Mosaic loss of chromosome Y promotes leukemogenesis and clonal hematopoiesis
Qi Zhang, … , Yu Liu, Chong Chen
Qi Zhang, … , Yu Liu, Chong Chen
Published February 8, 2022
Citation Information: JCI Insight. 2022;7(3):e153768. https://doi.org/10.1172/jci.insight.153768.
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Research Article Genetics Hematology

Mosaic loss of chromosome Y promotes leukemogenesis and clonal hematopoiesis

Abstract

Mosaic loss of chromosome Y (mLOY) in blood cells is one of the most frequent chromosome alterations in adult males. It is strongly associated with clonal hematopoiesis, hematopoietic malignancies, and other hematopoietic and nonhematopoietic diseases. However, whether there is a causal relationship between mLOY and human diseases is unknown. Here, we generated mLOY in murine hematopoietic stem and progenitor cells (HSPCs) with CRISPR/Cas9 genome editing. We found that mLOY led to dramatically increased DNA damage in HSPCs. Interestingly, HSPCs with mLOY displayed significantly enhanced reconstitution capacity and gave rise to clonal hematopoiesis in vivo. mLOY, which is associated with AML1-ETO translocation and p53 defects in patients with acute myeloid leukemia (AML), promoted AML in mice. Mechanistically, loss of KDM5D, a chromosome Y–specific histone 3 lysine 4 demethylase in both humans and mice, partially recapitulated mLOY in DNA damage and leukemogenesis. Thus, our study validates mLOY as a functional driver for clonal hematopoiesis and leukemogenesis.

Authors

Qi Zhang, Lei Zhao, Yi Yang, Shujun Li, Yu Liu, Chong Chen

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Figure 1

Generating mLOY in mouse HSPCs.

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Generating mLOY in mouse HSPCs. (A) Schematic of CRISPR/Cas9 genome edit...
(A) Schematic of CRISPR/Cas9 genome editing of chromosome Y in mouse HSPCs. sgSsty1-sgCas9 and sgSsty2-sgCas9 target repeat sequences located on chromosome Y, which causes chromosome Y elimination. (B) FISH analysis of chromosomes X and Y in sgScr-sgCas9, sgSsty1-sgCas9, and sgSsty2-sgCas9 HSPCs. Green, FITC-labeled whole-chromosome probe for Y chromosome; red, Texas red–labeled X chromosome probe for XqA7.3; blue, DAPI-labeled DNA. White arrows indicate chromosomes X and Y. Squares indicate single cells shown at a higher resolution in the right panels. Scale bars: 10 μm. (C) Heatmap showing the relative expression levels of the chromosome Y–specific genes in sgSsty1-sgCas9 and sgSsty2-sgCas9 HSPCs compared with sgScr-sgCas9 HSPCs, measured by RNA-seq 8 days after infection (n = 3 for each group). (D) Left: Representative immunofluorescence images of γH2AX foci in sgScr-sgCas9, sgSsty1-sgCas9, and sgSsty2-sgCas9 HSPCs. HSPCs were cultured 1 month in vitro until Cas9 elimination. Scale bars: 10 μm. Red, γH2AX; blue, DAPI-labeled DNA. Right: Plot of γH2AX foci per cell, shown as the mean ± SD. *FDR q < 0.05, **FDR q <0.01 (Kruskal-Wallis test). (E) Left: Representative images of comet assay of sgScr-sgCas9, sgSsty1-sgCas9, and sgSsty2-sgCas9 HSPCs. HSPCs were cultured 1 month in vitro until Cas9 elimination. Scale bars: 50 μm. Right: Results of comet assay of sgScr-sgCas9, sgSsty1-sgCas9, and sgSsty2-sgCas9 HSPCs. The tail moment is shown as the mean ± SD. *FDR q < 0.05, **FDR q < 0.01 (Kruskal-Wallis test). (F) GSEA showing the negative enrichment of the KEGG_MISMATCH_REPAIR gene set in sgSsty1-sgCas9 HSPCs (NES = –1.47; P = 0.04) (top) and sgSsty2-sgCas9 HSPCs (NES = –1.51; P = 0.03) (bottom).