Molecular Diagnosis & Therapy最新文献

Human endogenous retroviruses (HERVs) are remnants of ancient retroviral infections constituting nearly 8% of the human genome. Far from being inert genetic fossils, HERVs have co-evolved with their hosts, acquiring regulatory and functional roles that influence development, immunity and disease. In this comprehensive review, we examine the origin, evolutionary dynamics and structural diversity of HERVs, emphasising their integration, genomic organisation and mosaic recombination patterns. We further highlight how epigenetic modifications, intracellular factors such as transcriptional regulators, hormones and cytokines, and environmental influences regulate HERV expression. By synthesising current evidence linking HERV activity to immune modulation, tumourigenesis, autoimmunity and neurodegenerative processes, we provide an integrated perspective on their dual role as drivers of pathology and contributors to normal physiology. Our analysis underscores that HERVs are not only markers of past infections but also active genomic elements with potential clinical implications, from biomarker discovery to novel therapeutic targets, making their systematic investigation timely and highly relevant.

This review explores the transformative potential of salivary biomarkers in revolutionizing periodontal disease diagnostics and management through a non-invasive real-time analysis. A comprehensive literature search was performed using databases such as PubMed, Scopus, and Google Scholar, from inception to March 2025 employing keywords such as "salivary biomarkers," "periodontal disease," "biosensor," "precision dentistry," and "pro-inflammatory cytokines." Saliva, rich in inflammatory cytokines (e.g., interleukin-1β, interleukin-6, tumor necrosis factor-α), enzymatic markers (e.g., matrix metalloproteinase-8, matrix metalloproteinase-9), microbial DNA, oxidative stress indicators, and emerging biomarkers such as exosomes and microRNAs, offers a dynamic medium for early disease detection, risk stratification, and therapeutic monitoring. Technological advancements, including biosensors, lab-on-a-chip systems, and artificial intelligence-driven analytics, enable precise point-of-care applications that align with precision dentistry paradigms. These innovations facilitate personalized interventions by identifying subclinical inflammation, tracking disease progression, and tailoring treatments to individual molecular profiles. However, challenges such as biomarker variability, standardization of collection protocols, cost, and regulatory hurdles impede clinical translation. Future directions emphasize interdisciplinary collaboration, expanded biomarker panels, and rigorous clinical trials to validate diagnostic accuracy and scalability. By bridging gaps between laboratory research and clinical practice, salivary diagnostics promise to shift periodontal disease management from reactive to proactive care, enhancing patient outcomes and reducing systemic disease risks. This review underscores saliva's pivotal role in advancing oral health diagnostics while advocating for optimized methodologies to realize its full potential in precision dentistry.

Glioblastoma (GBM) is an aggressive primary brain tumor with a median survival of 14-15 months even with standard multimodality treatments. The effectiveness of surgical resection, chemotherapy, and radiation therapy are limited by resistance mechanisms including tumor heterogeneity, immunosuppression, presence of stem-like cells, and inhibited drug delivery due to the blood-brain barrier (BBB). The BBB is composed of endothelial cells with tight junctions and selective transport systems, which prevent drug delivery to the tumor at therapeutic levels. Amino acid (AA) transporters have emerged as promising therapeutic targets for overcoming these limitations and enhancing GBM treatment. This review highlights the role of AA transporters in GBM, emphasizing their potential in enhancing targeted therapy, diagnosis, and disease monitoring. We summarize and discuss the 22 AA transporters which are upregulated in GBM, as well as those that demonstrate prognostic correlation. Among these, LAT1 (SLC7A5) has garnered the most attention for its role in drug delivery and imaging, while other transporters exhibit potential as diagnostic and therapeutic targets. Furthermore, nanoparticle technology has emerged as an innovative strategy to enhance targeted therapy through AA transporters. They can enable extended drug circulation, enhanced BBB penetration, and target-specific localization, offering synergistic therapeutic effects. This review emphasizes the importance of AA transporters as multifaceted tools for improving GBM treatment outcomes and the potential of combining AA transporter-targeted therapies with emerging technologies to address the limitations of current GBM management strategies.

Background and objective: Primary ciliary dyskinesia (PCD) is a rare genetic condition characterised by abnormal ciliary motility, primarily affecting the respiratory tract. Despite its clinical significance, there is currently no gold standard for PCD diagnosis. This study aims to address this diagnostic challenge by evaluating a comprehensive approach in a large cohort of patients with suspected PCD.

Methods: We conducted a retrospective analysis of 128 patients with suspected PCD at a specialised clinical reference unit. A thorough anamnesis was performed, followed by a triad of diagnostic tests: (i) a high-speed video analysis of ciliary beat pattern; (ii) transmission electron microscopy for ciliary ultrastructure examination; and (iii) a genetic analysis, primarily through clinical exome sequencing. Correlations between the clinical, morphological and genetic findings were studied. Functional assays on RNA were performed to assess new splicing variants. Pearson's chi-square test was used to compare categorical variables and comparisons of means were performed using the Student's t-test.

Results: A definitive PCD diagnosis was established in 72% of the studied patients. Notably, only 58% of the diagnosed cases showed positive results across all three diagnostic tests. Patients with immotile cilia have a higher frequency of neonatal respiratory distress and had a higher likelihood of receiving a genetic diagnosis. A high-speed video analysis was altered in 116 patients, 53 of them with immotile cilia. A transmission electron microscopy revealed ultrastructural alterations in 67 patients, with class 1 defects being more common. DNAH5, RSPH1 and DNAH11 were the most represented genes among the 18 causal genes found. Among the 71 causal genetic variants found, we highlight the overrepresentation of the c.85G>T in RSPH1 and describe the aberrant effect on RNA of the splicing variants DNAH11:c.11497-6T>G, DNAH9:c.2596-2dup, CCDC40:c.2597A>G and CCDC40:c.2832G>A. Finally, we describe a severe phenotype associated with the RSPH1 gene, contrary to previously reported data.

Conclusions: This comprehensive analysis of a large cohort of patients with PCD underscores the challenges in achieving a definitive diagnosis and emphasises the need for a multi-faceted diagnostic approach. This study enhances our understanding of this rare condition, including the identification of new splicing variants and an unexpected severe phenotype associated with RSPH1, challenging previous assumptions about genotype-phenotype correlations in PCD.

Cardiovascular aging is a complex biological process involving progressive cellular and molecular changes that impair heart and vascular function. This review evaluates both fundamental mechanisms and therapeutic strategies, focusing on how recent advances in pharmacology, gene therapy, and regenerative medicine can be translated into clinical practice to mitigate age-related cardiovascular decline. We conducted a comprehensive analysis of peer-reviewed studies from 2000 to 2023, examining molecular pathways of cardiovascular aging and their modulation through pharmacological, genetic, and lifestyle interventions. The review prioritized clinical trials, translational research, and meta-analyses to assess therapeutic efficacy and safety. Current evidence highlights the effectiveness of senolytic drugs such as dasatinib and quercetin in reducing age-related cardiovascular dysfunction, while rapamycin and metformin show promise in improving cardiac longevity through metabolic regulation. Gene therapies, including clustered regularly interspaced short palindromic repeats (CRISPR)-based interventions, demonstrate potential in preclinical models for cardiac regeneration. Stem cell therapies and nanotechnology-based drug delivery systems are emerging as innovative approaches to enhance tissue repair. In addition, lifestyle modifications such as Mediterranean diet adherence and exercise significantly improve vascular health in aging populations. However, challenges remain in drug delivery, patient-specific responses, and long-term safety of novel therapies. The integration of targeted pharmacological treatments, advanced regenerative techniques, and personalized lifestyle interventions represents a transformative approach to managing cardiovascular aging. Future research should focus on optimizing therapeutic combinations, refining delivery methods, and validating biomarkers for clinical monitoring. A multidisciplinary strategy combining these advances will be essential to improve cardiovascular outcomes in aging populations.

Background and objective: Sacituzumab govitecan, an anti-trophoblast cell surface antigen 2 (TROP2) antibody-drug conjugate, has been approved by both the US Food and Drug Administration and European Medicines Agency for patients with metastatic triple-negative breast cancer who have received two or more prior systemic therapies, including at least one of them for advanced disease. Although TROP2 evaluation is not required for patient selection, survival data from the ASCENT trial show improved response rates in patients with high TROP2 expression by immunohistochemistry. However, there is no standardized testing assay for these patients. This study evaluated the consistency of TROP2 expression analysis across different immunohistochemistry assays.

Methods: Twenty-six triple-negative breast cancer samples were analyzed using three different immunohistochemistry assays on a Dako Omnis platform, according to manufacturer protocols. Specifically, ENZO-ABS380-0100 (assay A, used in ASCENT), Abcam SP295 (assay B, used in TROPiCS-02), and Santa Cruz B9-sc-376746 (assay C, used in cross-sectional studies). TROP2 expression on tumor cell membranes was quantified using the H-score, categorized as low (≤ 100), intermediate (> 101 to ≤ 200), and high (> 200). Assay agreement was evaluated using Cohen's κ and Gwet's AC2 statistics.

Results: Assay A showed a broader range of TROP2 expression, with 57.7% of samples (n = 15) classified as low, 34.6% (n = 9) as intermediate, and 7.7% (n = 2) as high expressors. Assay B identified only n = 5 (19.2%) low expressors, n = 11 (42.3%) intermediate, and n = 10 (38.4%) high. While assay C identified n = 4 (15.4%) low expressors, n = 12 (46.2%) intermediate, and n = 10 (38.4%) high. Not surprisingly, assays B and C exhibited substantial agreement, with 80.8% of cases showing consistent results (κ = 0.81; p < 0.0001), indicating similar staining outcomes for TROP2 expression. The overall concordance between Assay A, B, and C was fair to moderate (AC2 = 0.35, p = 0.0067).

Conclusions: Our hypothesis-generating study highlights significant variability among TROP2 assays, suggesting differences in sensitivity and specificity for triple-negative breast cancer. We demonstrate that TROP2 expression is both heterogeneous and dynamic across samples and assays, highlighting the need for methodological improvements in testing. Future research integrating computational pathology with standardized immunohistochemistry protocols and quantitative scoring systems may enhance the clinical utility of TROP2 as a biomarker in triple-negative breast cancer.

Background: Recurrent gastric or gastroesophageal junction cancers have poor prognoses and limited treatment options. While archival tumor tissue is commonly used for genomic profiling, it may not reflect molecular changes at recurrence.

Objective: We aimed to assess the utility of a circulating tumor DNA analysis in identifying actionable genomic alterations at recurrence and compare findings with archival primary tumor profiles.

Methods: This prospective multicenter observational study included 50 patients with recurrent stage II or III gastric or gastroesophageal junction adenocarcinoma who had undergone curative surgery and adjuvant chemotherapy. A circulating tumor DNA analysis was performed using the Guardant360 assay at recurrence, and results were compared with comprehensive genomic profiling from archival primary tumor tissue when available. The primary endpoint was the detection rate of actionable genomic alterations in circulating tumor DNA, defined as those with an OncoKB level ≥ 3 in any cancer type.

Results: Circulating tumor DNA was detected in 78% (39/50) of patients. Actionable genomic alterations were identified in 36%, meeting the primary endpoint. The most frequent actionable genomic alterations included PIK3CA mutations (18%), ARID1A mutations (10%), ERBB2 amplification (6%), and ATM mutations (6%). Among 18 patients with paired tissue and circulating tumor DNA data, 66.7% showed changes in actionable genomic alterations between the primary tumor and recurrence. New actionable alterations, including ERBB2 amplifications and PIK3CA mutations, were identified exclusively in circulating tumor DNA at recurrence.

Conclusions: A circulating tumor DNA analysis effectively captured genomic evolution at recurrence, identifying clinically relevant alterations not found in primary tumor tissue. These findings support the integration of a liquid biopsy into clinical practice to guide treatment for recurrent gastric or gastroesophageal junction cancer.

Background and objective: Colorectal cancer remains a major global health challenge, necessitating the development of accurate non-invasive diagnostic tools. Circulating and excretory microRNAs (miRNAs) are promising biomarkers owing to their stability and regulatory roles in tumorigenic pathways. While single miRNA assays often lack sufficient diagnostic accuracy, panels combining multiple miRNAs have shown enhanced performance. This systematic review and meta-analysis evaluated the diagnostic accuracy of multi-miRNA panels and explored their mechanistic relevance to colorectal cancer pathogenesis.

Methods: A comprehensive search of PubMed, Embase, Web of Science, and Scopus was conducted through March 2025 following Preferred Reporting Items for Systematic Reviews and Meta-Analyses guidelines. The study protocol was registered with PROSPERO (CRD420251060655). Eligible studies assessed the diagnostic accuracy of multi-miRNA panels for colorectal cancer using extractable data on sensitivity, specificity, and area under the curve. Data were extracted independently by two reviewers. A bivariate random-effects model was used to calculate pooled diagnostic estimates. Study quality was assessed with the Quality Assessment of Diagnostic Accuracy Studies 2 (QUADAS-2) tool, and heterogeneity was evaluated using I² statistics. Subgroup analyses were conducted by sample type (e.g., plasma, serum, stool) and panel size. Target genes of recurrent miRNAs were mapped to canonical colorectal cancer-related pathways.

Results: Twenty-nine studies comprising 5497 participants (3070 colorectal cancer cases and 2427 controls) and 35 multi-miRNA panels were included. Pooled sensitivity was 0.85 (95% confidence interval 0.80-0.88), specificity was 0.84 (95% confidence interval 0.80-0.88), and the area under the curve was 0.90, despite substantial heterogeneity (I² > 77%). Panels derived from plasma samples showed the highest balanced performance (sensitivity 0.88; specificity 0.87), while three-miRNA panels exhibited the best diagnostic trade-offs. Mechanistic analysis of 42 recurrent miRNAs revealed consistent involvement in key colorectal cancer pathways, including PI3K/AKT, Wnt/β-catenin, epithelial-mesenchymal transition, angiogenesis, and immune regulation.

Conclusions: Multi-miRNA panels derived from diverse biospecimen sources demonstrate high diagnostic accuracy for colorectal cancer and are mechanistically linked to fundamental oncogenic pathways. Future efforts should focus on panel standardization, biospecimen-specific validation, and integration into clinical workflows to advance precision oncology.

Gastric cancer is a highly heterogeneous disease, with substantial variations observed among patients in clinical manifestation, histological characteristics, and drug sensitivities. Achieving precision medicine necessitates a comprehensive understanding of the molecular mechanisms underlying gastric cancer and the establishment of robust preclinical models. Organoids, cultivated from cancer cells within tumor tissues, utilizing three-dimensional tissue culture techniques, faithfully replicate the features and heterogeneity of in vivo tumors and have emerged as a promising platform. This review explores the application of gastric cancer-derived organoids (GCOs) in preclinical research and clinical translation, highlighting current challenges and outlining future prospects. Although this technology remains some distance from direct clinical application, it holds tremendous potential for clinical utilization.

ESR1 gene mutations represent one of the main mechanisms of acquired resistance to endocrine therapy (ET) in estrogen receptor-positive (ER+) breast cancer. The introduction of liquid biopsy as a minimally invasive technique for analyzing circulating tumor DNA (ctDNA) has opened new avenues for real-time mutation monitoring and personalized treatment strategies. This review explores the clinical relevance of ESR1 mutations in endocrine resistance, the potential of liquid biopsy for early detection and monitoring, and the integration of advanced sequencing technologies and artificial intelligence to improve diagnostic accuracy. Preclinical and clinical studies on key mutations (D538G, Y537S) were analyzed, emerging technologies [(next-generation sequencing (NGS), digital droplet PCR (ddPCR), Cancer Personalized Profiling by deep Sequencing (CAPP-Seq), Targeted Digital Sequencing (TARDIS)] were compared, and survival data from seven major studies were summarized to assess the impact of ESR1 mutations on progression-free survival (PFS) and overall survival (OS). The results show that these mutations, particularly those affecting the ligand-binding domain, are associated with reduced efficacy of aromatase inhibitors and increased tumor aggressiveness. Liquid biopsy proves useful for early detection of resistance mutations and dynamic disease monitoring, but its clinical implementation is limited by low ctDNA levels, technological variability, and the lack of standardized clinical cut-offs. Integration with tissue biopsy, radiomics, and artificial intelligence (AI)-based platforms enhances its clinical utility and prognostic value. In conclusion, liquid biopsy, when combined with advanced technologies and predictive tools, represents an innovative resource for the personalized management of ER+ breast cancer, with the potential to guide timely therapeutic interventions and improve long-term survival.