Expression of Epithelial and Mesenchymal Markers in Plasmatic Extracellular Vesicles as a Diagnostic Tool for Neoplastic Processes

Abstract

Tumor-derived extracellular vesicles (TD-EVs) have active roles as cancer hallmark enablers. EVs RNA of epithelial and stromal cells carry information that facilitates the communication processes that contribute to oncological progression, so the objective of this work was to validate by RT-PCR the presence of epithelial (KRT19; CEA) and stromal (COL1A2; COL11A1) markers in RNA of plasmatic EVs in healthy and diverse-malignancy patients for the development of a non-invasive cancer diagnosis system using liquid biopsy. Ten asymptomatic controls and 20 cancer patients were included in the study, and results showed that the isolated plasmatic EVs by scanning transmission electron microscopy (STEM) andBiomedical Research Institute A Coruña nanoparticle tracking analysis (NTA) contained most exosome structures with also a considerable percentage of microvesicles. No differences were found in concentration and size distribution between the two cohorts of patients, but significant gene expression in epithelial and mesenchymal markers between healthy donors and patients with active oncological disease was shown. Results of quantitative RT-PCR are solid and reliable for KRT19, COL1A2, and COL11A1, so the analysis of RNA extracted from TD-EVs could be a correct approach to develop a diagnostic tool in oncological processes.

Unraveling the Metastatic Niche in Breast Cancer Bone Metastasis through Single-Cell RNA Sequencing

“…Consequently, clusters were grouped based on the expression of distinct cell type markers, leading to the classi cation into four clusters (epithelial cells, endothelial cells, immune cells, and broblasts). A similar analytical pipeline was applied to immune cells, where the identi cation of immune cell types was achieved by matching each cluster-speci c gene set with known signature genes of cell populations reported in previous literature [17][18][19][20][21][22][23][24] .…”

Unravelling the Metastatic Niche in Breast Cancer Patients with Bone Metastasis through Single-cell RNA Sequencing

“…Consequently, the clusters were grouped based on the expression of distinct cell type markers, leading to the classi cation of cells into four clusters (epithelial cells, endothelial cells, immune cells, and broblasts). A similar analytical pipeline was applied to immune cells, where the identi cation of immune cell types was achieved by matching each cluster-speci c gene set with known signature genes of cell populations reported in previous literature [17][18][19][20][21][22][23][24] .…”

Cancer screening with multicancer detection tests: A translational science review

“…Technologies to detect and measure cancer‐related changes in circulating tumor DNA (ctDNA) ( liquid biopsy ) and other analytes have supported MCD assay development (Table 2). 22–52 Proof of principle that ctDNA can detect occult malignancies was first documented by the incidental detection of cancer in women undergoing noninvasive prenatal screening for fetal aneuploidies 53 . Most MCD tests are designed to evaluate multiple features of ctDNA, such as mutation profiles, methylation signatures, and fragmentation patterns.…”

“…These biomarkers are components of several tumor‐related processes. For example, extracellular vesicles, which are secreted by living cells and may contain DNA, microRNAs, proteins, or lipids, are mediators of intercellular communication 44,45 …”

“…Assay developers continue to modify the biologic and algorithmic components of their tests. To categorize the readiness of MCD testing for clinical use, we mapped publicly reported MCD assays 22,23,25,27,28,30‐32,34‐36,39‐43,47,52,54,56‐74 to their phase of development (Figure 1) 26‐61,73,74 based on a five‐phase cancer biomarker framework developed by the NCI's Early Detection Research Network (EDRN) 5,75 . Most MCD tests are in early stages of development (Figure 1); their diagnostic performance is trained and validated based on retrospective studies using cohorts predominantly comprised of specimens obtained from individuals with a definitive diagnosis of cancer or from individuals with no known cancer (EDRN phase 2 or 3).…”