ADGRG1 enriches for functional human hematopoietic stem cells following ex vivo expansion–induced mitochondrial oxidative stress
Yandan Chen, … , Hal E. Broxmeyer, Bin Guo
Yandan Chen, … , Hal E. Broxmeyer, Bin Guo
Published August 31, 2021
Citation Information: J Clin Invest. 2021;131(20):e148329. https://doi.org/10.1172/JCI148329.
View: Text | PDF
Concise Communication

ADGRG1 enriches for functional human hematopoietic stem cells following ex vivo expansion–induced mitochondrial oxidative stress

Abstract

The heterogeneity of human hematopoietic stem cells (HSCs) and hematopoietic progenitor cells (HPCs) under stress conditions such as ex vivo expansion is poorly understood. Here, we report that the frequencies of SCID-repopulating cells were greatly decreased in cord blood (CB) CD34+ HSCs and HPCs upon ex vivo culturing. Transcriptomic analysis and metabolic profiling demonstrated that mitochondrial oxidative stress of human CB HSCs and HPCs notably increased, along with loss of stemness. Limiting dilution analysis revealed that functional human HSCs were enriched in cell populations with low levels of mitochondrial ROS (mitoROS) during ex vivo culturing. Using single-cell RNA-Seq analysis of the mitoROS low cell population, we demonstrated that functional HSCs were substantially enriched in the adhesion GPCR G1–positive (ADGRG1+) population of CD34+CD133+ CB cells upon ex vivo expansion stress. Gene set enrichment analysis revealed that HSC signature genes including MSI2 and MLLT3 were enriched in CD34+CD133+ADGRG1+ CB HSCs. Our study reveals that ADGRG1 enriches for functional human HSCs under oxidative stress during ex vivo culturing, which can be a reliable target for drug screening of agonists of HSC expansion.

Authors

Yandan Chen, Shuyi Fang, Qingwei Ding, Rongzhen Jiang, Jiefeng He, Qin Wang, Yuting Jin, Xinxin Huang, Sheng Liu, Maegan L. Capitano, Thao Trinh, Yincheng Teng, Qingyou Meng, Jun Wan, Hal E. Broxmeyer, Bin Guo

×

Figure 1

Functional CB HSCs are enriched in mitoROS low CD34+ cells upon ex vivo culture–induced mitochondrial oxidative stress.

Options: View larger image (or click on image) Download as PowerPoint
Functional CB HSCs are enriched in mitoROS low CD34+ cells upon ex vivo ...
(A and B) Frequency of human SRCs in freshly isolated CB CD34+ cells or ex vivo–cultured CB CD34+ cells, as determined by LDA. n = 4–5 mice per group. **P < 0.01; Poisson statistical analysis. (C) Histogram of mitoROS levels of fresh human CB CD34+ cells or cultured CB CD34+ cells. Representative histograms from 4 independent experiments are shown. (D) Strategy for SRC determination by LDA in mitoROS low CB CD34+ cells or mitoROS high CB CD34+ cells upon ex vivo culturing. (E and F) Percentage of hCD45+ cell chimerism in the BM of NSG mice 16 weeks after transplantation with 10,000 mitoROS low CB CD34+ cells or mitoROS high CB CD34+ cells upon ex vivo culturing. n = 10 mice per group. (F) Statistical data are shown as dot plots (mean ± SEM). **P < 0.01, by 2-tailed Student’s t test. (G and H) Frequency of human SRCs in mitoROS low CB CD34+ cells or mitoROS high CB CD34+ cells upon ex vivo culture stress. n = 4–15 mice per group. ***P < 0.001; Poisson statistical analysis. (I) Percentage of hCD45+ cell chimerism in the BM of NSG mice 4 months after transplantation with 50,000 mitoROS low CB CD34+ cells, mitoROS medium CB CD34+ cells, or mitoROS high CB CD34+ cells following ex vivo culturing. Representative data from 2 independent experiments are shown. n = 5 mice per group. ***P < 0.001, by 1-way ANOVA. NS, nonsignificant.