Herrero Carolina, Ferreirós Alba, Pérez-Fentes Daniel, León-Mateos Luis, López-López Rafael, Abal Miguel, Alonso-Alconada Lorena
Translational Medical Oncology Group (Oncomet), Health Research Institute of Santiago de Compostela (IDIS), University Hospital of Santiago de Compostela (SERGAS), CIBERONC, 15706 Santiago de Compostela, Spain.
Nasasbiotech, S.L., Canton Grande 9, 15003 A Coruña, Spain.
Biomedicines. 2023 Jan 30;11(2):404. doi: 10.3390/biomedicines11020404.
The two most developed biomarkers in liquid biopsy (LB)-circulating tumor cells and circulating tumor DNA-have been joined by the analysis of extracellular vesicles (EVs). EVs are lipid-bilayer enclosed structures released by all cell types containing a variety of molecules, including DNA, mRNA and miRNA. However, fast, efficient and a high degree of purity isolation technologies are necessary for their clinical routine implementation. In this work, the use of ExoGAG, a new easy-to-use EV isolation technology, was validated for the isolation of EVs from plasma and urine samples. After demonstrating its efficiency, an analysis of the genetic material contained in the EVs was carried out. Firstly, the sensitivity of the detection of point mutations in DNA from plasma EVs isolated by ExoGAG was analyzed. Then, a pilot study of mRNA expression using the nCounter NanoString platform in EV-mRNA from a healthy donor, a benign prostate hyperplasia patient and metastatic prostate cancer patient plasma and urine samples was performed, identifying the prostate cancer pathway as one of the main ones. This work provides evidence for the value of using ExoGAG for the isolation of EVs from plasma and urine samples, enabling downstream applications of the analysis of their genetic cargo.
液体活检（LB）中两种最成熟的生物标志物——循环肿瘤细胞和循环肿瘤DNA，现在又加入了细胞外囊泡（EVs）分析。EVs是由所有细胞类型释放的脂质双层包裹结构，包含多种分子，包括DNA、mRNA和miRNA。然而，其临床常规应用需要快速、高效且高纯度的分离技术。在这项工作中，新型易用的EV分离技术ExoGAG用于从血浆和尿液样本中分离EVs的方法得到了验证。在证明其效率后，对EVs中所含的遗传物质进行了分析。首先，分析了用ExoGAG从血浆EVs中分离的DNA中检测点突变的灵敏度。然后，使用nCounter NanoString平台对来自健康供体、良性前列腺增生患者和转移性前列腺癌患者血浆及尿液样本的EV-mRNA中的mRNA表达进行了一项初步研究，确定前列腺癌通路是主要通路之一。这项工作为使用ExoGAG从血浆和尿液样本中分离EVs的价值提供了证据，从而能够对其遗传物质进行下游分析应用。
