Bioengineering hemophilia A–specific microvascular grafts for delivery of full-length factor VIII into the bloodstream
Autor: | Tien Hua, Joseph Neumeyer, Ellis J. Neufeld, Ruei-Zeng Lin, Stacy E. Croteau, Kai Wang, Xuechong Hong, Juan M. Melero-Martin |
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Rok vydání: | 2019 |
Předmět: |
0301 basic medicine
Genetic enhancement Induced Pluripotent Stem Cells Gene Expression Bioengineering Mice SCID Hemophilia A Insert (molecular biology) Mice 03 medical and health sciences 0302 clinical medicine In vivo Animals Humans Medicine Induced pluripotent stem cell Blood Coagulation Embryonic Stem Cells Factor VIII business.industry Gene Transfer Techniques Genetic Therapy Gene Therapy Hematology medicine.disease Genetically modified organism Bleeding diathesis Disease Models Animal Phenotype Treatment Outcome 030104 developmental biology Coagulation 030220 oncology & carcinogenesis Microvessels Mutation Cancer research Blood Coagulation Tests business Ex vivo Stem Cell Transplantation |
Zdroj: | Blood Advances. 3:4166-4176 |
ISSN: | 2473-9537 2473-9529 |
Popis: | Hemophilia A (HA) is a bleeding disorder caused by mutations in the F8 gene encoding coagulation factor VIII (FVIII). Current treatments are based on regular infusions of FVIII concentrates throughout a patient’s life. Alternatively, viral gene therapies that directly deliver F8 in vivo have shown preliminary successes. However, hurdles remain, including lack of infection specificity and the inability to deliver the full-length version of F8 due to restricted viral cargo sizes. Here, we developed an alternative nonviral ex vivo gene-therapy approach that enables the overexpression of full-length F8 in patients’ endothelial cells (ECs). We first generated HA patient–specific induced pluripotent stem cells (HA-iPSCs) from urine epithelial cells and genetically modified them using a piggyBac DNA transposon system to insert multiple copies of full-length F8. We subsequently differentiated the modified HA-iPSCs into competent ECs with high efficiency, and demonstrated that the cells (termed HA-FLF8-iECs) were capable of producing high levels of FVIII. Importantly, following subcutaneous implantation into immunodeficient hemophilic (SCID-f8ko) mice, we demonstrated that HA-FLF8-iECs were able to self-assemble into vascular networks, and that the newly formed microvessels had the capacity to deliver functional FVIII directly into the bloodstream of the mice, effectively correcting the clotting deficiency. Moreover, our implant maintains cellular confinement, which reduces potential safety concerns and allows effective monitoring and reversibility. We envision that this proof-of-concept study could become the basis for a novel autologous ex vivo gene-therapy approach to treat HA. |
Databáze: | OpenAIRE |
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