Extracellular vesicles mediated exocytosis of antisense peptide nucleic acids.
Extracellular vesicles mediated exocytosis of antisense peptide nucleic acids.
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细胞外囊泡介导的反义肽核酸胞吐作用。
DOI：
10.1016/j.omtn.2021.07.018
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发表时间：
2021-09-03
期刊：
影响因子：
--
通讯作者：
Bahal R
中科院分区：
文献类型：
--
作者：
Malik S;Saltzman WM;Bahal R
肽核酸（PNAs），一种人工合成的DNA模拟物，已被广泛用于基于反义和抗原的生物医学应用。已作出重大努力，以增加细胞摄取的PNA，但在这里，我们研究了相对未开发的方面的细胞内运输和内吞回收的PNA。为了概念验证，我们使用了靶向miR-155的抗microRNA（miR）PNA。通过共聚焦和流式细胞术在HeLa细胞中研究PNA的亚细胞定位。含有PNA的细胞外囊泡的全面表征显示球形形态，负表面电荷密度，和四跨膜蛋白标记的存在。最重要的是，我们研究了rab 11 a和rab 27 b GTP酶在调节PNA胞吐中的作用。细胞器染色，然后通过共聚焦成像，显示较高的定位PNA在溶酶体。基因表达分析建立了增强的功能活性的PNA后，抑制内吞循环。多项研究报告了单链寡核苷酸、短干扰RNA（siRNA）和纳米载体的胞吐作用。据我们所知，这是第一个建立PNA经历内吞再循环和胞吐出肿瘤细胞的机制研究。这里提出的结果可以作为一个平台，以开发和优化策略，通过避免再循环途径提高PNA的治疗效果。作者在一系列基于细胞培养的功能测定中研究了肽核酸在癌细胞中经历胞吐作用。据指出，膜运输蛋白在肽核酸的胞吐和胞内运输中起重要作用。
Peptide nucleic acids (PNAs), a synthetic DNA mimic, have been extensively utilized for antisense- and antigene-based biomedical applications. Significant efforts have been made to increase the cellular uptake of PNAs, but here we examined relatively unexplored aspects of intracellular trafficking and endocytic recycling of PNAs. For proof-of-concept, we used anti-microRNA (miR) PNA targeting miR-155. The sub-cellular localization of PNA was studied via confocal and flow-cytometry-based assays in HeLa cells. A comprehensive characterization of PNA-containing extracellular vesicles revealed spherical morphology, negative surface charge density, and the presence of tetraspanin markers. Most importantly, we investigated rab11a and rab27b GTPases’ role in regulating the exocytosis of PNAs. Organelle staining, followed by confocal imaging, showed higher localization of PNA in lysosomes. Gene-expression analysis established the enhanced functional activity of PNA after inhibition of endocytic recycling. Multiple studies report the exocytosis of single-stranded oligonucleotides, short interfering RNAs (siRNAs), and nanocarriers. To our knowledge, this is the first mechanistic study to establish that PNA undergoes endocytic recycling and exocytosis out of tumor cells. The results presented here can serve as a platform to develop and optimize strategies for improving the therapeutic efficacy of PNAs by avoiding the recycling pathways. Authors investigated that peptide nucleic acid undergoes exocytosis in the cancer cells in a series of cell-culture-based functional assays. It was noted that membrane trafficking proteins play a significant role in the exocytosis and intracellular trafficking of peptide nucleic acid.
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影响因子：
46.9
作者：
Alvarez-Erviti, Lydia;Seow, Yiqi;Wood, Matthew J. A.
通讯作者：
Wood, Matthew J. A.
DOI：
10.1261/rna.058362.116
发表时间：
2017-01
期刊：
RNA (New York, N.Y.)
影响因子：
--
作者：
Hnedzko D;McGee DW;Karamitas YA;Rozners E
通讯作者：
Rozners E
影响因子：
4.7
作者：
Saleh, Amer F.;Arzumanov, Andrey;Abes, Rachida;Owen, David;Lebleu, Bernard;Gait, Michael J.
通讯作者：
Gait, Michael J.
影响因子：
15
作者：
He, Gaofei;Rapireddy, Sirinivas;Ly, Danith H.
通讯作者：
Ly, Danith H.
影响因子：
3.6
作者：
Bahal, Raman;Quijano, Elias;Glazer, Peter M.
通讯作者：
Glazer, Peter M.