Development of a Bispecific Antibody-Based Platform for Retargeting of Capsid Modified AAV Vectors
A major limiting factor for systemically delivered gene therapies is the lack of novel tissue specific AAV (Adeno-associated virus) derived vectors. Bispecific antibodies can be used to redirect AAVs to specific target receptors. Here, we demonstrate that the insertion of a short linear epitope “2E3...
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| Format: | Article |
| Language: | English |
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MDPI AG
2021-08-01
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| Series: | International Journal of Molecular Sciences |
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| Online Access: | https://www.mdpi.com/1422-0067/22/15/8355 |
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| author | Juliane Kuklik Stefan Michelfelder Felix Schiele Sebastian Kreuz Thorsten Lamla Philipp Müller John E. Park |
| author_facet | Juliane Kuklik Stefan Michelfelder Felix Schiele Sebastian Kreuz Thorsten Lamla Philipp Müller John E. Park |
| author_sort | Juliane Kuklik |
| collection | DOAJ |
| description | A major limiting factor for systemically delivered gene therapies is the lack of novel tissue specific AAV (Adeno-associated virus) derived vectors. Bispecific antibodies can be used to redirect AAVs to specific target receptors. Here, we demonstrate that the insertion of a short linear epitope “2E3” derived from human proprotein-convertase subtilisin/kexin type 9 (PCSK9) into different surface loops of the VP capsid proteins can be used for AAV de-targeting from its natural receptor(s), combined with a bispecific antibody-mediated retargeting. We chose to target a set of distinct disease relevant membrane proteins—fibroblast activation protein (FAP), which is upregulated on activated fibroblasts within the tumor stroma and in fibrotic tissues, as well as programmed death-ligand 1 (PD-L1), which is strongly upregulated in many cancers. Upon incubation with a bispecific antibody recognizing the 2E3 epitope and FAP or PD-L1, the bispecific antibody/rAAV complex was able to selectively transduce receptor positive cells. In summary, we developed a novel, rationally designed vector retargeting platform that can target AAVs to a new set of cellular receptors in a modular fashion. This versatile platform may serve as a valuable tool to investigate the role of disease relevant cell types and basis for novel gene therapy approaches. |
| first_indexed | 2024-03-10T09:13:44Z |
| format | Article |
| id | doaj.art-2803878cbc454b24ba7c1bf50f1d1bfb |
| institution | Directory Open Access Journal |
| issn | 1661-6596 1422-0067 |
| language | English |
| last_indexed | 2024-03-10T09:13:44Z |
| publishDate | 2021-08-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | International Journal of Molecular Sciences |
| spelling | doaj.art-2803878cbc454b24ba7c1bf50f1d1bfb2023-11-22T05:47:13ZengMDPI AGInternational Journal of Molecular Sciences1661-65961422-00672021-08-012215835510.3390/ijms22158355Development of a Bispecific Antibody-Based Platform for Retargeting of Capsid Modified AAV VectorsJuliane Kuklik0Stefan Michelfelder1Felix Schiele2Sebastian Kreuz3Thorsten Lamla4Philipp Müller5John E. Park6Division of Cancer Immunology and Immune Modulation, Boehringer Ingelheim Pharma GmbH & Co. KG, 88387 Biberach an der Riss, GermanyDivision of Research Beyond Borders, Boehringer Ingelheim Pharma GmbH & Co. KG, 88387 Biberach an der Riss, GermanyDivision of Biotherapeutics Discovery, Boehringer Ingelheim Pharma GmbH & Co. KG, 88387 Biberach an der Riss, GermanyDivision of Research Beyond Borders, Boehringer Ingelheim Pharma GmbH & Co. KG, 88387 Biberach an der Riss, GermanyDivision of Drug Discovery Sciences Biberach, Boehringer Ingelheim Pharma GmbH & Co. KG, 88387 Biberach an der Riss, GermanyBoehringer Ingelheim Venture Fund GmbH, 55216 Ingelheim am Rhein, GermanyDivision of Cancer Immunology and Immune Modulation, Boehringer Ingelheim Pharma GmbH & Co. KG, 88387 Biberach an der Riss, GermanyA major limiting factor for systemically delivered gene therapies is the lack of novel tissue specific AAV (Adeno-associated virus) derived vectors. Bispecific antibodies can be used to redirect AAVs to specific target receptors. Here, we demonstrate that the insertion of a short linear epitope “2E3” derived from human proprotein-convertase subtilisin/kexin type 9 (PCSK9) into different surface loops of the VP capsid proteins can be used for AAV de-targeting from its natural receptor(s), combined with a bispecific antibody-mediated retargeting. We chose to target a set of distinct disease relevant membrane proteins—fibroblast activation protein (FAP), which is upregulated on activated fibroblasts within the tumor stroma and in fibrotic tissues, as well as programmed death-ligand 1 (PD-L1), which is strongly upregulated in many cancers. Upon incubation with a bispecific antibody recognizing the 2E3 epitope and FAP or PD-L1, the bispecific antibody/rAAV complex was able to selectively transduce receptor positive cells. In summary, we developed a novel, rationally designed vector retargeting platform that can target AAVs to a new set of cellular receptors in a modular fashion. This versatile platform may serve as a valuable tool to investigate the role of disease relevant cell types and basis for novel gene therapy approaches.https://www.mdpi.com/1422-0067/22/15/8355adeno-associated viral vectorsAAVprotein engineeringretargetingbispecific antibodycapsid modification |
| spellingShingle | Juliane Kuklik Stefan Michelfelder Felix Schiele Sebastian Kreuz Thorsten Lamla Philipp Müller John E. Park Development of a Bispecific Antibody-Based Platform for Retargeting of Capsid Modified AAV Vectors International Journal of Molecular Sciences adeno-associated viral vectors AAV protein engineering retargeting bispecific antibody capsid modification |
| title | Development of a Bispecific Antibody-Based Platform for Retargeting of Capsid Modified AAV Vectors |
| title_full | Development of a Bispecific Antibody-Based Platform for Retargeting of Capsid Modified AAV Vectors |
| title_fullStr | Development of a Bispecific Antibody-Based Platform for Retargeting of Capsid Modified AAV Vectors |
| title_full_unstemmed | Development of a Bispecific Antibody-Based Platform for Retargeting of Capsid Modified AAV Vectors |
| title_short | Development of a Bispecific Antibody-Based Platform for Retargeting of Capsid Modified AAV Vectors |
| title_sort | development of a bispecific antibody based platform for retargeting of capsid modified aav vectors |
| topic | adeno-associated viral vectors AAV protein engineering retargeting bispecific antibody capsid modification |
| url | https://www.mdpi.com/1422-0067/22/15/8355 |
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