Obesity is a complex and incompletely understood disease,but current drug screening strategies mostly rely on immature in vitro adipose models which cannot recapitulate it properly.To address this issue,we developed a...Obesity is a complex and incompletely understood disease,but current drug screening strategies mostly rely on immature in vitro adipose models which cannot recapitulate it properly.To address this issue,we developed a statistically validated high-throughput screening model by seeding human mature adipocytes from patients,encapsulated in physiological collagen microfibers.These drop tissues ensured the maintenance of adipocyte viability and functionality for controlling glucose and fatty acids uptake,as well as glycerol release.As such,patients’BMI and insulin sensitivity displayed a strong inverse correlation:the healthy adipocytes were associated with the highest insulin-induced glucose uptake,while insulin resistance was confirmed in the underweight and severely obese adipocytes.Insulin sensitivity recovery was possible with two type 2 diabetes treatments,rosiglitazone and melatonin.Finally,the addition of blood vasculature to the model seemed to more accurately recapitulate the in vivo physiology,with particular respect to leptin secretion metabolism.展开更多
Human respiratory syncytial virus（RSV） is an important pediatric pathogen of lower respiratory tract worldwide. No vaccines and antiviral drugs are available. Herein the use of an RNA polymerase I-driven RSV minigen...Human respiratory syncytial virus（RSV） is an important pediatric pathogen of lower respiratory tract worldwide. No vaccines and antiviral drugs are available. Herein the use of an RNA polymerase I-driven RSV minigenome for analyzing RSV replication and screening anti-RSV drugs was investigated. The RNA polymerase I（Pol I） was used to transcribe RSV minigenome from the constructed plasmid, designated p HM-RSV-Gluc, of minigenome c DNA which comprised trailer region, gene start sequence（GS）, reverse complementary copy of Gaussia luciferase（Gluc） gene, gene end sequence（GE）, and leader region in the direction of 5＇–3＇end and was flanked by promoter and terminator of Pol I. The expression of Gluc was confirmed in p HM-RSV-Gluc transfected HEp-2 cells following RSV infection and had the characteristics of dose-dependent, which provided a rapid, sensitive, and quantitative method for quantifying virus titers and screening antiviral drugs.展开更多
基金The authors thank Nippon Ham Foods Ltd for their kind donation of collagen.This research was supported by a Kakenhi Grant-in-Aid for Early-Career Scientists(70838523)as well as a grant from the Japanese Ministry of Education,Culture,Sports,Science and Technology(18K09488).
文摘Obesity is a complex and incompletely understood disease,but current drug screening strategies mostly rely on immature in vitro adipose models which cannot recapitulate it properly.To address this issue,we developed a statistically validated high-throughput screening model by seeding human mature adipocytes from patients,encapsulated in physiological collagen microfibers.These drop tissues ensured the maintenance of adipocyte viability and functionality for controlling glucose and fatty acids uptake,as well as glycerol release.As such,patients’BMI and insulin sensitivity displayed a strong inverse correlation:the healthy adipocytes were associated with the highest insulin-induced glucose uptake,while insulin resistance was confirmed in the underweight and severely obese adipocytes.Insulin sensitivity recovery was possible with two type 2 diabetes treatments,rosiglitazone and melatonin.Finally,the addition of blood vasculature to the model seemed to more accurately recapitulate the in vivo physiology,with particular respect to leptin secretion metabolism.
基金supported by National Major Scientific and Technological Special Project for ‘‘AIDS and Viral Hepatitis and Other Major Infectious Diseases Prevention and Control’’ during the Twelfth Five-year Plan Period (No. 2013ZX10004-601)
文摘Human respiratory syncytial virus（RSV） is an important pediatric pathogen of lower respiratory tract worldwide. No vaccines and antiviral drugs are available. Herein the use of an RNA polymerase I-driven RSV minigenome for analyzing RSV replication and screening anti-RSV drugs was investigated. The RNA polymerase I（Pol I） was used to transcribe RSV minigenome from the constructed plasmid, designated p HM-RSV-Gluc, of minigenome c DNA which comprised trailer region, gene start sequence（GS）, reverse complementary copy of Gaussia luciferase（Gluc） gene, gene end sequence（GE）, and leader region in the direction of 5＇–3＇end and was flanked by promoter and terminator of Pol I. The expression of Gluc was confirmed in p HM-RSV-Gluc transfected HEp-2 cells following RSV infection and had the characteristics of dose-dependent, which provided a rapid, sensitive, and quantitative method for quantifying virus titers and screening antiviral drugs.
