Pathogenic human variant that dislocates GATA2 zinc fingers disrupts hematopoietic gene expression and signaling networks
Mabel Minji Jung, … , Sunduz Keles, Emery H. Bresnick
Mabel Minji Jung, … , Sunduz Keles, Emery H. Bresnick
Published February 21, 2023
Citation Information: J Clin Invest. 2023;133(7):e162685. https://doi.org/10.1172/JCI162685.
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Research Article Genetics Hematology

Pathogenic human variant that dislocates GATA2 zinc fingers disrupts hematopoietic gene expression and signaling networks

Abstract

Although certain human genetic variants are conspicuously loss of function, decoding the impact of many variants is challenging. Previously, we described a patient with leukemia predisposition syndrome (GATA2 deficiency) with a germline GATA2 variant that inserts 9 amino acids between the 2 zinc fingers (9aa-Ins). Here, we conducted mechanistic analyses using genomic technologies and a genetic rescue system with Gata2 enhancer–mutant hematopoietic progenitor cells to compare how GATA2 and 9aa-Ins function genome-wide. Despite nuclear localization, 9aa-Ins was severely defective in occupying and remodeling chromatin and regulating transcription. Variation of the inter–zinc finger spacer length revealed that insertions were more deleterious to activation than repression. GATA2 deficiency generated a lineage-diverting gene expression program and a hematopoiesis-disrupting signaling network in progenitors with reduced granulocyte-macrophage colony-stimulating factor (GM-CSF) and elevated IL-6 signaling. As insufficient GM-CSF signaling caused pulmonary alveolar proteinosis and excessive IL-6 signaling promoted bone marrow failure and GATA2 deficiency patient phenotypes, these results provide insight into mechanisms underlying GATA2-linked pathologies.

Authors

Mabel Minji Jung, Siqi Shen, Giovanni A. Botten, Thomas Olender, Koichi R. Katsumura, Kirby D. Johnson, Alexandra A. Soukup, Peng Liu, Qingzhou Zhang, Zena D. Jensvold, Peter W. Lewis, Robert A. Beagrie, Jason K.K. Low, Lihua Yang, Joel P. Mackay, Lucy A. Godley, Marjorie Brand, Jian Xu, Sunduz Keles, Emery H. Bresnick

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Figure 10

GATA2-mediated regulation of cellular signaling and differentiation.

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GATA2-mediated regulation of cellular signaling and differentiation. (A)...
(A) Western blot to detect GM-CSF–induced STAT5 phosphorylation (n = 6). (B) p-STAT5 quantification. Results were normalized to GM-CSF–treated hi–77+/+ empty (box-and-whisker plots with bounds from the 25th to the 75th percentiles, the median line, and whiskers ranging from minimum to maximum values) (n = 6). (C) Western blot to detect IL-6–induced STAT3 phosphorylation (p-STAT3) (n = 4). (D) p-STAT3 quantification. Results were normalized to IL-6–treated hi–77–/– empty (box-and-whisker plots with bounds from the 25th to the 75th percentiles, the median line, and whiskers ranging from minimum to maximum values) (n = 4). Statistical comparisons in B and D used paired 2-tailed Student’s t tests with Benjamini-Hochberg correction; *P < 0.05; ***P < 0.001; ****P < 0.0001. (E) Flow cytometric plots of CD11b+CD115– (granulocytic) and CD11b+CD115+ (monocytic) differentiated progenitors cultured for 3 days in control, GM-CSF–containing, or IL-6–containing media (n = 5). Plots (gated at CD11b+CD115–) of Ly6G+Ly6Chi and Ly6G+Ly6lo– differentiated progenitors cultured for 3 days in control, GM-CSF–containing, or IL-6–containing media (n = 5). (F) Quantification of CD11b+CD115–, CD11b+CD115+, CD11b+CD115–Ly6G+Ly6Ghi, and CD11b+CD115–Ly6G+Ly6lo– populations. Error bars represent mean ± SEM. For multiple comparisons with vehicle-treated control, statistics were calculated using 1-way ANOVA followed by Dunnett’s test; *P < 0.05; **P < 0.01; ***P < 0.001.