Therapeutic base editing to generate a gain-of-function <i>F9</i> variant for hemophilia B
The repair of pathological gene variants is an ultimate goal in treating genetic diseases; however, developing distinct therapeutic reagents for each of the numerous variants within a gene may not be scalable. Here, we investigated whether base editing to introduce a gain-of-function variant in blood coagulation factor IX (FIX) can increase FIX activity as a targeted therapeutic approach for hemophilia B. We engineered a G:C to A:T substitution at c.1151 of F9 by cytosine base editing to generate R338Q (the Shanghai F9 variant), which markedly increases coagulation factor activity. An adeno-associated virus vector harboring the base editor converted >60% of the target G:C to A:T and increased FIX activity in HEK293 cells harboring patient-derived F9 variants as well as in knock-in mice carrying a human F9 complementary DNA. Furthermore, administration of lipid nanoparticles containing the base-editor mRNA and guide RNA increased FIX activity in mice. These data indicate that cytosine base editing to generate R338Q in FIX is a broadly applicable genome-editing strategy for hemophilia B with residual FIX activity.
Authors
- Khishigjargal Batjargal (ORCID: https://orcid.org/0000-0001-5999-9910)
- Yuji Kashiwakura (ORCID: https://orcid.org/0000-0003-0585-3937)
- Taro Shimizu (ORCID: https://orcid.org/0000-0002-8326-2166)
- Tsukasa Ohmori (ORCID: https://orcid.org/0000-0001-5082-6394)
- Osamu Nureki (ORCID: https://orcid.org/0000-0003-1813-7008)
- Takafumi Hiramoto (ORCID: https://orcid.org/0000-0003-4054-6748)
- Yusuke Sato (ORCID: https://orcid.org/0000-0003-0913-7815)
- Rina Ito
- Tomoki Togashi (ORCID: https://orcid.org/0000-0002-2600-657X)
- Morisada Hayakawa (ORCID: https://orcid.org/0000-0002-1869-2450)
- Nemekhbayar Baatartsogt (ORCID: https://orcid.org/0000-0002-3004-8457)
- Tatsuhiro Ishida (ORCID: https://orcid.org/0000-0002-1333-6465)
- Haruka Takata (ORCID: https://orcid.org/0000-0001-7792-2239)
- Atsushi Hoshino (ORCID: https://orcid.org/0000-0002-4015-1319)
- Rikako Sato
- Yasumitsu Nagao (ORCID: https://orcid.org/0009-0001-3888-6682)
- Hina Naruoka
Institutions
- Sapporo City University (JP)
- Jichi Medical University (JP)
- Bunkyo University (JP)
- Tokyo University of Science (JP)
- Hokkaido University (JP)
- Hokkaido Pharmaceutical University (JP)
- Kyoto Prefectural University of Medicine (JP)
- Jichi Medical University Hospital (JP)
- The University of Tokyo (JP)
- Tokushima University (JP)
- The University of Osaka (JP)
Publication Details
- Journal
- Blood
- Published
- 2025-10-23
- DOI
- https://doi.org/10.1182/blood.2024027870
- Primary Topic
- Virus-based gene therapy research
- Type
- article
- Field-Weighted Citation Impact
- 0.00
Funders
- Naito Foundation
- Japan Agency for Medical Research and Development
- Jichi Medical University