Minimal Essential Human Factor VIII Alterations Enhance Secretion and Gene Therapy Efficiency
One important limitation for achieving therapeutic expression of human factor VIII (FVIII) in hemophilia A gene therapy is inefficient secretion of the FVIII protein. Substitution of five amino acids in the A1 domain of human FVIII with the corresponding porcine FVIII residues generated a secretion-...
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| Format: | Article |
| Language: | English |
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Elsevier
2020-12-01
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| Series: | Molecular Therapy: Methods & Clinical Development |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S2329050120302175 |
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doaj-db9d4486e2234e9ab30e776cd2f5ab10 |
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| record_format |
Article |
| collection |
DOAJ |
| language |
English |
| format |
Article |
| sources |
DOAJ |
| author |
Wenjing Cao Biao Dong Franziska Horling Jenni A. Firrman Johannes Lengler Matthias Klugmann Maurus de la Rosa Wenman Wu Qizhao Wang Hongying Wei Andrea R. Moore Sean A. Roberts Carmen J. Booth Werner Hoellriegl Dong Li Barbara Konkle Carol Miao Birgit M. Reipert Friedrich Scheiflinger Hanspeter Rottensteiner Weidong Xiao |
| spellingShingle |
Wenjing Cao Biao Dong Franziska Horling Jenni A. Firrman Johannes Lengler Matthias Klugmann Maurus de la Rosa Wenman Wu Qizhao Wang Hongying Wei Andrea R. Moore Sean A. Roberts Carmen J. Booth Werner Hoellriegl Dong Li Barbara Konkle Carol Miao Birgit M. Reipert Friedrich Scheiflinger Hanspeter Rottensteiner Weidong Xiao Minimal Essential Human Factor VIII Alterations Enhance Secretion and Gene Therapy Efficiency Molecular Therapy: Methods & Clinical Development factor VIII AAV gene therapy X5 secretion vector |
| author_facet |
Wenjing Cao Biao Dong Franziska Horling Jenni A. Firrman Johannes Lengler Matthias Klugmann Maurus de la Rosa Wenman Wu Qizhao Wang Hongying Wei Andrea R. Moore Sean A. Roberts Carmen J. Booth Werner Hoellriegl Dong Li Barbara Konkle Carol Miao Birgit M. Reipert Friedrich Scheiflinger Hanspeter Rottensteiner Weidong Xiao |
| author_sort |
Wenjing Cao |
| title |
Minimal Essential Human Factor VIII Alterations Enhance Secretion and Gene Therapy Efficiency |
| title_short |
Minimal Essential Human Factor VIII Alterations Enhance Secretion and Gene Therapy Efficiency |
| title_full |
Minimal Essential Human Factor VIII Alterations Enhance Secretion and Gene Therapy Efficiency |
| title_fullStr |
Minimal Essential Human Factor VIII Alterations Enhance Secretion and Gene Therapy Efficiency |
| title_full_unstemmed |
Minimal Essential Human Factor VIII Alterations Enhance Secretion and Gene Therapy Efficiency |
| title_sort |
minimal essential human factor viii alterations enhance secretion and gene therapy efficiency |
| publisher |
Elsevier |
| series |
Molecular Therapy: Methods & Clinical Development |
| issn |
2329-0501 |
| publishDate |
2020-12-01 |
| description |
One important limitation for achieving therapeutic expression of human factor VIII (FVIII) in hemophilia A gene therapy is inefficient secretion of the FVIII protein. Substitution of five amino acids in the A1 domain of human FVIII with the corresponding porcine FVIII residues generated a secretion-enhanced human FVIII variant termed B-domain-deleted (BDD)-FVIII-X5 that resulted in 8-fold higher FVIII activity levels in the supernatant of an in vitro cell-based assay system than seen with unmodified human BDD-FVIII. Analysis of purified recombinant BDD-FVIII-X5 and BDD-FVIII revealed similar specific activities for both proteins, indicating that the effect of the X5 alteration is confined to increased FVIII secretion. Intravenous delivery in FVIII-deficient mice of liver-targeted adeno-associated virus (AAV) vectors designed to express BDD-FVIII-X5 or BDD-FVIII achieved substantially higher plasma FVIII activity levels for BDD-FVIII-X5, even when highly efficient codon-optimized F8 nucleotide sequences were employed. A comprehensive immunogenicity assessment using in vitro stimulation assays and various in vivo preclinical models of hemophilia A demonstrated that the BDD-FVIII-X5 variant does not exhibit an increased immunogenicity risk compared to BDD-FVIII. In conclusion, BDD-FVIII-X5 is an effective FVIII variant molecule that can be further developed for use in gene- and protein-based therapeutics for patients with hemophilia A. |
| topic |
factor VIII AAV gene therapy X5 secretion vector |
| url |
http://www.sciencedirect.com/science/article/pii/S2329050120302175 |
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1724386955796414464 |
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doaj-db9d4486e2234e9ab30e776cd2f5ab102020-12-11T04:22:00ZengElsevierMolecular Therapy: Methods & Clinical Development2329-05012020-12-0119486495Minimal Essential Human Factor VIII Alterations Enhance Secretion and Gene Therapy EfficiencyWenjing Cao0Biao Dong1Franziska Horling2Jenni A. Firrman3Johannes Lengler4Matthias Klugmann5Maurus de la Rosa6Wenman Wu7Qizhao Wang8Hongying Wei9Andrea R. Moore10Sean A. Roberts11Carmen J. Booth12Werner Hoellriegl13Dong Li14Barbara Konkle15Carol Miao16Birgit M. Reipert17Friedrich Scheiflinger18Hanspeter Rottensteiner19Weidong Xiao20Sol Sherry Thrombosis Research Center, Temple University Medical School, 3400 North Broad Street, Philadelphia, PA, 19140, USASol Sherry Thrombosis Research Center, Temple University Medical School, 3400 North Broad Street, Philadelphia, PA, 19140, USADrug Discovery Austria, Baxalta Innovations GmbH (now part of Takeda), Donau-City Str. 7, Vienna 1220, AustriaDairy and Functional Foods Research Unit, ARS, USDA, 600 East Mermaid Lane, Wyndmoor, PA 19038, USADrug Discovery Austria, Baxalta Innovations GmbH (now part of Takeda), Donau-City Str. 7, Vienna 1220, AustriaDrug Discovery Austria, Baxalta Innovations GmbH (now part of Takeda), Donau-City Str. 7, Vienna 1220, AustriaDrug Discovery Austria, Baxalta Innovations GmbH (now part of Takeda), Donau-City Str. 7, Vienna 1220, AustriaSol Sherry Thrombosis Research Center, Temple University Medical School, 3400 North Broad Street, Philadelphia, PA, 19140, USASol Sherry Thrombosis Research Center, Temple University Medical School, 3400 North Broad Street, Philadelphia, PA, 19140, USASol Sherry Thrombosis Research Center, Temple University Medical School, 3400 North Broad Street, Philadelphia, PA, 19140, USASol Sherry Thrombosis Research Center, Temple University Medical School, 3400 North Broad Street, Philadelphia, PA, 19140, USASol Sherry Thrombosis Research Center, Temple University Medical School, 3400 North Broad Street, Philadelphia, PA, 19140, USADepartment of Comparative Medicine, Yale University School of Medicine, 310 Cedar St., BML 330, New Haven, CT 06510, USADrug Discovery Austria, Baxalta Innovations GmbH (now part of Takeda), Donau-City Str. 7, Vienna 1220, AustriaSol Sherry Thrombosis Research Center, Temple University Medical School, 3400 North Broad Street, Philadelphia, PA, 19140, USASeattle Children’s Research Institute, University of Washington, 1900 9th Ave, Seattle, WA 98195, USADepartment of Medicine/Hematology, University of Washington, 1900 9th Ave, Seattle, WA 98195, USADrug Discovery Austria, Baxalta Innovations GmbH (now part of Takeda), Donau-City Str. 7, Vienna 1220, AustriaDrug Discovery Austria, Baxalta Innovations GmbH (now part of Takeda), Donau-City Str. 7, Vienna 1220, AustriaDrug Discovery Austria, Baxalta Innovations GmbH (now part of Takeda), Donau-City Str. 7, Vienna 1220, Austria; Corresponding author: Hanspeter Rottensteiner, PhD, Baxalta Innovations GmbH (now part of Takeda), Donau-City Str. 7, Vienna 1220, Austria.Herman B Wells Center for Pediatric Research, Indiana University, Indianapolis, IN 46202, USA; Corresponding author: Weidong Xiao, Herman B Wells Center for Pediatric Research, Indiana University, Indianapolis, IN 46202, USA.One important limitation for achieving therapeutic expression of human factor VIII (FVIII) in hemophilia A gene therapy is inefficient secretion of the FVIII protein. Substitution of five amino acids in the A1 domain of human FVIII with the corresponding porcine FVIII residues generated a secretion-enhanced human FVIII variant termed B-domain-deleted (BDD)-FVIII-X5 that resulted in 8-fold higher FVIII activity levels in the supernatant of an in vitro cell-based assay system than seen with unmodified human BDD-FVIII. Analysis of purified recombinant BDD-FVIII-X5 and BDD-FVIII revealed similar specific activities for both proteins, indicating that the effect of the X5 alteration is confined to increased FVIII secretion. Intravenous delivery in FVIII-deficient mice of liver-targeted adeno-associated virus (AAV) vectors designed to express BDD-FVIII-X5 or BDD-FVIII achieved substantially higher plasma FVIII activity levels for BDD-FVIII-X5, even when highly efficient codon-optimized F8 nucleotide sequences were employed. A comprehensive immunogenicity assessment using in vitro stimulation assays and various in vivo preclinical models of hemophilia A demonstrated that the BDD-FVIII-X5 variant does not exhibit an increased immunogenicity risk compared to BDD-FVIII. In conclusion, BDD-FVIII-X5 is an effective FVIII variant molecule that can be further developed for use in gene- and protein-based therapeutics for patients with hemophilia A.http://www.sciencedirect.com/science/article/pii/S2329050120302175factor VIIIAAVgene therapyX5secretionvector |