Targeted erythropoietin selectively stimulates red blood cell expansion in vivo
The design of cell-targeted protein therapeutics can be informed by natural protein–protein interactions that use cooperative physical contacts to achieve cell type specificity. Here we applied this approach in vivo to the anemia drug erythropoietin (EPO), to direct its activity to EPO receptors (EP...
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| Format: | Article |
| Language: | en_US |
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National Academy of Sciences (U.S.)
2016
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| Online Access: | http://hdl.handle.net/1721.1/105542 https://orcid.org/0000-0002-5560-8246 |
| _version_ | 1826209507678617600 |
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| author | Burrill, Devin R. Vernet, Andyna Collins, James J. Silver, Pamela A. Way, Jeffrey C. |
| author2 | Institute for Medical Engineering and Science |
| author_facet | Institute for Medical Engineering and Science Burrill, Devin R. Vernet, Andyna Collins, James J. Silver, Pamela A. Way, Jeffrey C. |
| author_sort | Burrill, Devin R. |
| collection | MIT |
| description | The design of cell-targeted protein therapeutics can be informed by natural protein–protein interactions that use cooperative physical contacts to achieve cell type specificity. Here we applied this approach in vivo to the anemia drug erythropoietin (EPO), to direct its activity to EPO receptors (EPO-Rs) on red blood cell (RBC) precursors and prevent interaction with EPO-Rs on nonerythroid cells, such as platelets. Our engineered EPO molecule was mutated to weaken its affinity for EPO-R, but its avidity for RBC precursors was rescued via tethering to an antibody fragment that specifically binds the human RBC marker glycophorin A (huGYPA). We systematically tested the impact of these engineering steps on in vivo markers of efficacy, side effects, and pharmacokinetics. huGYPA transgenic mice dosed with targeted EPO exhibited elevated RBC levels, with only minimal platelet effects. This in vivo selectivity depended on the weakening EPO mutation, fusion to the RBC-specific antibody, and expression of huGYPA. The terminal plasma half-life of targeted EPO was ∼28.3 h in transgenic mice vs. ∼15.5 h in nontransgenic mice, indicating that huGYPA on mature RBCs acted as a significant drug sink but did not inhibit efficacy. In a therapeutic context, our targeting approach may allow higher restorative doses of EPO without platelet-mediated side effects, and also may improve drug pharmacokinetics. These results demonstrate how rational drug design can improve in vivo specificity, with potential application to diverse protein therapeutics. |
| first_indexed | 2024-09-23T14:23:34Z |
| format | Article |
| id | mit-1721.1/105542 |
| institution | Massachusetts Institute of Technology |
| language | en_US |
| last_indexed | 2024-09-23T14:23:34Z |
| publishDate | 2016 |
| publisher | National Academy of Sciences (U.S.) |
| record_format | dspace |
| spelling | mit-1721.1/1055422022-09-29T09:12:51Z Targeted erythropoietin selectively stimulates red blood cell expansion in vivo Burrill, Devin R. Vernet, Andyna Collins, James J. Silver, Pamela A. Way, Jeffrey C. Institute for Medical Engineering and Science MIT Synthetic Biology Center Massachusetts Institute of Technology. Department of Biological Engineering Collins, James J. The design of cell-targeted protein therapeutics can be informed by natural protein–protein interactions that use cooperative physical contacts to achieve cell type specificity. Here we applied this approach in vivo to the anemia drug erythropoietin (EPO), to direct its activity to EPO receptors (EPO-Rs) on red blood cell (RBC) precursors and prevent interaction with EPO-Rs on nonerythroid cells, such as platelets. Our engineered EPO molecule was mutated to weaken its affinity for EPO-R, but its avidity for RBC precursors was rescued via tethering to an antibody fragment that specifically binds the human RBC marker glycophorin A (huGYPA). We systematically tested the impact of these engineering steps on in vivo markers of efficacy, side effects, and pharmacokinetics. huGYPA transgenic mice dosed with targeted EPO exhibited elevated RBC levels, with only minimal platelet effects. This in vivo selectivity depended on the weakening EPO mutation, fusion to the RBC-specific antibody, and expression of huGYPA. The terminal plasma half-life of targeted EPO was ∼28.3 h in transgenic mice vs. ∼15.5 h in nontransgenic mice, indicating that huGYPA on mature RBCs acted as a significant drug sink but did not inhibit efficacy. In a therapeutic context, our targeting approach may allow higher restorative doses of EPO without platelet-mediated side effects, and also may improve drug pharmacokinetics. These results demonstrate how rational drug design can improve in vivo specificity, with potential application to diverse protein therapeutics. Wyss Institute for Biologically Inspired Engineering (Pilot Award 112475) United States. Defense Advanced Research Projects Agency (Grant W911NF- 11-2-0056) National Institutes of Health (U.S.) (Grant R01 GM036373) 2016-12-02T20:55:12Z 2016-12-02T20:55:12Z 2016-04 2015-12 Article http://purl.org/eprint/type/JournalArticle 0027-8424 1091-6490 http://hdl.handle.net/1721.1/105542 Burrill, Devin R. et al. “Targeted Erythropoietin Selectively Stimulates Red Blood Cell Expansion in Vivo.” Proceedings of the National Academy of Sciences 113.19 (2016): 5245–5250. © 2016 National Academy of Sciences https://orcid.org/0000-0002-5560-8246 en_US http://dx.doi.org/10.1073/pnas.1525388113 Proceedings of the National Academy of Sciences Article is made available in accordance with the publisher's policy and may be subject to US copyright law. Please refer to the publisher's site for terms of use. application/pdf National Academy of Sciences (U.S.) PNAS |
| spellingShingle | Burrill, Devin R. Vernet, Andyna Collins, James J. Silver, Pamela A. Way, Jeffrey C. Targeted erythropoietin selectively stimulates red blood cell expansion in vivo |
| title | Targeted erythropoietin selectively stimulates red blood cell expansion in vivo |
| title_full | Targeted erythropoietin selectively stimulates red blood cell expansion in vivo |
| title_fullStr | Targeted erythropoietin selectively stimulates red blood cell expansion in vivo |
| title_full_unstemmed | Targeted erythropoietin selectively stimulates red blood cell expansion in vivo |
| title_short | Targeted erythropoietin selectively stimulates red blood cell expansion in vivo |
| title_sort | targeted erythropoietin selectively stimulates red blood cell expansion in vivo |
| url | http://hdl.handle.net/1721.1/105542 https://orcid.org/0000-0002-5560-8246 |
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