한지훈 (성균관대학교, 현 Gladstone Institutes)

한빛사논문

조회300

성균관대학교, 현 Gladstone Institutes

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Sci. Transl. Med., Aug 07 2024, 16 (759) | https://doi.org/10.1126/scitranslmed.adi4830 Engineered exosomes with a photoinducible protein delivery system enable CRISPR-Cas-based epigenome editing in Alzheimer's disease

Jihoon Han ¹, Jae Hoon Sul ¹, Jeongmi Lee¹, Eunae Kim¹, Hark Kyun Kim ¹, Minshik Chae ¹, Jeein Lim ¹, Jongho Kim¹, Chanhee Kim ¹, Jun-Sik Kim ¹, Yoonsuk Cho ¹, Jae Hyung Park ^2,3,4,5, Yong Woo Cho ^4,6, Dong-Gyu Jo^1,3,4,5,7

¹School of Pharmacy, Sungkyunkwan University, Suwon 16419, Korea.

²School of Chemical Engineering, College of Engineering, Sungkyunkwan University, Suwon 16419, Korea.

³Biomedical Institute for Convergence, Sungkyunkwan University, Suwon 16419, Korea.

⁴ExoStem Tech Inc., Ansan 15588, Korea.

⁵Samsung Advanced Institute for Health Sciences and Technology, Sungkyunkwan University, Seoul 06351, Korea.

⁶Department of Chemical Engineering, Hanyang University, Ansan 15588, Korea.

⁷Institute of Quantum Biophysics, Sungkyunkwan University, Suwon 16419, Korea.

^*Corresponding author : Dong-Gyu Jo

Abstract

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.

논문정보

형식Research article
게재일2024년 08월 (BRIC 등록일 2024-08-08)
연구진국내 연구진
분야 의약학 > 유전 및 유전체의학

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