Lateral Flow Test (LFT) Detects Cell‐Free MicroRNAs Predictive of Preterm Birth Directly from Human Plasma

Extracellular Vesicles
/References

Despite extensive research toward the development of point‐of‐care nucleic acid tests (POC NATs) for the detection of microRNAs (miRs) from liquid biopsies, major hurdles remain including the strict requirement for extensive off‐chip sample preprocessing. Herein, a nucleic acid lateral flow test (NALFT) is reported on that enables the direct detection of endogenous miRs from as little as 3 μL of plasma without the requirement for any enzyme‐catalyzed target amplification or complex miR extraction steps. This is achieved through integration of a denaturing hydrogel composite material onto the LFT, allowing for near‐instantaneous on‐chip release of miRs from their carriers (extracellular vesicles or transport proteins) prior to detection. This next‐generation LFT is sensitive enough to detect endogenous concentrations of miR‐150‐5p, a predictive biomarker for preterm birth (PTB) found deregulated in maternal blood from as early as 12th week of pregnancy. Herein, a key step is represented toward a first bedside test for risk‐stratification during pregnancy by predicting true outcome at a very early stage. More generally, the universal and versatile nature of this novel sample preprocessing platform can further improve the robustness of existing NALFTs and facilitate their application at the POC. On‐chip microRNA release from carrier protein and extracellular vesicles using a denaturing hydrogel–cellulose pad enables direct detection of predictive circulating microRNA biomarkers for preterm birth directly from human plasma. This next‐generation bedside lateral flow test provides a unique tool for risk‐stratification during pregnancy by predicting true outcome at a very early stage.

View full article

Recent Publications

Cigarette smoke (CS) represents one of the most relevant environmental risk factors for several chronic pathologies. Tissue damage caused by CS exposure is mediated, at least in part, by oxidative stress induced by its toxic and pro-oxidant components. Evidence demonstrates that extracellular vesicles (EVs) released by various cell types exposed to CS extract (CSE) are characterized by altered biochemical cargo and gained pathological properties. In the present study, we evaluated the content of oxidized proteins and phospholipid fatty acid profiles of EVs released by human bronchial epithelial BEAS-2B cells treated with CSE. This specific molecular characterization has hitherto not been performed. After confirmation that CSE reduces viability of BEAS-2B cells and elevates intracellular ROS levels, in a dose-dependent manner, we demonstrated that 24 h exposure at 1% CSE, a concentration that only slight modifies cell viability but increases ROS levels, was able to increase carbonylated protein levels in cells and released EVs. The release of oxidatively modified proteins via EVs might represent a mechanism used by cells to remove toxic proteins in order to avoid their intracellular overloading. Moreover, 1% CSE induced only few changes in the fatty acid asset in BEAS-2B cell membrane phospholipids, whereas several rearrangements were observed in EVs released by CSE-treated cells. The impact of changes in acyl chain composition of CSE-EVs accounted for the increased saturation levels of phospholipids, a membrane parameter that might influence EV stability, uptake and, at least in part, EV-mediated biological effects. The present in vitro study adds new information concerning the biochemical composition of CSE-related EVs, useful to predict their biological effects on target cells. Furthermore, the information regarding the presence of oxidized proteins and the specific membrane features of CSE-related EVs can be useful to define the utilization of circulating EVs as marker for diagnosing of CS-induced lung damage and/or CS-related diseases.

2023
No items found.
No items found.
No items found.