Identification and characterisation of anti-IL-13 inhibitory single domain antibodies provides new insights into receptor selectivity and attractive opportunities for drug discovery
Interleukin-13 (IL-13) is a cytokine involved in T-cell immune responses and is a well validated therapeutic target for the treatment of asthma, along with other allergic and inflammatory diseases. IL-13 signals through a ternary signalling complex formed with the receptors IL-13Rα1 and IL-4Rα. This...
| Main Authors: | , , , , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
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Frontiers Media S.A.
2023-07-01
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| Series: | Frontiers in Immunology |
| Subjects: | |
| Online Access: | https://www.frontiersin.org/articles/10.3389/fimmu.2023.1216967/full |
| _version_ | 1797785617529044992 |
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| author | Kayleigh Walker Roberta Baravalle Rachel Holyfield Jacqueline Kalms Jacqueline Kalms Helena Wright Chitra Seewooruthun Frederick W. Muskett Anthony Scott-Tucker Andy Merritt Alistair Henry Alastair D. G. Lawson Gareth Hall Christine Prosser Christine Prosser Mark D. Carr |
| author_facet | Kayleigh Walker Roberta Baravalle Rachel Holyfield Jacqueline Kalms Jacqueline Kalms Helena Wright Chitra Seewooruthun Frederick W. Muskett Anthony Scott-Tucker Andy Merritt Alistair Henry Alastair D. G. Lawson Gareth Hall Christine Prosser Christine Prosser Mark D. Carr |
| author_sort | Kayleigh Walker |
| collection | DOAJ |
| description | Interleukin-13 (IL-13) is a cytokine involved in T-cell immune responses and is a well validated therapeutic target for the treatment of asthma, along with other allergic and inflammatory diseases. IL-13 signals through a ternary signalling complex formed with the receptors IL-13Rα1 and IL-4Rα. This complex is assembled by IL-13 initially binding IL-13Rα1, followed by association of the binary IL-13:IL-13Rα1 complex with IL-4Rα. The receptors are shared with IL-4, but IL-4 initially binds IL-4Rα. Here we report the identification and characterisation of a diverse panel of single-domain antibodies (VHHs) that bind to IL-13 (KD 40 nM-5.5 μM) and inhibit downstream IL-13 signalling (IC50 0.2-53.8 μM). NMR mapping showed that the VHHs recognise a number of epitopes on IL-13, including previously unknown allosteric sites. Further NMR investigation of VHH204 bound to IL-13 revealed a novel allosteric mechanism of inhibition, with the antibody stabilising IL-13 in a conformation incompatible with receptor binding. This also led to the identification of a conformational equilibrium for free IL-13, providing insights into differing receptor signalling complex assembly seen for IL-13 compared to IL-4, with formation of the IL-13:IL-13Rα1 complex required to stabilise IL-13 in a conformation with high affinity for IL-4Rα. These findings highlight new opportunities for therapeutic targeting of IL-13 and we report a successful 19F fragment screen of the IL-13:VHH204 complex, including binding sites identified for several hits. To our knowledge, these 19F containing fragments represent the first small-molecules shown to bind to IL-13 and could provide starting points for a small-molecule drug discovery programme. |
| first_indexed | 2024-03-13T00:56:34Z |
| format | Article |
| id | doaj.art-ed60b6efb669458b96fb34943581aadf |
| institution | Directory Open Access Journal |
| issn | 1664-3224 |
| language | English |
| last_indexed | 2024-03-13T00:56:34Z |
| publishDate | 2023-07-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Immunology |
| spelling | doaj.art-ed60b6efb669458b96fb34943581aadf2023-07-06T22:27:24ZengFrontiers Media S.A.Frontiers in Immunology1664-32242023-07-011410.3389/fimmu.2023.12169671216967Identification and characterisation of anti-IL-13 inhibitory single domain antibodies provides new insights into receptor selectivity and attractive opportunities for drug discoveryKayleigh Walker0Roberta Baravalle1Rachel Holyfield2Jacqueline Kalms3Jacqueline Kalms4Helena Wright5Chitra Seewooruthun6Frederick W. Muskett7Anthony Scott-Tucker8Andy Merritt9Alistair Henry10Alastair D. G. Lawson11Gareth Hall12Christine Prosser13Christine Prosser14Mark D. Carr15Leicester Institute of Structural and Chemical Biology, and Department of Molecular and Cell Biology, University of Leicester, Leicester, United KingdomLeicester Institute of Structural and Chemical Biology, and Department of Molecular and Cell Biology, University of Leicester, Leicester, United KingdomLeicester Institute of Structural and Chemical Biology, and Department of Molecular and Cell Biology, University of Leicester, Leicester, United KingdomLeicester Institute of Structural and Chemical Biology, and Department of Molecular and Cell Biology, University of Leicester, Leicester, United KingdomUCB Biopharma, UCB Pharma, Slough, United KingdomLeicester Institute of Structural and Chemical Biology, and Department of Molecular and Cell Biology, University of Leicester, Leicester, United KingdomLeicester Institute of Structural and Chemical Biology, and Department of Molecular and Cell Biology, University of Leicester, Leicester, United KingdomLeicester Institute of Structural and Chemical Biology, and Department of Molecular and Cell Biology, University of Leicester, Leicester, United KingdomUCB Biopharma, UCB Pharma, Slough, United KingdomLifeArc, Centre for Therapeutics Discovery, Stevenage Bioscience Catalyst, Stevenage, United KingdomUCB Biopharma, UCB Pharma, Slough, United KingdomUCB Biopharma, UCB Pharma, Slough, United KingdomLeicester Institute of Structural and Chemical Biology, and Department of Molecular and Cell Biology, University of Leicester, Leicester, United KingdomLeicester Institute of Structural and Chemical Biology, and Department of Molecular and Cell Biology, University of Leicester, Leicester, United KingdomUCB Biopharma, UCB Pharma, Slough, United KingdomLeicester Institute of Structural and Chemical Biology, and Department of Molecular and Cell Biology, University of Leicester, Leicester, United KingdomInterleukin-13 (IL-13) is a cytokine involved in T-cell immune responses and is a well validated therapeutic target for the treatment of asthma, along with other allergic and inflammatory diseases. IL-13 signals through a ternary signalling complex formed with the receptors IL-13Rα1 and IL-4Rα. This complex is assembled by IL-13 initially binding IL-13Rα1, followed by association of the binary IL-13:IL-13Rα1 complex with IL-4Rα. The receptors are shared with IL-4, but IL-4 initially binds IL-4Rα. Here we report the identification and characterisation of a diverse panel of single-domain antibodies (VHHs) that bind to IL-13 (KD 40 nM-5.5 μM) and inhibit downstream IL-13 signalling (IC50 0.2-53.8 μM). NMR mapping showed that the VHHs recognise a number of epitopes on IL-13, including previously unknown allosteric sites. Further NMR investigation of VHH204 bound to IL-13 revealed a novel allosteric mechanism of inhibition, with the antibody stabilising IL-13 in a conformation incompatible with receptor binding. This also led to the identification of a conformational equilibrium for free IL-13, providing insights into differing receptor signalling complex assembly seen for IL-13 compared to IL-4, with formation of the IL-13:IL-13Rα1 complex required to stabilise IL-13 in a conformation with high affinity for IL-4Rα. These findings highlight new opportunities for therapeutic targeting of IL-13 and we report a successful 19F fragment screen of the IL-13:VHH204 complex, including binding sites identified for several hits. To our knowledge, these 19F containing fragments represent the first small-molecules shown to bind to IL-13 and could provide starting points for a small-molecule drug discovery programme.https://www.frontiersin.org/articles/10.3389/fimmu.2023.1216967/fullinterleukin-13single domain antibodiesVHHreceptor signallingreceptor selectivityallosteric regulation |
| spellingShingle | Kayleigh Walker Roberta Baravalle Rachel Holyfield Jacqueline Kalms Jacqueline Kalms Helena Wright Chitra Seewooruthun Frederick W. Muskett Anthony Scott-Tucker Andy Merritt Alistair Henry Alastair D. G. Lawson Gareth Hall Christine Prosser Christine Prosser Mark D. Carr Identification and characterisation of anti-IL-13 inhibitory single domain antibodies provides new insights into receptor selectivity and attractive opportunities for drug discovery Frontiers in Immunology interleukin-13 single domain antibodies VHH receptor signalling receptor selectivity allosteric regulation |
| title | Identification and characterisation of anti-IL-13 inhibitory single domain antibodies provides new insights into receptor selectivity and attractive opportunities for drug discovery |
| title_full | Identification and characterisation of anti-IL-13 inhibitory single domain antibodies provides new insights into receptor selectivity and attractive opportunities for drug discovery |
| title_fullStr | Identification and characterisation of anti-IL-13 inhibitory single domain antibodies provides new insights into receptor selectivity and attractive opportunities for drug discovery |
| title_full_unstemmed | Identification and characterisation of anti-IL-13 inhibitory single domain antibodies provides new insights into receptor selectivity and attractive opportunities for drug discovery |
| title_short | Identification and characterisation of anti-IL-13 inhibitory single domain antibodies provides new insights into receptor selectivity and attractive opportunities for drug discovery |
| title_sort | identification and characterisation of anti il 13 inhibitory single domain antibodies provides new insights into receptor selectivity and attractive opportunities for drug discovery |
| topic | interleukin-13 single domain antibodies VHH receptor signalling receptor selectivity allosteric regulation |
| url | https://www.frontiersin.org/articles/10.3389/fimmu.2023.1216967/full |
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