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Microfluidic hemophilia models using blood from healthy donors
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 approache...
Autores principales: | , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
John Wiley and Sons Inc.
2019
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6971334/ https://www.ncbi.nlm.nih.gov/pubmed/31989085 http://dx.doi.org/10.1002/rth2.12286 |
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author | Yu, Xinren Panckeri, Karen A. Ivanciu, Lacramioara Camire, Rodney M. Coxon, Carmen H. Cuker, Adam Diamond, Scott L. |
author_facet | Yu, Xinren Panckeri, Karen A. Ivanciu, Lacramioara Camire, Rodney M. Coxon, Carmen H. Cuker, Adam Diamond, Scott L. |
author_sort | Yu, Xinren |
collection | PubMed |
description | 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. |
format | Online Article Text |
id | pubmed-6971334 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2019 |
publisher | John Wiley and Sons Inc. |
record_format | MEDLINE/PubMed |
spelling | pubmed-69713342020-01-27 Microfluidic hemophilia models using blood from healthy donors Yu, Xinren Panckeri, Karen A. Ivanciu, Lacramioara Camire, Rodney M. Coxon, Carmen H. Cuker, Adam Diamond, Scott L. Res Pract Thromb Haemost Methodological Articles 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. John Wiley and Sons Inc. 2019-12-02 /pmc/articles/PMC6971334/ /pubmed/31989085 http://dx.doi.org/10.1002/rth2.12286 Text en © 2019 The Authors. Research and Practice in Thrombosis and Haemostasis published by Wiley Periodicals, Inc on behalf of International Society on Thrombosis and Haemostasis. This is an open access article under the terms of the http://creativecommons.org/licenses/by-nc-nd/4.0/ License, which permits use and distribution in any medium, provided the original work is properly cited, the use is non‐commercial and no modifications or adaptations are made. |
spellingShingle | Methodological Articles Yu, Xinren Panckeri, Karen A. Ivanciu, Lacramioara Camire, Rodney M. Coxon, Carmen H. Cuker, Adam Diamond, Scott L. Microfluidic hemophilia models using blood from healthy donors |
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 | Methodological Articles |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6971334/ https://www.ncbi.nlm.nih.gov/pubmed/31989085 http://dx.doi.org/10.1002/rth2.12286 |
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