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.
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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

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Figure 3

ADGRG1 enriches for functional CB HSCs upon mitochondrial oxidative stress induced by ex vivo culturing.

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ADGRG1 enriches for functional CB HSCs upon mitochondrial oxidative stre...
(A and B) The percentage of hCD45+ cells in BM of NSG recipients transplanted with 50,000 CD34+CD133+ADGRG1+ CB cells or CD34+CD133+ADGRG1– CB cells was determined by FACS. Quantification data are shown as dot plots (mean ± SEM) in B. n = 10 mice per group. **P < 0.01, by 2-tailed Student’s t test. Ctrl, control. (C and D) Human SRCs in ADGRG1+ and ADGRG– CB CD34+CD133+ cells upon ex vivo culturing, as determined by LDA. n = 10 mice per group. ***P < 0.001, by Poisson statistical analysis. (E) The percentage of hCD45+ cells in PB of secondary recipient mice was determined 4 months after transplantation (mean ± SEM). n = 8–9 mice per group. *P < 0.05, by 2-tailed Student’s t test. (F and G) BM cells (5 × 106) from primary recipient mice transplanted with CD34+CD133+ADGRG1+ CB cells or CD34+CD133+ADGRG1– CB cells were infused into secondary recipient mice. The percentage of hCD45+ cells in BM was determined 4 months after transplantation (mean ± SEM). n = 8–9 mice per group. **P < 0.01, by 2-tailed Student’s t test. (H and I) Flow cytometric plots showing the homing efficiency of CD34+CD133+ADGRG1+ CB cells and CD34+CD133+ADGRG1– CB cells (mean ± SEM). n = 7 mice per group. Two-tailed Student’s t test.