Pub Date : 2024-06-24DOI: 10.1021/acsinfecdis.3c00728
Cheng Wang, Shaobo Wang, Xiangyu Ma, Xiaohong Yao, Kegang Zhan, Zai Wang, Di He, Wenting Zuo, Songling Han, Gaomei Zhao, Bin Cao, Jinghong Zhao, Xiuwu Bian, Junping Wang
SARS-CoV-2 infection starts from the association of its spike 1 (S1) subunit with sensitive cells. Vesicular endothelial cells and platelets are among the cell types that bind SARS-CoV-2, but the effectors that mediate viral attachment on the cell membrane have not been fully elucidated. Herein, we show that P-selectin (SELP), a biomarker for endothelial dysfunction and platelet activation, can facilitate the attachment of SARS-CoV-2 S1. Since we observe colocalization of SELP with S1 in the lung tissues of COVID-19 patients, we perform molecular biology experiments on human umbilical vein endothelial cells (HUVECs) to confirm the intermolecular interaction between SELP and S1. SELP overexpression increases S1 recruitment to HUVECs and enhances SARS-CoV-2 spike pseudovirion infection. The opposite results are determined after SELP downregulation. As S1 causes endothelial inflammatory responses in a dose-dependent manner, by activating the interleukin (IL)-17 signaling pathway, SELP-induced S1 recruitment may contribute to the development of a "cytokine storm" after viral infection. Furthermore, SELP also promotes the attachment of S1 to the platelet membrane. Employment of PSI-697, a small inhibitor of SELP, markedly decreases S1 adhesion to both HUVECs and platelets. In addition to the role of membrane SELP in facilitating S1 attachment, we also discover that soluble SELP is a prognostic factor for severe COVID-19 through a meta-analysis. In this study, we identify SELP as an adhesive site for the SARS-CoV-2 S1, thus providing a potential drug target for COVID-19 treatment.
{"title":"P-selectin Facilitates SARS-CoV-2 Spike 1 Subunit Attachment to Vesicular Endothelium and Platelets.","authors":"Cheng Wang, Shaobo Wang, Xiangyu Ma, Xiaohong Yao, Kegang Zhan, Zai Wang, Di He, Wenting Zuo, Songling Han, Gaomei Zhao, Bin Cao, Jinghong Zhao, Xiuwu Bian, Junping Wang","doi":"10.1021/acsinfecdis.3c00728","DOIUrl":"https://doi.org/10.1021/acsinfecdis.3c00728","url":null,"abstract":"<p><p>SARS-CoV-2 infection starts from the association of its spike 1 (S1) subunit with sensitive cells. Vesicular endothelial cells and platelets are among the cell types that bind SARS-CoV-2, but the effectors that mediate viral attachment on the cell membrane have not been fully elucidated. Herein, we show that P-selectin (SELP), a biomarker for endothelial dysfunction and platelet activation, can facilitate the attachment of SARS-CoV-2 S1. Since we observe colocalization of SELP with S1 in the lung tissues of COVID-19 patients, we perform molecular biology experiments on human umbilical vein endothelial cells (HUVECs) to confirm the intermolecular interaction between SELP and S1. SELP overexpression increases S1 recruitment to HUVECs and enhances SARS-CoV-2 spike pseudovirion infection. The opposite results are determined after SELP downregulation. As S1 causes endothelial inflammatory responses in a dose-dependent manner, by activating the interleukin (IL)-17 signaling pathway, SELP-induced S1 recruitment may contribute to the development of a \"cytokine storm\" after viral infection. Furthermore, SELP also promotes the attachment of S1 to the platelet membrane. Employment of PSI-697, a small inhibitor of SELP, markedly decreases S1 adhesion to both HUVECs and platelets. In addition to the role of membrane SELP in facilitating S1 attachment, we also discover that soluble SELP is a prognostic factor for severe COVID-19 through a meta-analysis. In this study, we identify SELP as an adhesive site for the SARS-CoV-2 S1, thus providing a potential drug target for COVID-19 treatment.</p>","PeriodicalId":17,"journal":{"name":"ACS Infectious Diseases","volume":null,"pages":null},"PeriodicalIF":4.0,"publicationDate":"2024-06-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141441699","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Fungi pose a global threat to humankind due to the increasing emergence of multi-drug-resistant fungi. There is a rising incidence of invasive fungal infections. Due to the structural complexity of fungal cell membranes, only a few classes of antifungal agents are effective and have been approved by the U.S. FDA. Hence, researchers globally are focusing on developing novel strategies to cure fungal infections. One of the potential strategies is the "Trojan horse" approach, which uses the siderophore-mediated iron acquisition (SIA) system to scavenge iron to deliver potent antifungal agents for therapeutics and diagnostics. These siderophore conjugates chelate to iron and are taken up through siderophore-iron transporters, which are overexpressed exclusively on microbes such as bacteria or fungi, but not mammalian cells. Our comprehensive review delves into recent advancements in the design of siderophore-conjugated antifungal agents to gain fungal cell entry. Notably, our focus extends to unraveling the intricate relationship between the structure of natural siderophores or siderophore-like molecules and the resulting antifungal activity. By exploring these design strategies, we aim to contribute to the ongoing discourse on combating drug-resistant fungal infections and advancing the landscape of antifungal theranostics.
由于耐多药真菌的不断出现,真菌对人类构成了全球性威胁。侵袭性真菌感染的发病率不断上升。由于真菌细胞膜结构复杂,只有几类抗真菌药物有效,并获得了美国 FDA 的批准。因此,全球研究人员都在致力于开发治疗真菌感染的新策略。其中一种潜在的策略是 "特洛伊木马 "方法,即利用嗜苷铁元素介导的铁获取(SIA)系统清除铁,从而将强效抗真菌剂用于治疗和诊断。这些嗜苷酸盐轭合物与铁螯合,并通过嗜苷酸盐-铁转运体被吸收,这些转运体只在细菌或真菌等微生物中过度表达,而不在哺乳动物细胞中表达。我们的综合综述深入探讨了设计嗜苷酸盐共轭抗真菌剂以进入真菌细胞的最新进展。值得注意的是,我们的研究重点延伸到揭示天然嗜苷酸盐或嗜苷酸盐类分子的结构与由此产生的抗真菌活性之间错综复杂的关系。通过探索这些设计策略,我们的目标是为正在进行的抗耐药性真菌感染和推进抗真菌治疗学的发展做出贡献。
{"title":"Siderophore-Conjugated Antifungals: A Strategy to Potentially Cure Fungal Infections.","authors":"Kalaiarasu Lakshminarayanan, Dhanashree Murugan, Janarthanan Venkatesan, Harashkumar Vasanthakumari Thirumalaiswamy, Charlène Gadais, Loganathan Rangasamy","doi":"10.1021/acsinfecdis.4c00046","DOIUrl":"https://doi.org/10.1021/acsinfecdis.4c00046","url":null,"abstract":"<p><p>Fungi pose a global threat to humankind due to the increasing emergence of multi-drug-resistant fungi. There is a rising incidence of invasive fungal infections. Due to the structural complexity of fungal cell membranes, only a few classes of antifungal agents are effective and have been approved by the U.S. FDA. Hence, researchers globally are focusing on developing novel strategies to cure fungal infections. One of the potential strategies is the \"Trojan horse\" approach, which uses the siderophore-mediated iron acquisition (SIA) system to scavenge iron to deliver potent antifungal agents for therapeutics and diagnostics. These siderophore conjugates chelate to iron and are taken up through siderophore-iron transporters, which are overexpressed exclusively on microbes such as bacteria or fungi, but not mammalian cells. Our comprehensive review delves into recent advancements in the design of siderophore-conjugated antifungal agents to gain fungal cell entry. Notably, our focus extends to unraveling the intricate relationship between the structure of natural siderophores or siderophore-like molecules and the resulting antifungal activity. By exploring these design strategies, we aim to contribute to the ongoing discourse on combating drug-resistant fungal infections and advancing the landscape of antifungal theranostics.</p>","PeriodicalId":17,"journal":{"name":"ACS Infectious Diseases","volume":null,"pages":null},"PeriodicalIF":4.0,"publicationDate":"2024-06-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141436381","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Endolysins, peptidoglycan hydrolases derived from bacteriophages (phages), are being developed as a promising alternative to conventional antibiotics. To obtain highly active endolysins, a diverse library of these endolysins is vital. We propose here microbial single-cell genome sequencing as an efficient tool to discover dozens of previously unknown endolysins, owing to its culture-independent sequencing method. As a proof of concept, we analyzed and recovered endolysin genes within prophage regions of Staphylococcus single-amplified genomes in human skin microbiome samples. We constructed a library of chimeric endolysins by shuffling domains of the natural endolysins and performed high-throughput screening against Staphylococcus aureus. One of the lead endolysins, bbst1027, exhibited desirable antimicrobial properties, such as rapid bactericidal activity, no detectable resistance development, and in vivo efficacy. We foresee that this endolysin discovery pipeline is in principle applicable to any bacterial target and boost the development of novel antimicrobial agents.
{"title":"Uncovering Endolysins against Methicillin-Resistant <i>Staphylococcus aureus</i> Using a Microbial Single-Cell Genome Database.","authors":"Takuya Yoda, Ayumi Matsuhashi, Ai Matsushita, Shohei Shibagaki, Yukie Sasakura, Kazuteru Aoki, Masahito Hosokawa, Soichiro Tsuda","doi":"10.1021/acsinfecdis.4c00039","DOIUrl":"https://doi.org/10.1021/acsinfecdis.4c00039","url":null,"abstract":"<p><p>Endolysins, peptidoglycan hydrolases derived from bacteriophages (phages), are being developed as a promising alternative to conventional antibiotics. To obtain highly active endolysins, a diverse library of these endolysins is vital. We propose here microbial single-cell genome sequencing as an efficient tool to discover dozens of previously unknown endolysins, owing to its culture-independent sequencing method. As a proof of concept, we analyzed and recovered endolysin genes within prophage regions of <i>Staphylococcus</i> single-amplified genomes in human skin microbiome samples. We constructed a library of chimeric endolysins by shuffling domains of the natural endolysins and performed high-throughput screening against <i>Staphylococcus aureus</i>. One of the lead endolysins, bbst1027, exhibited desirable antimicrobial properties, such as rapid bactericidal activity, no detectable resistance development, and in vivo efficacy. We foresee that this endolysin discovery pipeline is in principle applicable to any bacterial target and boost the development of novel antimicrobial agents.</p>","PeriodicalId":17,"journal":{"name":"ACS Infectious Diseases","volume":null,"pages":null},"PeriodicalIF":4.0,"publicationDate":"2024-06-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141436382","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-06-20DOI: 10.1021/acsinfecdis.4c00236
Pingli Qi, Xing Liu, Cui Li, Qiang Xu, Liting Hu, Huijin Duan, Guiqiu Zhao, Jing Lin
Fungal keratitis (FK) is a severe corneal condition caused by pathogenic fungi and is associated with the virulence of fungi and an excessive tissue inflammatory response. Progranulin (PGRN), functioning as a multifunctional growth factor, exerts a pivotal influence on the regulation of inflammation and autophagy. The aim of our research was to analyze the role of PGRN in Aspergillus fumigatus (A. fumigatus) keratitis. We found that PGRN expression was increased in the mouse cornea with A. fumigatus keratitis. In our experiments, corneas of mice with FK were treated with 100 ng/mL of PGRN. In vitro, RAW 264.7 cells were treated with 10 ng/mL of PGRN before A. fumigatus stimulation. The findings suggested that PGRN effectively alleviated corneal edema and decreased the expression of pro-inflammatory cytokines in mice. In stimulated RAW 264.7 cells, PGRN treatment suppressed the expression of pro-inflammatory cytokines IL-6 and TNF-α but promoted the expression of the anti-inflammatory cytokines IL-10. PGRN treatment significantly upregulated the expression of autophagy-related proteins LC3, Beclin-1, and Atg-7. 3-Methyladenine (3-MA, autophagy inhibitor) reversed the regulation of inflammatory cytokines by PGRN. In addition, our study demonstrated that PGRN also enhanced phagocytosis in RAW 264.7 cells. In summary, PGRN attenuated the inflammatory response of A. fumigatus keratitis by increasing autophagy and enhanced the phagocytic activity of RAW 264.7 cells. This showed that PGRN had a protective effect on A. fumigatus keratitis.
{"title":"Progranulin Protects against <i>Aspergillus fumigatus</i> Keratitis by Attenuating the Inflammatory Response through Enhancing Autophagy.","authors":"Pingli Qi, Xing Liu, Cui Li, Qiang Xu, Liting Hu, Huijin Duan, Guiqiu Zhao, Jing Lin","doi":"10.1021/acsinfecdis.4c00236","DOIUrl":"10.1021/acsinfecdis.4c00236","url":null,"abstract":"<p><p>Fungal keratitis (FK) is a severe corneal condition caused by pathogenic fungi and is associated with the virulence of fungi and an excessive tissue inflammatory response. Progranulin (PGRN), functioning as a multifunctional growth factor, exerts a pivotal influence on the regulation of inflammation and autophagy. The aim of our research was to analyze the role of PGRN in <i>Aspergillus fumigatus</i> (<i>A. fumigatus</i>) keratitis. We found that PGRN expression was increased in the mouse cornea with <i>A. fumigatus</i> keratitis. In our experiments, corneas of mice with FK were treated with 100 ng/mL of PGRN. In vitro, RAW 264.7 cells were treated with 10 ng/mL of PGRN before <i>A. fumigatus</i> stimulation. The findings suggested that PGRN effectively alleviated corneal edema and decreased the expression of pro-inflammatory cytokines in mice. In stimulated RAW 264.7 cells, PGRN treatment suppressed the expression of pro-inflammatory cytokines IL-6 and TNF-α but promoted the expression of the anti-inflammatory cytokines IL-10. PGRN treatment significantly upregulated the expression of autophagy-related proteins LC3, Beclin-1, and Atg-7. 3-Methyladenine (3-MA, autophagy inhibitor) reversed the regulation of inflammatory cytokines by PGRN. In addition, our study demonstrated that PGRN also enhanced phagocytosis in RAW 264.7 cells. In summary, PGRN attenuated the inflammatory response of <i>A. fumigatus</i> keratitis by increasing autophagy and enhanced the phagocytic activity of RAW 264.7 cells. This showed that PGRN had a protective effect on <i>A. fumigatus</i> keratitis.</p>","PeriodicalId":17,"journal":{"name":"ACS Infectious Diseases","volume":null,"pages":null},"PeriodicalIF":4.0,"publicationDate":"2024-06-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141430845","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Globally, there have been increasing reports of antimicrobial resistance in nontyphoidal Salmonella (NTS), which can develop into severe and potentially life-threatening diarrhea. This study focuses on the synergistic effects of DNA gyrase mutations and plasmid-mediated quinolone resistance (PMQR) genes, specifically qnrB19, on fluoroquinolone (FQ) resistance in Salmonella Typhimurium. By utilizing recombinant mutants, GyrAS83F and GyrAD87N, and QnrB19's, we discovered a significant increase in fluoroquinolones resistance when QnrB19 is present. Specifically, ciprofloxacin and moxifloxacin's inhibitory concentrations rose 10- and 8-fold, respectively. QnrB19 was found to enhance the resistance capacity of mutant DNA gyrases, leading to high-level FQ resistance. Additionally, we observed that the ratio of QnrB19 to DNA gyrase played a critical role in determining whether QnrB19 could protect DNA gyrase against FQ inhibition. Our findings underscore the critical need to understand these resistance mechanisms, as their coexistence enables bacteria to withstand therapeutic FQ levels, posing a significant challenge to treatment efficacy.
在全球范围内,有关非伤寒沙门氏菌(NTS)对抗菌药产生耐药性的报道越来越多,这种耐药性可发展成严重的腹泻,并可能危及生命。本研究的重点是 DNA 回旋酶突变和质粒介导的喹诺酮类药物耐药性(PMQR)基因(特别是 qnrB19)对伤寒沙门氏菌氟喹诺酮类药物(FQ)耐药性的协同作用。通过利用重组突变体 GyrAS83F 和 GyrAD87N 以及 QnrB19,我们发现当 QnrB19 存在时,对氟喹诺酮类药物的耐药性会显著增加。具体来说,环丙沙星和莫西沙星的抑制浓度分别上升了 10 倍和 8 倍。研究发现,QnrB19 能增强突变 DNA 回旋酶的抗药性,从而导致高水平的 FQ 抗药性。此外,我们还观察到,QnrB19 与 DNA 回旋酶的比例在决定 QnrB19 能否保护 DNA 回旋酶免受 FQ 抑制方面起着关键作用。我们的发现强调了了解这些抗性机制的迫切需要,因为它们的共存使细菌能够承受治疗性 FQ 水平,对治疗效果构成了重大挑战。
{"title":"Interplay between Amino Acid Substitution in GyrA and QnrB19: Elevating Fluoroquinolone Resistance in <i>Salmonella</i> Typhimurium.","authors":"Pondpan Suwanthada, Siriporn Kongsoi, Sasini Jayaweera, Mwangala Lonah Akapelwa, Jeewan Thapa, Chie Nakajima, Yasuhiko Suzuki","doi":"10.1021/acsinfecdis.4c00150","DOIUrl":"https://doi.org/10.1021/acsinfecdis.4c00150","url":null,"abstract":"<p><p>Globally, there have been increasing reports of antimicrobial resistance in nontyphoidal <i>Salmonella</i> (NTS), which can develop into severe and potentially life-threatening diarrhea. This study focuses on the synergistic effects of DNA gyrase mutations and plasmid-mediated quinolone resistance (PMQR) genes, specifically <i>qnrB19</i>, on fluoroquinolone (FQ) resistance in <i>Salmonella</i> Typhimurium. By utilizing recombinant mutants, GyrA<sup>S83F</sup> and GyrA<sup>D87N</sup>, and QnrB19's, we discovered a significant increase in fluoroquinolones resistance when QnrB19 is present. Specifically, ciprofloxacin and moxifloxacin's inhibitory concentrations rose 10- and 8-fold, respectively. QnrB19 was found to enhance the resistance capacity of mutant DNA gyrases, leading to high-level FQ resistance. Additionally, we observed that the ratio of QnrB19 to DNA gyrase played a critical role in determining whether QnrB19 could protect DNA gyrase against FQ inhibition. Our findings underscore the critical need to understand these resistance mechanisms, as their coexistence enables bacteria to withstand therapeutic FQ levels, posing a significant challenge to treatment efficacy.</p>","PeriodicalId":17,"journal":{"name":"ACS Infectious Diseases","volume":null,"pages":null},"PeriodicalIF":5.3,"publicationDate":"2024-06-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141425611","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-06-17DOI: 10.1021/acsinfecdis.4c00078
Joshua Grabeck, Jacob Mayer, Axel Miltz, Michele Casoria, Michael Quagliata, Denise Meinberger, Andreas R Klatt, Isabelle Wielert, Berenike Maier, Anna Maria Papini, Ines Neundorf
There are still no linear antimicrobial peptides (AMPs) available as a treatment option against bacterial infections. This is caused by several drawbacks that come with AMPs such as limited proteolytic stability and low selectivity against human cells. In this work, we screened a small library of rationally designed new peptides based on the cell-penetrating peptide sC18* toward their antimicrobial activity. We identified several effective novel AMPs and chose one out of this group to further increase its potency. Therefore, we introduced a triazole bridge at different positions to provide a preformed helical structure, assuming that this modification would improve (i) proteolytic stability and (ii) membrane activity. Indeed, placing the triazole bridge within the hydrophilic part of the linear analogue highly increased membrane activity as well as stability against enzymatic digestion. The new peptides, 8A and 8B, demonstrated high activity against several bacterial species tested including pathogenic N. gonorrhoeae and methicillin-resistant S. aureus. Since they exhibited significantly good tolerability against human fibroblast and blood cells, these novel peptides offer true alternatives for future clinical applications and are worth studying in more detail.
{"title":"Triazole-Bridged Peptides with Enhanced Antimicrobial Activity and Potency against Pathogenic Bacteria.","authors":"Joshua Grabeck, Jacob Mayer, Axel Miltz, Michele Casoria, Michael Quagliata, Denise Meinberger, Andreas R Klatt, Isabelle Wielert, Berenike Maier, Anna Maria Papini, Ines Neundorf","doi":"10.1021/acsinfecdis.4c00078","DOIUrl":"https://doi.org/10.1021/acsinfecdis.4c00078","url":null,"abstract":"<p><p>There are still no linear antimicrobial peptides (AMPs) available as a treatment option against bacterial infections. This is caused by several drawbacks that come with AMPs such as limited proteolytic stability and low selectivity against human cells. In this work, we screened a small library of rationally designed new peptides based on the cell-penetrating peptide sC18* toward their antimicrobial activity. We identified several effective novel AMPs and chose one out of this group to further increase its potency. Therefore, we introduced a triazole bridge at different positions to provide a preformed helical structure, assuming that this modification would improve (i) proteolytic stability and (ii) membrane activity. Indeed, placing the triazole bridge within the hydrophilic part of the linear analogue highly increased membrane activity as well as stability against enzymatic digestion. The new peptides, 8A and 8B, demonstrated high activity against several bacterial species tested including pathogenic <i>N. gonorrhoeae</i> and methicillin-resistant <i>S. aureus</i>. Since they exhibited significantly good tolerability against human fibroblast and blood cells, these novel peptides offer true alternatives for future clinical applications and are worth studying in more detail.</p>","PeriodicalId":17,"journal":{"name":"ACS Infectious Diseases","volume":null,"pages":null},"PeriodicalIF":5.3,"publicationDate":"2024-06-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141416569","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-06-14DOI: 10.1021/acsinfecdis.4c00224
Michael J Rudolph, Anastasiia M Tsymbal, Arkajyoti Dutta, Simon A Davis, Benjamin Algava, Jacques Y Roberge, Nilgun E Tumer, Xiao-Ping Li
Shiga toxins are the main virulence factors of Shiga toxin producing E. coli (STEC) and S. dysenteriae. There is no effective therapy to counter the disease caused by these toxins. The A1 subunits of Shiga toxins bind the C-termini of ribosomal P-stalk proteins to depurinate the sarcin/ricin loop. The ribosome binding site of Shiga toxin 2 has not been targeted by small molecules. We screened a fragment library against the A1 subunit of Shiga toxin 2 (Stx2A1) and identified a fragment, BTB13086, which bound at the ribosome binding site and mimicked the binding mode of the P-stalk proteins. We synthesized analogs of BTB13086 and identified a series of molecules with similar affinity and inhibitory activity. These are the first compounds that bind at the ribosome binding site of Stx2A1 and inhibit activity. These compounds hold great promise for further inhibitor development against STEC infection.
{"title":"Fragment Screening to Identify Inhibitors Targeting Ribosome Binding of Shiga Toxin 2.","authors":"Michael J Rudolph, Anastasiia M Tsymbal, Arkajyoti Dutta, Simon A Davis, Benjamin Algava, Jacques Y Roberge, Nilgun E Tumer, Xiao-Ping Li","doi":"10.1021/acsinfecdis.4c00224","DOIUrl":"https://doi.org/10.1021/acsinfecdis.4c00224","url":null,"abstract":"<p><p>Shiga toxins are the main virulence factors of Shiga toxin producing <i>E. coli</i> (STEC) and <i>S. dysenteriae</i>. There is no effective therapy to counter the disease caused by these toxins. The A1 subunits of Shiga toxins bind the C-termini of ribosomal P-stalk proteins to depurinate the sarcin/ricin loop. The ribosome binding site of Shiga toxin 2 has not been targeted by small molecules. We screened a fragment library against the A1 subunit of Shiga toxin 2 (Stx2A1) and identified a fragment, <b>BTB13086</b>, which bound at the ribosome binding site and mimicked the binding mode of the P-stalk proteins. We synthesized analogs of <b>BTB13086</b> and identified a series of molecules with similar affinity and inhibitory activity. These are the first compounds that bind at the ribosome binding site of Stx2A1 and inhibit activity. These compounds hold great promise for further inhibitor development against STEC infection.</p>","PeriodicalId":17,"journal":{"name":"ACS Infectious Diseases","volume":null,"pages":null},"PeriodicalIF":5.3,"publicationDate":"2024-06-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141315939","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-06-14Epub Date: 2024-05-17DOI: 10.1021/acsinfecdis.4c00062
Javier Chao-Pellicer, Iñigo Arberas-Jiménez, Ines Sifaoui, José E Piñero, Jacob Lorenzo-Morales
Primary amoebic meningoencephalitis (PAM) is a rare and fulminant neurodegenerative disease caused by the free-living amoeba Naegleria fowleri. Currently, there is a lack of standardized protocols for therapeutic action. In response to the critical need for effective therapeutic agents, we explored the Global Health Priority Box, a collection of 240 compounds provided by the Medicines for Malaria Venture (MMV). From this pool, flucofuron emerged as a promising candidate, exhibiting high efficacy against trophozoites of both N. fowleri strains (ATCC 30808 IC50 : 2.58 ± 0.64 μM and ATCC 30215 IC50: 2.47 ± 0.38 μM), being even active against the resistant cyst stage (IC50: 0.88 ± 0.07 μM). Moreover, flucofuron induced diverse metabolic events that suggest the triggering of apoptotic cell death. This study highlights the potential of repurposing medications for treating challenging diseases, such as PAM.
{"title":"Flucofuron as a Promising Therapeutic Agent against Brain-Eating Amoeba.","authors":"Javier Chao-Pellicer, Iñigo Arberas-Jiménez, Ines Sifaoui, José E Piñero, Jacob Lorenzo-Morales","doi":"10.1021/acsinfecdis.4c00062","DOIUrl":"10.1021/acsinfecdis.4c00062","url":null,"abstract":"<p><p>Primary amoebic meningoencephalitis (PAM) is a rare and fulminant neurodegenerative disease caused by the free-living amoeba <i>Naegleria fowleri</i>. Currently, there is a lack of standardized protocols for therapeutic action. In response to the critical need for effective therapeutic agents, we explored the Global Health Priority Box, a collection of 240 compounds provided by the Medicines for Malaria Venture (MMV). From this pool, flucofuron emerged as a promising candidate, exhibiting high efficacy against trophozoites of both <i>N. fowleri</i> strains (ATCC 30808 IC<sub>50</sub> : 2.58 ± 0.64 μM and ATCC 30215 IC<sub>50</sub>: 2.47 ± 0.38 μM), being even active against the resistant cyst stage (IC<sub>50</sub>: 0.88 ± 0.07 μM). Moreover, flucofuron induced diverse metabolic events that suggest the triggering of apoptotic cell death. This study highlights the potential of repurposing medications for treating challenging diseases, such as PAM.</p>","PeriodicalId":17,"journal":{"name":"ACS Infectious Diseases","volume":null,"pages":null},"PeriodicalIF":5.3,"publicationDate":"2024-06-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11184546/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140961498","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-06-14DOI: 10.1021/acsinfecdis.4c00116
Shannon Whiles, Quanzheng Zhang, Zach Chamberlain, Manish K Singh, Shaun Steele, Linda Zheng, Kristin Rosche, Weigang Huang, Huanyao Gao, Qisheng Zhang, Thomas Kawula
Francisella tularensis is a Gram-negative facultative intracellular bacterial pathogen that is classified by the Centers for Disease Control and Prevention as a Tier 1 Select Agent. F. tularensis infection causes the disease tularemia, also known as rabbit fever. Treatment of tularemia is limited to few effective antibiotics which are associated with high relapse rates, toxicity, and potential emergence of antibiotic-resistant strains. Consequently, new therapeutic options for tularemia are needed. Through screening a focused chemical library and subsequent structure-activity relationship studies, we have discovered a new and potent inhibitor of intracellular growth of Francisella tularensis, D8-03. Importantly, D8-03 effectively reduces bacterial burden in mice infected with F. tularensis. Preliminary mechanistic investigations suggest that D8-03 works through a potentially novel host-dependent mechanism and serves as a promising lead compound for further development.
{"title":"Discovery of D8-03 as an Inhibitor of Intracellular Growth of <i>Francisella tularensis</i>.","authors":"Shannon Whiles, Quanzheng Zhang, Zach Chamberlain, Manish K Singh, Shaun Steele, Linda Zheng, Kristin Rosche, Weigang Huang, Huanyao Gao, Qisheng Zhang, Thomas Kawula","doi":"10.1021/acsinfecdis.4c00116","DOIUrl":"https://doi.org/10.1021/acsinfecdis.4c00116","url":null,"abstract":"<p><p><i>Francisella tularensis</i> is a Gram-negative facultative intracellular bacterial pathogen that is classified by the Centers for Disease Control and Prevention as a Tier 1 Select Agent. <i>F. tularensis</i> infection causes the disease tularemia, also known as rabbit fever. Treatment of tularemia is limited to few effective antibiotics which are associated with high relapse rates, toxicity, and potential emergence of antibiotic-resistant strains. Consequently, new therapeutic options for tularemia are needed. Through screening a focused chemical library and subsequent structure-activity relationship studies, we have discovered a new and potent inhibitor of intracellular growth of <i>Francisella tularensis</i>, D8-03. Importantly, D8-03 effectively reduces bacterial burden in mice infected with <i>F. tularensis</i>. Preliminary mechanistic investigations suggest that D8-03 works through a potentially novel host-dependent mechanism and serves as a promising lead compound for further development.</p>","PeriodicalId":17,"journal":{"name":"ACS Infectious Diseases","volume":null,"pages":null},"PeriodicalIF":5.3,"publicationDate":"2024-06-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141320080","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-06-14Epub Date: 2024-05-16DOI: 10.1021/acsinfecdis.4c00009
Mirtha E Aguado, Sandra Carvalho, Mario E Valdés-Tresanco, De Lin, Norma Padilla-Mejia, Victoriano Corpas-Lopez, Martina Tesařová, Julius Lukeš, David Gray, Jorge González-Bacerio, Susan Wyllie, Mark C Field
Leishmaniasis is a neglected tropical disease; there is currently no vaccine and treatment is reliant upon a handful of drugs suffering from multiple issues including toxicity and resistance. There is a critical need for development of new fit-for-purpose therapeutics, with reduced toxicity and targeting new mechanisms to overcome resistance. One enzyme meriting investigation as a potential drug target in Leishmania is M17 leucyl-aminopeptidase (LAP). Here, we aimed to chemically validate LAP as a drug target in L. major through identification of potent and selective inhibitors. Using RapidFire mass spectrometry, the compounds DDD00057570 and DDD00097924 were identified as selective inhibitors of recombinant Leishmania major LAP activity. Both compounds inhibited in vitro growth of L. major and L. donovani intracellular amastigotes, and overexpression of LmLAP in L. major led to reduced susceptibility to DDD00057570 and DDD00097924, suggesting that these compounds specifically target LmLAP. Thermal proteome profiling revealed that these inhibitors thermally stabilized two M17 LAPs, indicating that these compounds selectively bind to enzymes of this class. Additionally, the selectivity of the inhibitors to act on LmLAP and not against the human ortholog was demonstrated, despite the high sequence similarities LAPs of this family share. Collectively, these data confirm LmLAP as a promising therapeutic target for Leishmania spp. that can be selectively inhibited by drug-like small molecules.
利什曼病是一种被忽视的热带疾病;目前还没有疫苗,治疗只能依靠少数几种存在毒性和抗药性等多种问题的药物。目前亟需开发新的适合目的的治疗药物,以降低毒性和克服抗药性的新机制为目标。M17亮氨酰肽酶(LAP)是一种值得研究的利什曼原虫潜在药物靶标酶。在此,我们旨在通过鉴定强效的选择性抑制剂,以化学方法验证 LAP 作为大利什曼病菌的药物靶点。利用 RapidFire 质谱技术,我们鉴定出 DDD00057570 和 DDD00097924 化合物是重组大利什曼原虫 LAP 活性的选择性抑制剂。这两种化合物都能抑制大利什曼病菌和唐诺瓦尼氏利什曼病菌胞内母细胞的体外生长,大利什曼病菌过表达 LmLAP 会降低对 DDD00057570 和 DDD00097924 的敏感性,这表明这两种化合物能特异性地靶向 LmLAP。热蛋白质组分析表明,这些抑制剂能使两种 M17 LAP 热稳定,表明这些化合物能选择性地与该类酶结合。此外,尽管 LmLAP 家族的序列高度相似,但这些抑制剂对 LmLAP 而非人类同源物具有选择性作用。总之,这些数据证实 LmLAP 是利什曼原虫的一个很有希望的治疗靶点,可被类药物小分子选择性地抑制。
{"title":"Identification and Validation of Compounds Targeting <i>Leishmania major</i> Leucyl-Aminopeptidase M17.","authors":"Mirtha E Aguado, Sandra Carvalho, Mario E Valdés-Tresanco, De Lin, Norma Padilla-Mejia, Victoriano Corpas-Lopez, Martina Tesařová, Julius Lukeš, David Gray, Jorge González-Bacerio, Susan Wyllie, Mark C Field","doi":"10.1021/acsinfecdis.4c00009","DOIUrl":"10.1021/acsinfecdis.4c00009","url":null,"abstract":"<p><p>Leishmaniasis is a neglected tropical disease; there is currently no vaccine and treatment is reliant upon a handful of drugs suffering from multiple issues including toxicity and resistance. There is a critical need for development of new fit-for-purpose therapeutics, with reduced toxicity and targeting new mechanisms to overcome resistance. One enzyme meriting investigation as a potential drug target in <i>Leishmania</i> is M17 leucyl-aminopeptidase (LAP). Here, we aimed to chemically validate LAP as a drug target in <i>L. major</i> through identification of potent and selective inhibitors. Using RapidFire mass spectrometry, the compounds DDD00057570 and DDD00097924 were identified as selective inhibitors of recombinant <i>Leishmania major</i> LAP activity. Both compounds inhibited <i>in vitro</i> growth of <i>L. major</i> and <i>L. donovani</i> intracellular amastigotes, and overexpression of <i>Lm</i>LAP in <i>L. major</i> led to reduced susceptibility to DDD00057570 and DDD00097924, suggesting that these compounds specifically target <i>Lm</i>LAP. Thermal proteome profiling revealed that these inhibitors thermally stabilized two M17 LAPs, indicating that these compounds selectively bind to enzymes of this class. Additionally, the selectivity of the inhibitors to act on <i>Lm</i>LAP and not against the human ortholog was demonstrated, despite the high sequence similarities LAPs of this family share. Collectively, these data confirm <i>Lm</i>LAP as a promising therapeutic target for <i>Leishmania</i> spp. that can be selectively inhibited by drug-like small molecules.</p>","PeriodicalId":17,"journal":{"name":"ACS Infectious Diseases","volume":null,"pages":null},"PeriodicalIF":5.3,"publicationDate":"2024-06-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11184559/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140961499","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}