Glia maturation factor-γ is required for initiation and maintenance of hematopoietic stem and progenitor cells
Abstract Background In vertebrates, hematopoietic stem and progenitor cells (HSPCs) emerge from hemogenic endothelium in the floor of the dorsal aorta and subsequently migrate to secondary niches where they expand and differentiate into committed lineages. Glia maturation factor γ (gmfg) is a key re...
| Main Authors: | , , , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
BMC
2023-04-01
|
| Series: | Stem Cell Research & Therapy |
| Subjects: | |
| Online Access: | https://doi.org/10.1186/s13287-023-03328-1 |
| _version_ | 1797832218028015616 |
|---|---|
| author | Honghu Li Qian Luo Shuyang Cai Ruxiu Tie Ye Meng Wei Shan Yulin Xu Xiangjun Zeng Pengxu Qian He Huang |
| author_facet | Honghu Li Qian Luo Shuyang Cai Ruxiu Tie Ye Meng Wei Shan Yulin Xu Xiangjun Zeng Pengxu Qian He Huang |
| author_sort | Honghu Li |
| collection | DOAJ |
| description | Abstract Background In vertebrates, hematopoietic stem and progenitor cells (HSPCs) emerge from hemogenic endothelium in the floor of the dorsal aorta and subsequently migrate to secondary niches where they expand and differentiate into committed lineages. Glia maturation factor γ (gmfg) is a key regulator of actin dynamics that was shown to be highly expressed in hematopoietic tissue. Our goal is to investigate the role and mechanism of gmfg in embryonic HSPC development. Methods In-depth bioinformatics analysis of our published RNA-seq data identified gmfg as a cogent candidate gene implicated in HSPC development. Loss and gain-of-function strategies were applied to study the biological function of gmfg. Whole-mount in situ hybridization, confocal microscopy, flow cytometry, and western blotting were used to evaluate changes in the number of various hematopoietic cells and expression levels of cell proliferation, cell apoptosis and hematopoietic-related markers. RNA-seq was performed to screen signaling pathways responsible for gmfg deficiency-induced defects in HSPC initiation. The effect of gmfg on YAP sublocalization was assessed in vitro by utilizing HUVEC cell line. Results We took advantage of zebrafish embryos to illustrate that loss of gmfg impaired HSPC initiation and maintenance. In gmfg-deficient embryos, the number of hemogenic endothelium and HSPCs was significantly reduced, with the accompanying decreased number of erythrocytes, myelocytes and lymphocytes. We found that blood flow modulates gmfg expression and gmfg overexpression could partially rescue the reduction of HSPCs in the absence of blood flow. Assays in zebrafish and HUVEC showed that gmfg deficiency suppressed the activity of YAP, a well-established blood flow mediator, by preventing its shuttling from cytoplasm to nucleus. During HSPC initiation, loss of gmfg resulted in Notch inactivation and the induction of Notch intracellular domain could partially restore the HSPC loss in gmfg-deficient embryos. Conclusions We conclude that gmfg mediates blood flow-induced HSPC maintenance via regulation of YAP, and contributes to HSPC initiation through the modulation of Notch signaling. Our findings reveal a brand-new aspect of gmfg function and highlight a novel mechanism for embryonic HSPC development. |
| first_indexed | 2024-04-09T14:04:16Z |
| format | Article |
| id | doaj.art-51da32aa5030432f8a0a8dd1ae6d4b54 |
| institution | Directory Open Access Journal |
| issn | 1757-6512 |
| language | English |
| last_indexed | 2024-04-09T14:04:16Z |
| publishDate | 2023-04-01 |
| publisher | BMC |
| record_format | Article |
| series | Stem Cell Research & Therapy |
| spelling | doaj.art-51da32aa5030432f8a0a8dd1ae6d4b542023-05-07T11:07:18ZengBMCStem Cell Research & Therapy1757-65122023-04-0114111910.1186/s13287-023-03328-1Glia maturation factor-γ is required for initiation and maintenance of hematopoietic stem and progenitor cellsHonghu Li0Qian Luo1Shuyang Cai2Ruxiu Tie3Ye Meng4Wei Shan5Yulin Xu6Xiangjun Zeng7Pengxu Qian8He Huang9Bone Marrow Transplantation Center, School of Medicine, The First Affiliated Hospital, Zhejiang UniversityBone Marrow Transplantation Center, School of Medicine, The First Affiliated Hospital, Zhejiang UniversityBone Marrow Transplantation Center, School of Medicine, The First Affiliated Hospital, Zhejiang UniversityBone Marrow Transplantation Center, School of Medicine, The First Affiliated Hospital, Zhejiang UniversityBone Marrow Transplantation Center, School of Medicine, The First Affiliated Hospital, Zhejiang UniversityBone Marrow Transplantation Center, School of Medicine, The First Affiliated Hospital, Zhejiang UniversityBone Marrow Transplantation Center, School of Medicine, The First Affiliated Hospital, Zhejiang UniversityBone Marrow Transplantation Center, School of Medicine, The First Affiliated Hospital, Zhejiang UniversityBone Marrow Transplantation Center, School of Medicine, The First Affiliated Hospital, Zhejiang UniversityBone Marrow Transplantation Center, School of Medicine, The First Affiliated Hospital, Zhejiang UniversityAbstract Background In vertebrates, hematopoietic stem and progenitor cells (HSPCs) emerge from hemogenic endothelium in the floor of the dorsal aorta and subsequently migrate to secondary niches where they expand and differentiate into committed lineages. Glia maturation factor γ (gmfg) is a key regulator of actin dynamics that was shown to be highly expressed in hematopoietic tissue. Our goal is to investigate the role and mechanism of gmfg in embryonic HSPC development. Methods In-depth bioinformatics analysis of our published RNA-seq data identified gmfg as a cogent candidate gene implicated in HSPC development. Loss and gain-of-function strategies were applied to study the biological function of gmfg. Whole-mount in situ hybridization, confocal microscopy, flow cytometry, and western blotting were used to evaluate changes in the number of various hematopoietic cells and expression levels of cell proliferation, cell apoptosis and hematopoietic-related markers. RNA-seq was performed to screen signaling pathways responsible for gmfg deficiency-induced defects in HSPC initiation. The effect of gmfg on YAP sublocalization was assessed in vitro by utilizing HUVEC cell line. Results We took advantage of zebrafish embryos to illustrate that loss of gmfg impaired HSPC initiation and maintenance. In gmfg-deficient embryos, the number of hemogenic endothelium and HSPCs was significantly reduced, with the accompanying decreased number of erythrocytes, myelocytes and lymphocytes. We found that blood flow modulates gmfg expression and gmfg overexpression could partially rescue the reduction of HSPCs in the absence of blood flow. Assays in zebrafish and HUVEC showed that gmfg deficiency suppressed the activity of YAP, a well-established blood flow mediator, by preventing its shuttling from cytoplasm to nucleus. During HSPC initiation, loss of gmfg resulted in Notch inactivation and the induction of Notch intracellular domain could partially restore the HSPC loss in gmfg-deficient embryos. Conclusions We conclude that gmfg mediates blood flow-induced HSPC maintenance via regulation of YAP, and contributes to HSPC initiation through the modulation of Notch signaling. Our findings reveal a brand-new aspect of gmfg function and highlight a novel mechanism for embryonic HSPC development.https://doi.org/10.1186/s13287-023-03328-1Glia maturation factor-γHSPCBlood flowYapNotchZebrafish |
| spellingShingle | Honghu Li Qian Luo Shuyang Cai Ruxiu Tie Ye Meng Wei Shan Yulin Xu Xiangjun Zeng Pengxu Qian He Huang Glia maturation factor-γ is required for initiation and maintenance of hematopoietic stem and progenitor cells Stem Cell Research & Therapy Glia maturation factor-γ HSPC Blood flow Yap Notch Zebrafish |
| title | Glia maturation factor-γ is required for initiation and maintenance of hematopoietic stem and progenitor cells |
| title_full | Glia maturation factor-γ is required for initiation and maintenance of hematopoietic stem and progenitor cells |
| title_fullStr | Glia maturation factor-γ is required for initiation and maintenance of hematopoietic stem and progenitor cells |
| title_full_unstemmed | Glia maturation factor-γ is required for initiation and maintenance of hematopoietic stem and progenitor cells |
| title_short | Glia maturation factor-γ is required for initiation and maintenance of hematopoietic stem and progenitor cells |
| title_sort | glia maturation factor γ is required for initiation and maintenance of hematopoietic stem and progenitor cells |
| topic | Glia maturation factor-γ HSPC Blood flow Yap Notch Zebrafish |
| url | https://doi.org/10.1186/s13287-023-03328-1 |
| work_keys_str_mv | AT honghuli gliamaturationfactorgisrequiredforinitiationandmaintenanceofhematopoieticstemandprogenitorcells AT qianluo gliamaturationfactorgisrequiredforinitiationandmaintenanceofhematopoieticstemandprogenitorcells AT shuyangcai gliamaturationfactorgisrequiredforinitiationandmaintenanceofhematopoieticstemandprogenitorcells AT ruxiutie gliamaturationfactorgisrequiredforinitiationandmaintenanceofhematopoieticstemandprogenitorcells AT yemeng gliamaturationfactorgisrequiredforinitiationandmaintenanceofhematopoieticstemandprogenitorcells AT weishan gliamaturationfactorgisrequiredforinitiationandmaintenanceofhematopoieticstemandprogenitorcells AT yulinxu gliamaturationfactorgisrequiredforinitiationandmaintenanceofhematopoieticstemandprogenitorcells AT xiangjunzeng gliamaturationfactorgisrequiredforinitiationandmaintenanceofhematopoieticstemandprogenitorcells AT pengxuqian gliamaturationfactorgisrequiredforinitiationandmaintenanceofhematopoieticstemandprogenitorcells AT hehuang gliamaturationfactorgisrequiredforinitiationandmaintenanceofhematopoieticstemandprogenitorcells |