定义RNA货物分选到细胞外囊泡中的参数。
Defining the Parameters for Sorting of RNA Cargo Into Extracellular Vesicles.
作者信息
Abdelgawad Ahmed, Huang Yiyao, Gololobova Olesia, Yu Yanbao, Witwer Kenneth W, Parashar Vijay, Batish Mona
机构信息
Department of Biological Sciences, University of Delaware, Newark, Delaware, USA.
Department of Medical and Molecular Sciences, University of Delaware, Newark, Delaware, USA.
出版信息
J Extracell Vesicles. 2025 Jul;14(7):e70113. doi: 10.1002/jev2.70113.
Extracellular vesicles (EVs) are small particles that are released by cells and mediate cell-cell communication by transferring bioactive molecules such as RNA. RNA cargo of EVs, including coding and non-coding RNAs, can change the behaviour of recipient cells, affecting processes including gene expression, proliferation, and Fapoptosis. CircRNAs are stable and resistant to degradation and have been shown to be enriched in EVs. They play key roles in gene regulation and are also emerging as promising biomarkers for disease diagnosis due to their stability and disease-specific expression. Although microRNAs (miRNAs) are the most well studied RNA cargo of EVs, very little is known about the mechanisms of enrichment of circular RNAs (circRNAs) as well as long linear RNAs. Here, we take a comprehensive genome-wide approach to investigate the role of structuredness and shape along with GC%, size, exon count and coding potential, in the sorting and enrichment of circular and long linear RNAs into EVs. We developed a model using these parameters to predict the likelihood of EV packaging of RNA and it was validated by using single molecule RNA imaging of EV bound RNAs. Furthermore, we found that structuredness could explain the relative enrichment of circRNAs over their linear counterparts. These results were validated on existing public databases of circular and linear RNAs in EVs. By identifying and analysing these factors, we aim to better understand the complex mechanisms behind EV-mediated RNA transfer and its impact on cell communication in both health and disease. This mechanistic understanding of RNA enrichment in EVs is crucial for engineering EVs with selective RNA cargo.
细胞外囊泡(EVs)是由细胞释放的小颗粒,通过转移RNA等生物活性分子介导细胞间通讯。EVs的RNA货物,包括编码RNA和非编码RNA,可改变受体细胞的行为,影响基因表达、增殖和凋亡等过程。环状RNA(circRNAs)稳定且抗降解,已被证明在EVs中富集。它们在基因调控中起关键作用,由于其稳定性和疾病特异性表达,也正成为疾病诊断中有前景的生物标志物。尽管微小RNA(miRNAs)是研究最深入的EVs的RNA货物,但对于环状RNA(circRNAs)以及长链线性RNA的富集机制知之甚少。在这里,我们采用全面的全基因组方法来研究结构和形状以及GC%、大小、外显子数量和编码潜能在环状和长链线性RNA分选和富集到EVs中的作用。我们利用这些参数开发了一个模型来预测RNA被包装到EVs中的可能性,并通过对与EV结合的RNA进行单分子RNA成像进行了验证。此外,我们发现结构可以解释circRNAs相对于其线性对应物的相对富集。这些结果在现有的EVs中环状和线性RNA的公共数据库上得到了验证。通过识别和分析这些因素,我们旨在更好地理解EV介导的RNA转移背后的复杂机制及其对健康和疾病中细胞通讯的影响。这种对EVs中RNA富集的机制理解对于构建具有选择性RNA货物的EVs至关重要。