Novel Leech Antimicrobial Peptides, Hirunipins: Real‐Time 3D Monitoring of Antimicrobial and Antibiofilm Mechanisms Using Optical Diffraction Tomography

Abstract Antimicrobial peptides (AMPs) are promising agents for treating antibiotic‐resistant bacterial infections. Although discovering novel AMPs is crucial for combating multidrug‐resistant bacteria and biofilm‐related infections, their clinical potential relies on precise, real‐time evaluation o...

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Main Authors: S. Dinesh Kumar, Jeongwon Park, Naveen Kumar Radhakrishnan, Yam Prasad Aryal, Geon‐Hwi Jeong, In‐Hyeok Pyo, Byambasuren Ganbaatar, Chul Won Lee, Sungtae Yang, Younhee Shin, Sathiyamoorthy Subramaniyam, Yu‐jin Lim, Sung‐Hak Kim, Seongsoo Lee, Song Yub Shin, Sung‐Jin Cho
Format: Article
Language:English
Published: Wiley 2025-03-01
Series:Advanced Science
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Online Access:https://doi.org/10.1002/advs.202409803
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author S. Dinesh Kumar
Jeongwon Park
Naveen Kumar Radhakrishnan
Yam Prasad Aryal
Geon‐Hwi Jeong
In‐Hyeok Pyo
Byambasuren Ganbaatar
Chul Won Lee
Sungtae Yang
Younhee Shin
Sathiyamoorthy Subramaniyam
Yu‐jin Lim
Sung‐Hak Kim
Seongsoo Lee
Song Yub Shin
Sung‐Jin Cho
author_facet S. Dinesh Kumar
Jeongwon Park
Naveen Kumar Radhakrishnan
Yam Prasad Aryal
Geon‐Hwi Jeong
In‐Hyeok Pyo
Byambasuren Ganbaatar
Chul Won Lee
Sungtae Yang
Younhee Shin
Sathiyamoorthy Subramaniyam
Yu‐jin Lim
Sung‐Hak Kim
Seongsoo Lee
Song Yub Shin
Sung‐Jin Cho
author_sort S. Dinesh Kumar
collection DOAJ
description Abstract Antimicrobial peptides (AMPs) are promising agents for treating antibiotic‐resistant bacterial infections. Although discovering novel AMPs is crucial for combating multidrug‐resistant bacteria and biofilm‐related infections, their clinical potential relies on precise, real‐time evaluation of efficacy, toxicity, and mechanisms. Optical diffraction tomography (ODT), a label‐free imaging technology, enables real‐time visualization of bacterial morphological changes, membrane damage, and biofilm formation over time. Here, a computational analysis of the leech transcriptome using an advanced AI‐based peptide screening strategy with ODT to identify potential AMPs is employed. Among the 19 potential AMPs identified, hirunipin 2 demonstrates potent antibacterial activity, low mammalian cytotoxicity, and minimal hemolytic effects. It demonstrates efficacy comparable to melittin, resistance to physiological salts and human serum, and a low likelihood of inducing bacterial resistance. Microscopy and 3D‐ODT confirm its disruption of bacterial membranes and intracellular aggregation, leading to cell death. Notably, hirunipin 2 effectively inhibits biofilm formation, eradicates preformed biofilms, and synergizes with antibiotics against multidrug‐resistant Acinetobacter baumannii (MDRAB) by enhancing membrane permeability. Additionally, hirunipin 2 significantly suppresses pro‐inflammatory cytokine expression in LPS‐stimulated macrophages, highlighting its anti‐inflammatory properties. These findings highlight hirunipin 2 as a strong candidate for developing novel antibacterial, anti‐inflammatory, and antibiofilm therapies, particularly against multidrug‐resistant bacterial infections.
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spelling doaj-art-7ec308b2f52c4a6e8117e710a18cfde82025-08-20T02:35:40ZengWileyAdvanced Science2198-38442025-03-011210n/an/a10.1002/advs.202409803Novel Leech Antimicrobial Peptides, Hirunipins: Real‐Time 3D Monitoring of Antimicrobial and Antibiofilm Mechanisms Using Optical Diffraction TomographyS. Dinesh Kumar0Jeongwon Park1Naveen Kumar Radhakrishnan2Yam Prasad Aryal3Geon‐Hwi Jeong4In‐Hyeok Pyo5Byambasuren Ganbaatar6Chul Won Lee7Sungtae Yang8Younhee Shin9Sathiyamoorthy Subramaniyam10Yu‐jin Lim11Sung‐Hak Kim12Seongsoo Lee13Song Yub Shin14Sung‐Jin Cho15Department of Cellular & Molecular Medicine School of Medicine Chosun University Gwangju 61452 Republic of KoreaGwangju Center Korea Basic Science Institute (KBSI) Gwangju 61751 Republic of KoreaDepartment of Biomedical Sciences Graduate School Chosun University Gwangju 61452 Republic of KoreaDepartment of Biological Sciences and Biotechnology College of Natural Sciences Chungbuk National University Cheongju Chungbuk 28644 Republic of KoreaDepartment of Biological Sciences and Biotechnology College of Natural Sciences Chungbuk National University Cheongju Chungbuk 28644 Republic of KoreaDepartment of Biological Sciences and Biotechnology College of Natural Sciences Chungbuk National University Cheongju Chungbuk 28644 Republic of KoreaDepartment of Chemistry Chonnam National University Gwangju 61186 Republic of KoreaDepartment of Chemistry Chonnam National University Gwangju 61186 Republic of KoreaInstitute of Well‐Aging Medicare & CSU G‐LAMP Project Group Chosun University Gwangju 61452 Republic of KoreaResearch and Development Center Insilicogen Inc Yongin‐si Gyeonggi‐do 16954 Republic of KoreaResearch and Development Center Insilicogen Inc Yongin‐si Gyeonggi‐do 16954 Republic of KoreaResearch and Development Center Insilicogen Inc Yongin‐si Gyeonggi‐do 16954 Republic of KoreaDepartment of Animal Science Chonnam National University Gwangju 61186 South KoreaGwangju Center Korea Basic Science Institute (KBSI) Gwangju 61751 Republic of KoreaDepartment of Cellular & Molecular Medicine School of Medicine Chosun University Gwangju 61452 Republic of KoreaDepartment of Biological Sciences and Biotechnology College of Natural Sciences Chungbuk National University Cheongju Chungbuk 28644 Republic of KoreaAbstract Antimicrobial peptides (AMPs) are promising agents for treating antibiotic‐resistant bacterial infections. Although discovering novel AMPs is crucial for combating multidrug‐resistant bacteria and biofilm‐related infections, their clinical potential relies on precise, real‐time evaluation of efficacy, toxicity, and mechanisms. Optical diffraction tomography (ODT), a label‐free imaging technology, enables real‐time visualization of bacterial morphological changes, membrane damage, and biofilm formation over time. Here, a computational analysis of the leech transcriptome using an advanced AI‐based peptide screening strategy with ODT to identify potential AMPs is employed. Among the 19 potential AMPs identified, hirunipin 2 demonstrates potent antibacterial activity, low mammalian cytotoxicity, and minimal hemolytic effects. It demonstrates efficacy comparable to melittin, resistance to physiological salts and human serum, and a low likelihood of inducing bacterial resistance. Microscopy and 3D‐ODT confirm its disruption of bacterial membranes and intracellular aggregation, leading to cell death. Notably, hirunipin 2 effectively inhibits biofilm formation, eradicates preformed biofilms, and synergizes with antibiotics against multidrug‐resistant Acinetobacter baumannii (MDRAB) by enhancing membrane permeability. Additionally, hirunipin 2 significantly suppresses pro‐inflammatory cytokine expression in LPS‐stimulated macrophages, highlighting its anti‐inflammatory properties. These findings highlight hirunipin 2 as a strong candidate for developing novel antibacterial, anti‐inflammatory, and antibiofilm therapies, particularly against multidrug‐resistant bacterial infections.https://doi.org/10.1002/advs.202409803antibiofilmantimicrobial peptideHirunipinLeechODT technology
spellingShingle S. Dinesh Kumar
Jeongwon Park
Naveen Kumar Radhakrishnan
Yam Prasad Aryal
Geon‐Hwi Jeong
In‐Hyeok Pyo
Byambasuren Ganbaatar
Chul Won Lee
Sungtae Yang
Younhee Shin
Sathiyamoorthy Subramaniyam
Yu‐jin Lim
Sung‐Hak Kim
Seongsoo Lee
Song Yub Shin
Sung‐Jin Cho
Novel Leech Antimicrobial Peptides, Hirunipins: Real‐Time 3D Monitoring of Antimicrobial and Antibiofilm Mechanisms Using Optical Diffraction Tomography
Advanced Science
antibiofilm
antimicrobial peptide
Hirunipin
Leech
ODT technology
title Novel Leech Antimicrobial Peptides, Hirunipins: Real‐Time 3D Monitoring of Antimicrobial and Antibiofilm Mechanisms Using Optical Diffraction Tomography
title_full Novel Leech Antimicrobial Peptides, Hirunipins: Real‐Time 3D Monitoring of Antimicrobial and Antibiofilm Mechanisms Using Optical Diffraction Tomography
title_fullStr Novel Leech Antimicrobial Peptides, Hirunipins: Real‐Time 3D Monitoring of Antimicrobial and Antibiofilm Mechanisms Using Optical Diffraction Tomography
title_full_unstemmed Novel Leech Antimicrobial Peptides, Hirunipins: Real‐Time 3D Monitoring of Antimicrobial and Antibiofilm Mechanisms Using Optical Diffraction Tomography
title_short Novel Leech Antimicrobial Peptides, Hirunipins: Real‐Time 3D Monitoring of Antimicrobial and Antibiofilm Mechanisms Using Optical Diffraction Tomography
title_sort novel leech antimicrobial peptides hirunipins real time 3d monitoring of antimicrobial and antibiofilm mechanisms using optical diffraction tomography
topic antibiofilm
antimicrobial peptide
Hirunipin
Leech
ODT technology
url https://doi.org/10.1002/advs.202409803
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