Zeb1 modulates hematopoietic stem cell fates required for suppressing acute myeloid leukemia
Alhomidi Almotiri, … , Florian A. Siebzehnrubl, Neil P. Rodrigues
Alhomidi Almotiri, … , Florian A. Siebzehnrubl, Neil P. Rodrigues
Published October 27, 2020
Citation Information: J Clin Invest. 2021;131(1):e129115. https://doi.org/10.1172/JCI129115.
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Zeb1 modulates hematopoietic stem cell fates required for suppressing acute myeloid leukemia

Abstract

Zeb1, a zinc finger E-box binding homeobox epithelial-mesenchymal transition (EMT) transcription factor, confers properties of “stemness,” such as self-renewal, in cancer. Yet little is known about the function of Zeb1 in adult stem cells. Here, we used the hematopoietic system as a well-established paradigm of stem cell biology to evaluate Zeb1-mediated regulation of adult stem cells. We employed a conditional genetic approach using the Mx1-Cre system to specifically knock out (KO) Zeb1 in adult hematopoietic stem cells (HSCs) and their downstream progeny. Acute genetic deletion of Zeb1 led to rapid-onset thymic atrophy and apoptosis-driven loss of thymocytes and T cells. A profound cell-autonomous self-renewal defect and multilineage differentiation block were observed in Zeb1-KO HSCs. Loss of Zeb1 in HSCs activated transcriptional programs of deregulated HSC maintenance and multilineage differentiation genes and of cell polarity consisting of cytoskeleton-, lipid metabolism/lipid membrane–, and cell adhesion–related genes. Notably, epithelial cell adhesion molecule (EpCAM) expression was prodigiously upregulated in Zeb1-KO HSCs, which correlated with enhanced cell survival, diminished mitochondrial metabolism, ribosome biogenesis, and differentiation capacity and an activated transcriptomic signature associated with acute myeloid leukemia (AML) signaling. ZEB1 expression was downregulated in AML patients, and Zeb1 KO in the malignant counterparts of HSCs — leukemic stem cells (LSCs) — accelerated MLL-AF9– and Meis1a/Hoxa9-driven AML progression, implicating Zeb1 as a tumor suppressor in AML LSCs. Thus, Zeb1 acts as a transcriptional regulator in hematopoiesis, critically coordinating HSC self-renewal, apoptotic, and multilineage differentiation fates required to suppress leukemic potential in AML.

Authors

Alhomidi Almotiri, Hamed Alzahrani, Juan Bautista Menendez-Gonzalez, Ali Abdelfattah, Badi Alotaibi, Lubaid Saleh, Adelle Greene, Mia Georgiou, Alex Gibbs, Amani Alsayari, Sarab Taha, Leigh-anne Thomas, Dhruv Shah, Sarah Edkins, Peter Giles, Marc P. Stemmler, Simone Brabletz, Thomas Brabletz, Ashleigh S. Boyd, Florian A. Siebzehnrubl, Neil P. Rodrigues

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Figure 1

Loss of Zeb1 affects effector and CM CD8+ T cells.

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Loss of Zeb1 affects effector and CM CD8+ T cells. (A) Q-PCR analysis of...
(A) Q-PCR analysis of mRNA Zeb1 expression in different hematopoietic populations (n = 6–7 except CLP n = 3). (B) Schematic of pIpC treatment to delete Zeb1 in Zeb1fl/fl;Mx1-Cre– (control) and Zeb1fl/fl;Mx1-Cre+ (Zeb1–/–) mice and analysis at day 14 after the last pIpC dose. (C) Representative gel electrophoresis analysis confirming Zeb1 deletion in BM cells and LSK population 14 days after the last dose of pIpC. (D) Representative gel electrophoresis analysis of Zeb1 deletion in BM C-KIT+ cells and spleen B (B220+) and T (CD3+) cells 14 days after the last dose of pIpC. (E) Frequency of differentiated cells in PB from control and Zeb1–/– mice 14 days after the last dose of pIpC from 4 independent experiments (n = 8–12 per group). (F) Gating strategy of naive, EM, and CM T cells using CD62L and CD44 markers along with T cell markers CD3, CD4, and CD8 in PB. Frequency of EM T cells (G) and CM T cells (H) within CD3+CD8+ T cells in PB, BM, and SP from control (n = 5 PB and BM, 6 SP) and Zeb1–/– (n = 5 PB and BM, 6–7 SP) mice from 2 independent experiments. Error bars show mean ± SEM. Mann-Whitney U test was used to calculate significance. *P < 0.05.