Pub Date : 2024-06-04DOI: 10.1021/acsinfecdis.4c00374
Zaiqiao Sun, Boxiao He, Zhifeng Yang, Yi Huang, Zhaoyu Duan, Chengyi Yu, Zhaokui Dan, Chonil Paek, Peng Chen, Jin Zhou, Jun Lei, Feng Wang*, Bing Liu* and Lei Yin*,
Tuberculosis (TB) is a prevalent and severe infectious disease that poses a significant threat to human health. However, it is frequently disregarded as there are not enough quick and accurate ways to diagnose tuberculosis. Here, we develop a strategy for tuberculosis detection to address the challenges, including an experimental strategy, namely, Double Adapter Directional Capture sequencing (DADCSeq), an easily operated and low-cost whole transcriptome sequencing method, and a computational method to identify hub differentially expressed genes as well as the diagnosis of TB based on whole transcriptome data using DADCSeq on peripheral blood mononuclear cells (PBMCs) from active TB and latent TB or healthy control. Applying our approach to create a robust and stable TB multi-mRNA risk probability model (TBMMRP) that can accurately distinguish active and latent TB patients, including active TB and healthy controls in clinical cohorts, this diagnostic biomarker was successfully validated by several independent cross-platform cohorts with favorable performance in differentiating active TB from latent TB or active TB from healthy controls and further demonstrated superior or similar diagnostic accuracy compared to previous diagnostic markers. Overall, we develop a low-cost and effective strategy for tuberculosis diagnosis; as the clinical cohort increases, we can expand to different disease kinds and learn new features through our disease diagnosis strategy.
{"title":"Cost-Effective Whole Transcriptome Sequencing Landscape and Diagnostic Potential Biomarkers in Active Tuberculosis","authors":"Zaiqiao Sun, Boxiao He, Zhifeng Yang, Yi Huang, Zhaoyu Duan, Chengyi Yu, Zhaokui Dan, Chonil Paek, Peng Chen, Jin Zhou, Jun Lei, Feng Wang*, Bing Liu* and Lei Yin*, ","doi":"10.1021/acsinfecdis.4c00374","DOIUrl":"10.1021/acsinfecdis.4c00374","url":null,"abstract":"<p >Tuberculosis (TB) is a prevalent and severe infectious disease that poses a significant threat to human health. However, it is frequently disregarded as there are not enough quick and accurate ways to diagnose tuberculosis. Here, we develop a strategy for tuberculosis detection to address the challenges, including an experimental strategy, namely, Double Adapter Directional Capture sequencing (DADCSeq), an easily operated and low-cost whole transcriptome sequencing method, and a computational method to identify hub differentially expressed genes as well as the diagnosis of TB based on whole transcriptome data using DADCSeq on peripheral blood mononuclear cells (PBMCs) from active TB and latent TB or healthy control. Applying our approach to create a robust and stable TB multi-mRNA risk probability model (TBMMRP) that can accurately distinguish active and latent TB patients, including active TB and healthy controls in clinical cohorts, this diagnostic biomarker was successfully validated by several independent cross-platform cohorts with favorable performance in differentiating active TB from latent TB or active TB from healthy controls and further demonstrated superior or similar diagnostic accuracy compared to previous diagnostic markers. Overall, we develop a low-cost and effective strategy for tuberculosis diagnosis; as the clinical cohort increases, we can expand to different disease kinds and learn new features through our disease diagnosis strategy.</p>","PeriodicalId":17,"journal":{"name":"ACS Infectious Diseases","volume":null,"pages":null},"PeriodicalIF":5.3,"publicationDate":"2024-06-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141236457","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}
Methicillin-resistant Staphylococcus aureus (MRSA) has become a serious threat to human public health and global economic development, and there is an urgent need to develop new antimicrobial agents. Flavonoids are the largest group of plant secondary metabolites, and the anti-S. aureus and anti-MRSA activities of flavonoids have now been widely reported. The aim of this Review is to describe plant-derived flavonoid active ingredients and their effects and mechanisms of inhibitory activity against MRSA in order to provide insights for screening novel antimicrobial agents. Here, 85 plant-derived flavonoids (14 flavones, 21 flavonols, 26 flavanones, 9 isoflavones, 12 chalcones, and 3 other classes) with anti-MRSA activity are reviewed. Among these flavonoids, flavones and isoflavones generally showed the most significant anti-MRSA activity (MICs: 1-8 μg/mL). The results of the present Review display that most of the flavonoids with excellent anti-MRSA activity were derived from Morus alba L. and Paulownia tomentosa (Thunb.) Steud. The antibacterial mechanism of flavonoids against MRSA is mainly achieved by disruption of membrane structures, inhibition of efflux pumps, and inhibition of β-lactamases and bacterial virulence factors. We hope this Review can provide insights into the development of novel antimicrobials based on natural products for treating MRSA infections.
{"title":"Plant Flavonoids with Antimicrobial Activity against Methicillin-Resistant <i>Staphylococcus aureus</i> (MRSA).","authors":"Shengnan Xu, Ayue Kang, Yue Tian, Xinhui Li, Shangshang Qin, Ruige Yang, Yong Guo","doi":"10.1021/acsinfecdis.4c00292","DOIUrl":"https://doi.org/10.1021/acsinfecdis.4c00292","url":null,"abstract":"<p><p>Methicillin-resistant <i>Staphylococcus aureus</i> (MRSA) has become a serious threat to human public health and global economic development, and there is an urgent need to develop new antimicrobial agents. Flavonoids are the largest group of plant secondary metabolites, and the anti-<i>S. aureus</i> and anti-MRSA activities of flavonoids have now been widely reported. The aim of this Review is to describe plant-derived flavonoid active ingredients and their effects and mechanisms of inhibitory activity against MRSA in order to provide insights for screening novel antimicrobial agents. Here, 85 plant-derived flavonoids (14 flavones, 21 flavonols, 26 flavanones, 9 isoflavones, 12 chalcones, and 3 other classes) with anti-MRSA activity are reviewed. Among these flavonoids, flavones and isoflavones generally showed the most significant anti-MRSA activity (MICs: 1-8 μg/mL). The results of the present Review display that most of the flavonoids with excellent anti-MRSA activity were derived from <i>Morus alba</i> L. and <i>Paulownia tomentosa</i> (Thunb.) Steud. The antibacterial mechanism of flavonoids against MRSA is mainly achieved by disruption of membrane structures, inhibition of efflux pumps, and inhibition of β-lactamases and bacterial virulence factors. We hope this Review can provide insights into the development of novel antimicrobials based on natural products for treating MRSA infections.</p>","PeriodicalId":17,"journal":{"name":"ACS Infectious Diseases","volume":null,"pages":null},"PeriodicalIF":5.3,"publicationDate":"2024-06-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141246874","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}
Leishmaniasis, one of the most overlooked tropical diseases, is a life-threatening illness caused by the parasite Leishmania donovani that is prevalent in underdeveloped nations. Over 350 million individuals in more than 90 different nations worldwide are at risk of contracting the disease, which has a current fatality rate of 50 000 mortalities each year. The administration of liposomal Amp B, pentavalent antimonials, and miltefosine are still considered integral components of the chemotherapy regimen. Antileishmanial medications fail to treat leishmaniasis because of their numerous drawbacks. These include inadequate effectiveness, toxicity, undesired side effects, drug resistance, treatment duration, and cost. Consequently, there is a need to overcome the limitations of conventional therapeutics. Nanotechnology has demonstrated promising outcomes in addressing these issues because of its small size and distinctive characteristics, such as enhanced bioavailability, lower toxicity, biodegradability, and targeted drug delivery. This review is an effort to highlight the recent progress in various nanodrug delivery systems (nDDSs) over the past five years for treating leishmaniasis. Although the preclinical outcomes of nDDSs have shown promising treatment for leishmaniasis, further research is needed for their clinical translation. Advancement in three primary priority domains─molecular diagnostics, clinical investigation, and knowledge dissemination and standardization─is imperative to propel the leishmaniasis field toward translational outcomes.
{"title":"Revolutionizing Leishmaniasis Treatment with Cutting Edge Drug Delivery Systems and Nanovaccines: An Updated Review","authors":"Srushti Tambe, Sagnik Nag, Shivani R. Pandya, Rohit Kumar, Kalpana Balakrishnan, Ranvijay Kumar, Sandeep Kumar, Purnima Amin and Piyush Kumar Gupta*, ","doi":"10.1021/acsinfecdis.4c00010","DOIUrl":"10.1021/acsinfecdis.4c00010","url":null,"abstract":"<p >Leishmaniasis, one of the most overlooked tropical diseases, is a life-threatening illness caused by the parasite <i>Leishmania donovani</i> that is prevalent in underdeveloped nations. Over 350 million individuals in more than 90 different nations worldwide are at risk of contracting the disease, which has a current fatality rate of 50 000 mortalities each year. The administration of liposomal Amp B, pentavalent antimonials, and miltefosine are still considered integral components of the chemotherapy regimen. Antileishmanial medications fail to treat leishmaniasis because of their numerous drawbacks. These include inadequate effectiveness, toxicity, undesired side effects, drug resistance, treatment duration, and cost. Consequently, there is a need to overcome the limitations of conventional therapeutics. Nanotechnology has demonstrated promising outcomes in addressing these issues because of its small size and distinctive characteristics, such as enhanced bioavailability, lower toxicity, biodegradability, and targeted drug delivery. This review is an effort to highlight the recent progress in various nanodrug delivery systems (nDDSs) over the past five years for treating leishmaniasis. Although the preclinical outcomes of nDDSs have shown promising treatment for leishmaniasis, further research is needed for their clinical translation. Advancement in three primary priority domains─molecular diagnostics, clinical investigation, and knowledge dissemination and standardization─is imperative to propel the leishmaniasis field toward translational outcomes.</p>","PeriodicalId":17,"journal":{"name":"ACS Infectious Diseases","volume":null,"pages":null},"PeriodicalIF":5.3,"publicationDate":"2024-06-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141198549","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 : 2024-06-03DOI: 10.1021/acsinfecdis.3c00562
Preeti Chand, Kritika Narula, Radhakrishnan VS, Shubham Sharma, Sangeeta Kumari, Neelima Mondal, Surinder P. Singh, Prashant Mishra and Tulika Prasad*,
Rationally designed multitargeted drugs, known as network therapeutics/multimodal drugs, have emerged as versatile therapeutic solutions to combat drug-resistant microbes. Here, we report novel mechanistic insights into cellular and molecular targets of ZnO quantum dots (QDs) against Candida albicans, a representative of fungal pathogens. Stable, monodispersed 4–6 nm ZnO QDs were synthesized using a wet chemical route, which exhibited dose-dependent inhibition on the growth dynamics of Candida. Treatment with 200 μg/mL ZnO QDs revealed an aberrant morphology and a disrupted cellular ultrastructure in electron microscopy and led to a 23% reduction in ergosterol content and a 53% increase in intracellular reactive oxygen species. Significant increase in steady-state fluorescence polarization and fluorescence lifetime decay of membrane probe 1,6-diphenyl-1,3,5-hexatriene (DPH) in treated cells, respectively, implied reduction in membrane fluidity and enhanced microviscosity. The observed reduction in passive diffusion of fluorescent Rhodamine 6G across the membrane validated the intricate relationship between ergosterol, membrane fluidity, and microviscosity. An inverse relationship existing between ergosterol biosynthetic genes, ERG11 and ERG3 in treated cells, related well with displayed higher susceptibilities. Furthermore, treated cells exhibited impaired functionality and downregulation of ABC drug efflux pumps. Multiple cellular targets of ZnO QDs in Candida were validated by in silico molecular docking. Thus, targeting ERG11, ERG3, and ABC drug efflux pumps might emerge as a versatile, nano-ZnO-based strategy in fungal therapeutics to address the challenges of drug resistance.
{"title":"Mechanistic Insights into Cellular and Molecular Targets of Zinc Oxide Quantum Dots (ZnO QDs) in Fungal Pathogen, Candida albicans: One Drug Multi-Targeted Therapeutic Approach","authors":"Preeti Chand, Kritika Narula, Radhakrishnan VS, Shubham Sharma, Sangeeta Kumari, Neelima Mondal, Surinder P. Singh, Prashant Mishra and Tulika Prasad*, ","doi":"10.1021/acsinfecdis.3c00562","DOIUrl":"10.1021/acsinfecdis.3c00562","url":null,"abstract":"<p >Rationally designed multitargeted drugs, known as network therapeutics/multimodal drugs, have emerged as versatile therapeutic solutions to combat drug-resistant microbes. Here, we report novel mechanistic insights into cellular and molecular targets of ZnO quantum dots (QDs) against <i>Candida albicans</i>, a representative of fungal pathogens. Stable, monodispersed 4–6 nm ZnO QDs were synthesized using a wet chemical route, which exhibited dose-dependent inhibition on the growth dynamics of <i>Candida</i>. Treatment with 200 μg/mL ZnO QDs revealed an aberrant morphology and a disrupted cellular ultrastructure in electron microscopy and led to a 23% reduction in ergosterol content and a 53% increase in intracellular reactive oxygen species. Significant increase in steady-state fluorescence polarization and fluorescence lifetime decay of membrane probe 1,6-diphenyl-1,3,5-hexatriene (DPH) in treated cells, respectively, implied reduction in membrane fluidity and enhanced microviscosity. The observed reduction in passive diffusion of fluorescent Rhodamine 6G across the membrane validated the intricate relationship between ergosterol, membrane fluidity, and microviscosity. An inverse relationship existing between ergosterol biosynthetic genes, <i>ERG11</i> and <i>ERG3</i> in treated cells, related well with displayed higher susceptibilities. Furthermore, treated cells exhibited impaired functionality and downregulation of ABC drug efflux pumps. Multiple cellular targets of ZnO QDs in <i>Candida</i> were validated by <i>in silico</i> molecular docking. Thus, targeting <i>ERG11</i>, <i>ERG3</i>, and ABC drug efflux pumps might emerge as a versatile, nano-ZnO-based strategy in fungal therapeutics to address the challenges of drug resistance.</p>","PeriodicalId":17,"journal":{"name":"ACS Infectious Diseases","volume":null,"pages":null},"PeriodicalIF":5.3,"publicationDate":"2024-06-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141236470","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 : 2024-05-31DOI: 10.1021/acsinfecdis.3c00682
Zheng Ma, Runqiu Liu, Jie Wang, Tao Yu, Yingqiu Zou, Fangfang Chen, Cui Cui, Huixin Yang and Hexin Xie*,
Bacterial resistance caused by β-lactamases has been a major threat to public health around the world, seriously weakening the efficacy of β-lactam antibiotics, the most widely used therapeutic agents against infectious diseases. To detect the bacterial resistance to β-lactam antibiotics, particularly specific type of β-lactam antibiotics, in a rapid manner, we report herein a relay-response chemiluminescence assay. This assay mainly consists of two reagents: a β-lactam-caged thiophenol and a thiophenol-sensitive chemiluminescence reporter, both of which are synthetically feasible. The selective hydrolysis of β-lactam by β-lactamase leads to the releasing of free thiophenol, which then triggers the emission of a chemiluminescence signal in a relay manner. Three thiophenol-caged β-lactams, structural analogues of cephalothin, cefotaxime, and meropenem, respectively, have been synthesized. And the application of this assay with these analogues of β-lactam antibiotics allows fast detection of β-lactamase-expressing resistant bacteria and, more impressively, provides detailed information on the resistant scope of bacteria.
{"title":"Rapid Detection of Bacterial Resistance to β-Lactam Antibiotics with a Relay-Response Chemiluminescence Assay","authors":"Zheng Ma, Runqiu Liu, Jie Wang, Tao Yu, Yingqiu Zou, Fangfang Chen, Cui Cui, Huixin Yang and Hexin Xie*, ","doi":"10.1021/acsinfecdis.3c00682","DOIUrl":"10.1021/acsinfecdis.3c00682","url":null,"abstract":"<p >Bacterial resistance caused by β-lactamases has been a major threat to public health around the world, seriously weakening the efficacy of β-lactam antibiotics, the most widely used therapeutic agents against infectious diseases. To detect the bacterial resistance to β-lactam antibiotics, particularly specific type of β-lactam antibiotics, in a rapid manner, we report herein a relay-response chemiluminescence assay. This assay mainly consists of two reagents: a β-lactam-caged thiophenol and a thiophenol-sensitive chemiluminescence reporter, both of which are synthetically feasible. The selective hydrolysis of β-lactam by β-lactamase leads to the releasing of free thiophenol, which then triggers the emission of a chemiluminescence signal in a relay manner. Three thiophenol-caged β-lactams, structural analogues of cephalothin, cefotaxime, and meropenem, respectively, have been synthesized. And the application of this assay with these analogues of β-lactam antibiotics allows fast detection of β-lactamase-expressing resistant bacteria and, more impressively, provides detailed information on the resistant scope of bacteria.</p>","PeriodicalId":17,"journal":{"name":"ACS Infectious Diseases","volume":null,"pages":null},"PeriodicalIF":5.3,"publicationDate":"2024-05-31","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141183057","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 : 2024-05-31DOI: 10.1021/acsinfecdis.4c00094
Conor J. Crawford, Livia Liporagi-Lopes, Carolina Coelho, Samuel R. Santos Junior, André Moraes Nicola, Maggie P. Wear, Raghav Vij, Stefan Oscarson* and Arturo Casadevall*,
Cryptococcus neoformans is a fungus classified by the World Health Organization as a critically important pathogen, which poses a significant threat to immunocompromised individuals. In this study, we present the chemical synthesis and evaluation of two semisynthetic vaccine candidates targeting the capsular polysaccharide glucuronoxylomannan (GXM) of C. neoformans. These semisynthetic glycoconjugate vaccines contain an identical synthetic decasaccharide (M2 motif) antigen. This antigen is present in serotype A strains, which constitute 95% of the clinical cryptococcosis cases. This synthetic oligosaccharide was conjugated to two proteins (CRM197 and Anthrax 63 kDa PA) and tested for immunogenicity in mice. The conjugates elicited a specific antibody response that bound to the M2 motif but also exhibited additional cross-reactivity toward M1 and M4 GXM motifs. Both glycoconjugates produced antibodies that bound to GXM in ELISA assays and to live fungal cells. Mice immunized with the CRM197 glycoconjugate produced weakly opsonic antibodies and displayed trends toward increased median survival relative to mice given a mock PBS injection (18 vs 15 days, p = 0.06). These findings indicate promise, achieving a successful vaccine demands further optimization of the glycoconjugate. This antigen could serve as a component in a multivalent GXM motif vaccine.
{"title":"Semisynthetic Glycoconjugate Vaccine Candidates against Cryptococcus neoformans","authors":"Conor J. Crawford, Livia Liporagi-Lopes, Carolina Coelho, Samuel R. Santos Junior, André Moraes Nicola, Maggie P. Wear, Raghav Vij, Stefan Oscarson* and Arturo Casadevall*, ","doi":"10.1021/acsinfecdis.4c00094","DOIUrl":"10.1021/acsinfecdis.4c00094","url":null,"abstract":"<p ><i>Cryptococcus neoformans</i> is a fungus classified by the World Health Organization as a critically important pathogen, which poses a significant threat to immunocompromised individuals. In this study, we present the chemical synthesis and evaluation of two semisynthetic vaccine candidates targeting the capsular polysaccharide glucuronoxylomannan (GXM) of <i>C. neoformans</i>. These semisynthetic glycoconjugate vaccines contain an identical synthetic decasaccharide (M2 motif) antigen. This antigen is present in serotype A strains, which constitute 95% of the clinical cryptococcosis cases. This synthetic oligosaccharide was conjugated to two proteins (CRM197 and Anthrax 63 kDa PA) and tested for immunogenicity in mice. The conjugates elicited a specific antibody response that bound to the M2 motif but also exhibited additional cross-reactivity toward M1 and M4 GXM motifs. Both glycoconjugates produced antibodies that bound to GXM in ELISA assays and to live fungal cells. Mice immunized with the CRM197 glycoconjugate produced weakly opsonic antibodies and displayed trends toward increased median survival relative to mice given a mock PBS injection (18 vs 15 days, <i>p</i> = 0.06). These findings indicate promise, achieving a successful vaccine demands further optimization of the glycoconjugate. This antigen could serve as a component in a multivalent GXM motif vaccine.</p>","PeriodicalId":17,"journal":{"name":"ACS Infectious Diseases","volume":null,"pages":null},"PeriodicalIF":5.3,"publicationDate":"2024-05-31","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://pubs.acs.org/doi/epdf/10.1021/acsinfecdis.4c00094","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141183063","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 : 2024-05-31DOI: 10.1021/acsinfecdis.4c00114
Apurva Pandey, Shalini Chopra, Simon J. Cleary, Marina López-Álvarez, Fiona M. Quimby, Aryn A.A. Alanizi, Sasank Sakhamuri, Ningjing Zhang, Mark R. Looney, Charles S. Craik, David M. Wilson* and Michael J. Evans*,
Understanding how the host immune system engages complex pathogens is essential to developing therapeutic strategies to overcome their virulence. While granzymes are well understood to trigger apoptosis in infected host cells or bacteria, less is known about how the immune system mobilizes individual granzyme species in vivo to combat diverse pathogens. Toward the goal of studying individual granzyme function directly in vivo, we previously developed a new class of radiopharmaceuticals termed “restricted interaction peptides (RIPs)” that detect biochemically active endoproteases using positron emission tomography (PET). In this study, we showed that secreted granzyme B proteolysis in response to diverse viral and bacterial pathogens could be imaged with [64Cu]Cu-GRIP B, a RIP that specifically targets granzyme B. Wild-type or germline granzyme B knockout mice were instilled intranasally with the A/PR/8/34 H1N1 influenza A strain to generate pneumonia, and granzyme B production within the lungs was measured using [64Cu]Cu-GRIP B PET/CT. Murine myositis models of acute bacterial (E. coli, P. aeruginosa, K. pneumoniae, and L. monocytogenes) infection were also developed and imaged using [64Cu]Cu-GRIP B. In all cases, the mice were studied in vivo using mPET/CT and ex vivo via tissue-harvesting, gamma counting, and immunohistochemistry. [64Cu]Cu-GRIP B uptake was significantly higher in the lungs of wild-type mice that received A/PR/8/34 H1N1 influenza A strain compared to mice that received sham or granzyme B knockout mice that received either treatment. In wild-type mice, [64Cu]Cu-GRIP B uptake was significantly higher in the infected triceps muscle versus normal muscle and the contralateral triceps inoculated with heat killed bacteria. In granzyme B knockout mice, [64Cu]Cu-GRIP B uptake above the background was not observed in the infected triceps muscle. Interestingly, live L. monocytogenes did not induce detectable granzyme B on PET, despite prior in vitro data, suggesting a role for granzyme B in suppressing their pathogenicity. In summary, these data show that the granzyme response elicited by diverse human pathogens can be imaged using PET. These results and data generated via additional RIPs specific for other granzyme proteases will allow for a deeper mechanistic study analysis of their complex in vivo biology.
了解宿主免疫系统如何与复杂的病原体打交道,对于制定克服病原体毒性的治疗策略至关重要。虽然人们对颗粒酶触发受感染宿主细胞或细菌凋亡的作用有了充分了解,但对免疫系统如何在体内调动单个颗粒酶种类来对抗各种病原体却知之甚少。为了实现在体内直接研究单个颗粒酶功能的目标,我们之前开发了一类新的放射性药物,称为 "受限相互作用肽(RIPs)",它能利用正电子发射断层扫描(PET)检测生化活性内切蛋白酶。我们在这项研究中发现,用[64Cu]Cu-GRIP B(一种特异性靶向颗粒酶 B 的 RIP)可以对颗粒酶 B 在多种病毒和细菌病原体作用下分泌的蛋白水解进行成像。用 A/PR/8/34 H1N1 甲型流感病毒株向野生型或基因型颗粒酶 B 基因敲除的小鼠鼻腔内灌注病毒以产生肺炎,然后用[64Cu]Cu-GRIP B PET/CT 测量肺内颗粒酶 B 的产生情况。此外,还开发了急性细菌(大肠杆菌、绿脓杆菌、肺炎双球菌和单核细胞增生性酵母菌)感染的小鼠肌炎模型,并使用[64Cu]Cu-GRIP B 进行成像。与接受假治疗或接受颗粒酶 B 基因敲除治疗的小鼠相比,接受 A/PR/8/34 H1N1 甲型流感病毒株治疗的野生型小鼠肺部的[64Cu]Cu-GRIP B 摄取量明显更高。在野生型小鼠中,受感染的肱三头肌对正常肌肉和接种了热致死细菌的对侧肱三头肌的[64Cu]Cu-GRIP B摄取量明显较高。在粒酶 B 基因敲除小鼠中,受感染的肱三头肌中未观察到高于背景的[64Cu]Cu-GRIP B 摄取。有趣的是,尽管之前有体外数据,但活的单核细胞增生梭状芽孢杆菌并未在 PET 上诱导出可检测到的颗粒酶 B,这表明颗粒酶 B 在抑制其致病性方面发挥了作用。总之,这些数据表明,各种人类病原体引起的颗粒酶反应可以用 PET 进行成像。这些结果以及通过其他颗粒酶蛋白酶特异性 RIPs 生成的数据将有助于对它们复杂的体内生物学进行更深入的机理研究分析。
{"title":"Imaging the Granzyme Mediated Host Immune Response to Viral and Bacterial Pathogens In Vivo Using Positron Emission Tomography","authors":"Apurva Pandey, Shalini Chopra, Simon J. Cleary, Marina López-Álvarez, Fiona M. Quimby, Aryn A.A. Alanizi, Sasank Sakhamuri, Ningjing Zhang, Mark R. Looney, Charles S. Craik, David M. Wilson* and Michael J. Evans*, ","doi":"10.1021/acsinfecdis.4c00114","DOIUrl":"10.1021/acsinfecdis.4c00114","url":null,"abstract":"<p >Understanding how the host immune system engages complex pathogens is essential to developing therapeutic strategies to overcome their virulence. While granzymes are well understood to trigger apoptosis in infected host cells or bacteria, less is known about how the immune system mobilizes individual granzyme species <i>in vivo</i> to combat diverse pathogens. Toward the goal of studying individual granzyme function directly <i>in vivo</i>, we previously developed a new class of radiopharmaceuticals termed “restricted interaction peptides (RIPs)” that detect biochemically active endoproteases using positron emission tomography (PET). In this study, we showed that secreted granzyme B proteolysis in response to diverse viral and bacterial pathogens could be imaged with [<sup>64</sup>Cu]Cu-GRIP B, a RIP that specifically targets granzyme B. Wild-type or germline granzyme B knockout mice were instilled intranasally with the A/PR/8/34 H1N1 influenza A strain to generate pneumonia, and granzyme B production within the lungs was measured using [<sup>64</sup>Cu]Cu-GRIP B PET/CT. Murine myositis models of acute bacterial (<i>E. coli</i>, <i>P. aeruginosa</i>, <i>K. pneumoniae</i>, and <i>L. monocytogenes</i>) infection were also developed and imaged using [<sup>64</sup>Cu]Cu-GRIP B. In all cases, the mice were studied <i>in vivo</i> using mPET/CT and <i>ex vivo</i> via tissue-harvesting, gamma counting, and immunohistochemistry. [<sup>64</sup>Cu]Cu-GRIP B uptake was significantly higher in the lungs of wild-type mice that received A/PR/8/34 H1N1 influenza A strain compared to mice that received sham or granzyme B knockout mice that received either treatment. In wild-type mice, [<sup>64</sup>Cu]Cu-GRIP B uptake was significantly higher in the infected triceps muscle versus normal muscle and the contralateral triceps inoculated with heat killed bacteria. In granzyme B knockout mice, [<sup>64</sup>Cu]Cu-GRIP B uptake above the background was not observed in the infected triceps muscle. Interestingly, live <i>L. monocytogenes</i> did not induce detectable granzyme B on PET, despite prior <i>in vitro</i> data, suggesting a role for granzyme B in suppressing their pathogenicity. In summary, these data show that the granzyme response elicited by diverse human pathogens can be imaged using PET. These results and data generated via additional RIPs specific for other granzyme proteases will allow for a deeper mechanistic study analysis of their complex <i>in vivo</i> biology.</p>","PeriodicalId":17,"journal":{"name":"ACS Infectious Diseases","volume":null,"pages":null},"PeriodicalIF":5.3,"publicationDate":"2024-05-31","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141178076","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 : 2024-05-30DOI: 10.1021/acsinfecdis.4c00401
Daniel E. Manson, Gene E. Ananiev, Song Guo, Spencer S. Ericksen, Emma E. Santa and Helen E. Blackwell*,
{"title":"Correction to “Abiotic Small Molecule Inhibitors and Activators of the LasR Quorum Sensing Receptor in Pseudomonas aeruginosa with Potencies Comparable or Surpassing N-Acyl Homoserine Lactones”","authors":"Daniel E. Manson, Gene E. Ananiev, Song Guo, Spencer S. Ericksen, Emma E. Santa and Helen E. Blackwell*, ","doi":"10.1021/acsinfecdis.4c00401","DOIUrl":"10.1021/acsinfecdis.4c00401","url":null,"abstract":"","PeriodicalId":17,"journal":{"name":"ACS Infectious Diseases","volume":null,"pages":null},"PeriodicalIF":5.3,"publicationDate":"2024-05-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141173831","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 : 2024-05-30DOI: 10.1021/acsinfecdis.4c00001
Kritika Singh, Shubham Vashishtha, Ankan Chakraborty, Ashish Kumar, Sheetal Thakur and Bishwajit Kundu*,
Conserved molecular signatures in multidrug-resistant Salmonella typhi can serve as novel therapeutic targets for mitigation of infection. In this regard, we present the S. typhi cell division activator protein (StCAP) as a conserved target across S. typhi variants. From in silico and fluorimetric assessments, we found that StCAP is a DNA-binding protein. Replacement of the identified DNA-interacting residue Arg34 of StCAP with Ala34 showed a dramatic (15-fold) increase in Kd value compared to the wild type (Kd 546 nm) as well as a decrease in thermal stability (10 °C shift). Out of the two screened molecules against the DNA-binding pocket of StCAP, eltrombopag, and nilotinib, the former displayed better binding. Eltrombopag inhibited the stand-alone S. typhi culture with an IC50 of 38 μM. The effect was much more pronounced on THP-1-derived macrophages (T1Mac) infected with S. typhi where colony formation was severely hindered with IC50 reduced further to 10 μM. Apoptotic protease activating factor1 (Apaf1), a key molecule for intrinsic apoptosis, was identified as an StCAP-interacting partner by pull-down assay against T1Mac. Further, StCAP-transfected T1Mac showed a significant increase in LC3 II (autophagy marker) expression and downregulation of caspase 3 protein. From these experiments, we conclude that StCAP provides a crucial survival advantage to S. typhi during infection, thereby making it a potent alternative therapeutic target.
{"title":"The Salmonella typhi Cell Division Activator Protein StCAP Impacts Pathogenesis by Influencing Critical Molecular Events","authors":"Kritika Singh, Shubham Vashishtha, Ankan Chakraborty, Ashish Kumar, Sheetal Thakur and Bishwajit Kundu*, ","doi":"10.1021/acsinfecdis.4c00001","DOIUrl":"10.1021/acsinfecdis.4c00001","url":null,"abstract":"<p >Conserved molecular signatures in multidrug-resistant <i>Salmonella typhi</i> can serve as novel therapeutic targets for mitigation of infection. In this regard, we present the <i>S. typhi</i> cell division activator protein (StCAP) as a conserved target across <i>S. typhi</i> variants. From <i>in silico</i> and fluorimetric assessments, we found that StCAP is a DNA-binding protein. Replacement of the identified DNA-interacting residue Arg<sup>34</sup> of StCAP with Ala<sup>34</sup> showed a dramatic (15-fold) increase in <i>K</i><sub>d</sub> value compared to the wild type (<i>K</i><sub>d</sub> 546 nm) as well as a decrease in thermal stability (10 °C shift). Out of the two screened molecules against the DNA-binding pocket of StCAP, eltrombopag, and nilotinib, the former displayed better binding. Eltrombopag inhibited the stand-alone <i>S. typhi</i> culture with an IC<sub>50</sub> of 38 μM. The effect was much more pronounced on THP-1-derived macrophages (T1Mac) infected with <i>S. typhi</i> where colony formation was severely hindered with IC<sub>50</sub> reduced further to 10 μM. Apoptotic protease activating factor1 (Apaf1), a key molecule for intrinsic apoptosis, was identified as an StCAP-interacting partner by pull-down assay against T1Mac. Further, StCAP-transfected T1Mac showed a significant increase in LC3 II (autophagy marker) expression and downregulation of caspase 3 protein. From these experiments, we conclude that StCAP provides a crucial survival advantage to <i>S. typhi</i> during infection, thereby making it a potent alternative therapeutic target.</p>","PeriodicalId":17,"journal":{"name":"ACS Infectious Diseases","volume":null,"pages":null},"PeriodicalIF":5.3,"publicationDate":"2024-05-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141178020","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 : 2024-05-29DOI: 10.1021/acsinfecdis.4c00232
Jennifer E. Collins, Tiantian Jiang, Jin Woo Lee, Karen Wendt, Flore Nardella, Jin Jeon, Raphaella Paes, Natalia Mojica Santos, Frances Rocamora, Maya Chang, Samuel Schaefer, Robert H. Cichewicz*, Elizabeth A. Winzeler* and Debopam Chakrabarti*,
Our previous work identified a series of 12 xanthoquinodin analogues and 2 emodin-dianthrones with broad-spectrum activities against Trichomonas vaginalis, Mycoplasma genitalium, Cryptosporidium parvum, and Plasmodium falciparum. Analyses conducted in this study revealed that the most active analogue, xanthoquinodin A1, also inhibits Toxoplasma gondii tachyzoites and the liver stage of Plasmodium berghei, with no cross-resistance to the known antimalarial targets PfACS, PfCARL, PfPI4K, or DHODH. In Plasmodium, inhibition occurs prior to multinucleation and induces parasite death following 12 h of compound exposure. This moderately fast activity has impeded resistance line generation, with xanthoquinodin A1 demonstrating an irresistible phenotype in both T. gondii and P. falciparum.
{"title":"Understanding the Antiplasmodial Action of Resistance-Refractory Xanthoquinodin A1","authors":"Jennifer E. Collins, Tiantian Jiang, Jin Woo Lee, Karen Wendt, Flore Nardella, Jin Jeon, Raphaella Paes, Natalia Mojica Santos, Frances Rocamora, Maya Chang, Samuel Schaefer, Robert H. Cichewicz*, Elizabeth A. Winzeler* and Debopam Chakrabarti*, ","doi":"10.1021/acsinfecdis.4c00232","DOIUrl":"10.1021/acsinfecdis.4c00232","url":null,"abstract":"<p >Our previous work identified a series of 12 xanthoquinodin analogues and 2 emodin-dianthrones with broad-spectrum activities against <i>Trichomonas vaginalis</i>, <i>Mycoplasma genitalium</i>, <i>Cryptosporidium parvum</i>, and <i>Plasmodium falciparum</i>. Analyses conducted in this study revealed that the most active analogue, xanthoquinodin A1, also inhibits <i>Toxoplasma gondii</i> tachyzoites and the liver stage of <i>Plasmodium berghei</i>, with no cross-resistance to the known antimalarial targets PfACS, PfCARL, PfPI4K, or DHODH. In <i>Plasmodium</i>, inhibition occurs prior to multinucleation and induces parasite death following 12 h of compound exposure. This moderately fast activity has impeded resistance line generation, with xanthoquinodin A1 demonstrating an irresistible phenotype in both <i>T. gondii</i> and <i>P. falciparum</i>.</p>","PeriodicalId":17,"journal":{"name":"ACS Infectious Diseases","volume":null,"pages":null},"PeriodicalIF":5.3,"publicationDate":"2024-05-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141173867","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}