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ETV6 represses inflammatory response genes and regulates HSPC function during stress hematopoiesis in mice
ETS variant 6 (ETV6) encodes a transcriptional repressor expressed in hematopoietic stem and progenitor cells (HSPCs), where it is required for adult hematopoiesis. Heterozygous pathogenic germline ETV6 variants are associated with thrombocytopenia 5 (T5), a poorly understood genetic condition resul...
Autores principales: | , , , , , , , , , , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
The American Society of Hematology
2023
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10514086/ https://www.ncbi.nlm.nih.gov/pubmed/37522715 http://dx.doi.org/10.1182/bloodadvances.2022009313 |
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author | Bloom, Mackenzie Oak, Ninad Baskin-Doerfler, Rebekah Feng, Ruopeng Iacobucci, Ilaria Baviskar, Pradyumna Zhao, Xujie Stroh, Alexa N. Li, Chunliang Ozark, Patrick Tillman, Heather S. Li, Yichao Verbist, Katherine C. Albeituni, Sabrin Scott, Danny C. King, Moeko T. McKinney-Freeman, Shannon L. Weiss, Mitchell J. Yang, Jun J. Nichols, Kim E. |
author_facet | Bloom, Mackenzie Oak, Ninad Baskin-Doerfler, Rebekah Feng, Ruopeng Iacobucci, Ilaria Baviskar, Pradyumna Zhao, Xujie Stroh, Alexa N. Li, Chunliang Ozark, Patrick Tillman, Heather S. Li, Yichao Verbist, Katherine C. Albeituni, Sabrin Scott, Danny C. King, Moeko T. McKinney-Freeman, Shannon L. Weiss, Mitchell J. Yang, Jun J. Nichols, Kim E. |
author_sort | Bloom, Mackenzie |
collection | PubMed |
description | ETS variant 6 (ETV6) encodes a transcriptional repressor expressed in hematopoietic stem and progenitor cells (HSPCs), where it is required for adult hematopoiesis. Heterozygous pathogenic germline ETV6 variants are associated with thrombocytopenia 5 (T5), a poorly understood genetic condition resulting in thrombocytopenia and predisposition to hematologic malignancies. To elucidate how germline ETV6 variants affect HSPCs and contribute to disease, we generated a mouse model harboring an Etv6(R355X) loss-of-function variant, equivalent to the T5-associated variant ETV6(R359X). Under homeostatic conditions, all HSPC subpopulations are present in the bone marrow (BM) of Etv6(R355X/+) mice; however, these animals display shifts in the proportions and/or numbers of progenitor subtypes. To examine whether the Etv6(R355X/+) mutation affects HSPC function, we performed serial competitive transplantation and observed that Etv6(R355X/+) lineage−sca1+cKit+ (LSK) cells exhibit impaired reconstitution, with near complete failure to repopulate irradiated recipients by the tertiary transplant. Mechanistic studies incorporating cleavage under target and release under nuclease assay, assay for transposase accessible chromatin sequencing, and high-throughput chromosome conformation capture identify ETV6 binding at inflammatory gene loci, including multiple genes within the tumor necrosis factor (TNF) signaling pathway in ETV6-sufficient mouse and human HSPCs. Furthermore, single-cell RNA sequencing of BM cells isolated after transplantation reveals upregulation of inflammatory genes in Etv6(R355X/+) progenitors when compared to Etv6(+/+) counterparts. Corroborating these findings, Etv6(R355X/+) HSPCs produce significantly more TNF than Etv6(+/+) cells post-transplantation. We conclude that ETV6 is required to repress inflammatory gene expression in HSPCs under conditions of hematopoietic stress, and this mechanism may be critical to sustain HSPC function. |
format | Online Article Text |
id | pubmed-10514086 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2023 |
publisher | The American Society of Hematology |
record_format | MEDLINE/PubMed |
spelling | pubmed-105140862023-09-23 ETV6 represses inflammatory response genes and regulates HSPC function during stress hematopoiesis in mice Bloom, Mackenzie Oak, Ninad Baskin-Doerfler, Rebekah Feng, Ruopeng Iacobucci, Ilaria Baviskar, Pradyumna Zhao, Xujie Stroh, Alexa N. Li, Chunliang Ozark, Patrick Tillman, Heather S. Li, Yichao Verbist, Katherine C. Albeituni, Sabrin Scott, Danny C. King, Moeko T. McKinney-Freeman, Shannon L. Weiss, Mitchell J. Yang, Jun J. Nichols, Kim E. Blood Adv Hematopoiesis and Stem Cells ETS variant 6 (ETV6) encodes a transcriptional repressor expressed in hematopoietic stem and progenitor cells (HSPCs), where it is required for adult hematopoiesis. Heterozygous pathogenic germline ETV6 variants are associated with thrombocytopenia 5 (T5), a poorly understood genetic condition resulting in thrombocytopenia and predisposition to hematologic malignancies. To elucidate how germline ETV6 variants affect HSPCs and contribute to disease, we generated a mouse model harboring an Etv6(R355X) loss-of-function variant, equivalent to the T5-associated variant ETV6(R359X). Under homeostatic conditions, all HSPC subpopulations are present in the bone marrow (BM) of Etv6(R355X/+) mice; however, these animals display shifts in the proportions and/or numbers of progenitor subtypes. To examine whether the Etv6(R355X/+) mutation affects HSPC function, we performed serial competitive transplantation and observed that Etv6(R355X/+) lineage−sca1+cKit+ (LSK) cells exhibit impaired reconstitution, with near complete failure to repopulate irradiated recipients by the tertiary transplant. Mechanistic studies incorporating cleavage under target and release under nuclease assay, assay for transposase accessible chromatin sequencing, and high-throughput chromosome conformation capture identify ETV6 binding at inflammatory gene loci, including multiple genes within the tumor necrosis factor (TNF) signaling pathway in ETV6-sufficient mouse and human HSPCs. Furthermore, single-cell RNA sequencing of BM cells isolated after transplantation reveals upregulation of inflammatory genes in Etv6(R355X/+) progenitors when compared to Etv6(+/+) counterparts. Corroborating these findings, Etv6(R355X/+) HSPCs produce significantly more TNF than Etv6(+/+) cells post-transplantation. We conclude that ETV6 is required to repress inflammatory gene expression in HSPCs under conditions of hematopoietic stress, and this mechanism may be critical to sustain HSPC function. The American Society of Hematology 2023-08-02 /pmc/articles/PMC10514086/ /pubmed/37522715 http://dx.doi.org/10.1182/bloodadvances.2022009313 Text en © 2023 by The American Society of Hematology. Licensed under Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International (CC BY-NC-ND 4.0), permitting only noncommercial, nonderivative use with attribution. All other rights reserved. https://creativecommons.org/licenses/by-nc-nd/4.0/This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/). |
spellingShingle | Hematopoiesis and Stem Cells Bloom, Mackenzie Oak, Ninad Baskin-Doerfler, Rebekah Feng, Ruopeng Iacobucci, Ilaria Baviskar, Pradyumna Zhao, Xujie Stroh, Alexa N. Li, Chunliang Ozark, Patrick Tillman, Heather S. Li, Yichao Verbist, Katherine C. Albeituni, Sabrin Scott, Danny C. King, Moeko T. McKinney-Freeman, Shannon L. Weiss, Mitchell J. Yang, Jun J. Nichols, Kim E. ETV6 represses inflammatory response genes and regulates HSPC function during stress hematopoiesis in mice |
title | ETV6 represses inflammatory response genes and regulates HSPC function during stress hematopoiesis in mice |
title_full | ETV6 represses inflammatory response genes and regulates HSPC function during stress hematopoiesis in mice |
title_fullStr | ETV6 represses inflammatory response genes and regulates HSPC function during stress hematopoiesis in mice |
title_full_unstemmed | ETV6 represses inflammatory response genes and regulates HSPC function during stress hematopoiesis in mice |
title_short | ETV6 represses inflammatory response genes and regulates HSPC function during stress hematopoiesis in mice |
title_sort | etv6 represses inflammatory response genes and regulates hspc function during stress hematopoiesis in mice |
topic | Hematopoiesis and Stem Cells |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10514086/ https://www.ncbi.nlm.nih.gov/pubmed/37522715 http://dx.doi.org/10.1182/bloodadvances.2022009313 |
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