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Single cell analyses demonstrate that a heme - GATA1 feedback loop regulates red cell differentiation

Raymond T. Doty, Xiaowei Yan, Christopher Lausted, Adam D. Munday, Zhantao Yang, Danielle Yi, Neda Jabbari, Li Liu, Siobán B. Keel, Qiang Tian and Janis L. Abkowitz

Key points

  • Heme rapidly upregulates ribosomal protein genes during early erythropoiesis to help ensure sufficient ribosomes for globin translation.

  • Heme later downregulates GATA1, GATA1-targets, and mitotic spindle genes to enable the normal termination of erythroid differentiation.

Abstract

Erythropoiesis is the complex, dynamic, and tightly-regulated process that generates all mature red blood cells. To understand this process, we mapped the developmental trajectories of progenitors from wildtype, erythropoietin-treated and Flvcr1-deleted mice at single cell resolution. Importantly, we linked the quantity of each cell's surface proteins to its total transcriptome, which is a novel method. Deletion of Flvcr1 results in high levels of intracellular heme, allowing us to identify heme-regulated circuitry. Our studies demonstrate that in early erythroid cells (CD71+Ter119neg-lo), heme increases ribosomal protein transcripts; suggesting that heme, in addition to upregulating globin transcription and translation, guarantees ample ribosomes for globin synthesis. In later erythroid cells (CD71+Ter119lo-hi), heme decreases GATA1, GATA1-target gene, and mitotic spindle gene expression. These changes occur quickly. For example, in confirmatory studies using human marrow erythroid cells, ribosomal protein transcripts and proteins increase, and GATA1 transcript and protein decrease, within 15-30 minutes of amplifying endogenous heme synthesis with ALA. Since GATA1 initiates heme synthesis, GATA1 and heme together direct red cell maturation, and heme stops GATA1 synthesis, our observations reveal a GATA1-heme autoregulatory loop and implicate GATA1 and heme as the co-master regulators of the normal erythroid differentiation program. In addition, since excessive heme could amplify ribosomal protein imbalance, prematurely lower GATA1 and impede mitosis, these data may help explain the ineffective (early termination of) erythropoiesis in Diamond Blackfan anemia and del(5q) myelodysplasia, disorders with excessive heme in CFU-E/proerythroblasts; explain why these anemias are macrocytic; and show why children with GATA1 mutations have DBA-like clinical phenotypes.

  • Submitted May 16, 2018.
  • Accepted December 1, 2018.