Ectopic Expression of FVIII in HPCs and MSCs Derived from hiPSCs with Site-Specific Integration of
| Autor: | Junya, Zhao, Miaojin, Zhou, Zujia, Wang, Lingqian, Wu, Zhiqing, Hu, Desheng, Liang |
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| Rok vydání: | 2021 |
| Předmět: |
Factor VIII
Base Sequence rDNA locus Genetic Vectors Induced Pluripotent Stem Cells Integrin alpha2 Mesenchymal Stem Cells platelet-targeted strategy Hematopoietic Stem Cells Hemophilia A DNA Ribosomal Article Ectopic Gene Expression Protein Domains Genetic Loci hemic and lymphatic diseases Transcription Activator-Like Effector Nucleases Gene Targeting Humans hematopoietic progenitor cell Promoter Regions Genetic nonviral vector Megakaryocytes mesenchymal stem cell Sequence Deletion |
| Zdroj: | International Journal of Molecular Sciences |
| ISSN: | 1422-0067 |
| Popis: | Hemophilia A (HA) is caused by mutations in the coagulation factor VIII (FVIII) gene (F8). Gene therapy is a hopeful cure for HA; however, FVIII inhibitors formation hinders its clinical application. Given that platelets promote coagulation via locally releasing α-granule, FVIII ectopically expressed in platelets has been attempted, with promising results for HA treatment. The B-domain-deleted F8 (BDDF8), driven by a truncated ITGA2B promoter, was targeted at the ribosomal DNA (rDNA) locus of HA patient-specific induced pluripotent stem cells (HA-iPSCs). The F8-modified, human induced pluripotent stem cells (2bF8-iPSCs) were differentiated into induced hematopoietic progenitor cells (iHPCs), induced megakaryocytes (iMKs), and mesenchymal stem cells (iMSCs), and the FVIII expression was detected. The ITGA2B promoter-driven BDDF8 was site-specifically integrated into the rDNA locus of HA-iPSCs. The 2bF8-iPSCs were efficiently differentiated into 2bF8-iHPCs, 2bF8-iMKs, and 2bF8-iMSCs. FVIII was 10.31 ng/106 cells in lysates of 2bF8-iHPCs, compared to 1.56 ng/106 cells in HA-iHPCs, and FVIII was 3.64 ng/106 cells in 2bF8-iMSCs lysates, while 1.31 ng/106 cells in iMSCs with CMV-driven BDDF8. Our results demonstrated a high expression of FVIII in iHPCs and iMSCs derived from hiPSCs with site-specific integration of ITGA2B promoter-driven BDDF8, indicating potential clinical prospects of this platelet-targeted strategy for HA gene therapy. |
| Databáze: | OpenAIRE |
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