TY - JOUR
T1 - Peptide Stability Is Important but Not a General Requirement for Antimicrobial and Antibiofilm Activity in Vitro and in Vivo
AU - Decker, Aaron P.
AU - Su, Yajuan
AU - Mishra, Biswajit
AU - Verma, Atul
AU - Lushnikova, Tamara
AU - Xie, Jingwei
AU - Wang, Guangshun
N1 - Funding Information:
This study was supported by the NIH grant GM138552 to J.X. and G.W. We thank Dr. Thomas McDonald (UNMC) for allowing us to use his speed-vac to dry the samples and Dr. Jayaram Lakshmaiah Narayana for formatting .
Publisher Copyright:
© 2023 American Chemical Society. All rights reserved.
PY - 2023/1/2
Y1 - 2023/1/2
N2 - Peptide stability to proteases has been a major requirement for developing peptide therapeutics. This study investigates the effects of peptide stability on antimicrobial and antibiofilm activity under various conditions. For this purpose, two human cathelicidin-derived peptides differing in stability to proteases were utilized. While GF-17, a peptide derived from the major antimicrobial region of human LL-37, can be rapidly cleaved by proteases, the engineered peptide 17BIPHE2 is resistant to multiple proteases. In the standard antimicrobial susceptibility, killing kinetics, and membrane permeabilization assays conducted in vitro using planktonic bacteria, these two peptides displayed similar potency. The two peptides were also similarly active against methicillin-resistant Staphylococcus aureus (MRSA) USA300 prior to biofilm formation. However, 17BIPHE2 was superior to GF-17 in disrupting preformed biofilms probably due to both enhanced stability and slightly higher DNA binding capacity. In a wax moth model, 17BIPHE2 better protected insects from MRSA infection-caused death than GF-17, consistent with the slower degradation of 17BIPHE2 than GF-17. Here, peptide antimicrobial activity was found to be critical for in vivo efficacy. When incorporated in the nanofiber/microneedle delivery device, GF-17 and 17BIPHE2 displayed a similar effect in eliminating MRSA in murine chronic wounds, underscoring the advantage of nanofibers in protecting the peptide from degradation. Since nanoformulation can ease the requirement of peptide stability, it opens the door to a direct use of natural peptides or their cocktails for antimicrobial treatment, accelerating the search of effective antibiofilm peptides to treat chronic wounds.
AB - Peptide stability to proteases has been a major requirement for developing peptide therapeutics. This study investigates the effects of peptide stability on antimicrobial and antibiofilm activity under various conditions. For this purpose, two human cathelicidin-derived peptides differing in stability to proteases were utilized. While GF-17, a peptide derived from the major antimicrobial region of human LL-37, can be rapidly cleaved by proteases, the engineered peptide 17BIPHE2 is resistant to multiple proteases. In the standard antimicrobial susceptibility, killing kinetics, and membrane permeabilization assays conducted in vitro using planktonic bacteria, these two peptides displayed similar potency. The two peptides were also similarly active against methicillin-resistant Staphylococcus aureus (MRSA) USA300 prior to biofilm formation. However, 17BIPHE2 was superior to GF-17 in disrupting preformed biofilms probably due to both enhanced stability and slightly higher DNA binding capacity. In a wax moth model, 17BIPHE2 better protected insects from MRSA infection-caused death than GF-17, consistent with the slower degradation of 17BIPHE2 than GF-17. Here, peptide antimicrobial activity was found to be critical for in vivo efficacy. When incorporated in the nanofiber/microneedle delivery device, GF-17 and 17BIPHE2 displayed a similar effect in eliminating MRSA in murine chronic wounds, underscoring the advantage of nanofibers in protecting the peptide from degradation. Since nanoformulation can ease the requirement of peptide stability, it opens the door to a direct use of natural peptides or their cocktails for antimicrobial treatment, accelerating the search of effective antibiofilm peptides to treat chronic wounds.
KW - LL-37
KW - MRSA
KW - antimicrobial susceptibility
KW - biofilms
KW - peptide stability
KW - wound healing
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U2 - 10.1021/acs.molpharmaceut.2c00918
DO - 10.1021/acs.molpharmaceut.2c00918
M3 - Article
C2 - 36485036
AN - SCOPUS:85143852349
SN - 1543-8384
VL - 20
SP - 738
EP - 749
JO - Molecular Pharmaceutics
JF - Molecular Pharmaceutics
IS - 1
ER -