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Engineered Fully Human Single-Chain Monoclonal Antibodies to PIM2 Kinase

Proviral integration site of Moloney virus-2 (PIM2) is overexpressed in multiple human cancer cells and high level is related to poor prognosis; thus, PIM2 kinase is a rational target of anti-cancer therapeutics. Several chemical inhibitors targeting PIMs/PIM2 or their downstream signaling molecules...

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Autores principales: Kaewchim, Kanasap, Glab-ampai, Kittirat, Mahasongkram, Kodchakorn, Chulanetra, Monrat, Seesuay, Watee, Chaicumpa, Wanpen, Sookrung, Nitat
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8588357/
https://www.ncbi.nlm.nih.gov/pubmed/34770845
http://dx.doi.org/10.3390/molecules26216436
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author Kaewchim, Kanasap
Glab-ampai, Kittirat
Mahasongkram, Kodchakorn
Chulanetra, Monrat
Seesuay, Watee
Chaicumpa, Wanpen
Sookrung, Nitat
author_facet Kaewchim, Kanasap
Glab-ampai, Kittirat
Mahasongkram, Kodchakorn
Chulanetra, Monrat
Seesuay, Watee
Chaicumpa, Wanpen
Sookrung, Nitat
author_sort Kaewchim, Kanasap
collection PubMed
description Proviral integration site of Moloney virus-2 (PIM2) is overexpressed in multiple human cancer cells and high level is related to poor prognosis; thus, PIM2 kinase is a rational target of anti-cancer therapeutics. Several chemical inhibitors targeting PIMs/PIM2 or their downstream signaling molecules have been developed for treatment of different cancers. However, their off-target toxicity is common in clinical trials, so they could not be advanced to official approval for clinical application. Here, we produced human single-chain antibody fragments (HuscFvs) to PIM2 by using phage display library, which was constructed in a way that a portion of phages in the library carried HuscFvs against human own proteins on their surface with the respective antibody genes in the phage genome. Bacterial derived-recombinant PIM2 (rPIM2) was used as an antigenic bait to fish out the rPIM2-bound phages from the library. Three E. coli clones transfected with the HuscFv genes derived from the rPIM2-bound phages expressed HuscFvs that bound also to native PIM2 from cancer cells. The HuscFvs presumptively interact with the PIM2 at the ATP binding pocket and kinase active loop. They were as effective as small chemical drug inhibitor (AZD1208, which is an ATP competitive inhibitor of all PIM isoforms for ex vivo use) in inhibiting PIM kinase activity. The HuscFvs should be engineered into a cell-penetrating format and tested further towards clinical application as a novel and safe pan-anti-cancer therapeutics.
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spelling pubmed-85883572021-11-13 Engineered Fully Human Single-Chain Monoclonal Antibodies to PIM2 Kinase Kaewchim, Kanasap Glab-ampai, Kittirat Mahasongkram, Kodchakorn Chulanetra, Monrat Seesuay, Watee Chaicumpa, Wanpen Sookrung, Nitat Molecules Article Proviral integration site of Moloney virus-2 (PIM2) is overexpressed in multiple human cancer cells and high level is related to poor prognosis; thus, PIM2 kinase is a rational target of anti-cancer therapeutics. Several chemical inhibitors targeting PIMs/PIM2 or their downstream signaling molecules have been developed for treatment of different cancers. However, their off-target toxicity is common in clinical trials, so they could not be advanced to official approval for clinical application. Here, we produced human single-chain antibody fragments (HuscFvs) to PIM2 by using phage display library, which was constructed in a way that a portion of phages in the library carried HuscFvs against human own proteins on their surface with the respective antibody genes in the phage genome. Bacterial derived-recombinant PIM2 (rPIM2) was used as an antigenic bait to fish out the rPIM2-bound phages from the library. Three E. coli clones transfected with the HuscFv genes derived from the rPIM2-bound phages expressed HuscFvs that bound also to native PIM2 from cancer cells. The HuscFvs presumptively interact with the PIM2 at the ATP binding pocket and kinase active loop. They were as effective as small chemical drug inhibitor (AZD1208, which is an ATP competitive inhibitor of all PIM isoforms for ex vivo use) in inhibiting PIM kinase activity. The HuscFvs should be engineered into a cell-penetrating format and tested further towards clinical application as a novel and safe pan-anti-cancer therapeutics. MDPI 2021-10-25 /pmc/articles/PMC8588357/ /pubmed/34770845 http://dx.doi.org/10.3390/molecules26216436 Text en © 2021 by the authors. https://creativecommons.org/licenses/by/4.0/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 (https://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Kaewchim, Kanasap
Glab-ampai, Kittirat
Mahasongkram, Kodchakorn
Chulanetra, Monrat
Seesuay, Watee
Chaicumpa, Wanpen
Sookrung, Nitat
Engineered Fully Human Single-Chain Monoclonal Antibodies to PIM2 Kinase
title Engineered Fully Human Single-Chain Monoclonal Antibodies to PIM2 Kinase
title_full Engineered Fully Human Single-Chain Monoclonal Antibodies to PIM2 Kinase
title_fullStr Engineered Fully Human Single-Chain Monoclonal Antibodies to PIM2 Kinase
title_full_unstemmed Engineered Fully Human Single-Chain Monoclonal Antibodies to PIM2 Kinase
title_short Engineered Fully Human Single-Chain Monoclonal Antibodies to PIM2 Kinase
title_sort engineered fully human single-chain monoclonal antibodies to pim2 kinase
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8588357/
https://www.ncbi.nlm.nih.gov/pubmed/34770845
http://dx.doi.org/10.3390/molecules26216436
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