Search Results (581)

Background/Objectives: Liquid biopsy (LB) is transforming cancer care by enabling minimally invasive tumor profiling. While current research and clinical pathways mostly focus on blood LB, sputum represents a non-invasive, readily available respiratory specimen that may offer unique advantages for lung cancer (LC) care. Despite its potential, the maturity, breadth, and clinical applicability of sputum-based LB remain elusive. Methods: We conducted a scoping review to systematically map the existing literature on sputum LB in LC. Electronic databases were searched for studies evaluating sputum-derived biomarkers—cytologic, genomic, epigenetic, transcriptomic, proteomic, metabolomic, metagenomic, and extracellular vesicle–derived products—across the LC care continuum. Study designs, technologies, clinical contexts, and reported outcomes were extracted and synthesized qualitatively. Results: The literature demonstrated substantial heterogeneity in sputum collection, processing, and analytical platforms. Early work focused on cytometry and genetic alterations, while recent studies increasingly explore DNA methylomics, microRNAs, extracellular vesicle-derived products, and multi-omics approaches. The evidence suggests potential utility of sputum biomarkers for early detection and risk stratification, particularly in high-risk populations, with emerging data supporting roles in molecular subtyping, response monitoring, prognostication, and surveillance. However, few studies report prospective validation, direct comparison with blood-based LB, or impact on actual patient outcomes. Conclusions: Sputum LB is a promising yet underdeveloped modality in LC care. This scoping review highlights technological innovations alongside significant methodological heterogeneity and translational gaps. Future research should focus on standardization, prospective validation, impact on patient outcomes, and integration with blood- and other body fluid–based LB, as well as imaging biomarkers. This will enable incorporation of sputum-based LB into actual clinical pathways of LC care. Full article

Multiple myeloma (MM) is a malignancy of plasma cells that is characterized by a complex and spatially heterogeneous genomic landscape. Despite this complexity, clinical monitoring remains largely dependent on localized bone marrow (BM) assessments. This dependence creates a significant diagnostic gap, as the primary monitoring tools fail to account for the spatial and temporal heterogeneity that drives tumor relapse. Liquid biopsy can serve as an adjunctive approach in assessing the pan-clonal landscape in MM through the molecular profiling of circulating tumor DNA (ctDNA) and circulating tumor cells (CTCs). In this review, we examine the clinical utility of liquid biopsy components in capturing mutational profiles, clonal evolution, treatment resistance mechanisms, and minimal residual disease (MRD), including early detection of relapse and extramedullary progression. We will further discuss current limitations, including variability in assay sensitivity, lack of standardization, and the need for prospective validation. Full article

Colorectal cancer (CRC) remains one of the leading causes of cancer-related mortality worldwide, with prognosis critically dependent on the stage at diagnosis. Traditional tissue biopsy presents well-known limitations, including tumor heterogeneity and invasiveness. Liquid biopsy, encompassing the analysis of circulating tumor DNA (ctDNA), circulating tumor cells (CTCs), exosomes, and other cell-free biomarkers, has emerged as a transformative approach for non-invasive tumor profiling. This comprehensive narrative review outlines the recent evidence published on the current state and future perspectives of liquid biopsy in CRC, with a focused emphasis on the role of artificial intelligence (AI), machine learning (ML), and deep learning (DL) in data analysis and clinical translation. Methods: A narrative review of the literature was conducted by searching PubMed/MEDLINE, EMBASE, and ClinicalTrials.gov for articles published between January 2020 and January 2026, using a predefined Boolean search string combining terms related to liquid biopsy biomarkers, colorectal cancer, and artificial intelligence methodologies. Filters were applied to include only English-language human studies. Additional relevant sources were consulted to ensure comprehensive coverage of the available literature. Liquid biopsy platforms, particularly ctDNA sequencing and methylation profiling, demonstrate increasing clinical utility across the CRC care continuum from population screening to post-surgical minimal residual disease (MRD) detection and real-time therapy monitoring. AI-driven analytical frameworks, including Random Forest, Convolutional Neural Networks, LSTM models, and more recently Large Language Models (LLMs), substantially augment the sensitivity and specificity of liquid biopsy interpretation, enabling multimodal data integration. The convergence of liquid biopsy technology and AI-driven analytics represents a paradigm shift toward precision oncology in CRC. Remaining challenges include analytical standardization, model explainability, regulatory harmonization, and equitable access. Future integration of federated learning frameworks and LLM-based clinical decision support tools will be essential for responsible clinical translation. Full article

Feto-maternal microchimerism (Mc) refers to the exchange of cells between the fetus and mother, and fetal–fetal Mc to the exchange between fetuses during pregnancy. This phenomenon occurs across mammalian species, including humans, mice, and cattle. Key data on Mc cells and theoretical considerations regarding the presence of fetal-derived material, such as trophoblast cells, cell-free fetal DNA (cffDNA), and exosomes in maternal blood are summarized. This review aims to first, synthesize current knowledge on feto-maternal and fetal–fetal Mc across mammals, second, address three core questions: how and where Mc has been demonstrated in animals, what techniques have been used over time to detect fetal-derived material and Mc, and how placental structures influence the frequency of Mc. Finally, it aims to identify gaps in the literature for species such as horses, goats, and pigs. This article concludes that Mc is a widespread phenomenon among mammals, but detection methods and reported frequencies vary significantly by species and placental type. A biological model is presented in this article in which multinucleated trophoblast cells undergo apoptosis, releasing cffDNA that enters the maternal blood circulation after multinucleated trophoblast invasion. Advances in molecular biology technology have improved the ability to detect fetal-derived material, cells, DNA, and exosomes in maternal blood. However, notable research gaps remain for Mc in horses, goats, and pigs, highlighting the need for targeted studies to better understand species-specific patterns or a general biological model. Full article

Liquid biopsy has emerged as a transformative tool in oncology, enabling minimally invasive and dynamic characterization of tumor biology. In prostate cancer, marked by high heterogeneity and frequent bone metastases, tissue biopsy is often challenging, highlighting the clinical value of circulating biomarkers. Circulating tumor DNA (ctDNA) is the most clinically advanced analyte, supporting detection of actionable alterations such as BRCA1/2 and ATM mutations, guiding targeted therapies, and enabling real-time monitoring of treatment response and resistance. Circulating tumor cells (CTCs) and extracellular vesicles (EVs) provide complementary insights into tumor biology and disease progression. However, challenges remain, including limited sensitivity in low tumor burden and biological confounders such as clonal hematopoiesis (CH), which can lead to false-positive findings. Emerging approaches, including fragmentomics and methylation profiling, offer improved tumor specificity and may help overcome these limitations. Together, these advances support the integration of liquid biopsy into clinical practice for personalized management and longitudinal monitoring in prostate cancer. Full article

Background/Objectives: Early detection through minimally invasive approaches is critical for timely patient stratification and optimal therapeutic decision-making in colorectal cancer (CRC). Liquid biopsy, based on the analysis of tumor-derived components in blood and other body fluids, has emerged as a promising strategy to overcome current limitations in CRC diagnosis and follow-up. This review evaluates the current landscape of liquid biopsy clinical trials in CRC, focusing on predictive biomarker detection, prognostic assessment, and disease monitoring. Methods: ClinicalTrials.gov was searched using the terms “colorectal cancer” and “liquid biopsy” yielding 153 registered trials. After manual screening, 44 trials were excluded for not using liquid biopsy for CRC management, leaving 109 trials for analysis. Of these, 25 were completed, and 13 had publicly available results related to liquid biopsy. Results: The included trials were conducted across 27 countries on four continents. Overall, 119 biomolecules assessments and 167 different endpoints were reported across 109 clinical trials. Because individual trials could evaluate multiple biomolecules and endpoints, counts exceed the total number of trials. Cell-free DNA (cfDNA) was evaluated in 92/109 trials (84%) and accounting for 77% of all biomolecule assessments. Circulatingtumor cells (CTCs) were analyzed in 9/109 trials (8%, representing 8% of all the biomolecules analyzed), and microRNAs (miRNAs) in 8/109 (7%, representing 7% of all the biomolecules analyzed). Treatment sensitivity was the most common endpoint (57/109, 52% of the clinical trials; representing 34% of all the 167 different endpoints analyzed), followed by disease progression (28/109, 26%; representing 17% of all the different endpoints analyzed) and diagnostic applications (21/109, 19%; representing 12% of all the different endpoints analyzed). Among the 25 completed studies, 10/25 (40%) were interventional and 15/25 (60%) observational, spanning 14 countries. The majority of completed trials (21/25, 84%) used cfDNA. Interventional studies were predominantly phase II (5/10), with fewer phase III trials (2/10), primarily evaluating treatment response, particularly in relation to EGFR inhibitors and RAS/BRAF mutation status. Four observational studies (4/15) investigated emerging biomarkers, including long noncoding RNAs and miRNAs. Conclusions: Current clinical trials highlight cfDNA as the dominant and most clinically advanced liquid biopsy biomarker in CRC, primarily used for treatment guidance and disease monitoring. In contrast, CTCs and RNA-based biomarkers remain underrepresented. The limited number of randomized late-phase trials, heterogeneity in study design, and technical challenges associated with emerging biomarkers underscore the need for standardized methodologies and robust validation before routine clinical implementation. Full article

Background: Small-cell lung cancer (SCLC) represents an aggressive malignancy associated with a poor prognosis, underscoring the critical demand for enhanced monitoring methodologies. Circulating tumor DNA (ctDNA) constitutes a promising non-invasive biomarker; however, reports employing highly sensitive tumor-informed assays in SCLC remain scarce. This investigation aimed to assess the clinical utility of a personalized ctDNA monitoring strategy for predicting therapeutic outcomes and resistance in SCLC patients. Methods: This prospective observational study enrolled patients diagnosed with unresectable SCLC. Whole exome sequencing was conducted on baseline tumor specimens to design customized 16-plex multiplex polymerase chain reaction (PCR) panels. Serial blood samples were obtained at baseline, at six-week intervals during treatment, and upon disease progression. Detection of ctDNA-based minimal residual disease (MRD) was performed using a tumor-informed assay (Huajianwei^® bespoke MRD) with ultra-deep sequencing. Results: Among seven evaluable patients, the baseline ctDNA-MRD positivity rate was 100%. A significant positive correlation was observed between the baseline ctDNA levels and radiographic tumor burden (r = 0.821, 95% confidence interval [CI] 0.179–0.973, p = 0.034). Longitudinal analysis indicated that patients exhibiting an early decline in MRD levels demonstrated a non-significant trend toward superior progression-free survival (PFS) compared to those with an MRD increase. Though this between-group difference did not reach conventional statistical significance, it represented a trend-level finding (p = 0.0665, hazard ratio [HR] = 0.24, 95% CI: 0.02–3.19), with no definitive prognostic association confirmed in this pilot cohort. Notably, an elevation in MRD preceded radiographic progression by as much as 135 days in certain instances. Conclusions: This study shows that dynamic tumor-informed ctDNA-based MRD monitoring reflects tumor burden changes and may correlate with clinical outcomes in SCLC, supporting its potential to guide personalized treatment and facilitate earlier therapeutic interventions compared to conventional imaging techniques. Prospective multicenter validation is needed to confirm its clinical utility. Full article

Background/Objectives: Circulating tumour DNA (ctDNA) assays enable non-invasive assessment of tumour burden and treatment response in oncology. However, quantitative ctDNA outputs (such as variant allele frequency, tumour fraction, and aggregate burden scores) remain difficult to interpret and compare across platforms. This evidence-mapping review evaluates current quantitative reporting approaches in pancreatic ductal adenocarcinoma (PDAC) and examines the potential role of KRAS mutant ctDNA as a biologically grounded reference metric. Methods: A systematic literature search was conducted across PubMed/MEDLINE and Scopus to identify studies reporting quantitative ctDNA metrics in PDAC. Eligible studies included those measuring plasma KRAS mutations and/or reporting variant allele frequency, tumour fraction, or multi-locus aggregate metrics. Additional relevant primary studies identified through broader manual searching of PubMed were assessed against the same prespecified eligibility and classification criteria before inclusion. Data were synthesised narratively, focusing on reporting frameworks, units of measurement, assay characteristics, and the interpretability of quantitative outputs across platforms. Results: Substantial heterogeneity was observed in ctDNA quantification methods and reporting standards. Ratio-based metrics such as variant allele frequency and tumour fraction were commonly used but varied according to assay design, plasma input volume, and background cell-free DNA levels. Few studies reported absolute mutant molecule counts per unit volume. Given that approximately 90–95% of PDACs harbour truncal activating KRAS mutations, plasma KRAS was consistently represented across platforms and demonstrated potential as a shared quantitative anchor. Limited standardisation was noted in distinguishing detectability from quantifiability based on sampling depth and counting statistics. Conclusions: Current ctDNA reporting in PDAC lacks a shared quantitative reference, limiting cross-study comparability. Reporting KRAS mutant molecules per millilitre and adopting an assay-agnostic framework distinguishing detection from quantification may improve interpretability, support harmonisation across platforms, and facilitate cumulative learning in pancreatic cancer ctDNA research. Full article

Exercise adaptation and training maladaptation arise from overlapping metabolic, redox, inflammatory, endocrine, and tissue-remodeling processes, so the translational question is not whether biomarkers change but when, where, and for which decision they become informative. This narrative review develops a decision-linked framework for minimally invasive biomarkers across the recovery–overload continuum and treats biomarker meaning as a molecule–matrix–time–decision relationship rather than as a stand-alone peak. The framework is organized around five coupled layers: stimulus architecture, signaling and release biology, sampling matrix and pre-analytics, bout-relative kinetics, and the monitoring decision to be supported. Current evidence indicates that no single biomarker reliably separates productive remodeling from delayed recovery, tissue strain, non-functional overreaching, or early maladaptation. Classical chemistry remains useful for bounded tasks, especially delayed tissue strain and stress reactivity; cfDNA appears promising for rapid load sensitivity; targeted metabolite panels are strongest for recovery phenotyping; and circulating RNAs and extracellular-vesicle cargo add mechanistic depth but remain constrained by pre-analytical fragility and incomplete standardization. The central practical implication is that overload is better interpreted as progressive loss of signal resolution than as threshold-crossing and that sparse temporally staggered panels are more likely to aid monitoring decisions than isolated markers or untimed high-dimensional profiles. Progress will depend on purpose-specific panels, transparent analytical standards, and prospective validation against symptoms, performance, and established measures across sex, hormonal, circadian, and training contexts. Full article

Background/Objectives: Cell-free DNA (cfDNA) is released into the circulation during inflammation-driven cellular injury and regulated cell death. Elevated cfDNA concentrations have been reported in several clinical settings, including chronic kidney disease, hemodialysis, and peritoneal dialysis (PD). We previously demonstrated that PD-related peritonitis induces an increase in circulating cfDNA; however, the mechanisms underlying cfDNA generation remained unclear. This study aimed (i) to confirm peritoneal cfDNA variation following peritonitis in PD patients, and (ii) to elucidate the apoptotic pathways responsible for cfDNA release. Methods: Fifty-four PD patients were enrolled and stratified into the following groups: Group A—no history of peritonitis (n = 25); Group B—remote peritonitis > 3 months prior (n = 21); Group C—recent peritonitis < 3 months prior (n = 8). cfDNA was quantified by qPCR. Apoptosis was assessed qualitatively by DNA laddering and quantitatively using ELISA assays for Caspase-3, Caspase-8 and Caspase-9. Results: cfDNA levels were significantly higher in patients with recent peritonitis compared to both other groups (p < 0.01). DNA laddering showed enhanced nucleosomal fragmentation, consistent with apoptosis. Caspase-3 concentrations were markedly increased in recent peritonitis (<3 months) and significantly correlated with cfDNA levels (ρ = 0.511, p < 0.01). Both Caspase-8 and Caspase-9 correlated with Caspase-3 (ρ = 0.57 and ρ = 0.47, respectively), indicating engagement of both extrinsic and intrinsic apoptotic pathways. Conclusions: In conclusion, peritoneal cfDNA in PD patients with peritonitis originates primarily from apoptosis and reflects dual-pathway caspase activation. cfDNA and Caspase-3 progressively decline with longer time elapsed from peritonitis, supporting their potential use as biomarkers for inflammatory activity and membrane recovery. Full article

Multiple myeloma is a complex hematologic malignancy characterized by significant biological heterogeneity, a relapsing–remission clinical course, and a continuously evolving therapeutic landscape. Accurate and timely laboratory assessment is central to disease management, supporting diagnosis, risk stratification, evaluation of treatment response, and long-term monitoring. Despite major advances in therapy, a critical need remains for laboratory tools that can detect disease with greater sensitivity, capture spatial and clonal tumor heterogeneity, and reflect the true depth of treatment response beyond conventional serological and bone marrow-based criteria. Recent laboratory innovations have the potential to transform myeloma care by enabling earlier detection, more accurate prognostication, and personalized therapeutic strategies. This review focuses specifically on innovative laboratory technologies for the diagnosis of multiple myeloma and the evaluation of treatment response. Within this scope, we examine the current diagnostic approaches and the role of high-throughput technologies for measurable residual disease assessment. We explore the emerging role of liquid biopsy approaches, including circulating tumor cells, cell-free DNA/RNA, and mass spectrometry for ultrasensitive detection of monoclonal proteins. We further discuss novel molecular biomarkers and the integration of artificial intelligence and machine learning tools to enhance data interpretation. The innovations reviewed here represent a shift in the contribution of laboratory medicine to myeloma care, offering a more precise, less invasive, and biologically informative framework for targeted and adaptive clinical decisions. Full article

Non-coding RNAs (ncRNAs) have emerged as important regulators of gene expression and cellular homeostasis, and their dysregulation is now recognized as a hallmark of cancer. Over the past decades, extensive research has demonstrated that diverse ncRNA classes, including microRNAs (miRNAs), long non-coding RNAs (lncRNAs), circular RNAs (circRNAs), and other small ncRNA species, participate in complex regulatory networks that influence tumor initiation, progression, metastasis, and therapy response. Through mechanisms such as transcriptional regulation, post-transcriptional gene silencing, epigenetic modulation, and competitive endogenous RNA interactions, ncRNAs shape the molecular circuitry underlying cancer development. In addition to their functional roles in tumor biology, many ncRNAs are released into biological fluids and can be detected as circulating molecules in blood, urine, saliva, and other biofluids. Their remarkable stability in extracellular environments has generated considerable interest in their use as minimally invasive biomarkers in liquid biopsy applications. Emerging evidence has shown that circulating ncRNAs (c-ncRNAs) can support cancer detection, disease stratification, and treatment monitoring. This narrative review provides an integrated view that links ncRNA-mediated regulatory networks with their application as liquid biopsy biomarkers, positioning ncRNAs as comprehensive indicators of tumor conditions. Particular emphasis is placed on c-ncRNA biomarkers, the integration of multiple ncRNA classes, and multi-analyte biomarker strategies that combine ncRNAs with complementary circulating molecules such as cell-free DNA and protein markers. Finally, we discuss the technical and clinical challenges that currently limit the translation of ncRNA-based diagnostics into clinical practice and highlight future directions for advancing ncRNA-guided liquid biopsy approaches in precision oncology. Full article

Preeclampsia (PE), pregnancy-associated high blood pressure linked to organ damage, affects 3–8% of all pregnancies and results worldwide in 70,000 maternal and 500,000 perinatal deaths each year. Untreated PE may progress to eclampsia with long-term health implications for both mother and child. Non-invasive prenatal diagnosis or screening applies cell-free DNA approaches and offers a less invasive and more economical method for early diagnosis and prediction of various pregnancy complications. Recently, cell-free assays, particularly blood-based cell-free DNA and RNA analysis, have shown great potential in early PE prediction and diagnosis. Here, we provide an updated review of the current understanding and discoveries of PE, focusing on recent publications (1 January 2019–30 December 2025) of liquid biopsy-derived circulating fetal cells (circulating trophoblasts and fetal nucleated red blood cells), cell-free DNA, cell-free RNA and small extracellular vesicles (i.e., exosomes). We aim to discuss the conceptual framework and technical evolution of liquid biopsy applications in preeclampsia pathogenesis, prediction and diagnosis. Progressing novel screening and diagnostic molecular biomarkers have high potential to facilitate early detection for patients at risk of PE. Liquid biopsy-based screening strategies may aid in providing timely intervention and treatment. Full article

Reproductive aging is characterized by progressive decline in ovarian reserve, reduced oocyte competence, and impaired endocrine coordination. Although these phenotypic changes are well documented, the molecular mechanisms that integrate aging-associated stress signals into coordinated ovarian dysfunction remain incompletely understood. Increasing evidence indicates that DNA methylation remodeling is closely associated with ovarian aging. Rather than representing isolated promoter-specific events, age-related methylation alterations may reflect progressive imbalance between DNA methyltransferases (DNMTs) and TET-mediated demethylation. Stress-responsive DNMT/TET dysregulation has been linked to distributed epigenetic remodeling across regulatory elements governing PI3K–AKT, TGF-β/SMAD, metabolic, and DNA damage response pathways in ovarian cell populations. We propose a network-level framework in which methylation drift preferentially affects highly connected regulatory hubs, potentially reducing transcriptional robustness and intercellular coordination within the follicular microenvironment. However, current human data remain largely correlative, and functional validation is required to determine whether methylation remodeling acts as a driver, amplifier, or biomarker of ovarian aging. Finally, we discuss translational implications, including circulating cell-free DNA signatures and epigenetic clock models, while emphasizing the importance of cell type-resolved and longitudinal studies. Collectively, the available evidence supports a model in which progressive DNMT/TET imbalance is associated with distributed pathway-level regulatory instability during ovarian aging. Full article

Background: Familial prostate cancer (PCa) accounts for nearly 20% of all PCa cases and is associated with increased genetic susceptibility and earlier disease onset. However, early detection and risk stratification in genetically predisposed individuals remain challenging. Circulating cell-free DNA (cfDNA) provides a minimally invasive source of tumor-derived genomic and epigenomic information. Integrating multi-omic cfDNA analyses may enhance the discovery of biomarkers relevant to familial PCa biology. Methods: We conducted a pilot feasibility study employing whole-genome, strand-specific sequencing of cfDNA from eight patients with familial PCa. A unified analytical pipeline was used to jointly profile genomic alterations and epigenomic features. Variant calling, methylation mapping, and allele-specific methylation (ASM) analysis were performed to identify somatic mutations, characterize epigenetic dysregulation, and explore potential interactions between genetic and epigenetic mechanisms. Results: Sequencing revealed 18,878 genetic variants, including 2276 potentially pathogenic alterations. We identified 26 recurrent high-impact mutations, such as stop-gain and start-loss variants, in genes including MUC4, MCM9, and SKA3. Epigenomic profiling demonstrated widespread gene-specific hypermethylation, consistent with transcriptional repression in these loci. ASM events were detected in TTC22, TEX51, WDR89, LAIR2, and SKA3, suggesting coordinated interactions between somatic variation and epigenetic regulation in familial PCa. Conclusions: This proof-of-concept study highlights the feasibility and potential of integrating whole-genome and epigenome profiling of cfDNA to decode the molecular architecture of familial prostate cancer. By jointly capturing genomic alterations and epigenetic signatures, including allele-specific methylation, this multi-omic liquid biopsy approach supports a high-resolution exploration of biologically relevant molecular features. Moreover, this integrated profiling strategy provides a minimally invasive and clinically scalable tool that may substantially improve risk assessment. These findings offer a promising foundation for future validation studies in larger cohorts, with the aim of advancing multi-omic cfDNA analysis as a next-generation technology in the field of precision oncologic epigenetics. Full article