Pub Date : 2020-04-01DOI: 10.1096/fasebj.2020.34.s1.09484
T. Stobdan, Huiwen W. Zhao, Dan Zhou, Arya Iranmehr, Lu-Bo Ying, V. Bafna, G. Haddad
The origin and evolution of life forms depended by and large on O2 availability over thousands of years. The mechanism of the adaptation process under strong selection pressure, however, is subject to some debate. For instance, it could be mediated by extant mutations or cryptic genetic variation that yield a fitness advantage in the new environment, or by mutations that arise de novo. For sexually reproducing organisms, multiple favored variants can also be acquired on a single haplotype via recombination to alleviate the effect of clonal interference and accelerate adaptation. It is however difficult to directly observe evolution in action. In the current study, we have identified the genomic intervals under selection in the high and low O2‐adapted Drosophila melanogaster. The adapted Drosophila melanogaster constitutes flies that could complete its life cycle in 90% and 4% O2 environments respectively, which otherwise is lethal for the naive flies. We used UAS‐RNAi x daGal4 system to functionally validate the role of specific genes of the top selected genomic interval. The top interval selected under high O2 environment consisted of 32 genes that includes four non‐protein‐coding genes. RNAi lines were available for 15 genes (15/28) that were evenly scattered across this interval. The high O2 tolerance assay indicate >98% eclosion rate in the adapted flies. Four genes i.e., CG15472, Klf15, CG2861 and CG42594, depicts a relatively higher number of pupation compared to the controls (12±0.6 to 14±2.5 versus <7 pupae counts per vial. The average pupae eclosed was higher for two of the genes, CG15472 and CG42594, although these numbers were significantly lower than the adapted flies. The percentage eclosion rate was significantly higher for CG15472 ‐(91.4 ±16.7%), which is close to the 98% eclosion seen in the adapted flies. Similarly, we have identified intervals under selection in the low O2 adapted flies. Remarkably, there was higher proportion of genes from the Notch signaling pathway in these selected intervals. Our results so far indicate that CG15472 from the top selected interval is one of the important gene playing critical role in the successful adaptation of flies to high O2 environment. Similarly, the Notch signaling pathway has an important role in the successful adaptation of flies to extremely low O2.
{"title":"Identification of Candidate Genes Involved in the Survival of Drosophila in Extreme O2 Environments","authors":"T. Stobdan, Huiwen W. Zhao, Dan Zhou, Arya Iranmehr, Lu-Bo Ying, V. Bafna, G. Haddad","doi":"10.1096/fasebj.2020.34.s1.09484","DOIUrl":"https://doi.org/10.1096/fasebj.2020.34.s1.09484","url":null,"abstract":"The origin and evolution of life forms depended by and large on O2 availability over thousands of years. The mechanism of the adaptation process under strong selection pressure, however, is subject to some debate. For instance, it could be mediated by extant mutations or cryptic genetic variation that yield a fitness advantage in the new environment, or by mutations that arise de novo. For sexually reproducing organisms, multiple favored variants can also be acquired on a single haplotype via recombination to alleviate the effect of clonal interference and accelerate adaptation. It is however difficult to directly observe evolution in action. In the current study, we have identified the genomic intervals under selection in the high and low O2‐adapted Drosophila melanogaster. The adapted Drosophila melanogaster constitutes flies that could complete its life cycle in 90% and 4% O2 environments respectively, which otherwise is lethal for the naive flies. We used UAS‐RNAi x daGal4 system to functionally validate the role of specific genes of the top selected genomic interval. The top interval selected under high O2 environment consisted of 32 genes that includes four non‐protein‐coding genes. RNAi lines were available for 15 genes (15/28) that were evenly scattered across this interval. The high O2 tolerance assay indicate >98% eclosion rate in the adapted flies. Four genes i.e., CG15472, Klf15, CG2861 and CG42594, depicts a relatively higher number of pupation compared to the controls (12±0.6 to 14±2.5 versus <7 pupae counts per vial. The average pupae eclosed was higher for two of the genes, CG15472 and CG42594, although these numbers were significantly lower than the adapted flies. The percentage eclosion rate was significantly higher for CG15472 ‐(91.4 ±16.7%), which is close to the 98% eclosion seen in the adapted flies. Similarly, we have identified intervals under selection in the low O2 adapted flies. Remarkably, there was higher proportion of genes from the Notch signaling pathway in these selected intervals. Our results so far indicate that CG15472 from the top selected interval is one of the important gene playing critical role in the successful adaptation of flies to high O2 environment. Similarly, the Notch signaling pathway has an important role in the successful adaptation of flies to extremely low O2.","PeriodicalId":22447,"journal":{"name":"The FASEB Journal","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2020-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141219338","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 : 2020-04-01DOI: 10.1096/fasebj.2020.34.s1.05439
O. Arishe, Jaine Mckenzie, F. Priviero, A. Ebeigbe, C. Webb
Aging is a major non‐modifiable risk factor for hypertension. Structural, functional and mechanical changes occur with aging. These changes are similar to those seen in hypertension in the vasculature. Also, aging increases the vascular dysfunction that occurs in hypertension. L‐arginase is a ureohydrolase enzyme that converts L‐arginine to L‐ornithine and urea. Arginase action reduces substrate (L‐arginine) availability for the formation of nitric oxide (NO). This reduces the level of NO and leads to reduced vasodilation and ultimately, vascular dysfunction.
{"title":"L‐Arginase induces Vascular Dysfunction in Old Spontaneously Hypertensive Rats","authors":"O. Arishe, Jaine Mckenzie, F. Priviero, A. Ebeigbe, C. Webb","doi":"10.1096/fasebj.2020.34.s1.05439","DOIUrl":"https://doi.org/10.1096/fasebj.2020.34.s1.05439","url":null,"abstract":"Aging is a major non‐modifiable risk factor for hypertension. Structural, functional and mechanical changes occur with aging. These changes are similar to those seen in hypertension in the vasculature. Also, aging increases the vascular dysfunction that occurs in hypertension. L‐arginase is a ureohydrolase enzyme that converts L‐arginine to L‐ornithine and urea. Arginase action reduces substrate (L‐arginine) availability for the formation of nitric oxide (NO). This reduces the level of NO and leads to reduced vasodilation and ultimately, vascular dysfunction.","PeriodicalId":22447,"journal":{"name":"The FASEB Journal","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2020-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141219497","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 : 2020-04-01DOI: 10.1096/fasebj.2020.34.s1.02759
lunawati lo bennett
The incidence of breast cancer mortality is one of the major challenges in the scientific community. Despite advances in diagnosis and treatment options, death among women worldwide are still high.
乳腺癌死亡率是科学界面临的主要挑战之一。尽管诊断和治疗方法不断进步，但全球妇女的死亡率仍然很高。
{"title":"Synergistic effect of sorafenib and resveratrol in human breast cancer MDA‐MB‐231 cells","authors":"lunawati lo bennett","doi":"10.1096/fasebj.2020.34.s1.02759","DOIUrl":"https://doi.org/10.1096/fasebj.2020.34.s1.02759","url":null,"abstract":"The incidence of breast cancer mortality is one of the major challenges in the scientific community. Despite advances in diagnosis and treatment options, death among women worldwide are still high.","PeriodicalId":22447,"journal":{"name":"The FASEB Journal","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2020-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141218089","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 : 2020-04-01DOI: 10.1096/fasebj.2020.34.s1.05640
T. Kiss, S. Tarantini, A. Yabluchanskiy, Tamás Csípő, Priya Balasubramanian, Á. Lipecz, D. Reglodi, E. Farkas, Xin A. Zhang, F. Bari, A. Csiszar, Z. Ungvari
Age‐related impairment of angiogenesis likely has a critical role in cerebromicrovascular rarefaction and development of vascular cognitive impairment and dementia (VCID) in the elderly. Recently, we demonstrated that aging is associated with NAD+ depletion in the vasculature and that administration of NAD+ precursors exerts potent anti‐aging vascular effects, rescuing endothelium‐mediated vasodilation in the cerebral circulation and improving cerebral blood supply. The present study was designed to elucidate how treatment with nicotinamide mononucleotide (NMN), a key NAD+ intermediate, impacts age‐related impairment of endothelial angiogenic processes. Using cerebromicrovascular endothelial cells (CMVECs) isolated from young and aged F344xBN rats, we demonstrated that compared with young cells, aged CMVECs exhibit impaired proliferation, cellular migration (measured by a wound‐healing assay using electric cell‐substrate impedance sensing [ECIS] technology), impaired ability to form capillary‐like structures, and increased oxidative stress. NMN treatment in aged CMVECs significantly improved angiogenic processes and attenuated H2O2 production. We also found that pre‐treatment with EX‐527, a pharmacological inhibitor of SIRT1, prevented NMN‐mediated restoration of angiogenic processes in aged CMVECs. Collectively, we find that normal cellular NAD+ levels are essential for normal endothelial angiogenic processes, suggesting that age‐related cellular NAD+ depletion and consequential SIRT1 dysregulation may be a potentially reversible mechanism underlying impaired angiogenesis and cerebromicrovascular rarefaction in aging. We recommend that pro‐angiogenic effects of NAD+ boosters should be considered in both preclinical and clinical studies.
{"title":"Nicotinamide mononucleotide (NMN) treatment attenuates oxidative stress and rescues angiogenic capacity in aged cerebromicrovascular endothelial cells: a potential mechanism for the prevention of vascular cognitive impairment.","authors":"T. Kiss, S. Tarantini, A. Yabluchanskiy, Tamás Csípő, Priya Balasubramanian, Á. Lipecz, D. Reglodi, E. Farkas, Xin A. Zhang, F. Bari, A. Csiszar, Z. Ungvari","doi":"10.1096/fasebj.2020.34.s1.05640","DOIUrl":"https://doi.org/10.1096/fasebj.2020.34.s1.05640","url":null,"abstract":"Age‐related impairment of angiogenesis likely has a critical role in cerebromicrovascular rarefaction and development of vascular cognitive impairment and dementia (VCID) in the elderly. Recently, we demonstrated that aging is associated with NAD+ depletion in the vasculature and that administration of NAD+ precursors exerts potent anti‐aging vascular effects, rescuing endothelium‐mediated vasodilation in the cerebral circulation and improving cerebral blood supply. The present study was designed to elucidate how treatment with nicotinamide mononucleotide (NMN), a key NAD+ intermediate, impacts age‐related impairment of endothelial angiogenic processes. Using cerebromicrovascular endothelial cells (CMVECs) isolated from young and aged F344xBN rats, we demonstrated that compared with young cells, aged CMVECs exhibit impaired proliferation, cellular migration (measured by a wound‐healing assay using electric cell‐substrate impedance sensing [ECIS] technology), impaired ability to form capillary‐like structures, and increased oxidative stress. NMN treatment in aged CMVECs significantly improved angiogenic processes and attenuated H2O2 production. We also found that pre‐treatment with EX‐527, a pharmacological inhibitor of SIRT1, prevented NMN‐mediated restoration of angiogenic processes in aged CMVECs. Collectively, we find that normal cellular NAD+ levels are essential for normal endothelial angiogenic processes, suggesting that age‐related cellular NAD+ depletion and consequential SIRT1 dysregulation may be a potentially reversible mechanism underlying impaired angiogenesis and cerebromicrovascular rarefaction in aging. We recommend that pro‐angiogenic effects of NAD+ boosters should be considered in both preclinical and clinical studies.","PeriodicalId":22447,"journal":{"name":"The FASEB Journal","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2020-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141218776","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 : 2020-04-01DOI: 10.1096/fasebj.2020.34.s1.04350
Laura C Miller Conrad
Pseudomonas aeruginosa is an emerging public health threat. A common Gram‐negative source of secondary infections, it has few effective treatment options, which are currently being undermined by the rise of resistance. New therapies are urgently needed. We had previously pursued an antivirulence strategy to block the production of pyocyanin, a redox‐active virulence factor. A photoaffinity analog of an antipyocyanin compound was developed to interrogate the inhibitor’s molecular mechanism of action. In the process, antibiotic adjuvant activity was suggested by the proteomics results. Using susceptibility assays, we found that these compounds amplify the bactericidal activity of colistin, a well‐characterized antibiotic, suggesting they may represent a first‐in‐class antibiotic adjuvant therapy. Analogs have the potential to not only widen the therapeutic index of cationic antibiotic peptides like colistin, but to be effective against colistin‐resistant strains. Multidrug‐resistant infections with P. aeruginosa are currently treated with colistin and the related polymyxin b, however, resistance to these drugs is also increasingly encountered. The adjuvants have the potential to preserve the life‐saving therapy of colistin when used in a combination therapy.
铜绿假单胞菌是一种新出现的公共卫生威胁。铜绿假单胞菌是一种常见的革兰氏阴性菌，可引起继发性感染，但它的有效治疗方案却很少，目前抗药性的增加又削弱了它的治疗效果。我们迫切需要新的疗法。此前，我们一直在寻求一种抗病毒策略，以阻断具有氧化还原作用的毒力因子--焦花青素的产生。我们开发了一种抗焦花青素化合物的光亲和类似物，以探究抑制剂的分子作用机制。在此过程中，蛋白质组学结果表明了抗生素的辅助活性。通过药敏试验，我们发现这些化合物放大了可乐定（一种特性良好的抗生素）的杀菌活性，这表明它们可能代表了第一类抗生素辅助疗法。类似物不仅有可能扩大阳离子抗生素肽（如秋水仙碱）的治疗指数，还能有效抑制耐秋水仙碱菌株。铜绿假单胞菌对多种药物产生耐药性，目前主要使用秋水仙素和相关的多粘菌素 b 进行治疗，但对这些药物产生耐药性的情况也越来越多。辅助剂在联合疗法中使用时，有可能保留可乐定的救命疗法。
{"title":"Photoaffinity Approach Reveals Antibiotic Adjuvant Activity toward Pseudomonas aeruginosa","authors":"Laura C Miller Conrad","doi":"10.1096/fasebj.2020.34.s1.04350","DOIUrl":"https://doi.org/10.1096/fasebj.2020.34.s1.04350","url":null,"abstract":"Pseudomonas aeruginosa is an emerging public health threat. A common Gram‐negative source of secondary infections, it has few effective treatment options, which are currently being undermined by the rise of resistance. New therapies are urgently needed. We had previously pursued an antivirulence strategy to block the production of pyocyanin, a redox‐active virulence factor. A photoaffinity analog of an antipyocyanin compound was developed to interrogate the inhibitor’s molecular mechanism of action. In the process, antibiotic adjuvant activity was suggested by the proteomics results. Using susceptibility assays, we found that these compounds amplify the bactericidal activity of colistin, a well‐characterized antibiotic, suggesting they may represent a first‐in‐class antibiotic adjuvant therapy. Analogs have the potential to not only widen the therapeutic index of cationic antibiotic peptides like colistin, but to be effective against colistin‐resistant strains. Multidrug‐resistant infections with P. aeruginosa are currently treated with colistin and the related polymyxin b, however, resistance to these drugs is also increasingly encountered. The adjuvants have the potential to preserve the life‐saving therapy of colistin when used in a combination therapy.","PeriodicalId":22447,"journal":{"name":"The FASEB Journal","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2020-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141218760","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 : 2020-04-01DOI: 10.1096/fasebj.2020.34.s1.02759
lunawati lo bennett
The incidence of breast cancer mortality is one of the major challenges in the scientific community. Despite advances in diagnosis and treatment options, death among women worldwide are still high.
乳腺癌死亡率是科学界面临的主要挑战之一。尽管诊断和治疗方法不断进步，但全球妇女的死亡率仍然很高。
{"title":"Synergistic effect of sorafenib and resveratrol in human breast cancer MDA‐MB‐231 cells","authors":"lunawati lo bennett","doi":"10.1096/fasebj.2020.34.s1.02759","DOIUrl":"https://doi.org/10.1096/fasebj.2020.34.s1.02759","url":null,"abstract":"The incidence of breast cancer mortality is one of the major challenges in the scientific community. Despite advances in diagnosis and treatment options, death among women worldwide are still high.","PeriodicalId":22447,"journal":{"name":"The FASEB Journal","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2020-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141217442","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 : 2020-04-01DOI: 10.1096/fasebj.2020.34.s1.09373
A. Pal, A. Al-Shaer, William Guesdon, Maria.J. Torres, Michael Armstrong, K. Quinn, N. Reisdorph, P. D. Neufer, S. Shaikh
Eicosapentaenoic acid (EPA) is a n‐3 polyunsaturated fatty acid that is poorly consumed in the western diet. Increased EPA consumption has been reported to improve glucose and insulin homeostasis in rodent models. We demonstrate that administration of pure EPA ethyl esters to C57BL/6J male mice improved obesity‐induced glucose intolerance, hyperinsulinemia, and hyperglycemia. Analyses of National Health and Nutrition Examination Survey data also revealed fasting glucose levels of obese adults to be inversely related with EPA intake in a sex‐dependent manner. To investigate potential mechanisms by which EPA improved glucose homeostasis, we compared rodent models consuming a control and a high fat diet in the presence or absence of EPA. 16S rRNA sequencing demonstrated that EPA supplementation was not associated with an improvement in the murine gut microbiome composition. Subsequent untargeted and targeted mass spectrometry analyses revealed distinct modifications to the lipidome. Notably, EPA dramatically enhanced the levels of 18‐hydroxyeicosapentaenoic acid (18‐HEPE) in the heart, white adipose tissue, and liver. Therefore, we investigated if administration of the downstream bioactive metabolite of 18‐HEPE, resolvin E1 (RvE1) could improve hyperinsulinemia and hyperglycemia. We also determined if these effects were mediated through the RvE1 receptor ERV1/ChemR23. RvE1 administration to obese mice mitigated hyperinsulinemia and hyperglycemia in a manner that was dependent on ERV1/ChemR23, as revealed with ERV1/ChemR23 knockout mice. Finally, we assessed if the host genome altered the metabolic outcomes of RvE1. Secondary SNP analyses revealed extensive genetic variation in human EPA and RvE1 metabolizing genes. RvE1’s effects on fasting insulin and glucose were divergent in diversity outbred mice that model human genetic variation. In conclusion, increased intake of EPA ethyl esters prevent obesity‐induced metabolic impairments through RvE1 binding to ChemR23. The data also underscore the critical need for precision prevention studies that account for host‐genetic variants in the EPA‐RvE1 axis.
{"title":"Eicosapentaenoic Acid Prevents Obesity‐induced Metabolic Impairments through The Host‐genetic Dependent Effects of Resolvin E1","authors":"A. Pal, A. Al-Shaer, William Guesdon, Maria.J. Torres, Michael Armstrong, K. Quinn, N. Reisdorph, P. D. Neufer, S. Shaikh","doi":"10.1096/fasebj.2020.34.s1.09373","DOIUrl":"https://doi.org/10.1096/fasebj.2020.34.s1.09373","url":null,"abstract":"Eicosapentaenoic acid (EPA) is a n‐3 polyunsaturated fatty acid that is poorly consumed in the western diet. Increased EPA consumption has been reported to improve glucose and insulin homeostasis in rodent models. We demonstrate that administration of pure EPA ethyl esters to C57BL/6J male mice improved obesity‐induced glucose intolerance, hyperinsulinemia, and hyperglycemia. Analyses of National Health and Nutrition Examination Survey data also revealed fasting glucose levels of obese adults to be inversely related with EPA intake in a sex‐dependent manner. To investigate potential mechanisms by which EPA improved glucose homeostasis, we compared rodent models consuming a control and a high fat diet in the presence or absence of EPA. 16S rRNA sequencing demonstrated that EPA supplementation was not associated with an improvement in the murine gut microbiome composition. Subsequent untargeted and targeted mass spectrometry analyses revealed distinct modifications to the lipidome. Notably, EPA dramatically enhanced the levels of 18‐hydroxyeicosapentaenoic acid (18‐HEPE) in the heart, white adipose tissue, and liver. Therefore, we investigated if administration of the downstream bioactive metabolite of 18‐HEPE, resolvin E1 (RvE1) could improve hyperinsulinemia and hyperglycemia. We also determined if these effects were mediated through the RvE1 receptor ERV1/ChemR23. RvE1 administration to obese mice mitigated hyperinsulinemia and hyperglycemia in a manner that was dependent on ERV1/ChemR23, as revealed with ERV1/ChemR23 knockout mice. Finally, we assessed if the host genome altered the metabolic outcomes of RvE1. Secondary SNP analyses revealed extensive genetic variation in human EPA and RvE1 metabolizing genes. RvE1’s effects on fasting insulin and glucose were divergent in diversity outbred mice that model human genetic variation. In conclusion, increased intake of EPA ethyl esters prevent obesity‐induced metabolic impairments through RvE1 binding to ChemR23. The data also underscore the critical need for precision prevention studies that account for host‐genetic variants in the EPA‐RvE1 axis.","PeriodicalId":22447,"journal":{"name":"The FASEB Journal","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2020-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141217803","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 : 2020-04-01DOI: 10.1096/fasebj.2020.34.s1.06068
Thiago Bruder do Nascimento, Eric J. Belin de Chantemèle
Thanks to the advent of highly active antiretroviral therapy (HAART), the life of people living with HIV has been extended to close to that of healthy people. However, patients with HIV are now presenting with accelerated development of cardiovascular disease (CVD) and CVD is currently the leading cause of death in patients with HIV on HAART. Herein, we tested the hypothesis that the protease inhibitor ritonavir contributes to HAART‐associated CVD by impairing vascular function via C‐C chemokine receptor type 5 (CCR5)‐ and reactive oxygen species‐mediated mechanisms. We treated 8–10‐week‐old male C57BL6/J with ritonavir for 4 consecutive weeks (5mg/kg/day, i.p). At the end of the treatment, blood was collected and aortas excised to measure H2O2 via Amplex Red and vascular function via wire myography. Aorta from ritonavir treated animals exhibited higher H2O2 levels, impaired relaxation to acetylcholine (Ach) and increased contractility to phenylephrine (Phe). Nox1 inhibition with GKT771, (10μM) fully restored endothelial function in rings from ritonavir treated animals. Ritonavir treatment also elevated RANTES (CCR5 ligand) plasma levels, as well as CCR5, NADPH oxidase 1 (Nox1) and NoxA1 transcript levels in the aorta. No difference was observed for others oxidases, such as Nox2 and Nox4. Repetition of the ritonavir treatment in CCR5 deficient mice revealed that CCR5 deficiency protects vessels from ritonavir induced‐endothelial dysfunction and upregulation of Nox1 and NoxA1 enzymes. To analyze the direct effects of RANTES on the vasculature, thoracic aorta, endothelial, and vascular smooth muscle cells were exposed to RANTES in different times (20ng/mL). RANTES elevated endothelial NOX1 expression and impaired ACh‐mediated relaxation. Concomitantly, Nox1 inhibition prevented RANTES‐mediated endothelial dysfunction. In addition, RANTES time‐dependently stimulated rat aortic smooth muscle cell proliferation, as measured by Ki67 expression, which was partially reverted by Nox1 inhibition. Our data suggest that the protease inhibitor ritonavir contributes to vascular dysfunction and cardiovascular disease by increasing RANTES‐mediated CCR5 activation ultimately leading to increases in Nox1 expression, ROS production, endothelial dysfunction and vascular smooth muscle cell proliferation. Taken together, these findings provide potential explanation for the increased risk of cardiovascular disease developed by patients living with HIV and present CCR5 as a potential target for treatment.
{"title":"The protease inhibition with Ritonavir impairs endothelial function and promotes vascular smooth cell proliferation via RANTES/C‐C chemokine receptor type 5 and Nox1‐derived reactive oxygen species pathway.","authors":"Thiago Bruder do Nascimento, Eric J. Belin de Chantemèle","doi":"10.1096/fasebj.2020.34.s1.06068","DOIUrl":"https://doi.org/10.1096/fasebj.2020.34.s1.06068","url":null,"abstract":"Thanks to the advent of highly active antiretroviral therapy (HAART), the life of people living with HIV has been extended to close to that of healthy people. However, patients with HIV are now presenting with accelerated development of cardiovascular disease (CVD) and CVD is currently the leading cause of death in patients with HIV on HAART. Herein, we tested the hypothesis that the protease inhibitor ritonavir contributes to HAART‐associated CVD by impairing vascular function via C‐C chemokine receptor type 5 (CCR5)‐ and reactive oxygen species‐mediated mechanisms. We treated 8–10‐week‐old male C57BL6/J with ritonavir for 4 consecutive weeks (5mg/kg/day, i.p). At the end of the treatment, blood was collected and aortas excised to measure H2O2 via Amplex Red and vascular function via wire myography. Aorta from ritonavir treated animals exhibited higher H2O2 levels, impaired relaxation to acetylcholine (Ach) and increased contractility to phenylephrine (Phe). Nox1 inhibition with GKT771, (10μM) fully restored endothelial function in rings from ritonavir treated animals. Ritonavir treatment also elevated RANTES (CCR5 ligand) plasma levels, as well as CCR5, NADPH oxidase 1 (Nox1) and NoxA1 transcript levels in the aorta. No difference was observed for others oxidases, such as Nox2 and Nox4. Repetition of the ritonavir treatment in CCR5 deficient mice revealed that CCR5 deficiency protects vessels from ritonavir induced‐endothelial dysfunction and upregulation of Nox1 and NoxA1 enzymes. To analyze the direct effects of RANTES on the vasculature, thoracic aorta, endothelial, and vascular smooth muscle cells were exposed to RANTES in different times (20ng/mL). RANTES elevated endothelial NOX1 expression and impaired ACh‐mediated relaxation. Concomitantly, Nox1 inhibition prevented RANTES‐mediated endothelial dysfunction. In addition, RANTES time‐dependently stimulated rat aortic smooth muscle cell proliferation, as measured by Ki67 expression, which was partially reverted by Nox1 inhibition. Our data suggest that the protease inhibitor ritonavir contributes to vascular dysfunction and cardiovascular disease by increasing RANTES‐mediated CCR5 activation ultimately leading to increases in Nox1 expression, ROS production, endothelial dysfunction and vascular smooth muscle cell proliferation. Taken together, these findings provide potential explanation for the increased risk of cardiovascular disease developed by patients living with HIV and present CCR5 as a potential target for treatment.","PeriodicalId":22447,"journal":{"name":"The FASEB Journal","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2020-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141217842","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 : 2020-04-01DOI: 10.1096/fasebj.2020.34.s1.05732
J. Jilek, Erica L. Toth, Kayla L. Frost, Kevyn A. Jacobus, Wenxi He, Michael J Goedken, N. Cherrington
Cisplatin is an alkylating antineoplastic agent that is indicated for the treatment of solid malignancies. Cisplatin is preferentially eliminated from systemic circulation via tubular secretion, whereby it exhibits dose‐limiting nephrotoxicity. Interindividual variability in xenobiotic transporter expression is a known contributor to differential cisplatin toxicity and efficacy, and may be the result of genetic, environmental, and pathological contributions. In this study, we aimed to determine if nonalcoholic steatohepatitis (NASH) alters cisplatin pharmacokinetics and if this change elicits differential nephrotoxicity. Sprague Dawley rats fed a control or methionine and choline deficient (MCD) diet to model NASH were given a single bolus dose of cisplatin and sacrificed after 72 h. The MCD diet resulted in a NASH hepatic phenotype that remained unchanged following cisplatin exposure. Drug‐naïve NASH rats also displayed no evidence of differential renal pathology relative to control rats. However, renal necrosis and inflammation were reduced in NASH by 40 and 63% following cisplatin treatment, respectively, relative to healthy controls. Furthermore, kidney weights of cisplatin‐treated control rats were increased by 31%, compared to an 18% increase in NASH. Plasma cisplatin clearance was reduced from 6.78 (control) to 4.04 mL/min in NASH, and cisplatin plasma AUC was significantly increased by 44% in NASH, relative to control. Cumulative urinary elimination of cisplatin was decreased from 73 to 34% of total dose and renal clearance was reduced from 4.64 to 1.49 mL/min in NASH, compared to control. Subsequently, renal intracellular accumulation of cisplatin after 6 h was reduced by 34% in NASH, relative to control. Supporting these findings, expression of proximal tubule cisplatin uptake transporters, Ctr1 and Ctr2, were reduced by 24 and 64%, respectively compared to healthy control rats, whereas expression of Oct1, Oct2, and Oct3 were unchanged. Interestingly, expression of cisplatin efflux transporters Mate1 and Atp7a were reduced by 52 and 31%, respectively, in NASH compared to control. Taken together, these data suggest that NASH alters renal uptake and efflux transporter expression, thereby attenuating cisplatin uptake and clearance in the kidney with a corresponding reduction in renal cell exposure and nephrotoxicity during NASH. As such, this study demonstrates that NASH can influence pharmacokinetics of drugs cleared by renal elimination, which may contribute to adverse drug reactions.
{"title":"Altered Cisplatin Pharmacokinetics during Nonalcoholic Steatohepatitis Contributes to Reduced Nephrotoxicity","authors":"J. Jilek, Erica L. Toth, Kayla L. Frost, Kevyn A. Jacobus, Wenxi He, Michael J Goedken, N. Cherrington","doi":"10.1096/fasebj.2020.34.s1.05732","DOIUrl":"https://doi.org/10.1096/fasebj.2020.34.s1.05732","url":null,"abstract":"Cisplatin is an alkylating antineoplastic agent that is indicated for the treatment of solid malignancies. Cisplatin is preferentially eliminated from systemic circulation via tubular secretion, whereby it exhibits dose‐limiting nephrotoxicity. Interindividual variability in xenobiotic transporter expression is a known contributor to differential cisplatin toxicity and efficacy, and may be the result of genetic, environmental, and pathological contributions. In this study, we aimed to determine if nonalcoholic steatohepatitis (NASH) alters cisplatin pharmacokinetics and if this change elicits differential nephrotoxicity. Sprague Dawley rats fed a control or methionine and choline deficient (MCD) diet to model NASH were given a single bolus dose of cisplatin and sacrificed after 72 h. The MCD diet resulted in a NASH hepatic phenotype that remained unchanged following cisplatin exposure. Drug‐naïve NASH rats also displayed no evidence of differential renal pathology relative to control rats. However, renal necrosis and inflammation were reduced in NASH by 40 and 63% following cisplatin treatment, respectively, relative to healthy controls. Furthermore, kidney weights of cisplatin‐treated control rats were increased by 31%, compared to an 18% increase in NASH. Plasma cisplatin clearance was reduced from 6.78 (control) to 4.04 mL/min in NASH, and cisplatin plasma AUC was significantly increased by 44% in NASH, relative to control. Cumulative urinary elimination of cisplatin was decreased from 73 to 34% of total dose and renal clearance was reduced from 4.64 to 1.49 mL/min in NASH, compared to control. Subsequently, renal intracellular accumulation of cisplatin after 6 h was reduced by 34% in NASH, relative to control. Supporting these findings, expression of proximal tubule cisplatin uptake transporters, Ctr1 and Ctr2, were reduced by 24 and 64%, respectively compared to healthy control rats, whereas expression of Oct1, Oct2, and Oct3 were unchanged. Interestingly, expression of cisplatin efflux transporters Mate1 and Atp7a were reduced by 52 and 31%, respectively, in NASH compared to control. Taken together, these data suggest that NASH alters renal uptake and efflux transporter expression, thereby attenuating cisplatin uptake and clearance in the kidney with a corresponding reduction in renal cell exposure and nephrotoxicity during NASH. As such, this study demonstrates that NASH can influence pharmacokinetics of drugs cleared by renal elimination, which may contribute to adverse drug reactions.","PeriodicalId":22447,"journal":{"name":"The FASEB Journal","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2020-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141218463","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 : 2020-04-01DOI: 10.1096/fasebj.2020.34.s1.06068
Thiago Bruder do Nascimento, Eric J. Belin de Chantemèle
Thanks to the advent of highly active antiretroviral therapy (HAART), the life of people living with HIV has been extended to close to that of healthy people. However, patients with HIV are now presenting with accelerated development of cardiovascular disease (CVD) and CVD is currently the leading cause of death in patients with HIV on HAART. Herein, we tested the hypothesis that the protease inhibitor ritonavir contributes to HAART‐associated CVD by impairing vascular function via C‐C chemokine receptor type 5 (CCR5)‐ and reactive oxygen species‐mediated mechanisms. We treated 8–10‐week‐old male C57BL6/J with ritonavir for 4 consecutive weeks (5mg/kg/day, i.p). At the end of the treatment, blood was collected and aortas excised to measure H2O2 via Amplex Red and vascular function via wire myography. Aorta from ritonavir treated animals exhibited higher H2O2 levels, impaired relaxation to acetylcholine (Ach) and increased contractility to phenylephrine (Phe). Nox1 inhibition with GKT771, (10μM) fully restored endothelial function in rings from ritonavir treated animals. Ritonavir treatment also elevated RANTES (CCR5 ligand) plasma levels, as well as CCR5, NADPH oxidase 1 (Nox1) and NoxA1 transcript levels in the aorta. No difference was observed for others oxidases, such as Nox2 and Nox4. Repetition of the ritonavir treatment in CCR5 deficient mice revealed that CCR5 deficiency protects vessels from ritonavir induced‐endothelial dysfunction and upregulation of Nox1 and NoxA1 enzymes. To analyze the direct effects of RANTES on the vasculature, thoracic aorta, endothelial, and vascular smooth muscle cells were exposed to RANTES in different times (20ng/mL). RANTES elevated endothelial NOX1 expression and impaired ACh‐mediated relaxation. Concomitantly, Nox1 inhibition prevented RANTES‐mediated endothelial dysfunction. In addition, RANTES time‐dependently stimulated rat aortic smooth muscle cell proliferation, as measured by Ki67 expression, which was partially reverted by Nox1 inhibition. Our data suggest that the protease inhibitor ritonavir contributes to vascular dysfunction and cardiovascular disease by increasing RANTES‐mediated CCR5 activation ultimately leading to increases in Nox1 expression, ROS production, endothelial dysfunction and vascular smooth muscle cell proliferation. Taken together, these findings provide potential explanation for the increased risk of cardiovascular disease developed by patients living with HIV and present CCR5 as a potential target for treatment.
{"title":"The protease inhibition with Ritonavir impairs endothelial function and promotes vascular smooth cell proliferation via RANTES/C‐C chemokine receptor type 5 and Nox1‐derived reactive oxygen species pathway.","authors":"Thiago Bruder do Nascimento, Eric J. Belin de Chantemèle","doi":"10.1096/fasebj.2020.34.s1.06068","DOIUrl":"https://doi.org/10.1096/fasebj.2020.34.s1.06068","url":null,"abstract":"Thanks to the advent of highly active antiretroviral therapy (HAART), the life of people living with HIV has been extended to close to that of healthy people. However, patients with HIV are now presenting with accelerated development of cardiovascular disease (CVD) and CVD is currently the leading cause of death in patients with HIV on HAART. Herein, we tested the hypothesis that the protease inhibitor ritonavir contributes to HAART‐associated CVD by impairing vascular function via C‐C chemokine receptor type 5 (CCR5)‐ and reactive oxygen species‐mediated mechanisms. We treated 8–10‐week‐old male C57BL6/J with ritonavir for 4 consecutive weeks (5mg/kg/day, i.p). At the end of the treatment, blood was collected and aortas excised to measure H2O2 via Amplex Red and vascular function via wire myography. Aorta from ritonavir treated animals exhibited higher H2O2 levels, impaired relaxation to acetylcholine (Ach) and increased contractility to phenylephrine (Phe). Nox1 inhibition with GKT771, (10μM) fully restored endothelial function in rings from ritonavir treated animals. Ritonavir treatment also elevated RANTES (CCR5 ligand) plasma levels, as well as CCR5, NADPH oxidase 1 (Nox1) and NoxA1 transcript levels in the aorta. No difference was observed for others oxidases, such as Nox2 and Nox4. Repetition of the ritonavir treatment in CCR5 deficient mice revealed that CCR5 deficiency protects vessels from ritonavir induced‐endothelial dysfunction and upregulation of Nox1 and NoxA1 enzymes. To analyze the direct effects of RANTES on the vasculature, thoracic aorta, endothelial, and vascular smooth muscle cells were exposed to RANTES in different times (20ng/mL). RANTES elevated endothelial NOX1 expression and impaired ACh‐mediated relaxation. Concomitantly, Nox1 inhibition prevented RANTES‐mediated endothelial dysfunction. In addition, RANTES time‐dependently stimulated rat aortic smooth muscle cell proliferation, as measured by Ki67 expression, which was partially reverted by Nox1 inhibition. Our data suggest that the protease inhibitor ritonavir contributes to vascular dysfunction and cardiovascular disease by increasing RANTES‐mediated CCR5 activation ultimately leading to increases in Nox1 expression, ROS production, endothelial dysfunction and vascular smooth muscle cell proliferation. Taken together, these findings provide potential explanation for the increased risk of cardiovascular disease developed by patients living with HIV and present CCR5 as a potential target for treatment.","PeriodicalId":22447,"journal":{"name":"The FASEB Journal","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2020-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141218780","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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