RNA干扰在癌症和其他多种疾病的治疗中具有广阔的应用前景,亟需发展高效、安全的递送手段来实现RNA干扰技术的临床应用。本研究使用瞬时纳米沉淀技术,制备系列稳定的、基于壳聚糖(CS)和三聚磷酸钠(TPP)的纳米粒子,系统研究了CS浓度、分...RNA干扰在癌症和其他多种疾病的治疗中具有广阔的应用前景,亟需发展高效、安全的递送手段来实现RNA干扰技术的临床应用。本研究使用瞬时纳米沉淀技术,制备系列稳定的、基于壳聚糖(CS)和三聚磷酸钠(TPP)的纳米粒子,系统研究了CS浓度、分子量及其与TPP的质量比对CS/TPP纳米粒子的物理化学性质的影响,并初步评价了纳米粒子的基因沉默效率。结果显示,CS浓度为0.3~0.6 g/L和CS/TPP质比为1/1~4/1时,可以形成较为稳定的复合物。其中,CS浓度为0.5g/L,质量比为2/1时可得到直径为70~100 nm的稳定纳米粒子。稳定的CS/TPP纳米粒子对siRNA的负载效率为75%~90%,细胞毒性低。在2.5 ng siRNA/μL浓度下,负载siRNA的CS/TPP纳米粒子可以引起约30%的目的基因的沉默。相关结果对制备稳定的、小尺寸CS纳米粒子及其siRNA递送研究具有一定的借鉴意义。展开更多
Efficient siRNA delivery is highly desirable for disease treatment.However,the application of conventional nanoparticles is limited by the inability to escape from endo-lysosomes.Herein,we report a strategy using smal...Efficient siRNA delivery is highly desirable for disease treatment.However,the application of conventional nanoparticles is limited by the inability to escape from endo-lysosomes.Herein,we report a strategy using small-molecule drugs to enhance siRNA endo-lysosomal release,addressing this challenge.We encapsulated gentamicin(GM)into the marketed Onpattro■ formulation to establish LNP-siRNA/GM nanoparticles that promote siRNA endo-lysosomal escape through endosomal disruption,mechanistically exhibiting unique functionality and synergistic effects of LNP-siRNA/GM to improve cancer therapy.Besides,GM induced reactive oxygen species(ROS)and phospholipids accumulation in endolysosomes,as well as the physical characteristics of lipid nanoparticles(LNPs)were preserved.We also revealed that GM causes endo-lysosomal swelling and disrupts the endosomal membrane to enable siRNA release,as confirmed by Galectin 3 recruitment and acridine orange release.This approach achieved∼81%mRNA-EGFR silencing,which is more than LNP-siEGFR(∼56.23%)by enhancing siRNA endo-lysosomal escape efficiency.Meanwhile,LNP-siEGFR/GM exhibited significant biological activities in HepG2 cells,driven by the synergistic effects of siEGFR and GM with the VEGF and CXCL12 downregulation of,and ROS and phospholipids upregulation.Furthermore,tumor growth was notably suppressed after intravenous injection of LNP-siEGFR/GM in tumor-bearing nude mice.The combination of EGFR-siRNA and GM could also greatly inhibit angiogenesis,be antiproliferative,and induce tumor cells apoptosis.Therefore,this GM and siRNA co-delivery system would provide an efficient strategy for siRNA endosomal escape,significantly improving knockdown in various LNPs based siRNA delivery systems and efficiently enhancing cancer therapy.展开更多
近日,北京大学药学院天然药物及仿生药物全国重点实验室杨振军教授团队在国际权威期刊Journal of Controlled Release在线发表了题为“A Mannosylated peptidyl lipid CManDA doped into cytidinyl/cationic lipids efficiently deliver...近日,北京大学药学院天然药物及仿生药物全国重点实验室杨振军教授团队在国际权威期刊Journal of Controlled Release在线发表了题为“A Mannosylated peptidyl lipid CManDA doped into cytidinyl/cationic lipids efficiently delivers siG12Ss to lung cancer in vivo”的研究工作。展开更多
Small interfering RNAs(siRNA)provide a novel and highly specific therapy due to their ability to effectively silence target genes,to date six siRNA therapeutics are approved for clinical use.Even so,some critical chal...Small interfering RNAs(siRNA)provide a novel and highly specific therapy due to their ability to effectively silence target genes,to date six siRNA therapeutics are approved for clinical use.Even so,some critical challenges remain to overcome in the therapeutic application of siRNAs,with delivery issues at the forefront.Among them,endo/lysosomal barrier is one of the important but often-neglected limitations hindering the delivery of siRNA therapeutics.In this review,we summarize the promising strategies that facilitate siRNAs overcoming endo/lysosomal barriers based on the cellular uptake and intracellular transport pathways,including promoting escape once endocytosis into the endo/lysosomes and bypassing lysosomes via endosome-Golgi-endoplasmic reticulum(ER)pathway or nonendocytosis pathway,and discuss the principal considerations and the future directions of promoting endo/lysosomal escape in the development of therapeutic siRNAs.展开更多
文摘RNA干扰在癌症和其他多种疾病的治疗中具有广阔的应用前景,亟需发展高效、安全的递送手段来实现RNA干扰技术的临床应用。本研究使用瞬时纳米沉淀技术,制备系列稳定的、基于壳聚糖(CS)和三聚磷酸钠(TPP)的纳米粒子,系统研究了CS浓度、分子量及其与TPP的质量比对CS/TPP纳米粒子的物理化学性质的影响,并初步评价了纳米粒子的基因沉默效率。结果显示,CS浓度为0.3~0.6 g/L和CS/TPP质比为1/1~4/1时,可以形成较为稳定的复合物。其中,CS浓度为0.5g/L,质量比为2/1时可得到直径为70~100 nm的稳定纳米粒子。稳定的CS/TPP纳米粒子对siRNA的负载效率为75%~90%,细胞毒性低。在2.5 ng siRNA/μL浓度下,负载siRNA的CS/TPP纳米粒子可以引起约30%的目的基因的沉默。相关结果对制备稳定的、小尺寸CS纳米粒子及其siRNA递送研究具有一定的借鉴意义。
基金supported by National Natural Science Foundation of China(81502688)Cooperation Research Funding of Capital Medical University(2020KJ000514)+1 种基金Cooperation Research Funding of Capital Medical University(2023KJ000814)R&D Program of Beijing Municipal Education Commission(KM202210025024).
文摘Efficient siRNA delivery is highly desirable for disease treatment.However,the application of conventional nanoparticles is limited by the inability to escape from endo-lysosomes.Herein,we report a strategy using small-molecule drugs to enhance siRNA endo-lysosomal release,addressing this challenge.We encapsulated gentamicin(GM)into the marketed Onpattro■ formulation to establish LNP-siRNA/GM nanoparticles that promote siRNA endo-lysosomal escape through endosomal disruption,mechanistically exhibiting unique functionality and synergistic effects of LNP-siRNA/GM to improve cancer therapy.Besides,GM induced reactive oxygen species(ROS)and phospholipids accumulation in endolysosomes,as well as the physical characteristics of lipid nanoparticles(LNPs)were preserved.We also revealed that GM causes endo-lysosomal swelling and disrupts the endosomal membrane to enable siRNA release,as confirmed by Galectin 3 recruitment and acridine orange release.This approach achieved∼81%mRNA-EGFR silencing,which is more than LNP-siEGFR(∼56.23%)by enhancing siRNA endo-lysosomal escape efficiency.Meanwhile,LNP-siEGFR/GM exhibited significant biological activities in HepG2 cells,driven by the synergistic effects of siEGFR and GM with the VEGF and CXCL12 downregulation of,and ROS and phospholipids upregulation.Furthermore,tumor growth was notably suppressed after intravenous injection of LNP-siEGFR/GM in tumor-bearing nude mice.The combination of EGFR-siRNA and GM could also greatly inhibit angiogenesis,be antiproliferative,and induce tumor cells apoptosis.Therefore,this GM and siRNA co-delivery system would provide an efficient strategy for siRNA endosomal escape,significantly improving knockdown in various LNPs based siRNA delivery systems and efficiently enhancing cancer therapy.
文摘近日,北京大学药学院天然药物及仿生药物全国重点实验室杨振军教授团队在国际权威期刊Journal of Controlled Release在线发表了题为“A Mannosylated peptidyl lipid CManDA doped into cytidinyl/cationic lipids efficiently delivers siG12Ss to lung cancer in vivo”的研究工作。
基金supported by National Natural Science Foundation of China(No.82173769)the National Key R&D Program of China(No.2021YFE0106900)+1 种基金Applied Basic Research Multiinvestment Foundation of Tianjin(No.21JCYBJC01540)the Science&Technology Development Fund of Tianjin Education Commission for Higher Education(No.2023ZD019)。
文摘Small interfering RNAs(siRNA)provide a novel and highly specific therapy due to their ability to effectively silence target genes,to date six siRNA therapeutics are approved for clinical use.Even so,some critical challenges remain to overcome in the therapeutic application of siRNAs,with delivery issues at the forefront.Among them,endo/lysosomal barrier is one of the important but often-neglected limitations hindering the delivery of siRNA therapeutics.In this review,we summarize the promising strategies that facilitate siRNAs overcoming endo/lysosomal barriers based on the cellular uptake and intracellular transport pathways,including promoting escape once endocytosis into the endo/lysosomes and bypassing lysosomes via endosome-Golgi-endoplasmic reticulum(ER)pathway or nonendocytosis pathway,and discuss the principal considerations and the future directions of promoting endo/lysosomal escape in the development of therapeutic siRNAs.
