Takahashi Masayuki, Seki Mineaki, Nashimoto Masayuki, Kabuta Tomohiro
Research Institute for Healthy Living, Niigata University of Pharmacy and Applied Life Sciences, Niigata, Niigata, Japan.
Department of Degenerative Neurological Diseases, National Institute of Neuroscience, National Center of Neurology and Psychiatry, Kodaira, Tokyo, Japan.
J Nucleic Acids. 2021 Nov 16;2021:2458470. doi: 10.1155/2021/2458470. eCollection 2021.
Although antisense oligonucleotide (ASO) therapeutics can be taken up by living cells without carrier molecules, a large part of incorporated ASOs are trapped in the endosomes and do not exert therapeutic effects. To improve their therapeutic effects, it would be important to elucidate the mechanism of cellular uptake and intracellular trafficking of ASOs. In this study, we investigated how SIDT1 affects cellular uptake and intracellular trafficking of ASOs. Fluorescence microscopic analysis suggested that most of naked ASOs are trafficked to the lysosomes via the endosomes. The data obtained from flow cytometry and fluorescence microscopy together showed that although the SIDT1 level barely affects the total cellular uptake of ASOs, it appears to affect the intracellular trafficking of ASOs. We also showed that SIDT1 exists mainly in the endoplasmic reticulum and that perturbing the normal level of SIDT1 suppresses the antisense effect of the naked ASO targeting miR-16.
尽管反义寡核苷酸（ASO）疗法无需载体分子就能被活细胞摄取，但大部分被摄取的ASO被困在内体中，无法发挥治疗作用。为了提高其治疗效果，阐明ASO的细胞摄取和细胞内运输机制至关重要。在本研究中，我们调查了SIDT1如何影响ASO的细胞摄取和细胞内运输。荧光显微镜分析表明，大多数裸露的ASO通过内体被运输到溶酶体。流式细胞术和荧光显微镜获得的数据共同表明，尽管SIDT1水平几乎不影响ASO的总细胞摄取，但它似乎会影响ASO的细胞内运输。我们还表明，SIDT1主要存在于内质网中，扰乱SIDT1的正常水平会抑制靶向miR-16的裸露ASO的反义效应。
