Brain Gene Silencing with Cationic Amino-Capped Poly(ethylene glycol) Polyplexes.
Brain Gene Silencing with Cationic Amino-Capped Poly(ethylene glycol) Polyplexes.
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DOI：
10.3390/biomedicines10092182
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发表时间：
2022-09-03
期刊：
影响因子：
4.7
通讯作者：
中科院分区：
文献类型：
--
作者：
大脑中的治疗性基因沉默通常使用高度侵入性的颅内给药方法和/或相对有毒的载体来实现。在这项工作中，我们使用了一种相对生物相容性的载体：聚（乙二醇）星形聚合物与胺基团（4APPA）通过鼻子到大脑的路线。4APPA复合抗瘙痒E3泛素蛋白连接酶（抗ITCH）siRNA以形成带正电荷（ζ电位+15 ± 5 mV）的150 nm纳米颗粒。siRNA-4APPA复合物在A431细胞系中表现出低细胞毒性（IC 50 = 13.92 ± 6 mg mL−1），并且在该细胞系中比Lipofectamine 2000（IC 50 = 0.033 ± 0.04 mg mL−1）的毒性低三个数量级。分别使用荧光激活细胞分选术（FACS）和共聚焦激光扫描显微镜（CLSM）证明了与siRNA-4APPA纳米颗粒结合的荧光标记的siRNA的细胞缔合和摄取。在体外A431细胞系中观察到ITCH基因的基因沉默（与单独使用抗ITCH siRNA相比，下调65%）。在鼻内给药荧光标记的siRNA-4APPA复合物时，在嗅球、大脑皮层和中脑区域的细胞中观察到荧光。最后，在健康大鼠模型中，鼻内给予siRNA-4APPA纳米颗粒（0.15 mg kg−1 siRNA，每日两次，持续3天）后，在脑细胞中观察到ITCH的下调（与未处理对照相比，ITCH剩余54 ± 13%）。脑中的基因沉默可以通过鼻内施用基于siRNA-聚（乙二醇）的聚合复合物来实现。
Therapeutic gene silencing in the brain is usually achieved using highly invasive intracranial administration methods and/or comparatively toxic vectors. In this work, we use a relatively biocompatible vector: poly(ethylene glycol) star-shaped polymer capped with amine groups (4APPA) via the nose to brain route. 4APPA complexes anti- itchy E3 ubiquitin protein ligase (anti-ITCH) siRNA to form positively charged (zeta potential +15 ± 5 mV) 150 nm nanoparticles. The siRNA-4APPA polyplexes demonstrated low cellular toxicity (IC50 = 13.92 ± 6 mg mL−1) in the A431 cell line and were three orders of magnitude less toxic than Lipofectamine 2000 (IC50 = 0.033 ± 0.04 mg mL−1) in this cell line. Cell association and uptake of fluorescently labelled siRNA bound to siRNA-4APPA nanoparticles was demonstrated using fluorescent activated cell sorting (FACS) and confocal laser scanning microscopy (CLSM), respectively. Gene silencing of the ITCH gene was observed in vitro in the A431 cell line (65% down regulation when compared to the use of anti-ITCH siRNA alone). On intranasal dosing with fluorescently labelled siRNA-4APPA polyplexes, fluorescence was seen in the cells of the olfactory bulb, cerebral cortex and mid-brain regions. Finally, down regulation of ITCH was seen in the brain cells (54 ± 13% ITCH remaining compared to untreated controls) in a healthy rat model, following intranasal dosing of siRNA-4APPA nanoparticles (0.15 mg kg−1 siRNA twice daily for 3 days). Gene silencing in the brain may be achieved by intranasal administration of siRNA- poly(ethylene glycol) based polyplexes.
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影响因子：
10.8
作者：
Malik, N;Wiwattanapatapee, R;Duncan, R
通讯作者：
Duncan, R
影响因子：
3.7
作者：
Chapman, Colin D.;Frey, William H., II;Craft, Suzanne;Danielyan, Lusine;Hallschmid, Manfred;Schithoth, Helgi B.;Benedict, Christian
通讯作者：
Benedict, Christian
DOI：
10.1056/nejmoa2110345
发表时间：
2021-11-04
期刊：
The New England journal of medicine
影响因子：
--
作者：
Thomas SJ;Moreira ED Jr;Kitchin N;Absalon J;Gurtman A;Lockhart S;Perez JL;Pérez Marc G;Polack FP;Zerbini C;Bailey R;Swanson KA;Xu X;Roychoudhury S;Koury K;Bouguermouh S;Kalina WV;Cooper D;Frenck RW Jr;Hammitt LL;Türeci Ö;Nell H;Schaefer A;Ünal S;Yang Q;Liberator P;Tresnan DB;Mather S;Dormitzer PR;Şahin U;Gruber WC;Jansen KU;C4591001 Clinical Trial Group
通讯作者：
C4591001 Clinical Trial Group
影响因子：
4.6
作者：
Rodriguez M;Lapierre J;Ojha CR;Kaushik A;Batrakova E;Kashanchi F;Dever SM;Nair M;El-Hage N
通讯作者：
El-Hage N
影响因子：
5.7
作者：
Osborn MF;Coles AH;Golebiowski D;Echeverria D;Moazami MP;Watts JK;Sena-Esteves M;Khvorova A
通讯作者：
Khvorova A