Improving hematopoietic differentiation from human induced pluripotent stem cells by the modulation of Hippo signaling with a diarylheptanoid derivative
Abstract Background The diarylheptanoid ASPP 049 has improved the quality of adult hematopoietic stem cell (HSC) expansion ex vivo through long-term reconstitution in animal models. However, its effect on hematopoietic regeneration from human induced pluripotent stem cells (hiPSCs) is unknown. Metho...
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| Format: | Article |
| Language: | English |
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BMC
2024-03-01
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| Series: | Stem Cell Research & Therapy |
| Subjects: | |
| Online Access: | https://doi.org/10.1186/s13287-024-03686-4 |
| _version_ | 1797275596267454464 |
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| author | Umnuaychoke Thongsa-ad Anongnat Wongpan Wasinee Wongkummool Phaewa Chaiwijit Kwanchanok Uppakara Gorawin Chaiyakitpattana Passanan Singpant Pirut Tong-ngam Amnat Chukhan Wachirachai Pabuprappap Sirapope Wongniam Apichart Suksamrarn Suradej Hongeng Usanarat Anurathapan Kasem Kulkeaw Alisa Tubsuwan Kanit Bhukhai |
| author_facet | Umnuaychoke Thongsa-ad Anongnat Wongpan Wasinee Wongkummool Phaewa Chaiwijit Kwanchanok Uppakara Gorawin Chaiyakitpattana Passanan Singpant Pirut Tong-ngam Amnat Chukhan Wachirachai Pabuprappap Sirapope Wongniam Apichart Suksamrarn Suradej Hongeng Usanarat Anurathapan Kasem Kulkeaw Alisa Tubsuwan Kanit Bhukhai |
| author_sort | Umnuaychoke Thongsa-ad |
| collection | DOAJ |
| description | Abstract Background The diarylheptanoid ASPP 049 has improved the quality of adult hematopoietic stem cell (HSC) expansion ex vivo through long-term reconstitution in animal models. However, its effect on hematopoietic regeneration from human induced pluripotent stem cells (hiPSCs) is unknown. Method We utilized a defined cocktail of cytokines without serum or feeder followed by the supplementation of ASPP 049 to produce hematopoietic stem/progenitor cells (HSPCs). Flow cytometry and trypan blue exclusion analysis were used to identify nonadherent and adherent cells. Nonadherent cells were harvested to investigate the effect of ASPP 049 on multipotency using LTC-IC and CFU assays. Subsequently, the mechanism of action was explored through transcriptomic profiles, which were validated by qRT-PCR, immunoblotting, and immunofluorescence analysis. Result The supplementation of ASPP 049 increased the number of phenotypically defined primitive HSPCs (CD34+CD45+CD90+) two-fold relative to seeded hiPSC colonies, indicating enhanced HSC derivation from hiPSCs. Under ASPP 049-supplemented conditions, we observed elevated HSPC niches, including CD144+CD73− hemogenic- and CD144+CD73+ vascular-endothelial progenitors, during HSC differentiation. Moreover, harvested ASPP 049-treated cells exhibited improved self-renewal and a significantly larger proportion of different blood cell colonies with unbiased lineages, indicating enhanced HSC stemness properties. Transcriptomics and KEGG analysis of sorted CD34+CD45+ cells-related mRNA profiles revealed that the Hippo signaling pathway is the most significant in responding to WWTR1/TAZ, which correlates with the validation of the protein expression. Interestingly, ASPP 049-supplemented HSPCs upregulated 11 genes similarly to umbilical cord blood-derived HSPCs. Conclusion These findings suggest that ASPP 049 can improve HSC-generating protocols with proliferative potentials, self-renewal ability, unbiased differentiation, and a definable mechanism of action for the clinical perspective of hematopoietic regenerative medicine. Graphical abstract |
| first_indexed | 2024-03-07T15:16:44Z |
| format | Article |
| id | doaj.art-1307d64570604af68cd78ca8bd7b61ed |
| institution | Directory Open Access Journal |
| issn | 1757-6512 |
| language | English |
| last_indexed | 2024-03-07T15:16:44Z |
| publishDate | 2024-03-01 |
| publisher | BMC |
| record_format | Article |
| series | Stem Cell Research & Therapy |
| spelling | doaj.art-1307d64570604af68cd78ca8bd7b61ed2024-03-05T17:52:14ZengBMCStem Cell Research & Therapy1757-65122024-03-0115112010.1186/s13287-024-03686-4Improving hematopoietic differentiation from human induced pluripotent stem cells by the modulation of Hippo signaling with a diarylheptanoid derivativeUmnuaychoke Thongsa-ad0Anongnat Wongpan1Wasinee Wongkummool2Phaewa Chaiwijit3Kwanchanok Uppakara4Gorawin Chaiyakitpattana5Passanan Singpant6Pirut Tong-ngam7Amnat Chukhan8Wachirachai Pabuprappap9Sirapope Wongniam10Apichart Suksamrarn11Suradej Hongeng12Usanarat Anurathapan13Kasem Kulkeaw14Alisa Tubsuwan15Kanit Bhukhai16Department of Physiology, Faculty of Science, Mahidol UniversityDepartment of Physiology, Faculty of Science, Mahidol UniversityStem Cell Research Group, Institute of Molecular Biosciences, Mahidol UniversityDepartment of Physiology, Faculty of Science, Mahidol UniversityChakri Naruebodindra Medical Institute, Faculty of Medicine Ramathibodi Hospital, Mahidol UniversityDepartment of Physiology, Faculty of Science, Mahidol UniversityStem Cell Research Group, Institute of Molecular Biosciences, Mahidol UniversityStem Cell Research Group, Institute of Molecular Biosciences, Mahidol UniversityPrima ScientificDepartment of Chemistry and Center of Excellence for Innovation in Chemistry, Faculty of Science, Ramkhamhaeng UniversityCenter for Scientific Instrumentation and Platform Services Unit, Faculty of Science, Mahidol UniversityDepartment of Chemistry and Center of Excellence for Innovation in Chemistry, Faculty of Science, Ramkhamhaeng UniversityDepartment of Pediatrics, Faculty of Medicine, Ramathibodi Hospital, Mahidol UniversityDepartment of Pediatrics, Faculty of Medicine, Ramathibodi Hospital, Mahidol UniversitySiriraj Integrative Center for Neglected Parasitic Diseases, Department of Parasitology, Faculty of Medicine Siriraj Hospital, Mahidol UniversityStem Cell Research Group, Institute of Molecular Biosciences, Mahidol UniversityDepartment of Physiology, Faculty of Science, Mahidol UniversityAbstract Background The diarylheptanoid ASPP 049 has improved the quality of adult hematopoietic stem cell (HSC) expansion ex vivo through long-term reconstitution in animal models. However, its effect on hematopoietic regeneration from human induced pluripotent stem cells (hiPSCs) is unknown. Method We utilized a defined cocktail of cytokines without serum or feeder followed by the supplementation of ASPP 049 to produce hematopoietic stem/progenitor cells (HSPCs). Flow cytometry and trypan blue exclusion analysis were used to identify nonadherent and adherent cells. Nonadherent cells were harvested to investigate the effect of ASPP 049 on multipotency using LTC-IC and CFU assays. Subsequently, the mechanism of action was explored through transcriptomic profiles, which were validated by qRT-PCR, immunoblotting, and immunofluorescence analysis. Result The supplementation of ASPP 049 increased the number of phenotypically defined primitive HSPCs (CD34+CD45+CD90+) two-fold relative to seeded hiPSC colonies, indicating enhanced HSC derivation from hiPSCs. Under ASPP 049-supplemented conditions, we observed elevated HSPC niches, including CD144+CD73− hemogenic- and CD144+CD73+ vascular-endothelial progenitors, during HSC differentiation. Moreover, harvested ASPP 049-treated cells exhibited improved self-renewal and a significantly larger proportion of different blood cell colonies with unbiased lineages, indicating enhanced HSC stemness properties. Transcriptomics and KEGG analysis of sorted CD34+CD45+ cells-related mRNA profiles revealed that the Hippo signaling pathway is the most significant in responding to WWTR1/TAZ, which correlates with the validation of the protein expression. Interestingly, ASPP 049-supplemented HSPCs upregulated 11 genes similarly to umbilical cord blood-derived HSPCs. Conclusion These findings suggest that ASPP 049 can improve HSC-generating protocols with proliferative potentials, self-renewal ability, unbiased differentiation, and a definable mechanism of action for the clinical perspective of hematopoietic regenerative medicine. Graphical abstracthttps://doi.org/10.1186/s13287-024-03686-4DiarylheptanoidHuman induced pluripotent stem cellsPrimitive hematopoietic stem and progenitor cellsHippo signaling pathway |
| spellingShingle | Umnuaychoke Thongsa-ad Anongnat Wongpan Wasinee Wongkummool Phaewa Chaiwijit Kwanchanok Uppakara Gorawin Chaiyakitpattana Passanan Singpant Pirut Tong-ngam Amnat Chukhan Wachirachai Pabuprappap Sirapope Wongniam Apichart Suksamrarn Suradej Hongeng Usanarat Anurathapan Kasem Kulkeaw Alisa Tubsuwan Kanit Bhukhai Improving hematopoietic differentiation from human induced pluripotent stem cells by the modulation of Hippo signaling with a diarylheptanoid derivative Stem Cell Research & Therapy Diarylheptanoid Human induced pluripotent stem cells Primitive hematopoietic stem and progenitor cells Hippo signaling pathway |
| title | Improving hematopoietic differentiation from human induced pluripotent stem cells by the modulation of Hippo signaling with a diarylheptanoid derivative |
| title_full | Improving hematopoietic differentiation from human induced pluripotent stem cells by the modulation of Hippo signaling with a diarylheptanoid derivative |
| title_fullStr | Improving hematopoietic differentiation from human induced pluripotent stem cells by the modulation of Hippo signaling with a diarylheptanoid derivative |
| title_full_unstemmed | Improving hematopoietic differentiation from human induced pluripotent stem cells by the modulation of Hippo signaling with a diarylheptanoid derivative |
| title_short | Improving hematopoietic differentiation from human induced pluripotent stem cells by the modulation of Hippo signaling with a diarylheptanoid derivative |
| title_sort | improving hematopoietic differentiation from human induced pluripotent stem cells by the modulation of hippo signaling with a diarylheptanoid derivative |
| topic | Diarylheptanoid Human induced pluripotent stem cells Primitive hematopoietic stem and progenitor cells Hippo signaling pathway |
| url | https://doi.org/10.1186/s13287-024-03686-4 |
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