Tailored collagen binding of albumin-fused hyperactive coagulation factor IX dictates in vivo distribution and functional properties
The efficacy of hemophilia B (HB) replacement therapy is evaluated by coagulation factor IX (FIX) activity in plasma, although FIX bound to extravascular type IV collagen (Col4) also contributes to efficient hemostasis. Here, we investigated the impact of engineering FIX for improved (K5R) or reduced (K5A) Col4 binding on the pharmacokinetic properties of FIX Padua, fused to human serum albumin (HSA<sub>QMP</sub>) engineered for favorable neonatal Fc receptor (FcRn) engagement. Hyperactive features and extended plasma half-life in human FcRn expressing mice, attributed to FIX Padua and HSA<sub>QMP</sub> engineering, respectively, was confirmed. In HB mice, Padua<sub>KA</sub>-HSA<sub>QMP</sub> exhibited negligible extravascular distribution and the highest plasma levels at early time points followed by the steepest decay. Conversely, Padua<sub>KR</sub>-HSA<sub>QMP</sub> showed increased extravascular distribution and a 3-fold longer functional half-life (80 hours). These findings support the use of Padua<sub>KA</sub>-HSA<sub>QMP</sub> and Padua<sub>KR</sub>-HSA<sub>QMP</sub> as hyperactive short- or long-term therapeutics, respectively, with opportunities for tailored HB replacement therapy.
Authors
- Maria Francesca Testa (ORCID: https://orcid.org/0000-0003-3818-3431)
- Jan Terje Andersen (ORCID: https://orcid.org/0000-0003-1710-1628)
- Kristin Hovden Aaen (ORCID: https://orcid.org/0000-0001-7181-1052)
- Mascia Benedusi (ORCID: https://orcid.org/0000-0002-4656-8609)
- Jeannette Nilsen (ORCID: https://orcid.org/0000-0001-7892-0892)
- Mirko Pinotti (ORCID: https://orcid.org/0000-0002-4114-7055)
- Giuseppe Valacchi (ORCID: https://orcid.org/0000-0002-8792-0947)
- Alessio Branchini (ORCID: https://orcid.org/0000-0002-6113-2694)
- Inger Sandlie (ORCID: https://orcid.org/0009-0005-8063-0145)
- Sopisa Benjakul (ORCID: https://orcid.org/0000-0003-3708-8125)
- Mari Nyquist-Andersen (ORCID: https://orcid.org/0000-0002-5248-6722)
- Alessio Cantore (ORCID: https://orcid.org/0000-0002-9741-997X)
- Cesare Canepari
- Marie Leangen Herigstad
- Francesco Bernardi
- Rebecca Tarantino
Institutions
- Oslo University Hospital (NO)
- North Carolina State University (US)
- Vita-Salute San Raffaele University (IT)
- University of Oslo (NO)
- Neurosciences Institute (US)
- University of Ferrara (IT)
- Kyung Hee University (KR)
- The San Raffaele Telethon Institute for Gene Therapy (IT)
- Istituti di Ricovero e Cura a Carattere Scientifico (IT)
- Istituto di Ricovero e Cura a Carattere Scientifico San Raffaele (IT)
Publication Details
- Journal
- Nature Communications
- Published
- 2025-09-29
- DOI
- https://doi.org/10.1038/s41467-025-62955-9
- Primary Topic
- Platelet Disorders and Treatments
- Type
- article
- Field-Weighted Citation Impact
- 0.00
Funders
- Novo Nordisk
- Norges Forskningsråd
- Università degli Studi di Ferrara
- Helse Sør-Øst RHF