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Generation and validation of structurally defined antibody–siRNA conjugates
Gene silencing by RNA interference (RNAi) has emerged as a powerful treatment strategy across a potentially broad range of diseases. Tailoring siRNAs to silence genes vital for cancer cell growth and function could be an effective treatment, but there are several challenges which must be overcome to...
Autores principales: | , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Oxford University Press
2020
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7261152/ https://www.ncbi.nlm.nih.gov/pubmed/32347936 http://dx.doi.org/10.1093/nar/gkaa286 |
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author | Nanna, Alex R Kel’in, Alexander V Theile, Christopher Pierson, Justin M Voo, Zhi Xiang Garg, Ashish Nair, Jayaprakash K Maier, Martin A Fitzgerald, Kevin Rader, Christoph |
author_facet | Nanna, Alex R Kel’in, Alexander V Theile, Christopher Pierson, Justin M Voo, Zhi Xiang Garg, Ashish Nair, Jayaprakash K Maier, Martin A Fitzgerald, Kevin Rader, Christoph |
author_sort | Nanna, Alex R |
collection | PubMed |
description | Gene silencing by RNA interference (RNAi) has emerged as a powerful treatment strategy across a potentially broad range of diseases. Tailoring siRNAs to silence genes vital for cancer cell growth and function could be an effective treatment, but there are several challenges which must be overcome to enable their use as a therapeutic modality, among which efficient and selective delivery to cancer cells remains paramount. Attempts to use antibodies for siRNA delivery have been reported but these strategies use either nonspecific conjugation resulting in mixtures, or site-specific methods that require multiple steps, introduction of mutations, or use of enzymes. Here, we report a method to generate antibody–siRNA (1:2) conjugates (ARCs) that are structurally defined and easy to assemble. This ARC platform is based on engineered dual variable domain (DVD) antibodies containing a natural uniquely reactive lysine residue for site-specific conjugation to β-lactam linker-functionalized siRNA. The conjugation is efficient, does not compromise the affinity of the parental antibody, and utilizes chemically stabilized siRNA. For proof-of-concept, we generated DVD-ARCs targeting various cell surface antigens on multiple myeloma cells for the selective delivery of siRNA targeting β-catenin (CTNNB1). A set of BCMA-targeting DVD-ARCs at concentrations as low as 10 nM revealed significant CTNNB1 mRNA and protein knockdown. |
format | Online Article Text |
id | pubmed-7261152 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2020 |
publisher | Oxford University Press |
record_format | MEDLINE/PubMed |
spelling | pubmed-72611522020-06-03 Generation and validation of structurally defined antibody–siRNA conjugates Nanna, Alex R Kel’in, Alexander V Theile, Christopher Pierson, Justin M Voo, Zhi Xiang Garg, Ashish Nair, Jayaprakash K Maier, Martin A Fitzgerald, Kevin Rader, Christoph Nucleic Acids Res Chemical Biology and Nucleic Acid Chemistry Gene silencing by RNA interference (RNAi) has emerged as a powerful treatment strategy across a potentially broad range of diseases. Tailoring siRNAs to silence genes vital for cancer cell growth and function could be an effective treatment, but there are several challenges which must be overcome to enable their use as a therapeutic modality, among which efficient and selective delivery to cancer cells remains paramount. Attempts to use antibodies for siRNA delivery have been reported but these strategies use either nonspecific conjugation resulting in mixtures, or site-specific methods that require multiple steps, introduction of mutations, or use of enzymes. Here, we report a method to generate antibody–siRNA (1:2) conjugates (ARCs) that are structurally defined and easy to assemble. This ARC platform is based on engineered dual variable domain (DVD) antibodies containing a natural uniquely reactive lysine residue for site-specific conjugation to β-lactam linker-functionalized siRNA. The conjugation is efficient, does not compromise the affinity of the parental antibody, and utilizes chemically stabilized siRNA. For proof-of-concept, we generated DVD-ARCs targeting various cell surface antigens on multiple myeloma cells for the selective delivery of siRNA targeting β-catenin (CTNNB1). A set of BCMA-targeting DVD-ARCs at concentrations as low as 10 nM revealed significant CTNNB1 mRNA and protein knockdown. Oxford University Press 2020-06-04 2020-04-29 /pmc/articles/PMC7261152/ /pubmed/32347936 http://dx.doi.org/10.1093/nar/gkaa286 Text en © The Author(s) 2020. Published by Oxford University Press on behalf of Nucleic Acids Research. http://creativecommons.org/licenses/by-nc/4.0/ This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/4.0/), which permits non-commercial re-use, distribution, and reproduction in any medium, provided the original work is properly cited. For commercial re-use, please contact journals.permissions@oup.com |
spellingShingle | Chemical Biology and Nucleic Acid Chemistry Nanna, Alex R Kel’in, Alexander V Theile, Christopher Pierson, Justin M Voo, Zhi Xiang Garg, Ashish Nair, Jayaprakash K Maier, Martin A Fitzgerald, Kevin Rader, Christoph Generation and validation of structurally defined antibody–siRNA conjugates |
title | Generation and validation of structurally defined antibody–siRNA conjugates |
title_full | Generation and validation of structurally defined antibody–siRNA conjugates |
title_fullStr | Generation and validation of structurally defined antibody–siRNA conjugates |
title_full_unstemmed | Generation and validation of structurally defined antibody–siRNA conjugates |
title_short | Generation and validation of structurally defined antibody–siRNA conjugates |
title_sort | generation and validation of structurally defined antibody–sirna conjugates |
topic | Chemical Biology and Nucleic Acid Chemistry |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7261152/ https://www.ncbi.nlm.nih.gov/pubmed/32347936 http://dx.doi.org/10.1093/nar/gkaa286 |
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