Microfluidic hemophilia models using blood from healthy donors
Abstract Background Microfluidic clotting assays permit drug action studies for hemophilia therapeutics under flow. However, limited availability of patient samples and Inter‐donor variability limit the application of such assays, especially with many patients on prophylaxis. Objective To develop ap...
| Main Authors: | , , , , , , |
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| Format: | Article |
| Language: | English |
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Elsevier
2020-01-01
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| Series: | Research and Practice in Thrombosis and Haemostasis |
| Subjects: | |
| Online Access: | https://doi.org/10.1002/rth2.12286 |
| _version_ | 1797725209512378368 |
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| author | Xinren Yu Karen A. Panckeri Lacramioara Ivanciu Rodney M. Camire Carmen H. Coxon Adam Cuker Scott L. Diamond |
| author_facet | Xinren Yu Karen A. Panckeri Lacramioara Ivanciu Rodney M. Camire Carmen H. Coxon Adam Cuker Scott L. Diamond |
| author_sort | Xinren Yu |
| collection | DOAJ |
| description | Abstract Background Microfluidic clotting assays permit drug action studies for hemophilia therapeutics under flow. However, limited availability of patient samples and Inter‐donor variability limit the application of such assays, especially with many patients on prophylaxis. Objective To develop approaches to phenocopy hemophilia using modified healthy blood in microfluidic assays. Methods Corn trypsin inhibitor (4 µg/mL)‐treated healthy blood was dosed with either anti–factor VIII (FVIII; hemophilia A model) or a recombinant factor IX (FIX) missense variant (FIX‐V181T; hemophilia B model). Treated blood was perfused at 100 s−1 wall shear rate over collagen/tissue factor (TF) or collagen/factor XIa (FXIa). Results Anti‐FVIII partially blocked fibrin production on collagen/TF, but completely blocked fibrin production on collagen/FXIa, a phenotype reversed with 1 µmol/L bispecific antibody (emicizumab), which binds FIXa and factor X. As expected, emicizumab had no significant effect on healthy blood (no anti‐FVIII present) perfused over collagen/FXIa. The efficacy of emicizumab in anti‐FVIII‐treated healthy blood phenocopied the action of emicizumab in the blood of a patient with hemophilia A perfused over collagen/FXIa. Interestingly, a patient‐derived FVIII‐neutralizing antibody reduced fibrin production when added to healthy blood perfused over collagen/FXIa. For low TF surfaces, reFIX‐V181T (50 µg/mL) fully blocked platelet and fibrin deposition, a phenotype fully reversed with anti‐TFPI. Conclusion Two new microfluidic hemophilia A and B models demonstrate the potency of anti‐TF pathway inhibitor, emicizumab, and a patient‐derived inhibitory antibody. Using collagen/FXIa‐coated surfaces resulted in reliable and highly sensitive hemophilia models. |
| first_indexed | 2024-03-12T10:27:52Z |
| format | Article |
| id | doaj.art-71da544d77aa43e085358a8ca40dc390 |
| institution | Directory Open Access Journal |
| issn | 2475-0379 |
| language | English |
| last_indexed | 2024-03-12T10:27:52Z |
| publishDate | 2020-01-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Research and Practice in Thrombosis and Haemostasis |
| spelling | doaj.art-71da544d77aa43e085358a8ca40dc3902023-09-02T09:31:45ZengElsevierResearch and Practice in Thrombosis and Haemostasis2475-03792020-01-0141546310.1002/rth2.12286Microfluidic hemophilia models using blood from healthy donorsXinren Yu0Karen A. Panckeri1Lacramioara Ivanciu2Rodney M. Camire3Carmen H. Coxon4Adam Cuker5Scott L. Diamond6Department of Chemical and Biomolecular Engineering Institute for Medicine and Engineering University of Pennsylvania Philadelphia PennsylvaniaPenn Comprehensive Hemophilia and Thrombosis Program Hospital of the University of Pennsylvania Philadelphia PennsylvaniaThe Raymond G. Perelman Center for Cellular and Molecular Therapeutics The Children’s Hospital of Philadelphia Philadelphia PennsylvaniaThe Raymond G. Perelman Center for Cellular and Molecular Therapeutics The Children’s Hospital of Philadelphia Philadelphia PennsylvaniaNational Institute for Biological Standards and Control Potters Bar UKPenn Comprehensive Hemophilia and Thrombosis Program Hospital of the University of Pennsylvania Philadelphia PennsylvaniaDepartment of Chemical and Biomolecular Engineering Institute for Medicine and Engineering University of Pennsylvania Philadelphia PennsylvaniaAbstract Background Microfluidic clotting assays permit drug action studies for hemophilia therapeutics under flow. However, limited availability of patient samples and Inter‐donor variability limit the application of such assays, especially with many patients on prophylaxis. Objective To develop approaches to phenocopy hemophilia using modified healthy blood in microfluidic assays. Methods Corn trypsin inhibitor (4 µg/mL)‐treated healthy blood was dosed with either anti–factor VIII (FVIII; hemophilia A model) or a recombinant factor IX (FIX) missense variant (FIX‐V181T; hemophilia B model). Treated blood was perfused at 100 s−1 wall shear rate over collagen/tissue factor (TF) or collagen/factor XIa (FXIa). Results Anti‐FVIII partially blocked fibrin production on collagen/TF, but completely blocked fibrin production on collagen/FXIa, a phenotype reversed with 1 µmol/L bispecific antibody (emicizumab), which binds FIXa and factor X. As expected, emicizumab had no significant effect on healthy blood (no anti‐FVIII present) perfused over collagen/FXIa. The efficacy of emicizumab in anti‐FVIII‐treated healthy blood phenocopied the action of emicizumab in the blood of a patient with hemophilia A perfused over collagen/FXIa. Interestingly, a patient‐derived FVIII‐neutralizing antibody reduced fibrin production when added to healthy blood perfused over collagen/FXIa. For low TF surfaces, reFIX‐V181T (50 µg/mL) fully blocked platelet and fibrin deposition, a phenotype fully reversed with anti‐TFPI. Conclusion Two new microfluidic hemophilia A and B models demonstrate the potency of anti‐TF pathway inhibitor, emicizumab, and a patient‐derived inhibitory antibody. Using collagen/FXIa‐coated surfaces resulted in reliable and highly sensitive hemophilia models.https://doi.org/10.1002/rth2.12286drug evaluationfibrinhemophiliahemostasismicrofluidics |
| spellingShingle | Xinren Yu Karen A. Panckeri Lacramioara Ivanciu Rodney M. Camire Carmen H. Coxon Adam Cuker Scott L. Diamond Microfluidic hemophilia models using blood from healthy donors Research and Practice in Thrombosis and Haemostasis drug evaluation fibrin hemophilia hemostasis microfluidics |
| title | Microfluidic hemophilia models using blood from healthy donors |
| title_full | Microfluidic hemophilia models using blood from healthy donors |
| title_fullStr | Microfluidic hemophilia models using blood from healthy donors |
| title_full_unstemmed | Microfluidic hemophilia models using blood from healthy donors |
| title_short | Microfluidic hemophilia models using blood from healthy donors |
| title_sort | microfluidic hemophilia models using blood from healthy donors |
| topic | drug evaluation fibrin hemophilia hemostasis microfluidics |
| url | https://doi.org/10.1002/rth2.12286 |
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