Improved intravenous lentiviral gene therapy based on endothelial-specific promoter-driven factor VIII expression for hemophilia A
Abstract Background Hemophilia A (HA) is an X-linked monogenic disorder caused by deficiency of the factor VIII (FVIII) gene in the intrinsic coagulation cascade. The current protein replacement therapy (PRT) of HA has many limitations including short term effectiveness, high cost, and life-time tre...
| Main Authors: | , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
BMC
2023-06-01
|
| Series: | Molecular Medicine |
| Subjects: | |
| Online Access: | https://doi.org/10.1186/s10020-023-00680-z |
| _version_ | 1797801381639225344 |
|---|---|
| author | Jie Gong Rui Yang Min Zhou Lung-Ji Chang |
| author_facet | Jie Gong Rui Yang Min Zhou Lung-Ji Chang |
| author_sort | Jie Gong |
| collection | DOAJ |
| description | Abstract Background Hemophilia A (HA) is an X-linked monogenic disorder caused by deficiency of the factor VIII (FVIII) gene in the intrinsic coagulation cascade. The current protein replacement therapy (PRT) of HA has many limitations including short term effectiveness, high cost, and life-time treatment requirement. Gene therapy has become a promising treatment for HA. Orthotopic functional FVIII biosynthesis is critical to its coagulation activities. Methods To investigate targeted FVIII expression, we developed a series of advanced lentiviral vectors (LVs) carrying either a universal promoter (EF1α) or a variety of tissue-specific promoters, including endothelial-specific (VEC), endothelial and epithelial-specific (KDR), and megakaryocyte-specific (Gp and ITGA) promoters. Results To examine tissue specificity, the expression of a B-domain deleted human F8 (F8BDD) gene was tested in human endothelial and megakaryocytic cell lines. Functional assays demonstrated FVIII activities of LV-VEC-F8BDD and LV-ITGA-F8BDD in the therapeutic range in transduced endothelial and megakaryocytic cells, respectively. In F8 knockout mice (F8 KO mice, F8 null mice), intravenous (iv) injection of LVs illustrated different degrees of phenotypic correction as well as anti-FVIII immune response for the different vectors. The iv delivery of LV-VEC-F8BDD and LV-Gp-F8BDD achieved 80% and 15% therapeutic FVIII activities over 180 days, respectively. Different from the other LV constructs, the LV-VEC-F8BDD displayed a low FVIII inhibitory response in the treated F8 null mice. Conclusions The LV-VEC-F8BDD exhibited high LV packaging and delivery efficiencies, with endothelial specificity and low immunogenicity in the F8 null mice, thus has a great potential for clinical applications. |
| first_indexed | 2024-03-13T04:49:36Z |
| format | Article |
| id | doaj.art-84321d1bbe18423281551686c5e11ad5 |
| institution | Directory Open Access Journal |
| issn | 1528-3658 |
| language | English |
| last_indexed | 2024-03-13T04:49:36Z |
| publishDate | 2023-06-01 |
| publisher | BMC |
| record_format | Article |
| series | Molecular Medicine |
| spelling | doaj.art-84321d1bbe18423281551686c5e11ad52023-06-18T11:17:03ZengBMCMolecular Medicine1528-36582023-06-0129111210.1186/s10020-023-00680-zImproved intravenous lentiviral gene therapy based on endothelial-specific promoter-driven factor VIII expression for hemophilia AJie Gong0Rui Yang1Min Zhou2Lung-Ji Chang3Chengdu Women’s and Children’s Central Hospital, School of Medicine, University of Electronic Science and Technology of ChinaSchool of Medicine, University of Electronic Science and Technology of ChinaChengdu Women’s and Children’s Central Hospital, School of Medicine, University of Electronic Science and Technology of ChinaSchool of Medicine, University of Electronic Science and Technology of ChinaAbstract Background Hemophilia A (HA) is an X-linked monogenic disorder caused by deficiency of the factor VIII (FVIII) gene in the intrinsic coagulation cascade. The current protein replacement therapy (PRT) of HA has many limitations including short term effectiveness, high cost, and life-time treatment requirement. Gene therapy has become a promising treatment for HA. Orthotopic functional FVIII biosynthesis is critical to its coagulation activities. Methods To investigate targeted FVIII expression, we developed a series of advanced lentiviral vectors (LVs) carrying either a universal promoter (EF1α) or a variety of tissue-specific promoters, including endothelial-specific (VEC), endothelial and epithelial-specific (KDR), and megakaryocyte-specific (Gp and ITGA) promoters. Results To examine tissue specificity, the expression of a B-domain deleted human F8 (F8BDD) gene was tested in human endothelial and megakaryocytic cell lines. Functional assays demonstrated FVIII activities of LV-VEC-F8BDD and LV-ITGA-F8BDD in the therapeutic range in transduced endothelial and megakaryocytic cells, respectively. In F8 knockout mice (F8 KO mice, F8 null mice), intravenous (iv) injection of LVs illustrated different degrees of phenotypic correction as well as anti-FVIII immune response for the different vectors. The iv delivery of LV-VEC-F8BDD and LV-Gp-F8BDD achieved 80% and 15% therapeutic FVIII activities over 180 days, respectively. Different from the other LV constructs, the LV-VEC-F8BDD displayed a low FVIII inhibitory response in the treated F8 null mice. Conclusions The LV-VEC-F8BDD exhibited high LV packaging and delivery efficiencies, with endothelial specificity and low immunogenicity in the F8 null mice, thus has a great potential for clinical applications.https://doi.org/10.1186/s10020-023-00680-zLentiviral vectorGene therapyHemophilia AFactor VIIIEndothelial-specific |
| spellingShingle | Jie Gong Rui Yang Min Zhou Lung-Ji Chang Improved intravenous lentiviral gene therapy based on endothelial-specific promoter-driven factor VIII expression for hemophilia A Molecular Medicine Lentiviral vector Gene therapy Hemophilia A Factor VIII Endothelial-specific |
| title | Improved intravenous lentiviral gene therapy based on endothelial-specific promoter-driven factor VIII expression for hemophilia A |
| title_full | Improved intravenous lentiviral gene therapy based on endothelial-specific promoter-driven factor VIII expression for hemophilia A |
| title_fullStr | Improved intravenous lentiviral gene therapy based on endothelial-specific promoter-driven factor VIII expression for hemophilia A |
| title_full_unstemmed | Improved intravenous lentiviral gene therapy based on endothelial-specific promoter-driven factor VIII expression for hemophilia A |
| title_short | Improved intravenous lentiviral gene therapy based on endothelial-specific promoter-driven factor VIII expression for hemophilia A |
| title_sort | improved intravenous lentiviral gene therapy based on endothelial specific promoter driven factor viii expression for hemophilia a |
| topic | Lentiviral vector Gene therapy Hemophilia A Factor VIII Endothelial-specific |
| url | https://doi.org/10.1186/s10020-023-00680-z |
| work_keys_str_mv | AT jiegong improvedintravenouslentiviralgenetherapybasedonendothelialspecificpromoterdrivenfactorviiiexpressionforhemophiliaa AT ruiyang improvedintravenouslentiviralgenetherapybasedonendothelialspecificpromoterdrivenfactorviiiexpressionforhemophiliaa AT minzhou improvedintravenouslentiviralgenetherapybasedonendothelialspecificpromoterdrivenfactorviiiexpressionforhemophiliaa AT lungjichang improvedintravenouslentiviralgenetherapybasedonendothelialspecificpromoterdrivenfactorviiiexpressionforhemophiliaa |