Summary

Fine-resolution differentiation trajectories of adult human hematopoietic stem cells (HSCs) involved in the generation of red cells is critical for understanding dynamic developmental changes that accompany human erythropoiesis. Using single-cell RNA sequencing (scRNA-seq) of primary human terminal erythroid cells (CD34-CD235a+) isolated directly from adult bone marrow (BM) and umbilical cord blood (UCB), we documented the transcriptome of terminally differentiated human erythroblasts at unprecedented resolution. The insights enabled us to distinguish polychromatic erythroblasts (PolyEs) at the early and late stages of development as well as the different development stages of orthochromatic erythroblasts (OrthoEs). We further identified a set of putative regulators of terminal erythroid differentiation and functionally validated three of the identified genes, AKAP8L, TERF2IP, and RNF10, by monitoring cell differentiation and apoptosis. We documented that knockdown of AKAP8L suppressed the commitment of HSCs to erythroid lineage and cell proliferation and delayed differentiation of colony-forming unit-erythroid (CFU-E) to the proerythroblast stage (ProE). In contrast, the knockdown of TERF2IP and RNF10 delayed differentiation of PolyE to OrthoE stage. Taken together, the convergence and divergence of the transcriptional continuums at single-cell resolution underscore the transcriptional regulatory networks that underlie human fetal and adult terminal erythroid differentiation.

Overall design

CD235a+ cells were isolated from 3 adult bone marrow and 3 umbilical cord blood samples and further sequenced by 10x Genomics platform to obtain transcriptomic profiles.UMI Count matrices were generated by cellranger.

Contributors

Peng Huang 1 2 3 4, Yongzhong Zhao 5, Jianmei Zhong 1 2 4, Xinhua Zhang 6, Qifa Liu 7, Xiaoxia Qiu 3, Shaoke Chen 3, Hongxia Yan 8, Christopher Hillyer 8, Narla Mohandas 8, Xinghua Pan 9 10, Xiangmin Xu 11 2 4

Contact

To be supplemented.