Pluripotent stem cells reveal erythroid-specific activities of the GATA1 N-terminus
Marta Byrska-Bishop, … , Mitchell J. Weiss, Stella T. Chou
Marta Byrska-Bishop, … , Mitchell J. Weiss, Stella T. Chou
Published January 26, 2015
Citation Information: J Clin Invest. 2015;125(3):993-1005. https://doi.org/10.1172/JCI75714.
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Pluripotent stem cells reveal erythroid-specific activities of the GATA1 N-terminus

Abstract

Germline GATA1 mutations that result in the production of an amino-truncated protein termed GATA1s (where s indicates short) cause congenital hypoplastic anemia. In patients with trisomy 21, similar somatic GATA1s-producing mutations promote transient myeloproliferative disease and acute megakaryoblastic leukemia. Here, we demonstrate that induced pluripotent stem cells (iPSCs) from patients with GATA1-truncating mutations exhibit impaired erythroid potential, but enhanced megakaryopoiesis and myelopoiesis, recapitulating the major phenotypes of the associated diseases. Similarly, in developmentally arrested GATA1-deficient murine megakaryocyte-erythroid progenitors derived from murine embryonic stem cells (ESCs), expression of GATA1s promoted megakaryopoiesis, but not erythropoiesis. Transcriptome analysis revealed a selective deficiency in the ability of GATA1s to activate erythroid-specific genes within populations of hematopoietic progenitors. Although its DNA-binding domain was intact, chromatin immunoprecipitation studies showed that GATA1s binding at specific erythroid regulatory regions was impaired, while binding at many nonerythroid sites, including megakaryocytic and myeloid target genes, was normal. Together, these observations indicate that lineage-specific GATA1 cofactor associations are essential for normal chromatin occupancy and provide mechanistic insights into how GATA1s mutations cause human disease. More broadly, our studies underscore the value of ESCs and iPSCs to recapitulate and study disease phenotypes.

Authors

Marta Byrska-Bishop, Daniel VanDorn, Amy E. Campbell, Marisol Betensky, Philip R. Arca, Yu Yao, Paul Gadue, Fernando F. Costa, Richard L. Nemiroff, Gerd A. Blobel, Deborah L. French, Ross C. Hardison, Mitchell J. Weiss, Stella T. Chou

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Figure 6

Single-cell gene expression analysis predicts erythroid-to-myeloid fate bias in GATA1s mutant progenitors.

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Single-cell gene expression analysis predicts erythroid-to-myeloid fate ...
(A) Projections of expression patterns of iPSC-derived committed cells (filled circles in both plots) and CD43+CD41+CD235+ progenitors expressing WT GATA1 (left, open diamonds) or GATA1s (right, open diamonds) onto linear discriminants 1 and 2 (LD1 and LD2). Projections for each cell are colored according to the classification obtained from LDA using a probability threshold of greater than 0.90. Classifications of committed cells: erythroid (red circles), megakaryocytic (blue circles), myeloid (green circles); classifications of WT GATA1 (left) or GATA1s (right) progenitors: predicted erythroid (pEry) (red diamonds), predicted megakaryocytic (pMeg) (blue diamonds), predicted myeloid (pMyelo) (green diamonds). Gray diamonds represent progenitors that were unclassified at a probability threshold of greater than 0.90. (B) Hierarchical clustering on probabilities of belonging to erythroid, megakaryocytic, or myeloid class assigned by LDA to progenitor cells. WT GATA1 or GATA1s progenitors that were assigned to a given class with a probability of greater than 0.90 are represented on 2 heat maps on the left, while cells that were unclassified (probability < 0.90) are shown on 2 heat maps on the right. (C) Classification of CD43+CD41+CD235+ progenitors using LDA. Plotted are fractions of WT GATA1 and GATA1s progenitor populations (consisting of a total of 311 and 274 single cells, respectively) classified into predicted cell fate with a probability of greater than 0.90. The remaining cells are plotted as unclassified. *P < 0.05 (Fisher’s exact test).