具有光诱导蛋白递送系统的工程外泌体可在阿尔茨海默病中实现基于 CRISPR-Cas 的表观基因组编辑。

IF 15.8 1区 医学 Q1 CELL BIOLOGY Science Translational Medicine Pub Date : 2024-08-07 DOI:10.1126/scitranslmed.adi4830
Jihoon Han, Jae Hoon Sul, Jeongmi Lee, Eunae Kim, Hark Kyun Kim, Minshik Chae, Jeein Lim, Jongho Kim, Chanhee Kim, Jun-Sik Kim, Yoonsuk Cho, Jae Hyung Park, Yong Woo Cho, Dong-Gyu Jo
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引用次数: 0

摘要

有效地在细胞内输送治疗蛋白质有可能治疗多种疾病。然而,高效地跨细胞膜递送功能性蛋白质仍具有挑战性。外泌体是各类细胞自然分泌的纳米级囊泡,可作为治疗性生物分子的纳米载体。在这里,我们设计了一种装有光诱导货物蛋白释放系统的外泌体,该系统被称为 mMaple3 介导的蛋白装入外泌体并从外泌体释放(MAPLEX),在该系统中,货物蛋白可通过与光可裂解蛋白(mMaple3)结合的外泌体膜标记物融合而装入外泌体,随后通过蓝光照射诱导光裂解而从外泌体膜释放。利用这一系统,我们首先在体外向成纤维细胞输送八聚体结合转录因子 4 和 SRY-box 转录因子 2,从而诱导转录调控。其次,我们通过递送 Cre 重组酶,在 Cre 报告小鼠体内诱导基因重组。最后,我们在 5xFAD 和 3xTg-AD 两种阿尔茨海默病模型小鼠的大脑中实现了靶向表观基因组编辑。给小鼠注射装有β位点淀粉样前体蛋白裂解酶1(Bace1)靶向单导核糖核酸的MAPLEXs,再加上DNA甲基转移酶3A的催化域,成功地甲基化了Bace1启动子内的靶向CpG位点。这种方法导致 5xFAD 和 3xTg-AD 小鼠中 Bace1 的表达明显减少,识别记忆障碍得到改善,淀粉样病理减少。这些结果表明,MAPLEX 是一种高效的细胞内蛋白质递送系统,可以递送治疗多种疾病的各种蛋白质。
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Engineered exosomes with a photoinducible protein delivery system enable CRISPR-Cas–based epigenome editing in Alzheimer’s disease
Effective intracellular delivery of therapeutic proteins can potentially treat a wide array of diseases. However, efficient delivery of functional proteins across the cell membrane remains challenging. Exosomes are nanosized vesicles naturally secreted by various types of cells and may serve as promising nanocarriers for therapeutic biomolecules. Here, we engineered exosomes equipped with a photoinducible cargo protein release system, termed mMaple3-mediated protein loading into and release from exosome (MAPLEX), in which cargo proteins can be loaded into the exosomes by fusing them with photocleavable protein (mMaple3)–conjugated exosomal membrane markers and subsequently released from the exosomal membrane by inducing photocleavage with blue light illumination. Using this system, we first induced transcriptional regulation by delivering octamer-binding transcription factor 4 and SRY-box transcription factor 2 to fibroblasts in vitro. Second, we induced in vivo gene recombination in Cre reporter mice by delivering Cre recombinase. Last, we achieved targeted epigenome editing in the brains of 5xFAD and 3xTg-AD mice, two models of Alzheimer’s disease. Administration of MAPLEXs loaded with β-site amyloid precursor protein cleaving enzyme 1 (Bace1)–targeting single guide RNA–incorporated dCas9 ribonucleoprotein complexes, coupled with the catalytic domain of DNA methyltransferase 3A, resulted in successful methylation of the targeted CpG sites within the Bace1 promoter. This approach led to a significant reduction in Bace1 expression, improved recognition memory impairment, and reduced amyloid pathology in 5xFAD and 3xTg-AD mice. These results suggest that MAPLEX is an efficient intracellular protein delivery system that can deliver diverse therapeutic proteins for multiple diseases.
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来源期刊
Science Translational Medicine
Science Translational Medicine CELL BIOLOGY-MEDICINE, RESEARCH & EXPERIMENTAL
CiteScore
26.70
自引率
1.20%
发文量
309
审稿时长
1.7 months
期刊介绍: Science Translational Medicine is an online journal that focuses on publishing research at the intersection of science, engineering, and medicine. The goal of the journal is to promote human health by providing a platform for researchers from various disciplines to communicate their latest advancements in biomedical, translational, and clinical research. The journal aims to address the slow translation of scientific knowledge into effective treatments and health measures. It publishes articles that fill the knowledge gaps between preclinical research and medical applications, with a focus on accelerating the translation of knowledge into new ways of preventing, diagnosing, and treating human diseases. The scope of Science Translational Medicine includes various areas such as cardiovascular disease, immunology/vaccines, metabolism/diabetes/obesity, neuroscience/neurology/psychiatry, cancer, infectious diseases, policy, behavior, bioengineering, chemical genomics/drug discovery, imaging, applied physical sciences, medical nanotechnology, drug delivery, biomarkers, gene therapy/regenerative medicine, toxicology and pharmacokinetics, data mining, cell culture, animal and human studies, medical informatics, and other interdisciplinary approaches to medicine. The target audience of the journal includes researchers and management in academia, government, and the biotechnology and pharmaceutical industries. It is also relevant to physician scientists, regulators, policy makers, investors, business developers, and funding agencies.
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