Brain-targeted delivery of therapeutic molecules by exosomes derived from engineered human iPS cells: a potential therapeutic approach for Huntington's disease

项目总结 亨廷顿病(HD)是一种由CAG重复扩增引起的毁灭性的神经退行性疾病。 亨廷顿蛋白(HTT)基因。在突变体HTT中，CAG重复序列被翻译成多聚谷氨酰胺(PolyQ)区域 具有神经毒性的蛋白质。目前的治疗努力集中在抑制血管内皮生长因子的表达上。 突变的HTT蛋白或靶向神经毒性的下游通路。我们和其他人已经证明，CAG 重复靶向磷二酸吗啉低聚物(PMOS)和神经营养蛋白BDNF 对HD的治疗益处。然而，PMO和BDNF通常不会越过血脑屏障(BBB)， 阻碍了临床应用的翻译。因此，更高效的送货车辆能够BBB 渗透对于开发治疗HD和其他神经退行性疾病的有效疗法至关重要 总体而言。Exosome(Exo)是一种细胞分泌的细胞外载体，具有穿透血脑屏障的潜力，能够 提供外源治疗分子。IPSC被认为是Exo的最佳来源之一 基于高Exo产量的制造，可获得与cGMP兼容的临床级制造平台 IPSC生产和外源制造，以及进行基因组编辑以建立工程化IPSC的可行性 产生改良的Exo，以便更有效地进行脑靶向。因此，我们建议制定战略，以 从经过改造的ipscs中生产脑靶向Exo，作为PMO和BDNF的输送载体，以及进一步 严密研究PMOS和BDNF脑靶向Exo的治疗效果 HD神经元和小鼠模型。该项目的成功将为未来更大规模的调查奠定基础 旨在将Huma IPSC衍生的Exo用于靶向输送治疗分子(如PMO、化学物质 化合物和神经营养因子)治疗各种神经退行性疾病。

PROJECT SUMMARY Huntington’s disease (HD) is a devastating neurodegenerative disease caused by a CAG repeat expansion in the gene huntingtin (HTT). The CAG repeat is translated into a polyglutamine (polyQ) tract in the mutant HTT protein that has neurotoxic properties. Current therapeutic efforts are focused at suppressing the expression of the mutant HTT protein or targeting downstream pathways of neurotoxicity. We and others have shown that CAG repeat-targeting phosphorodiamidate morpholino oligomers (PMOs) and neurotrophic protein BDNF have therapeutic benefits for HD. However, PMOs and BDNF do not usually cross the blood-brain barrier (BBB), hindering the translation of clinical application. Therefore, more efficient delivery vehicles capable of BBB penetration are critical for the development of effective therapies for HD and other neurodegenerative disorders in general. Exosomes (Exo) are cell-secreted extracellular vehicles with BBB penetration potential, capable of delivering exogeneous therapeutic molecules. iPSCs have been considered as one of the best sources for Exo manufacture, based on high Exo yield, availability of cGMP-compatible clinical-grade manufacture platform for iPSC production and Exo manufacture, and feasibility to perform genome editing to establish engineered iPSCs that produce modified Exo for more efficient brain targeting. We therefore propose to develop strategies to produce brain-targeting Exo from engineered iPSCs as delivery vehicles for PMOs as well as BDNF, and further rigorously study the therapeutic efficacy of the brain-targeting Exo loaded with PMOs and BDNF in a panel of HD neuron and mouse models. Success of this project will set the stage for future larger scale investigations aimed at using huma iPSC-derived Exo for targeted delivery of therapeutic molecules (such as PMOs, chemical compounds and neurotrophic factors) for various neurodegenerative disorders.