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Cited by 3Hemophilia Treatment and ResearchVirus-based gene therapy researchPlatelet Disorders and Treatments

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Engineered coagulation factor VIII with enhanced secretion and coagulation potential for hemophilia A gene therapy

Open access
Kashiwakura, Yuji, Nakajima, Yuto, Horinaka, Kio, Lopes, Tiago J. S., Furuta, Yuma, Yamaguchi, Yuki, Baatartsogt, Nemekhbayar, Hayakawa, Morisada, Katakai, Yuko, Uchiyama, Susumu, Nureki, Osamu, Nogami, Keiji, Ohmori, Tsukasa·2026·Blood
DOI 10.1182/blood.2025028481PMC PMC12883863

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Abstract

The major challenges of gene therapy for hemophilia A using adeno-associated virus (AAV) vectors are reducing vector doses and the long-term maintenance of stable factor VIII (FVIII). In this study, we developed engineered human B-domain–deleted FVIIIs (FVIIISQ) with enhanced secretion and coagulation potential. Intracellular accumulation was markedly reduced in some engineered FVIIISQ, resulting in reduced unfolded protein responses. The administration of AAV vectors carrying engineered FVIIISQ to hemophilia A mice resulted in ∼8-fold higher FVIII activity and 4-fold higher FVIII antigen levels compared with wild-type FVIIISQ administration. The specific FVIII activity of the engineered FVIIISQ was 3.6 times higher than that of the wild-type FVIIISQ, and its binding to activated coagulation factor IX was significantly enhanced, which is supported by the structural analysis. In macaques, the administration of AAV5 vector carrying the engineered FVIIISQ without CpG sequences resulted in a supraphysiological increase in plasma FVIII activity at a dose one-thirtieth that of valoctocogene roxaparvovec (2 × 1012 vector genome per kg). The engineered FVIIISQ may thus provide stable, long-term therapeutic efficacy in AAV-mediated hemophilia A gene therapy even at low doses.

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Metadata

Authors
Kashiwakura, Y., Nakajima, Y., Horinaka, K., Lopes, T. J. S., Furuta, Y., Yamaguchi, Y., Baatartsogt, N., Hayakawa, M., Katakai, Y., Uchiyama, S., Nureki, O., Nogami, K., & Ohmori, T.
Affiliations
  • Jichi Medical University (JP)
  • Nara Medical University Hospital (JP)
  • The University of Tokyo (JP)
  • 5Nezu Life Sciences, Heidelberg, Germany
  • The University of Osaka (JP)
  • Corporation for Production and Research of Laboratory Primates (JP)
Corresponding author
Ohmori, Tsukasa
ORCID
Venue
Blood
Publisher
American Society of Hematology
Published
2026-01-22
Volume
147
Issue
4
Pages
402-415
ISSN
  • 0006-4971 (ISSN-L)
  • 1528-0020
DOI
10.1182/blood.2025028481
PMC
PMC12883863
OpenAlex
W4415977576
Cited by
3
References
6
Citations by year
2026: 3
FWCI
2.20
Type
journal article
Language
en
Open access
Licence
cc-by-nc-nd
Locations
  • Blood (publishedVersion, cc-by-nc-nd)
  • bioRxiv (Cold Spring Harbor Laboratory) (acceptedVersion, cc-by-nc-nd)
  • Europe PMC (PubMed Central) (publishedVersion, cc-by-nc-nd)
  • PubMed (publishedVersion)
  • PubMed Central (publishedVersion, cc-by-nc-nd)
Funders
  • Japan Agency for Medical Research and Development (21fk0410037)
  • Jichi Medical University
  • SENSHIN Medical Research Foundation
Topics
  • Health Sciences › Medicine › Hematology › Hemophilia Treatment and Research
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Genetics › Virus-based gene therapy research
  • Health Sciences › Medicine › Hematology › Platelet Disorders and Treatments
Keywords
hemophilia A, gene therapy, adeno-associated virus vectors, factor VIII, B-domain-deleted factor VIII, intracellular accumulation, unfolded protein response, AAV5, macaque model, valoctocogene roxaparvovec
MeSH terms
Dependovirus, Animals, Blood Coagulation, Factor VIII, Genetic Vectors, Hemophilia A, Humans, Male, Mice, Inbred C57BL, Protein Engineering, Genetic Therapy, Mice

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