mRNA is a novel class of therapeutic modality that holds great promise in vaccination,protein replacement therapy,cancer immunotherapy,immune cell engineering etc.However,optimization of mRNA molecules and efficient i...mRNA is a novel class of therapeutic modality that holds great promise in vaccination,protein replacement therapy,cancer immunotherapy,immune cell engineering etc.However,optimization of mRNA molecules and efficient in vivo delivery are quite important but challenging for its broad application.Here we present an ionizable lipid nanoparticle(iLNP)based on iBL0713 lipid for in vitro and in vivo expression of desired proteins using codon-optimized mRNAs.mRNAs encoding luciferase or erythropoietin(EPO)were prepared by in vitro transcription and formulated with proposed iLNP,to form iLP171/mRNA formulations.It was revealed that both luciferase and EPO proteins were successfully expressed by human hepatocellular carcinoma cells and hepatocytes.The maximum amount of protein expression was found at 6 h post-administration.The expression efficiency of EPO with codon-optimized mRNA was significantly higher than that of unoptimized mRNA.Moreover,no toxicity or immunogenicity was observed for these mRNA formulations.Therefore,our study provides a useful and promising platform for mRNA therapeutic development.展开更多
Small interfering RNA(siRNA)constitutes a promising therapeutic modality supporting the potential functional cure of hepatitis B.A novel ionizable lipidoid nanoparticle(RBP131)and a state-of-the-art lyophilization tec...Small interfering RNA(siRNA)constitutes a promising therapeutic modality supporting the potential functional cure of hepatitis B.A novel ionizable lipidoid nanoparticle(RBP131)and a state-of-the-art lyophilization technology were developed in this study,enabling to deliver siRNA targeting apolipoprotein B(APOB)into the hepatocytes with an ED_(50)of 0.05 mg/kg after intravenous injection.In addition,according to the requirements of Investigational New Drug(IND)application,a potent siRNA targeting hepatitis B virus(HBV)was selected and encapsulated with RBP131 to fabricate a therapeutic formulation termed RB-HBV008.展开更多
It was recognized that a majority of the whole human genome is transcribed but only about 2% of genome actually encode all the proteins that were supposed, according to the central dogma, executing most of the biologi...It was recognized that a majority of the whole human genome is transcribed but only about 2% of genome actually encode all the proteins that were supposed, according to the central dogma, executing most of the biological functions of an organism. At an age proteins dominate, over 90% nonprotein coding regions were long regarded as trash, but nevertheless it was puzzling why god would allocate such a big portion of a genome to things without obvious meanings, and such curiosity has heightened when it was shown among various genome sequencing projects that the percentage of non-coding sequences is in almost strict correlation with the complexity of the organisms.展开更多
Type 2 diabetes mellitus（T2 DM） is a common metabolic disease influenced by both genetic and environmental factors. In this study, we performed an in-house genotyping and meta-analysis study using three independent ...Type 2 diabetes mellitus（T2 DM） is a common metabolic disease influenced by both genetic and environmental factors. In this study, we performed an in-house genotyping and meta-analysis study using three independent GWAS datasets of T2 DM and found that rs3743121, located 1 kb downstream of AQR,was a novel susceptibility SNP associated with T2 DM. The risk allele C of rs3743121 was correlated with the increased expression of AQR in white blood cells, similar to that observed in T2 DM models. The knockdown of AQR in HepG2 facilitated the glucose uptake, decreased the expression level of PCK2,increased the phosphorylation of GSK-3β, and restored the insulin sensitivity. Furthermore, the suppression of AQR inhibited the mTOR pathway and the protein ubiquitination process. Our study suggests that AQR is a novel type 2 diabetes-associated gene that regulates signaling pathways critical for glucose metabolism.展开更多
To modulate gene expression in research studies or in potential clinical therapies,transfection of exogenous nucleic acids including plasmid DNA and small interference RNA(siRNA)are generally performed.However,the cel...To modulate gene expression in research studies or in potential clinical therapies,transfection of exogenous nucleic acids including plasmid DNA and small interference RNA(siRNA)are generally performed.However,the cellular processing and the fate of these nucleic acids remain elusive.By investigating the cellular behavior of transfected nucleic acids using confocal imaging,here we show that when siRNA was cotransfected into cultured cells with other nucleic acids,including single-stranded RNA oligonucleotides,single and double-stranded DNA oligonucleotides,as well as long double-stranded plasmid DNA,they all aggregate in the same cytoplasmic granules.Interestingly,the amount of siRNA aggregating in granules was found not to correlate with the gene silencing activity,suggesting that assembly of cytoplasmic granules triggered by siRNA transfection may be separable from the siRNA silencing event.Our results argue against the claim that the siRNAaggregating granules are the functional site of RNA interference(RNAi).Taken together,our studies suggest that,independent of their types or forms,extraneously transfected nucleic acids are processed through a common cytoplasmic pathway and trigger the formation of a new type of cytoplasmic granules“transfection granules”.展开更多
基金supported by the National Natural Science Foundation of China(31871003,81402863,31901053)the Beijing Institute of Technology Research Fund Program for Young Scholars and the Fundamental Research Funds for the Central Universities(3052018065,1870012222004)+2 种基金the Natural Science Foundation of Guangdong Province(2019A1515010776)the Hunan Provincial Natural Science Foundation of China(2018JJ1019,2019JJ50196)the Hu-Xiang Young Talent Program(2018RS3094).
文摘mRNA is a novel class of therapeutic modality that holds great promise in vaccination,protein replacement therapy,cancer immunotherapy,immune cell engineering etc.However,optimization of mRNA molecules and efficient in vivo delivery are quite important but challenging for its broad application.Here we present an ionizable lipid nanoparticle(iLNP)based on iBL0713 lipid for in vitro and in vivo expression of desired proteins using codon-optimized mRNAs.mRNAs encoding luciferase or erythropoietin(EPO)were prepared by in vitro transcription and formulated with proposed iLNP,to form iLP171/mRNA formulations.It was revealed that both luciferase and EPO proteins were successfully expressed by human hepatocellular carcinoma cells and hepatocytes.The maximum amount of protein expression was found at 6 h post-administration.The expression efficiency of EPO with codon-optimized mRNA was significantly higher than that of unoptimized mRNA.Moreover,no toxicity or immunogenicity was observed for these mRNA formulations.Therefore,our study provides a useful and promising platform for mRNA therapeutic development.
基金This work was supported by the National Drug Program of China(2012ZX09102301-006,2015ZX09102-023-002,2014ZX09304313-001,and 2009ZX09103732)the National Natural Science Foundation of China(31871003)+1 种基金the Beijing-Tianjin-Hebei Basic Research Cooperation Project(19JCZDJC64100)the Beijing Nova Program from Beijing Municipal Science&Technology Commission(Z201100006820005).
文摘Small interfering RNA(siRNA)constitutes a promising therapeutic modality supporting the potential functional cure of hepatitis B.A novel ionizable lipidoid nanoparticle(RBP131)and a state-of-the-art lyophilization technology were developed in this study,enabling to deliver siRNA targeting apolipoprotein B(APOB)into the hepatocytes with an ED_(50)of 0.05 mg/kg after intravenous injection.In addition,according to the requirements of Investigational New Drug(IND)application,a potent siRNA targeting hepatitis B virus(HBV)was selected and encapsulated with RBP131 to fabricate a therapeutic formulation termed RB-HBV008.
文摘It was recognized that a majority of the whole human genome is transcribed but only about 2% of genome actually encode all the proteins that were supposed, according to the central dogma, executing most of the biological functions of an organism. At an age proteins dominate, over 90% nonprotein coding regions were long regarded as trash, but nevertheless it was puzzling why god would allocate such a big portion of a genome to things without obvious meanings, and such curiosity has heightened when it was shown among various genome sequencing projects that the percentage of non-coding sequences is in almost strict correlation with the complexity of the organisms.
基金supported by the National Basic Research Program of the Chinese Ministry of Science and Technology (2013CB530700)Key Programs from the National Natural Science Foundation of China(Nos. 81630034 and 81130003)
文摘Type 2 diabetes mellitus（T2 DM） is a common metabolic disease influenced by both genetic and environmental factors. In this study, we performed an in-house genotyping and meta-analysis study using three independent GWAS datasets of T2 DM and found that rs3743121, located 1 kb downstream of AQR,was a novel susceptibility SNP associated with T2 DM. The risk allele C of rs3743121 was correlated with the increased expression of AQR in white blood cells, similar to that observed in T2 DM models. The knockdown of AQR in HepG2 facilitated the glucose uptake, decreased the expression level of PCK2,increased the phosphorylation of GSK-3β, and restored the insulin sensitivity. Furthermore, the suppression of AQR inhibited the mTOR pathway and the protein ubiquitination process. Our study suggests that AQR is a novel type 2 diabetes-associated gene that regulates signaling pathways critical for glucose metabolism.
文摘To modulate gene expression in research studies or in potential clinical therapies,transfection of exogenous nucleic acids including plasmid DNA and small interference RNA(siRNA)are generally performed.However,the cellular processing and the fate of these nucleic acids remain elusive.By investigating the cellular behavior of transfected nucleic acids using confocal imaging,here we show that when siRNA was cotransfected into cultured cells with other nucleic acids,including single-stranded RNA oligonucleotides,single and double-stranded DNA oligonucleotides,as well as long double-stranded plasmid DNA,they all aggregate in the same cytoplasmic granules.Interestingly,the amount of siRNA aggregating in granules was found not to correlate with the gene silencing activity,suggesting that assembly of cytoplasmic granules triggered by siRNA transfection may be separable from the siRNA silencing event.Our results argue against the claim that the siRNAaggregating granules are the functional site of RNA interference(RNAi).Taken together,our studies suggest that,independent of their types or forms,extraneously transfected nucleic acids are processed through a common cytoplasmic pathway and trigger the formation of a new type of cytoplasmic granules“transfection granules”.
