microRNAs are small oligonucleotides involved in post-transcriptional gene regulation whose alteration is found in several diseases, including cancer, and therefore their detection is crucial for diagnosis, prognosis, and treatment purposes. Field-Effect Transistor-based biosensors (bioFETs) represent a promising technology for the clinical detection of microRNAs. However, one of the main challenges associated with this technology is the Debye screening, becoming significant at the high ionic strengths required for effective hybridization. We aimed at detecting oncogenic microRNA-155 by using a bioFET system using as capture element a complementary RNA probe (antimiR-155) combined with the introduction of PEG molecules (20 kDa, PEG20), at an ionic strength of 300 mM. We optimized the co-immobilization ratio between antimiR-155 and PEG20 and assessed its impact on the interactions between the oligonucleotides. The kinetics can be well described by the Langmuir-Freundlich isotherm with an affinity constant within the range typical of nucleic acid interactions. The introduction of PEG20 significantly enhanced the detection sensitivity of miR-155 by reaching a level of less than 200 pM, together with excellent discrimination against other clinically relevant microRNAs. Our findings demonstrate that the incorporation of PEG20 constitutes an effective strategy to mitigate the Debye screening effects and facilitates bioFET-based clinical applications at physiological ionic strengths.

The reproducibility of the analysis of oligonucleotides using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) remains a significant challenge. This is mainly attributed to factors such as the choice and application of the matrix, as well as the inhomogeneity of the sample spots. Although previous studies have explored various parameters, such as ionic and sugar-based matrices, and different spotting methods, none have comprehensively integrated these factors into a unified approach.

Various matrices for the analysis of oligonucleotides and the effects of diverse variables, including different matrices, solvent compositions, additives, and application techniques, on the analytical performance of these matrices were investigated. Mass spectrometry analysis was conducted by a MALDI-TOF MS in linear negative mode.

Our research systematically evaluates the combined effects of diverse variables to enhance the analytical performance of MALDI-TOF MS in oligonucleotide analysis. We focused on the standard deviation of mass-to-charge ratios and the signal-to-noise (S/N) ratios. Out of 48 samples, only 19 met the S/N criteria, which is that the signals must be detectable over the whole mass range of interest (4-10 kDa).

The ionic matrix 6-aza-2-thiothymine (ATT) with 1-methylimidazole resulted consistently in a reduced standard deviation and achieved high mass precisions. Additionally, we observed that the S/N ratios and mass precision of 3-hydroxypicolinic acid (3-HPA) varied significantly depending on the solvent composition and the presence of additives.

MicroRNAs (miRNAs) have emerged as critical biomarkers for early cancer diagnosis and monitoring. However, their isolation from clinical samples typically yields only trace amounts, significantly limiting the sensitivity and efficiency of cancer detection. To address this challenge, we present a octangular reconfigurable acoustic tweezer (ORAT) as an integrated platform for precise tumor cell tracking and in-situ detection of trace miRNAs. By simultaneously modulating multidirectional acoustic signals and parameters, the ORAT dynamically reshapes the acoustic field, enabling precise control over manipulation areas, particle spacing, array angles, distribution patterns, and node rotation. This device allows selective particle manipulation across entire regions or specific areas through adaptive adjustments of the microchamber boundary. Notably, the ORAT achieves rapid and accurate localization and labeling of rare tumor cells within a large population of normal cells. Furthermore, it enhances the sensitivity of CRISPR/Cas-based miRNA detection in digital microdroplets by three orders of magnitude, if compared to that of the conventional tube-based method. With its versatile capabilities, the ORAT holds remarkable promise for advancing nucleic acid analysis in a wide range of cancers and related diseases.

MicroRNA (miRNA) is a common tumor marker, whose abnormal expression is often closely related to the occurrence of various diseases. However, the conventional method for detecting miRNA is qRT-PCR, requiring additional reverse transcription steps, well-trained professionals, and expensive thermal cycling equipment. In this work, we propose a novel isothermal amplification technique (exponential rolling circle amplification-hybridization chain reaction, EXRCA-HCR) for AgNPs@gel-enhanced fluorescence specific and ultrasensitive detection of miRNA-21. This novel technique consists of rolling circle amplification (RCA), exponential isothermal amplification reaction (EXPAR) and hybridization chain reaction (HCR). Combining these three amplification methods, EXRCA-HCR provides a unique cascade amplification strategy, inheriting the advantages of linear amplification and exponential amplification. Under optimal conditions, this novel EXRCA-HCR exhibits a wide fluorescent detection range from 200 fM to 200 nM for miRNA-21, with low detection limit of 21.47 fM. By introducing AgNPs@gel, the fabricated paper-based fluorosensor based on EXRCA-HCR provides a simple and rapid visual detection of miRNA-21. This research puts forward a promising approach for detecting miRNA-21, which can be applied for early diagnosis.

Human settlements on the Moon, crewed missions to Mars and space tourism will become a reality in the next few decades. Human presence in space, especially for extended periods of time, will therefore steeply increase. However, despite more than 60 years of spaceflight, the mechanisms underlying the effects of the space environment on human physiology are still not fully understood. Animals, ranging in complexity from flies to monkeys, have played a pioneering role in understanding the (patho)physiological outcome of critical environmental factors in space, in particular altered gravity and cosmic radiation. The use of animals in biomedical research is increasingly being criticized because of ethical reasons and limited human relevance. Driven by the 3Rs concept, calling for replacement, reduction and refinement of animal experimentation, major efforts have been focused in the past decades on the development of alternative methods that fully bypass animal testing or so-called new approach methodologies. These new approach methodologies range from simple monolayer cultures of individual primary or stem cells all up to bioprinted 3D organoids and microfluidic chips that recapitulate the complex cellular architecture of organs. Other approaches applied in life sciences in space research contribute to the reduction of animal experimentation. These include methods to mimic space conditions on Earth, such as microgravity and radiation simulators, as well as tools to support the processing, analysis or application of testing results obtained in life sciences in space research, including systems biology, live-cell, high-content and real-time analysis, high-throughput analysis, artificial intelligence and digital twins. The present paper provides an in-depth overview of such methods to replace or reduce animal testing in life sciences in space research.

Progestin and adipoQ receptor family member 4 (PAQR4) gene is a recently discovered seven-transmembrane protein-coding gene that belongs to the PAQR family. An increasing amount of evidence suggests that PAQR4 is upregulated in multiple tumors and participates in tumor progression and chemotherapy resistance via different signaling pathways; PAQR4 regulates cellular ceramide homeostasis by influencing sphingolipid metabolism and glycerol metabolism, and plays a significant role in adipose tissue remodeling. Meanwhile, it is known that the differential expression of PAQR4 is associated with the occurrence of various diseases and is a potential biomarker and therapeutic target. This article conducts a systematic review of the subcellular localization of PAQR4, its topological structure characteristics, and its functions in cancer occurrence, metabolic diseases, and fertility, and provides clues for the future research and translational application of PAQR4.

Hair follicle stem cells (HFSCs) can quickly activate and migrate to the wound site, differentiating into epidermal stem cells to facilitate early epithelialization. During the wound healing process, microRNAs (miRNAs) function coordinately. Chinese medicine borneol is derived from the Cinnamomum camphora plant, and its principal component, L-borneol, is renowned for its potential in facilitating skin wound healing. However, it remains unclear whether L-borneol can stimulate HFSCs to differentiate into epidermal cells or whether miRNAs are involved in this process. In the current study, HFSCs were isolated from the vibrissae of rats and identified based on the expression of CD34, Integrin-β1 and keratin type 1 cytoskeletal 15(CK15). We observed that stimulation with L-borneol significantly increased the differentiation marker K14 in HFSCs, suggesting that L-borneol could promote the differentiation of HFSCs into basal layer cells. On this basis, we transfected and confirmed that rno-miR-127 inhibitor could promote the differentiation of HFSCs. Furthermore, we demonstrated that PODXL2 is a target gene of rno-miR-127 through dual-luciferase reporter assays and confirmed that the rno-miR-127 mimic could inhibit the expression of PODXL2. To further elucidate the targeting relationship, we constructed the siPODXL2 fragment using siRNA technology, demonstrating that reducing PODXL2 expression can inhibit the differentiation of HFSCs into basal layer cells. Finally, a rat full-thickness skin defect model illustrated L-borneol-mediated negative regulation of PODXL2 by rno-miR-127, promoting skin injury repair through HFSCs.

RNA activation (RNAa), a gene regulatory mechanism mediated by small activating RNAs (saRNAs) and microRNAs (miRNAs), has significant implications for therapeutic applications. Unlike small interfering RNA (siRNA), which is known for gene silencing in RNA interference (RNAi), synthetic saRNAs can stably upregulate target gene expression at the transcriptional level through the assembly of the RNA-induced transcriptional activation (RITA) complex. Moreover, the dual functionality of endogenous miRNAs in RNAa (hereafter referred to as mi-RNAa) reveals their complex role in cellular processes and disease pathology. Emerging studies suggest saRNAs' potential as a novel therapeutic modality for diseases such as metabolic disorders, hearing loss, tumors, and Alzheimer's. Notably, MTL-CEBPA, the first saRNA drug candidate, shows promise in hepatocellular carcinoma treatment, while RAG-01 is being explored for non-muscle-invasive bladder cancer, highlighting clinical advancements in RNAa. This review synthesizes our current understanding of the mechanisms of RNAa and highlights recent advancements in the study of mi-RNAa and the therapeutic development of saRNAs.

Age estimation is a critical aspect of human identification. Traditional methods, reliant on morphological examinations, are often suitable for living subjects. However, there are relatively few studies on age estimation based on biological samples, such as blood. Recent advancements have concentrated on DNA methylation for forensic age prediction. However, to explore further possibilities, this study investigated microRNAs (miRNAs) as alternative molecular markers for age estimation. Peripheral blood samples from 127 healthy individuals were analyzed for miRNA expression using small RNA sequencing. Lasso regression selected 103 candidate miRNAs, and Shapley additive explanations (SHAP) analysis identified 38 key miRNAs significant for age prediction. Five machine learning models were developed, with the elastic net model achieving the best performance (MAE of 4.08 years) on the testing set, surpassing current miRNA age estimation results. Additionally, we observed significant changes in the expression levels of miRNAs in healthy individuals aged 48-52 years. This study demonstrated the potential of blood miRNA biomarkers in age prediction and provides a set of miRNA markers for developing more accurate age prediction methods.

Melanoma is a devastating form of skin cancer characterized by a high mutational burden, limited treatment success, and dismal prognosis. Although immunotherapy and targeted therapies have significantly revolutionized melanoma treatment, the majority of patients fail to achieve durable responses, highlighting the urgent need for novel therapeutic strategies. Ferroptosis, an iron-dependent form of regulated cell death driven by the overwhelming accumulation of lipid peroxides, has emerged as a promising therapeutic approach in preclinical melanoma models. A deeper understanding of the ferroptosis landscape in melanoma based on its biology characteristics, including phenotypic plasticity, metabolic state, genomic alterations, and epigenetic changes, as well as the complex role and mechanisms of ferroptosis in immune cells could provide a foundation for developing effective treatments. In this review, we outline the molecular mechanisms of ferroptosis, decipher the role of melanoma biology in ferroptosis regulation, reveal the therapeutic potential of ferroptosis in melanoma, and discuss the pressing questions that should guide future investigations into ferroptosis in melanoma.

MicroRNAs hold great potential as biomarkers for assessing the progression of acute pancreatitis (AP). This study aimed to explore the value of miR-146b-5p in the diagnosis and prognosis of AP patients.

110 AP patients were included and divided into 40 severe AP (SAP) patients and 70 non-SAP patients based on disease severity. Serum miR-146b-5p levels were measured using RT-qPCR. The diagnostic value of miR-146b-5p was evaluated utilizing ROC curves. Pearson correlation coefficient was employed to analyze the correlations between APACHEII, BISAP, and MCTSI scores and miR-146b-5p levels. The AP cell model was constructed by treating AR42J cells with deoxycholic acid (DCA), the proliferative capacity of cells was measured with CCK-8, apoptosis was measured by flow cytometry, and IL-6 and IL-8 protein levels were analyzed by ELISA.

Serum miR-146b-5p levels were decreased in SAP and unfavorable patients. Serum miR-146b-5p was able to effectively differentiate between SAP and non-SAP patients, and also effectively differentiate between unfavorable and favorable patients. MiR-146b-5p levels were significantly negatively correlated with APACHEII score (r=-0.6676), BISAP score (r=-0.5696), and MCTSI score (r=-0.5857). Furthermore, in the AP cell model, miR-146b-5p expression was down-regulated, proliferative capacity was diminished, apoptosis was increased, and IL-6 and IL-8 levels were elevated, but overexpression of miR-146b-5p partially reversed these changes.

miR-146b-5p expression is down-regulated in the serum of SAP patients and cells, and it has a good diagnostic effect. It may be a potential biomarker and therapeutic target for AP.

The precise identification and differentiation of multiple microRNAs (miRNAs) with high spatial resolution in specific cells remain a significant challenge, primarily due to the limited availability of spectrally distinguishable fluorophores and the absence of cell-specific recognition capabilities. In this study, we introduce a programmed fluorescence-encoding DNA nanoflower (CNFs) system based on the self-assembly of rolling circle amplification (RCA), enabling multiplexed miRNA imaging in living cells. The CNFs system is rationally designed to consist of three key components: a CD63 aptamer region, dual fluorophore encoding regions, and an miRNA recognition region. The polyvalent tandem CD63 aptamer enhances the cellular targeting specificity and endocytic uptake efficiency. By controlling dual fluorophores and three levels of intensity within encoding regions, it generates 9 distinct barcodes for labeling multiple targets. Additionally, when conjugated with molecular beacons (MBs), CNFs facilitate the simultaneous detection of multiplexed intracellular miRNAs. Using this CNFs system, we successfully evaluated the expression profiles of nine miRNAs in breast cancer. Overall, we expect that this CNFs system will be a valuable tool for disease-related multiplex miRNAs biomarker imaging in specific cells and the exploration of miRNAs' molecular regulation mechanisms.

The hippocampus is the main target of glucocorticoids (GCs) in the brain since it contains the greatest concentration of the specific receptors. GCs are among the factors modulating adult hippocampal neurogenesis (AHN), which occurs in mammalians, including humans. Prolonged exposure to high GC levels triggers AHN impairment and induces affective and cognitive deficits, consistently with hippocampal neurogenesis functions. Cushing's syndrome (CS) is a rare endocrine disorder characterized by persistently elevated GC levels, namely, cortisol, that also results in affective disorders and impairment of hippocampus-associated memory, suggesting a disruption of hippocampal neurogenesis. Players of adult neurogenesis process, such as Neural Stem/Progenitor Cells and differentiating neuronal cells, release exosomes able to cross brain blood barrier, reaching the peripheral blood. MicroRNAs are known to be selectively enriched in neuronal exosomes and to play a crucial role in adult neurogenesis regulation. The main question addressed in this exploratory study was whether neuroplasticity-related microRNAs (miRNAs), carried by neuronal-derived exosomes in peripheral blood, could reflect alterations in neurogenic processes associated with Cushing's syndrome. Hence, in the present work, we measured the content in selected miRNAs of neuronally derived exosomes in peripheral blood of patients affected by endogenous and active CS and age and sex-matched healthy subjects. The human miRNAs (miR-126, miR-9, miR-223, miR-34a, miR-124a, and miR-146a) were quantified by RT-qPCR. All the miRNAs analyzed were significantly differentially expressed in CS patients as compared to healthy subjects. Our findings support the following: (i) patients with Cushing's syndrome (CS) may exhibit a putative dysregulation of neurogenesis that could underlie the early-onset impairment of affective and cognitive functions; (ii) the exosomal cargo may represent a potential biomarker for monitoring functional and dysfunctional neuroplasticity processes in adult humans. Additional studies are needed to confirm and expand upon the findings across a wider cohort of patients.

Wound healing in diabetic patients is mainly hindered by a combination of long-term glycosylation, persistent inflammatory response, and immunosuppressive state. The interaction of these factors not only results in considerable prolongation of the wound healing process but also elevates the likelihood of recurrent ulcer development, profoundly affecting patients' quality of life. Traditional treatments, including surgical debridement, anti-infection, dressing application, vascular intervention, and glycaemic control, can only relieve some symptoms. However, they are often ineffective in addressing the underlying cause of impaired wound healing. It is of concern that the importance of the immune microenvironment in diabetic wound healing has not yet been fully appreciated and investigated, and the homeostasis of the immune microenvironment is crucial for promoting cell proliferation, angiogenesis, and tissue repair. However, this microenvironment is often dysregulated in the diabetic state. This paper reviews the key factors leading to dysregulation of the immune microenvironment, including immune cell dysfunction, abnormal cytokine expression, and disruption of key signalling pathways, and introduces an innovative silicone-based microneedle drug delivery method, which takes advantage of microneedle's precise targeting and highly efficient drug loading capacity to deliver drugs with immunomodulatory functions directly to the wound in a sustained manner, activate the corresponding signalling pathways, promote the polarization of M1 macrophages into the M2 phenotype, and stimulate neovascularization, providing a low inflammatory and pro-angiogenic immune microenvironment for diabetic wound healing, which provides a new therapeutic idea and means for diabetic wound healing.

Progressive multifocal leukoencephalopathy (PML) is a demyelinating disease caused by JC polyomavirus (JCV). The histopathology of PML is morphologically diverse and characterized by the classical triad of demyelination, enlarged oligodendroglial nuclei, and bizarre astrocytes. Pathological diagnostic criteria for PML require both the classical triad and viral detection in brain tissue. However, the frequency of this triad in surgical pathology specimens and its correlation with disease progression and viral loads remain unclear. In this study, 117 brain tissues from 91 pathologically confirmed PML patients were investigated. PML histopathology was found to be spatially and temporally pleomorphic, and not all brain tissues exhibited the complete classical triad. The sensitivity of quantitative PCR for detecting JCV in brain tissues was 100%, whereas that of immunohistochemistry (IHC) was 83.5-87.8%. Viral loads in biopsy samples were significantly higher than those in autopsy samples and decreased over time after disease onset. To systematically characterize PML lesions from the outer border to the demyelinated center, we developed a histological classification based on the classical triad and macrophage infiltration. This classification correlated with viral loads, with subtypes characterized by abundant enlarged oligodendroglial nuclei at the demyelination border exhibiting the highest levels of JCV DNA. Pathological variability was influenced by spatial and temporal factors rather than by underlying diseases, although PML associated with acquired immunodeficiency syndrome exhibited more severe demyelination. In conclusion, histomorphological variability in PML reflects viral replication activity, emphasizing the importance of comprehensive pathological evaluation. Combining histomorphology, tissue-based PCR for viral DNA detection, and IHC for viral antigens is crucial for assessing disease progression. Early brain biopsy from the demyelination border offers the best opportunity for a definitive diagnosis of PML and may guide therapy targeting active lesions.

PIWI-interacting RNAs (piRNAs) are a class of small, non-coding RNAs predominantly expressed in the germ cells of animals and play a crucial role in maintaining genomic integrity, mediating transposon suppression, and ensuring gene stability. Beyond their functions in reproductive cells, piRNAs also play roles in various human diseases, including cancer, suggesting their potential as significant biomarkers critical for disease diagnosis and treatment. Wet-lab methods to identify piRNA-disease associations require substantial resources and are often hit-or-miss. With advancements in computational technologies, an increasing number of researchers are employing computational methods to efficiently predict potential piRNA-disease associations. The sparsity of data in piRNA-disease association studies significantly limits model performance improvement. In this study, we propose a novel computational model, iPiDA_CL, to predict potential piRNA-disease associations through contrastive learning methods, which do not require negative samples. The model represents piRNA-disease association pairs as a bipartite graph and computes the initial embeddings of piRNAs and diseases using Gaussian kernel similarity, with features updated via LightGCN. Based on the siamese network framework, iPiDA_CL constructs online and target networks and employs data augmentation in the target network to build a contrastive learning objective that optimizes model parameters without introducing negative samples. Finally, cross-prediction methods are used to calculate specific piRNA-disease association scores. A series of experimental results demonstrate that iPiDA_CL surpasses state-of-the-art methods in both performance and computational efficiency. The application of iPiDA_CL to the miRNA-disease association dataset underscores its versatility across various ncRNA-disease association task. Furthermore, a case study highlights iPiDA_CL as an efficient and promising tool for predicting piRNA-disease associations.

Barramundi (Lates calcarifer) are emerging as a key species in warm-water aquaculture worldwide; however, disease outbreaks caused by Vibrio spp. are impeding industry expansion. Climate change is expected to exacerbate this issue by intensifying extreme weather events, including unusually cold temperatures, thereby increasing the risk of disease. In this study, we investigated the combined effect of cold stress and V. harveyi infection on the early transcriptome (mRNA) and microRNA responses of juvenile barramundi to enhance our understanding of host-pathogen interactions. High levels of differential gene expression were observed in fish subjected to cold stress (22 °C) post-infection with V. harveyi, with 3231 differentially expressed genes and an extensive pro-inflammatory immune response. In contrast, most differentially expressed microRNAs were associated with fish infected with V. harveyi housed under optimal temperature conditions (30 °C). MicroRNAs play a crucial role in regulating gene expression, typically through downregulation of target mRNAs. The significant upregulation of miRNAs in barramundi kept at 30 °C, and the lack of miRNA upregulation in cold stressed fish, suggests that cold stress impaired the immune-regulatory capacity of affected fish, resulting in a hyper-inflammatory response that may account for the increased mortality observed. This study is the first dual study of the transcriptome and microRNA response of barramundi to V. harveyi infection and expands understanding of the innate immune response in barramundi and the regulatory role of microRNAs in teleost fish.

MicroRNAs (miRNAs) are small, non-coding RNA molecules that play crucial roles in regulating gene expression, and their dysregulation is implicated in various human diseases. Over the years, several research groups have identified miRNAs as promising therapeutic targets for intervention. Therapeutic strategies involve either overexpression or knockdown of specific miRNAs. This review aims to provide a comprehensive overview of synthetic poly-miRNAs and miRNA sponges, highlighting their therapeutic applications. It begins with an introduction to miRNAs and their role in human diseases, followed by a detailed discussion on synthetic poly-miRNAs and miRNA sponges by exploring their application in cardiovascular, inflammatory, autoimmune, and metabolic disorders, as well as in cancer therapy. Additionally, strategies for targeted delivery, challenges, and limitations of these therapies are addressed.

DNA walkers are powerful nanomachines for biosensing and bioimaging, yet their application in microRNA detection faces significant challenges. Current DNA walker designs often involve complex structures with separate walker and track components, leading to reduced efficiency and potential interference in cellular environments. Here, we developed a self-propelled multi-leg walker (SMW) where autocatalytic hairpin assembly (ACHA) components are anchored on a DNA nanowire to create an initial track structure. Upon target recognition, catalytic hairpin assembly (CHA) reactions dynamically transform sections of track into multi-leg walkers with newly generated triggers. ACHA reactions are then initiated by these triggers, driving the walker to move autonomously along the unreacted track while generating amplified fluorescent signals. The SMW demonstrated excellent performance with a detection limit of 2.9 pM for miR-21, successfully distinguishing cancer cells from normal cells and enabling microRNA detection in clinical tissue samples. This innovative design strategy opens new possibilities for developing simplified yet efficient DNA nanomachines for broader biomedical applications.

The first encounter I ever had with Fritz Eckstein was in 1969 at Stanford University to discuss a polyribonucleotide in which the phosphate was replaced by thiophosphate groups, engendering increased interferon induction (i.e. antiviral activity). His research work then focused on the versatility of oligonucleotides as potential therapeutics. Spanning a period of several decades, various other leads of research were undertaken, i.e. 2'- and 3'-amino or -azido-substituted deoxyribonucleoside analogs, hammerhead ribozymes, small non-coding mRNAs (siRNAs, miRNAs) for monitoring gene therapy, and thiophosphate-substituted nucleotide analogs to be used in RNA and DNA sequencing. This exemplary scientific career generated not one but a multitude of magic bullets for biomedical research and application.

Nephrotic syndrome is a complex renal condition characterized by abnormal protein permeability into the urine space, leading to edema and renal failure. Recent research suggests that deregulation of microRNAs contributes to the pathogenesis of this disease. MicroRNAs are small, non-coding RNA molecules that regulate gene expression by binding to complementary messenger RNA sequences. In this study, we employed bioinformatics techniques to analyze microRNA expression in urine samples from nephrotic syndrome patients and healthy control participants. Our results revealed a significant disruption of microRNA expression profiles in patients with nephrotic syndrome, indicating that these microRNAs may play a crucial role in the disease. This study highlights the potential of urinary microRNAs as biomarkers for nephrotic syndrome and warrants further investigation into their functional significance in the disease pathogenesis.

Cancer is a formidable adversary in contemporary healthcare. Routine screening and early diagnosis are crucial for favourable therapeutic outcomes. Publications, clinical trials, and patent landscape analysis suggest miRNA as promising biomarkers for diagnosis and prognosis of various cancers. This review intends to shed a holistic view of the current and futuristic methods for electrochemical biosensing platforms, using miRNA as biomarkers, coupled with microfluidics, machine learning techniques, and portable electronic devices. Electrochemical biosensors are thoroughly reviewed as they are promising candidate in the design and development of such devices where there is an in-depth exploration of the existing molecular techniques and sophisticated electrochemical biosensing strategies developed for the detection of miRNAs. Additionally, the review will critically analyze diverse signal enhancement strategies and microfluidic platforms specifically tailored for the detection of miRNA. Practical examples of such integrated electrochemical microfluidic biosensors are thoroughly cited along with the prospect of integration of these techniques with portable electronics, highlighting the future potential of highly integrated and accessible diagnostic solutions. Furthermore, the review will also encompass an assessment of the ongoing clinical trials investigating the utility of miRNA as cancer biomarker in diagnostic settings. Moreover, by assessing existing patents, the review shall provide a nuanced understanding of the intellectual property landscape, identifying key players, emerging technologies, and potential future directions. Our review with a 360-degree updated view on molecular biology components, electrochemical biosensors, engineering device design, clinical trials and patent landscape would appeal to researchers, engineers and clinicians working in the area of cancer molecular diagnosis.

Dysregulated microRNAs (miRNAs) have significant potential as diagnostic tools for various chronic diseases; however, their therapeutic applications remain largely unexplored. Given their capacity to regulate multiple pathways, miRNAs are promising candidates for treating pleiotropic diseases, such as inflammatory bowel disease (IBD). In our study, we conducted a comprehensive review of the literature of miRNA-146 levels in the inflamed tissues of IBD patients and murine colitis models. Initially, we quantified the expression of miRNA-146a and miRNA-146b in the colons of mice using the dextran sodium sulfate (DSS)-inducedacute model of IBD. We selected miRNA-146a for further study due to its anti-inflammatory properties and potential relevance in IBD treatment. We hypothesized that a macrophage model of inflammation would be well-suited to studying the effects of this miRNA. Subsequently, we investigated the use of lipid nanoparticles (LNPs) for the targeted delivery of miRNA-146a to macrophages, which play a key role in IBD. Our results indicated that miRNA-146a levels increased in the DSS model and LNP-mediated delivery effectively downregulated genes associated with inflammation. These findings highlight the critical role of miRNA-146a in modulating IBD and suggest that LNP-based delivery could be a promising therapeutic strategy for managing inflammatory responses.

Uveitis associated with juvenile idiopathic arthritis (JIA-U) is a clinically silent vision-impairing disease. Early detection and aggressive treatment are crucial for optimal visual outcome. Alterations in levels of microRNAs (miRNAs) are characteristic of autoimmune diseases. The present clinical study sought to explore the expression of miRNAs in JIA-U and their potential role as a predictive biomarker.

MiRNA expression profiling was performed on peripheral blood mononuclear cells derived from pediatric patients with JIA, JIA-U, or other types of uveitis using the high-throughput small-RNA sequencing (on Next Generation Sequencing (NGS)). Patient- and disease-related data were retrieved from the medical files. Main outcome measure was the differential expression of miRNAs among the groups.

The cohort included 35 patients; 20 children with JIA-U (8 with active disease), 10 with JIA without ocular involvement, and 5 with other types of uveitis (4 with active disease). Mean age was 8.6 years; 83% were female. Nineteen patients (54%) received immunomodulatory treatment. The expression of miR-4485-3p was significantly increased in patients with JIA-U compared to patients with JIA alone (p < 0.05), with no difference between patients with active or inactive uveitis. The expression in patients with uveitis of other etiologies was similar to the expression in JIA-U patients.

This study demonstrates a differential expression profile of a specific miRNA in JIA patients with and without uveitis. If verified in larger studies, the findings may assist to identify JIA patients at risk to develop uveitis and to improve early detection of disease activity.

The growth and development of duck skeletal muscle significantly affect duck meat production, making it essential to understand the molecular mechanisms underlying these processes. Circular RNAs (circRNAs) and microRNAs (miRNAs) are identified in many species and play essential roles in the regulation of myogenic processes; however, research on circRNAs and miRNAs involved in the duck skeletal muscle development is limited. In prior whole-transcriptome RNA sequencing study, we identified differential expression of miR-22-5p and the novel circular RNA circFBLN2, which arises from the second exon of the FBLN2 gene, in duck primary myoblasts (DPMs). In this study, we confirmed the circular structure of circFBLN2 and explored its expression patterns and functional implications in myogenesis. To elucidate the function of circFBLN2 in the myogenic processes of duck, we conducted experiments involving both the silencing and overexpression of circFBLN2 in DPMs. Our findings indicated that circFBLN2 inhibits DPM proliferation while promoting their differentiation. Conversely, when miR-22-5p was silenced and overexpressed, it exhibited opposing effects by promoting the proliferation of DPMs and inhibiting their differentiation. These results suggest a complex dynamic interplay between circFBLN2 and miR-22-5p in the regulation of DPMs proliferation and differentiation. Additionally, our results revealed that both circFBLN2 and myocyte enhancer factor 2 C (MEF2C) act as sponges for miR-22-5p, as demonstrated by binding predictions and dual-luciferase reporter assays. These results offer novel perspectives on the regulatory pathways underlying the duck embryonic skeletal muscle development, underscoring the pivotal function of circFBLN2 in the regulation of miR-22-5p expression. This research deepens our comprehension of the molecular underpinnings of avian myogenesis, potentially paving the way for more effective approaches to bolster growth and development of livestock.

Globally, lung adenocarcinoma (LUAD) remains the leading cause of cancer‑related mortality, highlighting the urgent need for innovative therapeutic approaches. Necroptosis has been recognized as a crucial mechanism for inhibiting cancer progression. Research has revealed a significant association between microRNA (miRNA)‑mediated necroptosis and tumor progression. The present study aimed to elucidate the role and underlying mechanisms of miR‑10b‑5p in regulating necroptosis in the context of LUAD. In an investigation of LUAD, miRNA sequencing was conducted on both LUAD and adjacent non‑tumor tissues, followed by the integration of external database information to identify specific target miRNAs. The expression of miR‑10b‑5p was verified in LUAD tissues and corresponding adjacent non‑cancerous tissues using immunohistochemistry. In vitro experiments, utilizing LUAD cell lines engineered to modulate miR‑10b‑5p levels, assessed its effects on cellular activities and necroptosis. The inhibition of PKP3 by miR‑10b‑5p was determined using a dual luciferase reporter system. Furthermore, alterations in miR‑10b‑5p levels were found to affect PKP3 expression and inhibit the RIPK3/MLKL signaling pathway, as evidenced by western blot analysis in LUAD cell lines. The effect of PKP3 knockdown on cell activity and necroptosis in LUAD cell lines with low miR‑10b‑5p expression levels was assessed using cell function assays. Finally, a nude mouse xenograft model was used to investigate the effect of miR‑10b‑5p on LUAD growth in vivo and its specific mechanism of action. It has been revealed that miR‑10b‑5p levels are significantly elevated in LUAD specimens. Further investigations demonstrated that an increase in miR‑10b‑5p enhances the proliferation of LUAD cells and suppresses the progression of necroptosis, as evidenced by in vitro experiments. Through dual luciferase reporter assays, PKP3 was confirmed as a direct target negatively regulated by miR‑10b‑5p, leading to reduced expression levels. Western blot analysis indicated that miR‑10b‑5p inhibits the RIPK3/MLKL pathway activation through downregulation of PKP3, which leads to increased cell proliferation and decreased necroptosis. However, knockdown of PKP3 reversed the inhibitory effect of miR‑10b‑5p inhibitors on cellular activity and inhibited necrosis by suppressing the RIPK3/MLKL signalling pathway. In addition, animal model studies demonstrated that inhibition of miR‑10b‑5p activated the RIPK3/MLKL pathway by promoting PKP3 expression and significantly reduced LUAD growth by promoting necroptosis. In conclusion, our studies have revealed that the miR‑10b‑5p functions as a tumorigenic factor, enhancing various cellular activities in LUAD cells and suppressing necroptosis by specifically targeting PKP3, thereby inhibiting activation of the RIPK3/MLKL pathway. Importantly, interventions using inhibitors that specifically target miR‑10b‑5p have shown significant success in impeding the progression of LUAD by promoting necroptosis in both cellular and animal models. Thus, targeting miR‑10b‑5p holds considerable potential as a therapeutic strategy for LUAD.

The sublethal effect of insecticides can affect the population dynamics of pests by changing the physiological or behavioral changes, which poses a serious threat to the sustainable control of crop pests in the field. However, the molecular regulation mechanism that mediates the sublethal effect of insecticides on crop pests remains unsolved. Here, we show that the sublethal effect of spinetoram and cyantraniliprole on Spodoptera frugiperda is mediated by the molecular module of microRNA-9993/microRNA-2a-3p - farnesyl diphosphate synthase gene (FPPS)/juvenile hormone (JH) acid methyltransferase gene (JHAMT) - JH. Spinetoram prolonged the duration of larvae and pupae, decreased the weight of pupae, while cyantraniliprole prolonged the duration of larvae and decreased the emergence rate. Similarly, injection of the juvenile hormone analogue (JHA) methoprene significantly prolonged the developmental duration of larvae and pupae, decreased the pupal weight and emergence rate. This sublethal phenotypic change was due to the upregulation of key JH synthesis genes, including FPPS and JHAMT, mediated by spinetoram and cyantraniliprole, resulting in an increase in JH titer. Furthermore, it was confirmed by small RNA sequencing, dual luciferase analysis and agomir-miRNA injection, miR-9993 and miR-2a-3p that it could bind FPPS and JHAMT respectively, and regulated the expression level of FPPS and JHAMT to affect the titer of JH, thus changing the phenotype of S. frugiperda. Collectively, these results provide insights into the mechanism of insecticide regulation of sublethal effects of pests, expand our understanding of development-related miRNAs, and reveal key factors involved in JH signaling pathways that support sublethal effects of insecticides.

Excessive deposition of abdominal fat in chickens has a significant impact on the poultry industry, and there is increasing evidence that non-coding RNAs play a crucial role in fat development. In our previous RNA-seq study, miR-200b-3p was found to be differentially expressed during different developmental periods of fat in Gushi chickens. In this study, we report that miR-200b-3p can directly bind to the 3'UTR region of SESN1 to promote proliferation and inhibit differentiation of preadipocytes. Overexpression of SESN1 inhibits preadipocyte proliferation and promotes differentiation. In contrast, inhibition of SESN1 resulted in the opposite outcome. Interestingly, we also identified the circADGRF5/miR-200b-3p/SESN1 ceRNA network involved in the developmental process of preadipocytes. Furthermore, we validated a novel circRNA, circADGRF5, in this report and found that it regulates SESN1 expression through competitive binding with miR-200b-3p. In conclusion, these data suggest that miR-200b-3p directly targets SESN1 to regulate the proliferation and differentiation of preadipocytes, and circADGRF5 regulates SESN1 expression through competitive binding with miR-200b-3p.

Helicobacter pylori infection and the resulting inflammation of the stomach are widely recognized as the primary risk factors for the development of gastric cancer (human health). Despite numerous attempts, the correlation between various virulence factors of H. pylori and stomach cancer remains mainly unexplained. The cag pathogenicity island (cagPAI) is a widely recognized indicator of virulence in H. pylori. MicroRNAs play crucial roles in a wide range of biological and pathological processes and dysregulated expressions of miRNAs have been detected in numerous cancer types. However, research on the correlation between H. pylori infection and its cagPAI, as well as the differential expression of microRNAs in gastric cancer, is lacking.

The aim of this study was to examine the differential expression of miRNAs in 80 patients with gastric cancer, specifically in connection to the presence of H. pylori and its cag pathogenicity island (cagPAI).

Biopsies of 80 gastric cancer patients were collected and used for H. pylori DNA isolation and tissue miRNA isolation, and further analyzed for cagPAI and miRNA expression and their association.

Elevated levels of miR-21, miR-155, and miR-223 were detected in malignant tissues. The expression of miR-21 and miR-223 was considerably elevated in biopsies that tested positive for H. pylori, whereas the expression of miR-34a was reduced. H. pylori cagPAI samples that are functionally intact exhibit greater expression of miR-21 and miR-223 compared to cagPAI samples that are partially deleted, in both normal and malignant tissues.

Thus, the novelty of our study lies in its focus on the differential expression of specific miRNAs in relation to the functional integrity of the cagPAI in H. pylori-infected gastric cancer patients, offering a more detailed understanding of the interplay between H. pylori virulence factors and miRNA regulation than previous studies.

CVDs is a leading cause of morbidity, mortality, and healthcare expenditure worldwide. Identifying individuals at risk or in the incipient stages of disease is instrumental in enabling timely interventions, preventive measures, and tailored treatment regimens. The landscape of CVDs is complicated by their heterogeneity, encompassing a spectrum of conditions such as coronary artery disease, heart failure, arrhythmias, and valvular disorders. In recent years, the integration of biomarkers into cardiovascular medicine has emerged as a paradigm-shifting approach with the potential to revolutionize early detection and risk stratification. By synthesizing a multitude of studies, we aim to provide a comprehensive resource that illuminates the transformative potential of biomarkers in ushering in a new era of precision cardiovascular medicine.

To identify the biomarkers for the detection and diagnosis of CVDs.

This review examines key studies from 2015 to the present that investigate the impact of cardiac biomarkers on cardiovascular outcomes. Data were gathered from PubMed, Cochrane Library, and Embase to ensure a comprehensive analysis. The review focuses on various cardiac biomarkers, assessing their levels and changes in relation to cardiovascular health, with special emphasis on advanced biomarkers such as proteomic and metabolomic markers in cardiovascular disease (CVD) diagnosis. Peer-reviewed studies published in English that evaluated the diagnostic, prognostic, or therapeutic role of cardiac biomarkers were included, with priority given to clinical trials, cohort studies, systematic reviews, and meta-analyses providing quantitative biomarker data. Studies unrelated to cardiac biomarkers, case reports, editorials, conference abstracts, and those with small sample sizes or insufficient methodological rigor were excluded. The review also accounts for potential confounding factors and research limitations, ensuring a balanced assessment of the literature. By synthesizing data from academic papers, clinical reports, and research articles, this study provides a comprehensive evaluation of the evolving role of cardiac biomarkers in CVD diagnosis and risk stratification.

Biomarkers play a pivotal role in cardiovascular disease risk prediction, diagnosis, and treatment by providing dynamic biological insights. High-sensitivity cardiac troponins (hs-cTn) enhance myocardial injury detection, while circulating microRNAs (miR-208, miR-499) serve as early indicators of myocardial infarction and heart failure. Lipoprotein(a) [Lp(a)] predicts long-term cardiovascular risk, and inflammatory biomarkers such as C-reactive protein (CRP) and interleukin-6 (IL-6) are linked to adverse outcomes. Multi-biomarker panels, such as hs-cTn with B-type natriuretic peptide (BNP), improve heart failure prognosis, while metabolomic profiling enables precision medicine. Additionally, biomarkers like BNP and NT-proBNP facilitate real-time therapeutic monitoring. These findings underscore the critical role of biomarkers in refining risk stratification, improving diagnostic accuracy, and enabling personalized treatment strategies in cardiovascular medicine.

The advancement of cardiovascular biomarkers has significantly enhanced early detection, risk stratification, and personalized treatment. Emerging biomarkers, including genetic variants, metabolomics, microRNAs, and imaging-based markers, provide deeper insights into disease mechanisms. Integrating multi-omic approaches with artificial intelligence may further refine predictive accuracy and therapeutic decision-making. However, clinical translation requires rigorous validation through large-scale, multicenter studies to ensure reliability and applicability across diverse populations. Standardization, cost-effectiveness assessments, and the development of biomarker panels are essential for clinical adoption. Future research should focus on bridging discovery and implementation, advancing precision medicine to improve cardiovascular outcomes.

The tumor microenvironment (TME) plays a crucial role in the development, progression, and metastasis of oral squamous cell carcinoma (OSCC). The TME comprises various cellular and acellular components, including immune cells, stromal cells, cytokines, extracellular matrix, and the oral microbiome, all of which dynamically interact with tumor cells to influence their behavior. Immunosuppression is a key feature of the OSCC TME, with regulatory T cells (Tregs), myeloid-derived suppressor cells (MDSCs), and tumor-associated macrophages (TAMs) contributing to an environment that allows tumor cells to evade immune surveillance and supports angiogenesis. The oral microbiome also plays a pivotal role in OSCC pathogenesis, as dysbiosis, or imbalances in the microbiota, can lead to chronic inflammation, which promotes carcinogenesis through the production of pro-inflammatory cytokines and reactive oxygen species (ROS). Pathogens like Porphyromonas gingivalis and Fusobacterium nucleatum have, hence, been implicated in OSCC-driven tumor progression, as they induce inflammation, activate oncogenic pathways, and modulate immune responses. In this review, we discuss how the interplay between immunosuppression and microbiome-driven inflammation creates a tumor-promoting environment in OSCC, leading to treatment resistance and poor patient outcomes, and explore the potential therapeutic implication of a better understanding of OSCC etiology and molecular changes.

PSA is widely used in prostate cancer screening. However, false-positive PSA results can lead to misdiagnosis and wrong puncture biopsy, while false-negative PSA results can result in missed diagnosis and delayed treatment. There is an urgent need to find convenient, economical and non-invasive diagnostic methods to reduce the false-negative and false-positive rates of PSA. The aim of this study was to discover new miRNA panels to detect prostate cancer.

We selected 10 miRNAs in the literature that were associated with prostate cancer. Afterwards, we measured the expression levels of these miRNAs in serum of 112 prostate cancer patients and healthy controls through a training phase and a validation phase. By plotting receiver operating characteristic curve, the miRNAs with the highest diagnosis value were chosen. Then, a set of miRNAs with the top diagnostic value was identified using stepwise logistic regression.

The findings showed that 5 kinds of miRNAs (let-7b-5p, miR-15a-5p, miR-133a-3p, miR-15b-5p, miR-144-3p) were abnormally expressed in the serum of prostate cancer patients. The diagnostic panel constructed with these 3 miRNAs including let-7b-5p, miR-15a-5p miR-15b-5p and which have high specificity and sensitivity in detecting prostate cancer (area under the curve (AUC) = 0.899). Furthermore, the genes FAM107A and TAF1C may be potential therapeutic targets for prostate cancer.

A three-microRNA panel has an important diagnostic value in prostate cancer and is expected to serve as diagnostic biomarker for prostate cancer. Furthermore, the genes FAM107A and TAF1C may be potential therapeutic targets for prostate cancer.

MicroRNA-seq data is produced by aligning small RNA sequencing reads of different microRNA transcript isoforms, called isomiRs, to known microRNAs. Aggregation to microRNA-level counts discards information and violates core assumptions of differential expression methods developed for mRNA-seq data. We establish miRglmm, a differential expression method for microRNA-seq data, that uses a generalized linear mixed model of isomiR-level counts, facilitating detection of miRNA with differential expression or differential isomiR usage. We demonstrate that miRglmm outperforms current differential expression methods in estimating differential expression for miRNA, whether or not there is differential isomiR usage, and simultaneously provides estimates of isomiR-level differential expression.

Since microRNAs are released into the bloodstream and miRNA profiles are supposed to differ between healthy and tumour patients, miRNAs seem to have potential as biomarkers. An essential prerequisite for biomarkers in a routine diagnostic setup is their stability in serum over time. In this study, serum samples from 10 healthy dogs were analysed at different time points and under various temperature conditions (after 24 and 48 h, at 4° or 20 °C) for the copy number of eight miRNAs (miR-20b, 21, 122, 126, 192, 214, 222, 494) using ddPCR. The miR-21 had the highest copy number, whereas miR-494 had the lowest copy number in canine blood samples. The values of each miRNA varied individually between the dogs, showing a 5 to 10-fold range. Stability differed between the miRNAs, with miR-192 having the best stability. The copy number of miR-20b, miR-126 and miR-214 decreased not significantly during 48 h storage time. In contrast, miR-21, miR-122 and miR-222 were stable for 24 h only but decreased significantly after 48 h. The (in)stability of individual canine miRNAs must be considered when transferring study results into veterinary routine diagnostics, as the transport and storage conditions are variable. As far as possible, standardisation of sampling, storage and quantification of miRNAs is needed.

Nanoparticle technology, particularly gold nanoparticles (AuNPs), is being developed for a wide range of applications, including as a delivery system of peptides or nucleic acids (NA). Their use in precision medicine requires detailed engineering of NP functionalization to optimize their function and minimize off-target toxicity. Two main routes can be found in the literature for the attachment of NA strands to AuNPs: covalent binding via a thiol group or passive adsorption onto a specially adapted coating previously applied to the metallic core. In this latter case, the coating is often a positively charged polymer, as polyethylenimine, which due to its high positive charge can induce cytotoxicity. Here, we investigated an innovative strategy based on the initial coating of the particles using calix[4]arene macrocycles bearing polyethylene glycol chains as an interesting alternative to polyethylenimine for NA adsorption. Because any molecular modification of AuNPs may affect the cytotoxicity and cellular uptake, we compared the behavior of these AuNPs to that of particles obtained via a classical thiol covalent attachment in MCF-7 and GC-1 spg cell lines. We showed a high biocompatibility of both AuNPs-NA internalized in vitro. The difference in subcellular localization of both AuNPs-NA in MCF-7 cells compared to GC-1 spg cells suggests that their subcellular target is cell- and coating-dependent. This finding provides valuable insights for developing alternative NA delivery systems with a high degree of tunability.

Aberrant microRNA expression is associated with tumor progression in various organs. Detecting microRNAs as clinical cancer biomarkers can facilitate early cancer diagnosis and monitoring. However, the rapid and accurate quantification of microRNAs from biological samples remains a significant challenge. Here we developed a one-pot isothermal assay utilizing a molecular circuit with CRISPR/Cas13a (CRISPR-circuit) to rapidly convert, amplify and report different microRNAs within 15 min at the attomolar (aM) level. Then the full process was performed on an active centrifugal microfluidic chip and its corresponding portable equipment for parallel detection of multiple microRNAs, including miR-21, miR-141, miR-196a, and miR-1246. We also demonstrated its application for identifying cell lines and clinical samples of cancer patients with varying microRNA levels, which showed a strong correlation with the RT-qPCR. The assay can be easily adapted for the detection of any microRNA by simply modifying the converter primer, thereby holding significant potential for accurate disease detection and clinical diagnosis.

This study aimed to investigate the effect of miR-325 on KIF20B expression and its role in the invasion and proliferation of colorectal cancer cells.

Colorectal cancer HCT116 cells were cultured and transfected with a miR-325 inhibitor. KIF20B expression was assessed using quantitative polymerase chain reaction (qPCR) and western blotting. Cell proliferation was assessed with the Cell Counting Kit-8 (CCK8) assay, while invasion was evaluated using Transwell and scratch wound healing assays. The expression levels of the invasion-related proteins Matrix Metalloproteinase-2 (MMP-2) and MMP-9 were also analyzed.

The q-PCR and western blot results demonstrated that KIF20B expression was significantly reduced by miR-325 inhibition. The CCK8 assay revealed that miR-325 inhibition decreased cell proliferation. Furthermore, Transwell and Scratch Wound Healing assays showed that miR-325 inhibition suppressed the invasive capacity of colorectal cancer cells. The inhibition of miR-325 also led to decreased expression levels of MMP-2 and MMP-9.

miR-325 inhibition effectively suppresses KIF20B expression, reducing the invasion and proliferation of colorectal cancer cells, suggesting miR-325 as a potential therapeutic target.

Breast cancer is the primary cause of cancer-related death and the most frequent malignancy among women in Western countries. Although there have been advancements in combination treatments and targeted therapies for the metastatic diseases management, metastatic breast cancer is still the second most common cause of cancer-related deaths among U.S. women. The routes of metastasis encompass invasion, intravasation, circulation, extravasation, infiltration into a remote location to establish a metastatic niche, and the formation of micro-metastases in a new environment. Each of these processes is regulated by changes in gene expression. MicroRNAs (miRNAs) are widely expressed by a variety of organisms and have a key role in cell activities including suppressing or promoting cancer through regulating various pathways. Target gene expression is post-transcriptionally regulated by miRNAs, which contribute to the development, spread, and metastasis of breast cancer. In this study, we comprehensively discussed the role of miRNAs as predictors of breast cancer metastasis, their correlation with the spread of the disease to certain organs, and their potential application as targets for breast cancer treatment. We also provided molecular mechanisms of miRNAs in the progression of breast cancer, as well as current challenges in miRNA-based therapeutic approaches. Furthermore, as one of the primary issues with the treatment of solid malignancies is the efficient delivery of miRNAs, we examined a number of cutting-edge carriers for miRNA-based therapies and CRISPR/Cas9 as a targeted therapy for breast cancer.

The phosphatases of regenerating liver (PRLs) are members of the protein tyrosine phosphatase (PTP) superfamily that play pro-oncogenic roles in cell proliferation, migration, and survival. We previously demonstrated that PRLs can post-translationally down-regulate PTEN, a tumor suppressor frequently inactivated in human cancers, by dephosphorylating PTEN at Tyr336, which promotes the NEDD4-mediated PTEN ubiquitination and proteasomal degradation. Here, we report that PRLs can also reduce PTEN expression by up-regulating microRNA-21 (miR-21), which is one of the most frequently overexpressed miRNAs in solid tumors. We observe a broad correlation between PRL and miR-21 levels in multiple human cancers. Mechanistically, PRL2, the most abundant and ubiquitously expressed PRL family member, promotes the JAK2/STAT3 pathway-mediated miR-21 expression by directly dephosphorylating JAK2 at Tyr570. Finally, we confirm that the PRL2-mediated miR-21 expression contributes to its oncogenic potential in breast cancer cells. Our study defines a new functional role of PRL2 in PTEN regulation through a miR-21-dependent post-transcriptional mechanism, in addition to our previously reported NEDD4-dependent post-translational PTEN regulation. Together, these studies further establish the PRLs as negative regulators of PTEN.