A hemophilia A mouse model for the in vivo assessment of emicizumab function

Blood. 2020 Aug 6;136(6):740-748. doi: 10.1182/blood.2019004334.

Abstract

The bispecific antibody emicizumab is increasingly used for hemophilia A treatment. However, its specificity for human factors IX and X (FIX and FX) has limited its in vivo functional analysis to primate models of acquired hemophilia. Here, we describe a novel mouse model that allows emicizumab function to be examined. Briefly, FVIII-deficient mice received IV emicizumab 24 hours before tail-clip bleeding was performed. A second infusion with human FIX and FX, administered 5 minutes before bleeding, generated consistent levels of emicizumab (0.7-19 mg/dL for 0.5-10 mg/kg doses) and of both FIX and FX (85 and 101 U/dL, respectively, after dosing at 100 U/kg). Plasma from these mice display FVIII-like activity in assays (diluted activated partial thromboplastin time and thrombin generation), similar to human samples containing emicizumab. Emicizumab doses of 1.5 mg/kg and higher significantly reduced blood loss in a tail-clip-bleeding model using FVIII-deficient mice. However, reduction was incomplete compared with mice treated with human FVIII concentrate, and no difference in efficacy between doses was observed. From this model, we deducted FVIII-like activity from emicizumab that corresponded to a dose of 4.5 U of FVIII per kilogram (ie, 9.0 U/dL). Interestingly, combined with a low FVIII dose (5 U/kg), emicizumab provided enough additive activity to allow complete bleeding arrest. This model could be useful for further in vivo analysis of emicizumab.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Antibodies, Bispecific / administration & dosage
  • Antibodies, Bispecific / immunology
  • Antibodies, Bispecific / therapeutic use*
  • Antibodies, Monoclonal, Humanized / administration & dosage
  • Antibodies, Monoclonal, Humanized / immunology
  • Antibodies, Monoclonal, Humanized / therapeutic use*
  • Drug Therapy, Combination
  • Factor IX / administration & dosage*
  • Factor IX / analysis
  • Factor IX / immunology
  • Factor VIII / administration & dosage
  • Factor VIII / analysis
  • Factor VIII / therapeutic use
  • Factor X / administration & dosage*
  • Factor X / analysis
  • Factor X / immunology
  • Factor XIa / pharmacology
  • Female
  • Hemophilia A / blood
  • Hemophilia A / complications
  • Hemophilia A / drug therapy*
  • Hemophilia A / immunology
  • Hemorrhage / drug therapy*
  • Hemorrhage / etiology
  • Infusions, Intravenous
  • Male
  • Mice
  • Mice, Inbred C57BL
  • Mice, Knockout
  • Models, Animal*
  • Partial Thromboplastin Time
  • Tail / injuries
  • Thrombin / biosynthesis

Substances

  • Antibodies, Bispecific
  • Antibodies, Monoclonal, Humanized
  • emicizumab
  • F8 protein, human
  • Factor VIII
  • Factor IX
  • Factor X
  • Factor XIa
  • Thrombin