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조미영Mi Young Cho
한국기초과학지원연구원
J. Extracell. Vesicles, Jul 2024, 13 (7) | https://doi.org/10.1002/jev2.12489
Evaluation of unmodified human cell-derived extracellular vesicle mitochondrial deoxyribonucleic acid-based biodistribution in rodents
Young-Woo Cho1,2, Mi Young Cho3, Jaehyeon Yoon1, Da Eun Hong1, Ju-young Lee1,2, Hye Sun Park3, Hyunseung Lee3, Kwan Soo Hong3,4, Lee Won-Kyu1, Choi Saehae1, Suk-Gil Song2, Young-Woock Noh1
1Division of Drug Safety Evaluation, NDDC, Osong Medical Innovation Foundation, Cheongju, South Korea
2College of Pharmacy, Chungbuk National University, Cheongju, South Korea
3Biopharmaceutical Research Center, Korea Basic Science Institute, Cheongju, South Korea
4Department of Chemistry, Chung-Ang University, Seoul, South Korea
CorrespondenceYoung-Woo Cho and Young-Woock Noh
Young-Woo Cho and Mi Young Cho contributed equally to this study.
Abstract
Recently, extracellular vesicles (EVs) have been developed as therapeutic targets for various diseases. Biodistribution is crucial for EVs intended for therapeutic purposes because it can determine the degree of on- and off-target effects. This study aimed to explore techniques to evaluate the biodistribution of unmodified EVs. We devised a novel quantitative polymerase chain reaction (qPCR)-based assay to detect unmodified EVs by targeting mitochondrial deoxyribonucleic acid (mtDNA), a constituent of EVs. We focused on specific mtDNA regions that exhibited homologous variations distinct from their rodent mtDNA counterparts to establish this analytical approach. Herein, we successfully designed primers and probes targeting human and rodent mtDNA sequences and developed a highly specific and sensitive qPCR method. Furthermore, the quantification range of EVs isolated from various cells differed based on the manufacturer and cell source. IRDye 800CW-labelled Expi293F EV mimetics were administered to the animals via the tail vein to compare the imaging test and mtDNA-qPCR results. The results obtained from imaging tests and mtDNA-qPCR to investigate EV biodistribution patterns revealed differences. The results revealed that our newly developed method effectively determined the biodistribution of unmodified EVs with high sensitivity and reproducibility.
논문정보
- Research article
- 2024년 07월 (BRIC 등록일 2024-08-06)
- 국내 연구진
- 바이오·의료융합 > 바이오센싱 및 나노바이오물질
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