帕金森病(PD)是第二常见的神经退行性疾病,其特征是称为路易体的α-突触核蛋白(α-syn)聚集体的积累,导致黑质中多巴胺能(DA)神经元的逐渐丧失。虽然α-syn表达可以通过反义寡核苷酸(ASO)和异源双链寡核苷酸(HDO)通过侧脑室(ICV)注射减弱,安全有效地将寡核苷酸外周靶向递送到DA神经元中的挑战仍未解决。这里,我们设计了一种新的DNA/DNA双链(互补DNA,coDNA)基于α-syn-ASO序列的具有胆固醇缀合的分子(Chol-HDO(coDNA)),并评估了其沉默效率。Further,Chol-HDO@LMNPs,Chol-HDO加载,具有DSPE-PEG2000-左旋多巴修饰(L-DOPA-CECm)包被的纳米颗粒(NPs)的脑血管内皮细胞膜,被开发用于通过尾静脉注射靶向治疗PD。CECm促进NPs的血脑屏障(BBB)渗透,与从网状内皮系统摄取中逸出的胆固醇一起,以及L-DOPA脱羧成多巴胺,促进NP向PD位点进行DA神经元再生。行为测试表明,纳米诱饵改善了HDO对PD小鼠的功效。这些发现为开发加载HDO的仿生纳米诱饵以精确治疗PD提供了见解。重要性声明:α-突触核蛋白(α-syn)聚集体的积累是PD的标志。我们先前的研究设计了一种靶向人类SNCA的特异性反义寡核苷酸(ASO),但创伤性脑室内(ICV)不利于临床应用。这里,我们进一步优化ASO通过创建一个DNA/DNA双链分子与胆固醇结合,命名为Chol-HDO(coDNA),并通过使用DSPE-PEG2000和L-DOPA改造脑血管内皮细胞膜(CECm),开发了DA靶向的仿生纳米诱饵Chol-HDO@LMNPs。体内结果表明,尾静脉注射Chol-HDO@LMNPs可以靶向大脑中的DA神经元,并改善PD小鼠模型的运动缺陷。这项研究提供了一个有前途的外周递送平台L-DOPA-CECm纳米诱饵装载了一种新的Chol-HDO(coDNA)靶向PD治疗中的DA神经元。
Parkinson\'s disease (PD) is the second most common neurodegenerative disorder characterized by the accumulation of α-synuclein (α-syn) aggregates called Lewy bodies leading to the gradual loss of dopaminergic (DA) neurons in the substantia nigra. Although α-syn expression can be attenuated by antisense oligonucleotides (ASOs) and heteroduplex oligonucleotide (HDO) by intracerebroventricular (ICV) injection, the challenge to peripheral targeted delivery of oligonucleotide safely and effectively into DA neurons remains unresolved. Here, we designed a new DNA/DNA double-stranded (complementary DNA, coDNA) molecule with cholesterol conjugation (Chol-HDO (coDNA)) based on an α-syn-ASO sequence and evaluated its silence efficiency. Further, Chol-HDO@LMNPs, Chol-HDO-loaded, cerebrovascular endothelial cell membrane with DSPE-PEG2000-levodopa modification (L-DOPA-CECm)-coated nanoparticles (NPs), were developed for the targeted treatment of PD by tail intravenous injection. CECm facilitated the blood-brain barrier (BBB) penetration of NPs, together with cholesterol escaped from reticuloendothelial system uptake, as well as L-DOPA was decarboxylated into dopamine which promoted the NPs toward the PD site for DA neuron regeneration. The behavioral tests demonstrated that the
nanodecoys improved the efficacy of HDO on PD mice. These findings provide insights into the development of biomimetic
nanodecoys loading HDO for precise therapy of PD. STATEMENT OF SIGNIFICANCE: The accumulation of α-synuclein (α-syn) aggregates is a hallmark of PD. Our previous study designed a specific antisense oligonucleotide (ASO) targeting human SNCA, but the traumatic intracerebroventricular (ICV) is not conducive to clinical application. Here, we further optimize the ASO by creating a DNA/DNA double-stranded molecule with cholesterol-conjugated, named Chol-HDO (coDNA), and develop a DA-targeted biomimetic nanodecoy Chol-HDO@LMNPs by engineering cerebrovascular endothelial cells membranes (CECm) with DSPE-PEG2000 and L-DOPA. The in vivo results demonstrated that tail vein injection of Chol-HDO@LMNPs could target DA neurons in the brain and ameliorate motor deficits in a PD mouse model. This investigation provides a promising peripheral delivery platform of L-DOPA-CECm nanodecoy loaded with a new Chol-HDO (coDNA) targeting DA neurons in PD therapy.