Zbtb14 regulates monocyte and macrophage development through inhibiting pu.1 expression in zebrafish
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Evaluation Summary:
This manuscript by Deng et al., is a valuable evaluation of zbtb14 and its role in normal myelopoiesis. The authors provided convincing data supporting the role played by zbtb14 in monocyte and macrophage development and its regulation involving the modulation of PU.1 expression. The finding that a mutation in ZBTB14 exists in AML patients also implies how important this gene product is in normal human myelopoiesis.
(This preprint has been reviewed by eLife. We include the public reviews from the reviewers here; the authors also receive private feedback with suggested changes to the manuscript. The reviewers remained anonymous to the authors.)
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Abstract
Macrophages and their precursor cells, monocytes, are the first line of defense of the body against foreign pathogens and tissue damage. Although the origins of macrophages are diverse, some common transcription factors (such as PU.1) are required to ensure proper development of monocytes/macrophages. Here, we report that the deficiency of zbtb14 , a transcription repressor gene belonging to ZBTB family, leads to an aberrant expansion of monocyte/macrophage population in zebrafish. Mechanistically, Zbtb14 functions as a negative regulator of pu.1 , and SUMOylation on a conserved lysine is essential for the repression activity of Zbtb14. Moreover, a serine to phenylalanine mutation found in an acute myeloid leukemia (AML) patient could target ZBTB14 protein to autophagic degradation. Hence, ZBTB14 is a newly identified gene implicated in both normal and malignant myelopoiesis.
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Evaluation Summary:
This manuscript by Deng et al., is a valuable evaluation of zbtb14 and its role in normal myelopoiesis. The authors provided convincing data supporting the role played by zbtb14 in monocyte and macrophage development and its regulation involving the modulation of PU.1 expression. The finding that a mutation in ZBTB14 exists in AML patients also implies how important this gene product is in normal human myelopoiesis.
(This preprint has been reviewed by eLife. We include the public reviews from the reviewers here; the authors also receive private feedback with suggested changes to the manuscript. The reviewers remained anonymous to the authors.)
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Reviewer #1 (Public Review):
In this manuscript, the authors generated a zbtb14 mutant zebrafish strain via CRISPR-Cas9. In the mutant fish, they found an abnormal expansion of primitive macrophages during early development and adult macrophages in the kidney marrow. The abnormal expansion of macrophages in the mutants was confirmed to be caused by the loss of zbtb14 function, as over-expressing either zebrafish zbtb14 or human ZBTB14 could rescue the phenotype. To explore the underlying molecular mechanism, the authors performed RNA-seq analysis and found that the expression of pu.1 was up-regulation in the mutant macrophages. They further showed that the injection of mpeg1.1:pu.1-DBD construct into the mutant embryos to suppress the Pu.1 activity was able to rescue the mutant phenotype. The authors then went on to show that the …
Reviewer #1 (Public Review):
In this manuscript, the authors generated a zbtb14 mutant zebrafish strain via CRISPR-Cas9. In the mutant fish, they found an abnormal expansion of primitive macrophages during early development and adult macrophages in the kidney marrow. The abnormal expansion of macrophages in the mutants was confirmed to be caused by the loss of zbtb14 function, as over-expressing either zebrafish zbtb14 or human ZBTB14 could rescue the phenotype. To explore the underlying molecular mechanism, the authors performed RNA-seq analysis and found that the expression of pu.1 was up-regulation in the mutant macrophages. They further showed that the injection of mpeg1.1:pu.1-DBD construct into the mutant embryos to suppress the Pu.1 activity was able to rescue the mutant phenotype. The authors then went on to show that the SUMOylation of Zbtb14 plays an essential role in the transcriptional repression activity of the proteins. Finally, the authors documented that over-expressing the S8F mutant form of human ZBTB14, an AML associated mutation, failed to rescue the macrophage phenotype in zbtb14 mutants, suggesting that the loss of ZBTB14 function may be associated with the development of AML. Overall, the findings are interesting in developmental biology and gene regulation, especially in normal and malignant myelopoiesis.
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Reviewer #2 (Public Review):
The authors of this paper show that zbtb14 negatively regulates pu.1, an essential transcription factor for myelopoiesis. This ablation of zbtb14 caused an increase in macrophages in developing zebrafish. This activity appears to be due to the SUMOylation of zbtb14. Interestingly, they also describe a mutation found in an AML patient that renders ZBTB14 incapable of repressing pu.1.
The manuscript is well written and shows strong data. The lack of macrophages in the developing zebrafish looks very solid, as are the data that show zbtb14 represses pu.1. Finally, the data indicating that the human ZBTB14 mutant functions as a loss-of-function transcription factor is solid.
I agree that the manuscript addresses the aims of the study. The results support the author's conclusions. Importantly, this is interesting …
Reviewer #2 (Public Review):
The authors of this paper show that zbtb14 negatively regulates pu.1, an essential transcription factor for myelopoiesis. This ablation of zbtb14 caused an increase in macrophages in developing zebrafish. This activity appears to be due to the SUMOylation of zbtb14. Interestingly, they also describe a mutation found in an AML patient that renders ZBTB14 incapable of repressing pu.1.
The manuscript is well written and shows strong data. The lack of macrophages in the developing zebrafish looks very solid, as are the data that show zbtb14 represses pu.1. Finally, the data indicating that the human ZBTB14 mutant functions as a loss-of-function transcription factor is solid.
I agree that the manuscript addresses the aims of the study. The results support the author's conclusions. Importantly, this is interesting information for the field. The regulation of hematopoiesis, while studied for 50+ years, has still not been completely elucidated; knowing that zbtb14 is an essential gene for this process is critical. Furthermore, the finding that this gene product is dysregulated in AML elucidates more information about how to use these mechanistic studies to disciover new genes and positively affect human health.
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