal Liver Terminal Erythropoiesis

Genomics Proteomics Bioinformatics. 2023 Aug 30:S1672-0229(23)00102-X. doi: 10.1016/j.gpb.2023.07.001. Online ahead of print.

Authors

Yongshuai Han¹, Shihui Wang², Yaomei Wang³, Yumin Huang⁴, Chengjie Gao⁵, Xinhua Guo⁵, Lixiang Chen⁶, Huizhi Zhao⁷, Xiuli An⁸

Affiliations

¹ School of Life Sciences, Zhengzhou University, Zhengzhou 450001, China; Laboratory of Membrane Biology, New York Blood Center, New York, NY 10065, USA.
² Laboratory of Membrane Biology, New York Blood Center, New York, NY 10065, USA; Institute of Hematology, People's Hospital of Zhengzhou University, Zhengzhou 450003, China.
³ Laboratory of Membrane Biology, New York Blood Center, New York, NY 10065, USA; Department of Hematology, the Affiliated Cancer Hospital of Zhengzhou University, Zhengzhou 450000, China.
⁴ Laboratory of Membrane Biology, New York Blood Center, New York, NY 10065, USA; Department of Hematology, the First Affiliated Hospital of Zhengzhou University, Zhengzhou 450002, China.
⁵ Laboratory of Membrane Biology, New York Blood Center, New York, NY 10065, USA.
⁶ School of Life Sciences, Zhengzhou University, Zhengzhou 450001, China.
⁷ School of Life Sciences, Zhengzhou University, Zhengzhou 450001, China. Electronic address: hzzhao@zzu.edu.cn.
⁸ Laboratory of Membrane Biology, New York Blood Center, New York, NY 10065, USA. Electronic address: xan@nybc.org.

PMID: 37657739
DOI: 10.1016/j.gpb.2023.07.001

Abstract

The fetal liver (FL) is the key erythropoietic organ during fetal development, but knowledge on human FL erythropoiesis is very limited. In this study, we sorted primary erythroblasts from FL cells and performed RNA sequencing (RNA-seq) analyses. We found that temporal gene expression patterns reflected changes in function during primary human FL terminal erythropoiesis. Notably, the expression of genes enriched in proteolysis and autophagy was up-regulated in orthochromatic erythroblasts (OrthoEs), suggesting the involvement of these pathways in enucleation. We also performed RNA-seq of in vitro cultured erythroblasts derived from FL CD34⁺ cells. Comparison of transcriptomes between the primary and cultured erythroblasts revealed significant differences, indicating impacts of the culture system on gene expression. Notably, the expression of lipid metabolism-related genes was increased in cultured erythroblasts. We further immortalized erythroid cell lines from FL and cord blood (CB) CD34⁺ cells (FL-iEry and CB-iEry, respectively). FL-iEry and CB-iEry were immortalized at the proerythroblast stage and can be induced to differentiate into OrthoEs, but their enucleation ability was very low. Comparison of the transcriptomes between OrthoEs with and without enucleation capability revealed the down-regulation of pathways involved in chromatin organization and mitophagy in OrthoEs without enucleation capacity, indicating that defects in chromatin organization and mitophagy contribute to the inability of OrthoEs to enucleate. Additionally, the expression of HBE1, HBZ, and HBG2 was up-regulated in FL-iEry compared with CB-iEry, and this up-regulation was accompanied by down-regulated expression of BCL11A and up-regulated expression of LIN28B and IGF2BP1. Our study provides new insights into human FL erythropoiesis and rich resources for future studies.

Keywords: Enucleation; Human fetal liver; Immortalized erythroid cell line; Terminal erythropoiesis; Transcriptome.