Ectopic Expression of FVIII in HPCs and MSCs Derived from hiPSCs with Site-Specific Integration of <i>ITGA2B</i> Promoter-Driven <i>BDDF8</i> Gene in Hemophilia A

Ectopic Expression of FVIII in HPCs and MSCs Derived from hiPSCs with Site-Specific Integration of <i>ITGA2B</i> Promoter-Driven <i>BDDF8</i> Gene in Hemophilia A

Hemophilia A (HA) is caused by mutations in the coagulation factor VIII (FVIII) gene <i>(F8)</i>. 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 e...

Full description

Bibliographic Details
Main Authors: Junya Zhao, Miaojin Zhou, Zujia Wang, Lingqian Wu, Zhiqing Hu, Desheng Liang
Format: Article
Language:English
Published: MDPI AG 2022-01-01
Series:International Journal of Molecular Sciences
Subjects:
hemophilia A
platelet-targeted strategy
nonviral vector
rDNA locus
hematopoietic progenitor cell
mesenchymal stem cell
Online Access:https://www.mdpi.com/1422-0067/23/2/623
Description
Summary:Hemophilia A (HA) is caused by mutations in the coagulation factor VIII (FVIII) gene <i>(F8)</i>. 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 <i>F8</i> (<i>BDDF8</i>), driven by a truncated <i>ITGA2B</i> promoter, was targeted at the ribosomal DNA (rDNA) locus of HA patient-specific induced pluripotent stem cells (HA-iPSCs). The <i>F8</i>-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 <i>ITGA2B</i> promoter-driven <i>BDDF8</i> 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/10<sup>6</sup> cells in lysates of 2bF8-iHPCs, compared to 1.56 ng/10<sup>6</sup> cells in HA-iHPCs, and FVIII was 3.64 ng/10<sup>6</sup> cells in 2bF8-iMSCs lysates, while 1.31 ng/10<sup>6</sup> cells in iMSCs with CMV-driven <i>BDDF8</i>. Our results demonstrated a high expression of FVIII in iHPCs and iMSCs derived from hiPSCs with site-specific integration of <i>ITGA2B</i> promoter-driven <i>BDDF8</i>, indicating potential clinical prospects of this platelet-targeted strategy for HA gene therapy.
ISSN:1661-6596
1422-0067

Similar Items