Pub Date : 2024-09-04DOI: 10.1101/2024.09.03.609146
Anna-Mari Haapanen-Saaristo, Noora Virtanen, Elena Tcarenkova, Katri Mataleena Vaparanta, Minna Ampuja, Eeva-Riikka Vehniäinen, Ilkka Paatero
Global warming increases the risk of dangerous heat waves, which may have deleterious effects on humans and wildlife. Here, we have utilized zebrafish embryos as a model to analyse heat stress and effect of chemical compounds on responses to heat stress. The temperature adaptation limit of zebrafish embryos was 37°C in behavioural test and 38°C in cardiac test. Polyaromatic hydrocarbon phenanthrene completely blocked the behavioural adaptation to heat stress. Interestingly, the cardiotoxic effects of lapatinib, phenanthrene and paclitaxel were induced by heat stress. Taken together, our data indicates that motility and cardiac function of zebrafish embryos can be utilized as a model to analyze modulatory effects of compounds on heat-stress.
{"title":"Heat stress sensitizes zebrafish embryos to neurological and cardiac toxicity.","authors":"Anna-Mari Haapanen-Saaristo, Noora Virtanen, Elena Tcarenkova, Katri Mataleena Vaparanta, Minna Ampuja, Eeva-Riikka Vehniäinen, Ilkka Paatero","doi":"10.1101/2024.09.03.609146","DOIUrl":"https://doi.org/10.1101/2024.09.03.609146","url":null,"abstract":"Global warming increases the risk of dangerous heat waves, which may have deleterious effects on humans and wildlife. Here, we have utilized zebrafish embryos as a model to analyse heat stress and effect of chemical compounds on responses to heat stress. The temperature adaptation limit of zebrafish embryos was 37°C in behavioural test and 38°C in cardiac test. Polyaromatic hydrocarbon phenanthrene completely blocked the behavioural adaptation to heat stress. Interestingly, the cardiotoxic effects of lapatinib, phenanthrene and paclitaxel were induced by heat stress. Taken together, our data indicates that motility and cardiac function of zebrafish embryos can be utilized as a model to analyze modulatory effects of compounds on heat-stress.","PeriodicalId":501518,"journal":{"name":"bioRxiv - Pharmacology and Toxicology","volume":"49 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2024-09-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142184231","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-09-02DOI: 10.1101/2024.08.30.610527
Cara F. Smith, Mamadou Alpha Balde, Lilyrose Bahrabadi, Merilyn Amponsah-Asamoah, Keira Y. Larson, Sean P. Maroney, David Ceja-Galindo, Martin Millimouno, Naby Camara, Jordan Benjamin, Nicklaus P. Brandehoff, Cassandra M. Modahl, Maxwell C. McCabe, Mitchell J. Cohen, Todd A. Castoe, Cellou Balde, Kate Jackson, Stephen P. Mackessy, Kirk C. Hansen, Anthony J. Saviola
Identification and characterization of snake venom toxins that interfere with hemostasis have important implications for the treatment of snake envenomation, the bioprospecting of therapeutically useful molecules, and the development of research tools for investigating hematologic disorders. Many venoms have been shown to possess thrombolytic activity. However, it remains unclear if actions on other clot-stabilizing proteins beyond fibrin chains contribute significantly to venom-induced thrombolysis because the clot-wide targets of venom proteases and the mechanisms responsible for thrombolysis are not well understood. Here, we utilize a high-throughput time-based thrombolysis assay in combination with untargeted peptidomics to provide comprehensive insight into the effects of venom from six snake species on blood clot degradation. We compare thrombolytic profiles across venoms with variable levels of proteases and generate venom-specific fingerprints of cleavage specificity. We also compare the specific effects of venoms that possess a range of thrombolytic activity on fibrin subunits and other clot-bound proteins involved in clot structure. Venoms with higher thrombolytic activity demonstrated an enhanced ability to target multiple sites across fibrin chains critical to clot stability and structure, as well as clot-stabilizing proteins including fibronectin and vitronectin. Collectively, this study significantly expands our understanding of the thrombolytic and fibrinolytic effects of snake venom by determining the full suite of clot-specific venom targets that are involved in clot formation and stability.
{"title":"Discerning Specific Thrombolytic Activities and Blood Clot Degradomes of Diverse Snake Venoms with Untargeted Peptidomics","authors":"Cara F. Smith, Mamadou Alpha Balde, Lilyrose Bahrabadi, Merilyn Amponsah-Asamoah, Keira Y. Larson, Sean P. Maroney, David Ceja-Galindo, Martin Millimouno, Naby Camara, Jordan Benjamin, Nicklaus P. Brandehoff, Cassandra M. Modahl, Maxwell C. McCabe, Mitchell J. Cohen, Todd A. Castoe, Cellou Balde, Kate Jackson, Stephen P. Mackessy, Kirk C. Hansen, Anthony J. Saviola","doi":"10.1101/2024.08.30.610527","DOIUrl":"https://doi.org/10.1101/2024.08.30.610527","url":null,"abstract":"Identification and characterization of snake venom toxins that interfere with hemostasis have important implications for the treatment of snake envenomation, the bioprospecting of therapeutically useful molecules, and the development of research tools for investigating hematologic disorders. Many venoms have been shown to possess thrombolytic activity. However, it remains unclear if actions on other clot-stabilizing proteins beyond fibrin chains contribute significantly to venom-induced thrombolysis because the clot-wide targets of venom proteases and the mechanisms responsible for thrombolysis are not well understood. Here, we utilize a high-throughput time-based thrombolysis assay in combination with untargeted peptidomics to provide comprehensive insight into the effects of venom from six snake species on blood clot degradation. We compare thrombolytic profiles across venoms with variable levels of proteases and generate venom-specific fingerprints of cleavage specificity. We also compare the specific effects of venoms that possess a range of thrombolytic activity on fibrin subunits and other clot-bound proteins involved in clot structure. Venoms with higher thrombolytic activity demonstrated an enhanced ability to target multiple sites across fibrin chains critical to clot stability and structure, as well as clot-stabilizing proteins including fibronectin and vitronectin. Collectively, this study significantly expands our understanding of the thrombolytic and fibrinolytic effects of snake venom by determining the full suite of clot-specific venom targets that are involved in clot formation and stability.","PeriodicalId":501518,"journal":{"name":"bioRxiv - Pharmacology and Toxicology","volume":"67 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2024-09-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142184255","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-09-02DOI: 10.1101/2024.08.31.610588
Shubhra Bhattacharjee, Miguel A Chacon-Teran, Michael Findlater, Stacey M Louie, Jeremy D Bailoo, Amrika Deonarine
There has been limited research into arsenolipid toxicological risks and health-related outcomes due to challenges with their separation, identification, and quantification within complex biological matrices (e.g., fish, seaweed). Analytical approaches for arsenolipid identification such as suspect screening have not been well documented and there are no certified standard reference materials, leading to issues with reproducibility and uncertainty regarding the accuracy of results. In this study, a detailed workflow for the identification of arsenolipids utilizing suspect screening coupled with data independent analysis is presented and applied to three commercially available standard reference materials (Hijiki seaweed, dogfish liver, and tuna). Hexane and dichloromethane/methanol extraction, followed by reversed-phase high-performance liquid chromatography-inductively coupled plasma mass spectrometry and liquid chromatography-electrospray ionization-quadrupole time-of-flight mass spectrometry. Using the workflow developed, mass fragmentation matching, mass error calculations, and retention time matching were performed to identify suspect arsenolipids. Arseno-fatty acids (AsFAs), arsenohydrocarbons (AsHCs), and arsenosugar phospholipids (AsSugPLs) were identified with high confidence; AsHC332, AsHC360, and AsSugPL720 in seaweed, AsHC332 in tuna, and AsFA474 and AsFA502 in the dogfish liver. AsHC332, AsHC360, and AsFA502 were identified as promising candidates for further work on synthesis, quantification using MS/MS, and toxicity testing.
{"title":"Suspect screening-data independent analysis workflow for the identification of arsenolipids in marine standard reference materials","authors":"Shubhra Bhattacharjee, Miguel A Chacon-Teran, Michael Findlater, Stacey M Louie, Jeremy D Bailoo, Amrika Deonarine","doi":"10.1101/2024.08.31.610588","DOIUrl":"https://doi.org/10.1101/2024.08.31.610588","url":null,"abstract":"There has been limited research into arsenolipid toxicological risks and health-related outcomes due to challenges with their separation, identification, and quantification within complex biological matrices (e.g., fish, seaweed). Analytical approaches for arsenolipid identification such as suspect screening have not been well documented and there are no certified standard reference materials, leading to issues with reproducibility and uncertainty regarding the accuracy of results. In this study, a detailed workflow for the identification of arsenolipids utilizing suspect screening coupled with data independent analysis is presented and applied to three commercially available standard reference materials (Hijiki seaweed, dogfish liver, and tuna). Hexane and dichloromethane/methanol extraction, followed by reversed-phase high-performance liquid chromatography-inductively coupled plasma mass spectrometry and liquid chromatography-electrospray ionization-quadrupole time-of-flight mass spectrometry. Using the workflow developed, mass fragmentation matching, mass error calculations, and retention time matching were performed to identify suspect arsenolipids. Arseno-fatty acids (AsFAs), arsenohydrocarbons (AsHCs), and arsenosugar phospholipids (AsSugPLs) were identified with high confidence; AsHC332, AsHC360, and AsSugPL720 in seaweed, AsHC332 in tuna, and AsFA474 and AsFA502 in the dogfish liver. AsHC332, AsHC360, and AsFA502 were identified as promising candidates for further work on synthesis, quantification using MS/MS, and toxicity testing.","PeriodicalId":501518,"journal":{"name":"bioRxiv - Pharmacology and Toxicology","volume":"11 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2024-09-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142184260","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-09-02DOI: 10.1101/2024.08.31.610658
Dmitry Lifanov, Dulamsuren Zorigt, Evgenya Shabalina, Abdullah Khalil, Konstantin Gorbunov, Elena Petersen
This paper describes a method for determining the cytotoxicity of chemical compounds based on the detection of fluorescent proteins - in this case, green fluorescent protein (GFP) and red fluorescent protein (RFP), which are released into the medium from dead cells. This method is similar in principle to the lactate dehydrogenase test (LDH test), but it does not require a reaction with a chromogenic substrate. This method also makes it possible to independently determine the viability of different lines when used in cocultures. Experiments were preformed on a classical monolayer, spheroids and 3D cultures in alginate hydrogel. Capecitabine was used as a model cytotoxic agent. We included liver cells (Huh7) in a coculture model and determined changes in the cytotoxicity levels of capecitabine against NCI-H1299 cells. The experimental part also found that there were differences in sensitivity to capecitabine depending on the type of 3D cultures used.
{"title":"Method for determining of cytotoxicity based on the release of fluorescent proteins","authors":"Dmitry Lifanov, Dulamsuren Zorigt, Evgenya Shabalina, Abdullah Khalil, Konstantin Gorbunov, Elena Petersen","doi":"10.1101/2024.08.31.610658","DOIUrl":"https://doi.org/10.1101/2024.08.31.610658","url":null,"abstract":"This paper describes a method for determining the cytotoxicity of chemical compounds based on the detection of fluorescent proteins - in this case, green fluorescent protein (GFP) and red fluorescent protein (RFP), which are released into the medium from dead cells. This method is similar in principle to the lactate dehydrogenase test (LDH test), but it does not require a reaction with a chromogenic substrate. This method also makes it possible to independently determine the viability of different lines when used in cocultures. Experiments were preformed on a classical monolayer, spheroids and 3D cultures in alginate hydrogel. Capecitabine was used as a model cytotoxic agent. We included liver cells (Huh7) in a coculture model and determined changes in the cytotoxicity levels of capecitabine against NCI-H1299 cells. The experimental part also found that there were differences in sensitivity to capecitabine depending on the type of 3D cultures used.","PeriodicalId":501518,"journal":{"name":"bioRxiv - Pharmacology and Toxicology","volume":"6 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2024-09-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142184254","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-08-30DOI: 10.1101/2024.08.29.610278
Hannah Roe, Han-Hsuan D Tsai, Nicholas Ball, Fred A Wright, Weihsueh Chiu, Ivan Rusyn
An important element of the European Unions Registration, Evaluation, Authorisation and Restriction of Chemicals (REACH) regulation is the evaluation by the European Chemicals Agency (ECHA) of testing proposals submitted by the registrants to address data gaps in standard REACH information requirements. The registrants may propose adaptations, and ECHA evaluates the reasoning and issues a written decision. Read-across is a common adaptation type, yet it is widely assumed that ECHA often does not agree that the justifications are adequate to waive standard testing requirements. From 2008 to August 2023, a total of 2,630 Testing Proposals were submitted to ECHA; of these, 1,538 had published decisions that were systematically evaluated in this study. Each document was manually reviewed, and information extracted for further analyses. Read-across hypotheses were standardized into 17 assessment elements (AEs); each submission was classified as to the AEs relied upon by the registrants and by ECHA. Data was analyzed for patterns and associations. Testing Proposal Evaluations (TPEs) with adaptations comprised 23% (353) of the total; analogue (168) or group (136) read-across adaptations were most common. Of 304 read-across-containing TPEs, 49% were accepted; the odds of acceptance were significantly greater for group read-across submissions. The data was analyzed by Annex (i.e., tonnage), test guideline study, read-across hypothesis AEs, as well as target and source substance types and their structural similarity. While most ECHA decisions with both positive and negative decisions on whether the proposed read-across was adequate were context-specific, a number of significant associations were identified that influence the odds of acceptance. Overall, this analysis provides an unbiased overview of 15 years of experience with testing proposal-specific read-across adaptations by both registrants and ECHA. These data will inform future submissions as they identify most critical AEs to increase the odds of read-across acceptance.
{"title":"A Systematic Analysis of Read-Across Adaptations in Testing Proposal Evaluations by the European Chemicals Agency","authors":"Hannah Roe, Han-Hsuan D Tsai, Nicholas Ball, Fred A Wright, Weihsueh Chiu, Ivan Rusyn","doi":"10.1101/2024.08.29.610278","DOIUrl":"https://doi.org/10.1101/2024.08.29.610278","url":null,"abstract":"An important element of the European Unions Registration, Evaluation, Authorisation and Restriction of Chemicals (REACH) regulation is the evaluation by the European Chemicals Agency (ECHA) of testing proposals submitted by the registrants to address data gaps in standard REACH information requirements. The registrants may propose adaptations, and ECHA evaluates the reasoning and issues a written decision. Read-across is a common adaptation type, yet it is widely assumed that ECHA often does not agree that the justifications are adequate to waive standard testing requirements. From 2008 to August 2023, a total of 2,630 Testing Proposals were submitted to ECHA; of these, 1,538 had published decisions that were systematically evaluated in this study. Each document was manually reviewed, and information extracted for further analyses. Read-across hypotheses were standardized into 17 assessment elements (AEs); each submission was classified as to the AEs relied upon by the registrants and by ECHA. Data was analyzed for patterns and associations. Testing Proposal Evaluations (TPEs) with adaptations comprised 23% (353) of the total; analogue (168) or group (136) read-across adaptations were most common. Of 304 read-across-containing TPEs, 49% were accepted; the odds of acceptance were significantly greater for group read-across submissions. The data was analyzed by Annex (i.e., tonnage), test guideline study, read-across hypothesis AEs, as well as target and source substance types and their structural similarity. While most ECHA decisions with both positive and negative decisions on whether the proposed read-across was adequate were context-specific, a number of significant associations were identified that influence the odds of acceptance. Overall, this analysis provides an unbiased overview of 15 years of experience with testing proposal-specific read-across adaptations by both registrants and ECHA. These data will inform future submissions as they identify most critical AEs to increase the odds of read-across acceptance.","PeriodicalId":501518,"journal":{"name":"bioRxiv - Pharmacology and Toxicology","volume":"34 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2024-08-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142184257","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-08-30DOI: 10.1101/2024.08.29.610279
Sara Ficai, Andrea Papait, Marta Magatti, Alice Masserdotti, Michael Gasik, Antonietta Rosa Silini, Ornella Parolini
In today's context, microplastics pollution has become an increasingly pressing issue not only for the environmental fallout but also for the assumed negative effects on human health. It is now well-established that microplastics (>1 mm in size) can enter the human body through ingestion, inhalation, dermal contact and also maternal-fetal transmission. Alarming was the recent findings of microplastics within the human term placenta. Among the degradation by-products of microplastics, Bisphenol A (BPA) has emerged as a hazardous chemical, with potential toxicity at multisystemic level, particularly on the earliest stages of human development. Based on these findings, our study focuses on assessing the impact of BPA on properties and functions of mesenchymal stromal cells isolated from the amniotic membrane (hAMSC) of the human term placenta. The amniotic membrane surrounds the fetus, playing a fundamental protective role toward toxic chemicals and pollutants that the mother may encounter. Our research revealed how exposure to increasing concentrations of BPA compromise mitochondrial functionality in hAMSC, resulting in enhanced production of reactive oxygen species at mitochondrial level (mtROS). This, in turn, leads to the stabilization of p53, which triggers an increased expression of p21 and p27 encoding genes and an imbalance in the genetic expression of Bax and Bcl-2. Additionally, we observed upregulated expression of cytokines and chemokines associated with the senescence-associated secretory phenotype (SASP). The increased oxidative stress, which plays a central role in BPA-mediated toxicity, can trigger the activation of the senescence pathways, or culminate in cell death, due to the overwhelming stress conditions. Therefore, our results provide novel insights into the mechanism of action of BPA and elucidates its impact on the functionality of hAMSC. This underscores the pressing need to reconsider the use of BPA as a plastic additive, mitigating the potential adverse effects on babies.
在当今背景下，微塑料污染已成为一个日益紧迫的问题，这不仅是因为其对环境造成的影响，还因为其可能对人类健康造成的负面影响。微塑料（1 毫米大小）可通过摄入、吸入、皮肤接触和母胎传播进入人体，这一点现已得到充分证实。令人震惊的是，最近在人类胎盘中发现了微塑料。在微塑料的降解副产品中，双酚 A（BPA）已成为一种危险化学品，在多系统水平上具有潜在毒性，尤其是在人类发育的最初阶段。基于这些发现，我们的研究重点是评估双酚 A 对从人类足月胎盘羊膜中分离出来的间充质基质细胞（hAMSC）的特性和功能的影响。羊膜包围着胎儿，对母亲可能遇到的有毒化学物质和污染物起着基本的保护作用。我们的研究揭示了暴露于浓度不断增加的双酚 A 如何损害羊膜间充质干细胞线粒体的功能，从而导致线粒体水平活性氧（mtROS）的产生增加。这反过来又导致 p53 的稳定，从而引发 p21 和 p27 编码基因表达的增加以及 Bax 和 Bcl-2 基因表达的失衡。此外，我们还观察到与衰老相关分泌表型（SASP）有关的细胞因子和趋化因子的表达上调。氧化应激在双酚 A 介导的毒性中起着核心作用，氧化应激的增加会触发衰老通路的激活，或在压倒性应激条件下最终导致细胞死亡。因此，我们的研究结果提供了有关双酚 A 作用机制的新见解，并阐明了它对 hAMSC 功能的影响。这凸显了重新考虑使用双酚 A 作为塑料添加剂、减轻其对婴儿的潜在不利影响的迫切需要。
{"title":"Bisphenol A Disrupts Mitochondrial Functionality Leading to Senescence and Apoptosis in Human Amniotic Mesenchymal Stromal Cells","authors":"Sara Ficai, Andrea Papait, Marta Magatti, Alice Masserdotti, Michael Gasik, Antonietta Rosa Silini, Ornella Parolini","doi":"10.1101/2024.08.29.610279","DOIUrl":"https://doi.org/10.1101/2024.08.29.610279","url":null,"abstract":"In today's context, microplastics pollution has become an increasingly pressing issue not only for the environmental fallout but also for the assumed negative effects on human health. It is now well-established that microplastics (>1 mm in size) can enter the human body through ingestion, inhalation, dermal contact and also maternal-fetal transmission. Alarming was the recent findings of microplastics within the human term placenta. Among the degradation by-products of microplastics, Bisphenol A (BPA) has emerged as a hazardous chemical, with potential toxicity at multisystemic level, particularly on the earliest stages of human development. Based on these findings, our study focuses on assessing the impact of BPA on properties and functions of mesenchymal stromal cells isolated from the amniotic membrane (hAMSC) of the human term placenta. The amniotic membrane surrounds the fetus, playing a fundamental protective role toward toxic chemicals and pollutants that the mother may encounter. Our research revealed how exposure to increasing concentrations of BPA compromise mitochondrial functionality in hAMSC, resulting in enhanced production of reactive oxygen species at mitochondrial level (mtROS). This, in turn, leads to the stabilization of p53, which triggers an increased expression of p21 and p27 encoding genes and an imbalance in the genetic expression of Bax and Bcl-2. Additionally, we observed upregulated expression of cytokines and chemokines associated with the senescence-associated secretory phenotype (SASP). The increased oxidative stress, which plays a central role in BPA-mediated toxicity, can trigger the activation of the senescence pathways, or culminate in cell death, due to the overwhelming stress conditions. Therefore, our results provide novel insights into the mechanism of action of BPA and elucidates its impact on the functionality of hAMSC. This underscores the pressing need to reconsider the use of BPA as a plastic additive, mitigating the potential adverse effects on babies.","PeriodicalId":501518,"journal":{"name":"bioRxiv - Pharmacology and Toxicology","volume":"52 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2024-08-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142184335","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-08-30DOI: 10.1101/2024.08.29.610289
Xinliu Zeng, Yongjie Wang, Karina Gisselle Farias, Andrew Rappa, Christine Darko, Anthony A Sauve, Yue Yang
NAD+ deficiency underlies obesity-induced metabolic disturbances. Here we evaluated the treatment effect of a new and potent NAD+ enhancer, dihydronicotinamide riboside (NRH), in diet-induced obese mice with hyperglycemia and hyperlipidemia. Administering NRH for 7 weeks improved glucose homeostasis by enhancing pancreatic beta-cell functional mass, increasing muscle insulin sensitivity, and reducing hepatic gluconeogenesis. NRH treatment also mobilized fat deposition, reduced circulating lipid, and improved white adipose function. Significant elevation in multi-tissue NAD+ levels and sirtuin (SIRT) activities, especially SIRT3, mediated these metabolic improvements. Inhibiting adenosine kinase (ADK), a newly recognized enzyme in the NRH-induced NAD+ synthesis pathway, blocked NRH effect in improving glucose and lipid metabolism. ADK inhibition also reduced tissue NAD+ elevation and the subsequent activation of SIRT3, suggesting an active ADK pathway is necessary for NRH-induced metabolic benefits. These observations, for the first time, establish NRH as a promising intervention for correcting obesity-induced metabolic syndrome.
{"title":"NRH, a potent NAD+ booster, improves glucose homeostasis and lipid metabolism in diet-induced obese mice though an active adenosine kinase pathway.","authors":"Xinliu Zeng, Yongjie Wang, Karina Gisselle Farias, Andrew Rappa, Christine Darko, Anthony A Sauve, Yue Yang","doi":"10.1101/2024.08.29.610289","DOIUrl":"https://doi.org/10.1101/2024.08.29.610289","url":null,"abstract":"NAD+ deficiency underlies obesity-induced metabolic disturbances. Here we evaluated the treatment effect of a new and potent NAD+ enhancer, dihydronicotinamide riboside (NRH), in diet-induced obese mice with hyperglycemia and hyperlipidemia. Administering NRH for 7 weeks improved glucose homeostasis by enhancing pancreatic beta-cell functional mass, increasing muscle insulin sensitivity, and reducing hepatic gluconeogenesis. NRH treatment also mobilized fat deposition, reduced circulating lipid, and improved white adipose function. Significant elevation in multi-tissue NAD+ levels and sirtuin (SIRT) activities, especially SIRT3, mediated these metabolic improvements. Inhibiting adenosine kinase (ADK), a newly recognized enzyme in the NRH-induced NAD+ synthesis pathway, blocked NRH effect in improving glucose and lipid metabolism. ADK inhibition also reduced tissue NAD+ elevation and the subsequent activation of SIRT3, suggesting an active ADK pathway is necessary for NRH-induced metabolic benefits. These observations, for the first time, establish NRH as a promising intervention for correcting obesity-induced metabolic syndrome.","PeriodicalId":501518,"journal":{"name":"bioRxiv - Pharmacology and Toxicology","volume":"9 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2024-08-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142223912","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-08-29DOI: 10.1101/2024.08.28.609965
Alexandre Joushomme, Antoine Désilets, William Champagne, Malihe Hassanzadeh, Gabriel Lemieux, Alice Gravel-Trudeau, Matthieu Lepage, Sabrina Lafrenière, Ulrike Froehlich, Karin List, Pierre-Luc Boudreault, Richard Leduc
TMPRSS13, a member of the Type II Transmembrane Serine Proteases (TTSP) family, is involved in cancer progression and in cell entry of respiratory viruses. To date, no inhibitors have been specifically developed toward this protease. In this study, a chemical library of 65 ketobenzothiazole-based peptidomimetic molecules was screened against a proteolytically active form of recombinant TMPRSS13 to identify novel inhibitors. Following an initial round of screening, subsequent synthesis of additional derivatives supported by molecular modelling, uncovered important molecular determinants involved in TMPRSS13 inhibition. One inhibitor, N-0430, achieved low nanomolar affinity towards TMPRSS13 activity in a cellular context. Using a SARS-CoV-2 pseudovirus cell entry model, we further show the ability of N-0430 to block TMPRSS13-dependent entry of the pseudovirus. The identified peptidomimetic inhibitors and the molecular insights of their potency gained from this study will aid in the development of specific TMPRSS13 inhibitors.
{"title":"Development of ketobenzothiazole-based peptidomimetic TMPRSS13 inhibitors with low nanomolar potency","authors":"Alexandre Joushomme, Antoine Désilets, William Champagne, Malihe Hassanzadeh, Gabriel Lemieux, Alice Gravel-Trudeau, Matthieu Lepage, Sabrina Lafrenière, Ulrike Froehlich, Karin List, Pierre-Luc Boudreault, Richard Leduc","doi":"10.1101/2024.08.28.609965","DOIUrl":"https://doi.org/10.1101/2024.08.28.609965","url":null,"abstract":"TMPRSS13, a member of the Type II Transmembrane Serine Proteases (TTSP) family, is involved in cancer progression and in cell entry of respiratory viruses. To date, no inhibitors have been specifically developed toward this protease. In this study, a chemical library of 65 ketobenzothiazole-based peptidomimetic molecules was screened against a proteolytically active form of recombinant TMPRSS13 to identify novel inhibitors. Following an initial round of screening, subsequent synthesis of additional derivatives supported by molecular modelling, uncovered important molecular determinants involved in TMPRSS13 inhibition. One inhibitor, N-0430, achieved low nanomolar affinity towards TMPRSS13 activity in a cellular context. Using a SARS-CoV-2 pseudovirus cell entry model, we further show the ability of N-0430 to block TMPRSS13-dependent entry of the pseudovirus. The identified peptidomimetic inhibitors and the molecular insights of their potency gained from this study will aid in the development of specific TMPRSS13 inhibitors.","PeriodicalId":501518,"journal":{"name":"bioRxiv - Pharmacology and Toxicology","volume":"27 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2024-08-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142184258","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-08-29DOI: 10.1101/2024.08.28.610162
Thomas H. Hampton, Roxanna Barnaby, Carolyn Roche, Amanda Nymon, Kiyoshi Ferreira Fukutani, Todd A. MacKenzie, Bruce A. Stanton
The combination of elexacaftor/tezacaftor/ivacaftor (ETI, Trikafta) reverses the primary defect in Cystic Fibrosis (CF) by improving CFTR mediated Cl- and HCO3- secretion by airway epithelial cells (AEC), leading to improved lung function and less frequent exacerbations and hospitalizations. However, studies have shown that CFTR modulators like ivacaftor, a component of ETI, has numerous effects on CF cells beyond improved CFTR channel function. Because little is known about the effect of ETI on CF AEC gene expression we exposed primary human AEC to ETI for 48 hours and interrogated the transcriptome by RNA-seq and qPCR. ETI increased defensin gene expression (DEFB1) an observation consistent with reports of decreased bacterial burden in the lungs of people with CF (pwCF). ETI also decreased MMP10 and MMP12 gene expression, suggesting that ETI may reduce proteolytic induced lung destruction in CF. ETI also reduced the expression of the stress response gene heme oxygenase (HMOX1). qPCR analysis confirmed DEFB1, HMOX1, MMP10 and MMP12 gene expression results observed by RNA-seq. Gene pathway analysis revealed that ETI decreased inflammatory signaling, cellular proliferation and MHC Class II antigen presentation. Collectively, these findings suggest that the clinical observation that ETI reduces lung infections in pwCF is related in part to drug induced increases in DEFB1, and that ETI may reduce lung damage by reducing MMP10 and MMP12 gene expression, which is predicted to reduce matrix metalloprotease activity. Moreover, pathway analysis also identified several genes responsible for the ETI induced reduction in inflammation observed in people with CF
eplexacaftor/tezacaftor/ivacaftor（ETI，Trikafta）联合疗法通过改善气道上皮细胞（AEC）在CFTR介导下分泌Cl-和HCO3-的情况，逆转了囊性纤维化（CF）的主要缺陷，从而改善了肺功能，减少了病情恶化和住院次数。然而，研究表明，CFTR 调节剂（如 ETI 的成分 ivacaftor）除了改善 CFTR 通道功能外，还对 CF 细胞有许多影响。由于人们对 ETI 对 CF AEC 基因表达的影响知之甚少，因此我们将原代人类 AEC 暴露于 ETI 48 小时，并通过 RNA-seq 和 qPCR 分析转录组。ETI 增加了防御素基因（DEFB1）的表达，这一观察结果与有关减少 CF 患者（pwCF）肺部细菌负担的报道一致。ETI 还降低了 MMP10 和 MMP12 基因的表达，这表明 ETI 可能会减少蛋白水解引起的 CF 患者肺部破坏。qPCR 分析证实了 RNA-seq 观察到的 DEFB1、HMOX1、MMP10 和 MMP12 基因表达结果。基因通路分析表明，ETI 减少了炎症信号传导、细胞增殖和 MHC II 类抗原呈递。总之，这些研究结果表明，临床观察发现 ETI 可减少 pwCF 的肺部感染，这在一定程度上与药物诱导 DEFB1 的增加有关，而且 ETI 可能会通过减少 MMP10 和 MMP12 基因表达来减轻肺部损伤，而 MMP10 和 MMP12 基因表达预计会降低基质金属蛋白酶的活性。此外，通路分析还确定了几个基因，它们对 ETI 诱导的 CF 患者炎症反应的减轻负有责任。
{"title":"Gene expression responses of CF airway epithelial cells exposed to elexacaftor/tezacaftor/ivacaftor (ETI) suggest benefits beyond improved CFTR channel function","authors":"Thomas H. Hampton, Roxanna Barnaby, Carolyn Roche, Amanda Nymon, Kiyoshi Ferreira Fukutani, Todd A. MacKenzie, Bruce A. Stanton","doi":"10.1101/2024.08.28.610162","DOIUrl":"https://doi.org/10.1101/2024.08.28.610162","url":null,"abstract":"The combination of elexacaftor/tezacaftor/ivacaftor (ETI, Trikafta) reverses the primary defect in Cystic Fibrosis (CF) by improving CFTR mediated Cl- and HCO3- secretion by airway epithelial cells (AEC), leading to improved lung function and less frequent exacerbations and hospitalizations. However, studies have shown that CFTR modulators like ivacaftor, a component of ETI, has numerous effects on CF cells beyond improved CFTR channel function. Because little is known about the effect of ETI on CF AEC gene expression we exposed primary human AEC to ETI for 48 hours and interrogated the transcriptome by RNA-seq and qPCR. ETI increased defensin gene expression (DEFB1) an observation consistent with reports of decreased bacterial burden in the lungs of people with CF (pwCF). ETI also decreased MMP10 and MMP12 gene expression, suggesting that ETI may reduce proteolytic induced lung destruction in CF. ETI also reduced the expression of the stress response gene heme oxygenase (HMOX1). qPCR analysis confirmed DEFB1, HMOX1, MMP10 and MMP12 gene expression results observed by RNA-seq. Gene pathway analysis revealed that ETI decreased inflammatory signaling, cellular proliferation and MHC Class II antigen presentation. Collectively, these findings suggest that the clinical observation that ETI reduces lung infections in pwCF is related in part to drug induced increases in DEFB1, and that ETI may reduce lung damage by reducing MMP10 and MMP12 gene expression, which is predicted to reduce matrix metalloprotease activity. Moreover, pathway analysis also identified several genes responsible for the ETI induced reduction in inflammation observed in people with CF","PeriodicalId":501518,"journal":{"name":"bioRxiv - Pharmacology and Toxicology","volume":"23 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2024-08-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142184259","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Background: Sodium-glucose cotransporter 2 inhibitors (SGLT2i) can benefit patients with type 2 diabetes mellitus by reducing hazardous renal outcomes. This study aimed to evaluate whether the combination of sodium-glucose cotransporter 2 inhibitors (SGLT2i) and conventional renin-angiotensin system blockers (RASB) provides a synergistic effect on renal outcomes in patients with type 2 diabetes mellitus, compared to the combination of RASB and dipeptidyl peptidase 4 inhibitors (DPP4i). Methods: This is a retrospective cohort study. The study utilized data from the Taiwan National Health Insurance Research Database (NHIRD), including patients with type 2 diabetes mellitus enrolled between January 1, 2016, and December 31, 2016. Participants were divided into two groups: the case group (n = 3,622) receiving RASB plus SGLT2i and the comparison group (n = 3,622) receiving RASB plus DPP4i. The groups were matched 1:1 based on gender, age, and Charlson comorbidity index. Additionally, TriNetX was used for external validation. Results: Prior to matching, unadjusted hazard ratios (HRs) showed significant differences favoring the SGLT2i group for chronic kidney disease (CKD) (HR: 0.66; 95% CI, 0.58–0.74), advanced kidney failure (HR: 0.64; 95% CI, 0.44–0.93), and initiation of long-term dialysis (HR: 0.61; 95% CI, 0.38–0.97). These differences remained significant post-matching: CKD (HR: 0.74; 95% CI, 0.65–0.84), advanced kidney failure (HR: 0.62; 95% CI, 0.42–0.92), and commencement of long-term dialysis (HR: 0.53; 95% CI, 0.32–0.87). The renal benefits of the combination therapy were consistently observed in the TriNetX dataset. Limitations: NHIRD lacks key clinical factors (e.g., physical features, lab data), potential baseline disparities due to retrospective design, and limited generalizability beyond Taiwanese patients, despite TriNexT validation. Conclusions: In patients with type 2 diabetes mellitus, combination therapy with SGLT2i and RASB yielded better renal outcomes.
{"title":"Renal outcomes of combination therapy with sodium-glucose cotransporter 2 inhibitors plus renin-angiotensin system blockers in patients with type 2 diabetes mellitus: A population-based cohort study","authors":"Ming-Hsien Tsai, Ming-chih Chen, Yen-Chun Huang, Wei-Shan Chang, Kai-Yuan Hsiao, Hung-Hsiang Liou, Yu-Wei Fang","doi":"10.1101/2024.08.28.610101","DOIUrl":"https://doi.org/10.1101/2024.08.28.610101","url":null,"abstract":"Background: Sodium-glucose cotransporter 2 inhibitors (SGLT2i) can benefit patients with type 2 diabetes mellitus by reducing hazardous renal outcomes. This study aimed to evaluate whether the combination of sodium-glucose cotransporter 2 inhibitors (SGLT2i) and conventional renin-angiotensin system blockers (RASB) provides a synergistic effect on renal outcomes in patients with type 2 diabetes mellitus, compared to the combination of RASB and dipeptidyl peptidase 4 inhibitors (DPP4i).\u0000Methods: This is a retrospective cohort study. The study utilized data from the Taiwan National Health Insurance Research Database (NHIRD), including patients with type 2 diabetes mellitus enrolled between January 1, 2016, and December 31, 2016. Participants were divided into two groups: the case group (n = 3,622) receiving RASB plus SGLT2i and the comparison group (n = 3,622) receiving RASB plus DPP4i. The groups were matched 1:1 based on gender, age, and Charlson comorbidity index. Additionally, TriNetX was used for external validation.\u0000Results: Prior to matching, unadjusted hazard ratios (HRs) showed significant differences favoring the SGLT2i group for chronic kidney disease (CKD) (HR: 0.66; 95% CI, 0.58–0.74), advanced kidney failure (HR: 0.64; 95% CI, 0.44–0.93), and initiation of long-term dialysis (HR: 0.61; 95% CI, 0.38–0.97). These differences remained significant post-matching: CKD (HR: 0.74; 95% CI, 0.65–0.84), advanced kidney failure (HR: 0.62; 95% CI, 0.42–0.92), and commencement of long-term dialysis (HR: 0.53; 95% CI, 0.32–0.87). The renal benefits of the combination therapy were consistently observed in the TriNetX dataset.\u0000Limitations: NHIRD lacks key clinical factors (e.g., physical features, lab data), potential baseline disparities due to retrospective design, and limited generalizability beyond Taiwanese patients, despite TriNexT validation.\u0000Conclusions: In patients with type 2 diabetes mellitus, combination therapy with SGLT2i and RASB yielded better renal outcomes.","PeriodicalId":501518,"journal":{"name":"bioRxiv - Pharmacology and Toxicology","volume":"62 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2024-08-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142223938","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
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