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Fragments of gD Protein as Inhibitors of BTLA/HVEM Complex Formation - Design, Synthesis, and Cellular Studies

One of the major current trends in cancer immunotherapy is the blockade of immune checkpoint proteins that negatively regulate the immune response. This has been achieved through antibodies blocking PD-1/PD-L1 and CTLA-4/CD80/CD86 interactions. Such antibodies have revolutionized oncological therapy...

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Autores principales: Kuncewicz, Katarzyna, Battin, Claire, Sieradzan, Adam, Karczyńska, Agnieszka, Orlikowska, Marta, Wardowska, Anna, Pikuła, Michał, Steinberger, Peter, Rodziewicz-Motowidło, Sylwia, Spodzieja, Marta
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7700651/
https://www.ncbi.nlm.nih.gov/pubmed/33238640
http://dx.doi.org/10.3390/ijms21228876
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author Kuncewicz, Katarzyna
Battin, Claire
Sieradzan, Adam
Karczyńska, Agnieszka
Orlikowska, Marta
Wardowska, Anna
Pikuła, Michał
Steinberger, Peter
Rodziewicz-Motowidło, Sylwia
Spodzieja, Marta
author_facet Kuncewicz, Katarzyna
Battin, Claire
Sieradzan, Adam
Karczyńska, Agnieszka
Orlikowska, Marta
Wardowska, Anna
Pikuła, Michał
Steinberger, Peter
Rodziewicz-Motowidło, Sylwia
Spodzieja, Marta
author_sort Kuncewicz, Katarzyna
collection PubMed
description One of the major current trends in cancer immunotherapy is the blockade of immune checkpoint proteins that negatively regulate the immune response. This has been achieved through antibodies blocking PD-1/PD-L1 and CTLA-4/CD80/CD86 interactions. Such antibodies have revolutionized oncological therapy and shown a new way to fight cancer. Additional (negative) immune checkpoints are also promising targets in cancer therapy and there is a demand for inhibitors for these molecules. Our studies are focused on BTLA/HVEM complex, which inhibits T-cell proliferation and cytokine production and therefore has great potential as a new target for cancer treatment. The goal of the presented studies was the design and synthesis of compounds able to block BTLA/HVEM interactions. For that purpose, the N-terminal fragment of glycoprotein D (gD), which interacts with HVEM, was used. Based on the crystal structure of the gD/HVEM complex and MM/GBSA analysis performed on it, several peptides were designed and synthesized as potential inhibitors of the BTLA/HVEM interaction. Affinity tests, ELISA tests, and cellular-based reporter assays were performed on these compounds to check their ability to bind to HVEM and to inhibit BTLA/HVEM complex formation. For leading peptides candidates, all-atom and subsequent docking simulations with a coarse-grained force field were performed to determine their binding modes. To further evaluate their potential as drug candidates, their stability in plasma and their cytotoxicity effects on PBMCs were assessed. Our data indicate that the peptide gD(1-36)(K10C-T29C) is the best candidate as a future drug. It interacts with HVEM protein, blocks the BTLA/HVEM interaction, and is nontoxic to cells. The present study provides a new perspective on the development of BTLA/HVEM inhibitors that disrupt protein interactions.
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spelling pubmed-77006512020-11-30 Fragments of gD Protein as Inhibitors of BTLA/HVEM Complex Formation - Design, Synthesis, and Cellular Studies Kuncewicz, Katarzyna Battin, Claire Sieradzan, Adam Karczyńska, Agnieszka Orlikowska, Marta Wardowska, Anna Pikuła, Michał Steinberger, Peter Rodziewicz-Motowidło, Sylwia Spodzieja, Marta Int J Mol Sci Article One of the major current trends in cancer immunotherapy is the blockade of immune checkpoint proteins that negatively regulate the immune response. This has been achieved through antibodies blocking PD-1/PD-L1 and CTLA-4/CD80/CD86 interactions. Such antibodies have revolutionized oncological therapy and shown a new way to fight cancer. Additional (negative) immune checkpoints are also promising targets in cancer therapy and there is a demand for inhibitors for these molecules. Our studies are focused on BTLA/HVEM complex, which inhibits T-cell proliferation and cytokine production and therefore has great potential as a new target for cancer treatment. The goal of the presented studies was the design and synthesis of compounds able to block BTLA/HVEM interactions. For that purpose, the N-terminal fragment of glycoprotein D (gD), which interacts with HVEM, was used. Based on the crystal structure of the gD/HVEM complex and MM/GBSA analysis performed on it, several peptides were designed and synthesized as potential inhibitors of the BTLA/HVEM interaction. Affinity tests, ELISA tests, and cellular-based reporter assays were performed on these compounds to check their ability to bind to HVEM and to inhibit BTLA/HVEM complex formation. For leading peptides candidates, all-atom and subsequent docking simulations with a coarse-grained force field were performed to determine their binding modes. To further evaluate their potential as drug candidates, their stability in plasma and their cytotoxicity effects on PBMCs were assessed. Our data indicate that the peptide gD(1-36)(K10C-T29C) is the best candidate as a future drug. It interacts with HVEM protein, blocks the BTLA/HVEM interaction, and is nontoxic to cells. The present study provides a new perspective on the development of BTLA/HVEM inhibitors that disrupt protein interactions. MDPI 2020-11-23 /pmc/articles/PMC7700651/ /pubmed/33238640 http://dx.doi.org/10.3390/ijms21228876 Text en © 2020 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Kuncewicz, Katarzyna
Battin, Claire
Sieradzan, Adam
Karczyńska, Agnieszka
Orlikowska, Marta
Wardowska, Anna
Pikuła, Michał
Steinberger, Peter
Rodziewicz-Motowidło, Sylwia
Spodzieja, Marta
Fragments of gD Protein as Inhibitors of BTLA/HVEM Complex Formation - Design, Synthesis, and Cellular Studies
title Fragments of gD Protein as Inhibitors of BTLA/HVEM Complex Formation - Design, Synthesis, and Cellular Studies
title_full Fragments of gD Protein as Inhibitors of BTLA/HVEM Complex Formation - Design, Synthesis, and Cellular Studies
title_fullStr Fragments of gD Protein as Inhibitors of BTLA/HVEM Complex Formation - Design, Synthesis, and Cellular Studies
title_full_unstemmed Fragments of gD Protein as Inhibitors of BTLA/HVEM Complex Formation - Design, Synthesis, and Cellular Studies
title_short Fragments of gD Protein as Inhibitors of BTLA/HVEM Complex Formation - Design, Synthesis, and Cellular Studies
title_sort fragments of gd protein as inhibitors of btla/hvem complex formation - design, synthesis, and cellular studies
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7700651/
https://www.ncbi.nlm.nih.gov/pubmed/33238640
http://dx.doi.org/10.3390/ijms21228876
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