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Using a new HSPC senescence model in vitro to explore the mechanism of cellular memory in aging HSPCs
BACKGROUND: Age-associated changes attenuate human blood system functionality through the aging of hematopoietic stem and progenitor cells (HSPCs), manifested in human populations an increase in myeloproliferative disease and even leukemia; therefore, study on HSPC senescence bears great significanc...
Autores principales: | , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
BioMed Central
2021
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8351417/ https://www.ncbi.nlm.nih.gov/pubmed/34365970 http://dx.doi.org/10.1186/s13287-021-02455-x |
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author | Dong, Yongpin Guo, Chunni Zhou, Wuxiong Li, Wenfang Zhang, Lina |
author_facet | Dong, Yongpin Guo, Chunni Zhou, Wuxiong Li, Wenfang Zhang, Lina |
author_sort | Dong, Yongpin |
collection | PubMed |
description | BACKGROUND: Age-associated changes attenuate human blood system functionality through the aging of hematopoietic stem and progenitor cells (HSPCs), manifested in human populations an increase in myeloproliferative disease and even leukemia; therefore, study on HSPC senescence bears great significance to treat hematopoietic-associated disease. Furthermore, the mechanism of HSPC aging is lacking, especially the cellular memory mechanism. Here, we not only reported a new HSPC senescence model in vitro, but also propose and verify the cellular memory mechanism of HSPC aging of the Polycomb/Trithorax system. METHODS: HSPCs (Lin(−)c-kit(+) cells) were isolated and purified by magnetic cell sorting (MACS). The proportions and cell cycle distribution of cells were determined by flow cytometry; senescence-related β-galactosidase assay, transmission electron microscope (TEM), and colony-forming unit (CFU)-mix assay were detected for identification of the old HSPC model. Proteomic tests and RNA-seq were applied to analyze differential pathways and genes in the model cells. qPCR, Western blot (WB), and chromatin immunoprecipitation PCR (CHIP-PCR) were used to detect the gene expression of cell memory-related proteins. Knockdown of cell memory-related key genes was performed with shRNA interference. RESULTS: In the model old HSPCs, β-gal activity, cell cycle, colony-forming ability, aging-related cell morphology, and metabolic pathway were significantly changed compared to the young HSPCs. Furthermore, we found the model HSPCs have more obvious aging manifestations than those of natural mice, and IL3 is the major factor contributing to HSPC aging in the model. We also observed dramatic changes in the expression level of PRC/TrxG complexes. After further exploring the downstream molecules of PRC/TrxG complexes, we found that Uhrf1 and TopII played critical roles in HSPC aging based on the HSPC senescence model. CONCLUSIONS: These findings proposed a new HSPC senescence model in vitro which we forecasted could be used to preliminary screen the drugs of the HSPC aging-related hemopathy and suggested cellular memory mechanism of HSPC aging. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1186/s13287-021-02455-x. |
format | Online Article Text |
id | pubmed-8351417 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2021 |
publisher | BioMed Central |
record_format | MEDLINE/PubMed |
spelling | pubmed-83514172021-08-09 Using a new HSPC senescence model in vitro to explore the mechanism of cellular memory in aging HSPCs Dong, Yongpin Guo, Chunni Zhou, Wuxiong Li, Wenfang Zhang, Lina Stem Cell Res Ther Research BACKGROUND: Age-associated changes attenuate human blood system functionality through the aging of hematopoietic stem and progenitor cells (HSPCs), manifested in human populations an increase in myeloproliferative disease and even leukemia; therefore, study on HSPC senescence bears great significance to treat hematopoietic-associated disease. Furthermore, the mechanism of HSPC aging is lacking, especially the cellular memory mechanism. Here, we not only reported a new HSPC senescence model in vitro, but also propose and verify the cellular memory mechanism of HSPC aging of the Polycomb/Trithorax system. METHODS: HSPCs (Lin(−)c-kit(+) cells) were isolated and purified by magnetic cell sorting (MACS). The proportions and cell cycle distribution of cells were determined by flow cytometry; senescence-related β-galactosidase assay, transmission electron microscope (TEM), and colony-forming unit (CFU)-mix assay were detected for identification of the old HSPC model. Proteomic tests and RNA-seq were applied to analyze differential pathways and genes in the model cells. qPCR, Western blot (WB), and chromatin immunoprecipitation PCR (CHIP-PCR) were used to detect the gene expression of cell memory-related proteins. Knockdown of cell memory-related key genes was performed with shRNA interference. RESULTS: In the model old HSPCs, β-gal activity, cell cycle, colony-forming ability, aging-related cell morphology, and metabolic pathway were significantly changed compared to the young HSPCs. Furthermore, we found the model HSPCs have more obvious aging manifestations than those of natural mice, and IL3 is the major factor contributing to HSPC aging in the model. We also observed dramatic changes in the expression level of PRC/TrxG complexes. After further exploring the downstream molecules of PRC/TrxG complexes, we found that Uhrf1 and TopII played critical roles in HSPC aging based on the HSPC senescence model. CONCLUSIONS: These findings proposed a new HSPC senescence model in vitro which we forecasted could be used to preliminary screen the drugs of the HSPC aging-related hemopathy and suggested cellular memory mechanism of HSPC aging. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1186/s13287-021-02455-x. BioMed Central 2021-08-09 /pmc/articles/PMC8351417/ /pubmed/34365970 http://dx.doi.org/10.1186/s13287-021-02455-x Text en © The Author(s) 2021 https://creativecommons.org/licenses/by/4.0/Open AccessThis article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) . The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/ (https://creativecommons.org/publicdomain/zero/1.0/) ) applies to the data made available in this article, unless otherwise stated in a credit line to the data. |
spellingShingle | Research Dong, Yongpin Guo, Chunni Zhou, Wuxiong Li, Wenfang Zhang, Lina Using a new HSPC senescence model in vitro to explore the mechanism of cellular memory in aging HSPCs |
title | Using a new HSPC senescence model in vitro to explore the mechanism of cellular memory in aging HSPCs |
title_full | Using a new HSPC senescence model in vitro to explore the mechanism of cellular memory in aging HSPCs |
title_fullStr | Using a new HSPC senescence model in vitro to explore the mechanism of cellular memory in aging HSPCs |
title_full_unstemmed | Using a new HSPC senescence model in vitro to explore the mechanism of cellular memory in aging HSPCs |
title_short | Using a new HSPC senescence model in vitro to explore the mechanism of cellular memory in aging HSPCs |
title_sort | using a new hspc senescence model in vitro to explore the mechanism of cellular memory in aging hspcs |
topic | Research |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8351417/ https://www.ncbi.nlm.nih.gov/pubmed/34365970 http://dx.doi.org/10.1186/s13287-021-02455-x |
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