![]() Here, we evaluate how different blood anticoagulation options and processing times affect platelet function and protein content ex vivo. Our study mainly highlights the merits of liquid biopsy as a promising biomarker for non-invasive detection in the future, particularly for the early detection of NSCLC, thereby benefitting human health.Įx vivo assays of platelet function critically inform mechanistic and clinical hematology studies, where effects of divergent blood processing methods on platelet composition are apparent, but unspecified. Additionally, we outline the existing challenges and possible solutions for liquid-biopsy biomarkers. This review summarizes some latest research on the diagnosis of early-stage NSCLC via liquid biopsy, including circulating DNA, circulating tumor cells, exosomes, and tumor-educated platelets, as well as their detection technologies, such as fluorescence in-situ hybridization-based, polymerase chain reaction-based, next-generation sequencing-based, Chip-based, and microfluidic methods. Owing to new detection technologies, liquid biopsy reduces the need for invasive treatments and enhances the accuracy and specificity of early detection of cancer in clinical settings. In comparison, liquid biopsy taken from body fluid reflects extensive malignant features nonexistent in primary tumors. Invasive tissue biopsy is a common diagnostic tool that is usually extracted from the primary tumor to indicate molecular composition. To date, various reports emphasize the need for non-small cell lung carcinoma (NSCLC), both with higher incidence and mortality and less effective treatments thus, emphasizing the need for early detection of NSCLC for improved patient outcomes. Liquid biopsy, an innovative method for early diagnosis of cancer, has changed the traditional method of diagnosing lung cancer and is considered a feasible auxiliary diagnostic tool. In this review, we will identify the main challenges and biases introduced during the different stages of biomarker discovery in liquid biopsies with cfRNA and propose solutions to minimize them. One of the main drawbacks in the field of RNA-based liquid biopsies is the low reproducibility of the results, often caused by technical and biological variability, a lack of standardized protocols and insufficient cohorts. Since 2020, a handful of cfDNA tests have been approved for therapy selection by the FDA, however, no cfRNA tests are approved to date. Historically, studies have aimed at the detection of specific mutations on cell-free DNA (cfDNA), but recently, the study of cell-free RNA (cfRNA) has gained traction. Although it can target any biomolecule, most liquid biopsy studies have focused on circulating nucleic acids. ![]() The analysis of biomarkers in biological fluids, also known as liquid biopsies, is seen with great potential to diagnose complex diseases such as cancer with a high sensitivity and minimal invasiveness. Our results suggest that dissecting the complexity of EVs subpopulations illuminates their biological significance and offers a promising liquid biopsy approach. ![]() The EV-enriched differentiating genes were associated with specific biological pathways, such as immune systems, liver function, and toxic substance regulation in lung cancer, liver cancer, and multiple myeloma, respectively. We observed specific enrichment patterns of cancer-associated cf-mRNA in each vesicular and non-vesicular subpopulation. We found that the majority of cf-mRNA was enriched and protected in EVs with remarkable stability in RNase-rich environments. We developed a strategy to purify and sequence ultra-low amounts of cf-mRNA from vesicular and non-vesicular subpopulations with the implementation of RNA spike-ins to control for technical variability and to normalize for intrinsic drastic differences in the amount of cf-mRNA carried in each plasma fraction. Morphology and size distribution analysis showed the successful separation of medium and small EVs and non-vesicular carriers. Here, we performed a transcriptomic analysis of size-fractionated plasma from lung cancer, liver cancer, multiple myeloma, and healthy donors. However, carriers of cell-free messenger RNA (cf-mRNA) in human biofluid and their association with cancer remain poorly understood. Extracellular vesicles (EVs) have been shown as key mediators of extracellular small RNA transport. ![]()
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