Molecular Diagnosis & Therapy最新文献

{"title":"Liquid Biopsy in Primary CNS Tumors: Bridging Biology, Technology, and Clinical Care.","authors":"Catalina Trejo-Becerril, Ariela Souroujon-Torun, Lucia Taja-Chayeb, Alexandra Díaz-Alba, Enrique Caballé-Pérez, David Aguilar-Cardoso, Sylvia Harari-Arakindji, Alejandro Rodríguez-Camacho, Sergio Moreno-Jimenez, Rogelio Trejo, Alipio González-Vázquez, José Omar Navarro-Fernández, Bernardo Cacho-Díaz, Talia Wegman-Ostrosky","doi":"10.1007/s40291-026-00849-2","DOIUrl":"https://doi.org/10.1007/s40291-026-00849-2","url":null,"abstract":"<p><p>Liquid biopsy has emerged as a minimally invasive approach for the molecular characterization and longitudinal monitoring of primary central nervous system (CNS) tumors. Although extensively validated in systemic malignancies, its clinical application in CNS tumors is challenged by the blood-brain barrier, low analyte abundance, and heterogeneous assay performance. Recent advances have expanded the spectrum of detectable tumor-derived components, including circulating tumor DNA (ctDNA), cell-free DNA, extracellular vesicles, RNA species, nucleosomes, metabolites, and lipids, across multiple biofluids such as cerebrospinal fluid (CSF), plasma, serum, urine, saliva, and tears. The aim of this study was to review the biological foundations, analytes, biofluids, clinical applications, and technical limitations of liquid biopsy in primary CNS tumors, with emphasis on diagnostic, prognostic, and surveillance value. We synthesized current evidence on tumor-derived analytes detectable through liquid biopsy, their molecular correlates, and their performance across biofluids. CSF is consistently the most informative biofluid for CNS tumors, with higher analyte concentration and superior concordance with tumor tissue compared with plasma. CtDNA in CSF reliably identifies hallmark alterations, including IDH1/2, H3K27M, TERT, BRAF, ATRX, TP53, 1p/19q codeletion, and MYCN amplification, thereby enabling better diagnosis, molecular classification, and therapeutic stratification. Plasma-based assays are less sensitive but remain valuable for longitudinal monitoring, especially when combined with ultrasensitive sequencing or fragmentomic approaches. Emerging biomarkers, including nucleosome footprints, exosomes, proteins, microRNA (miRNA)/long noncoding RNA (lncRNA)/circular RNA (circRNA) signatures, lipidomic panels, and metabolites such as D-2-hydroxyglutarate, show potential for integration into multimodal diagnostics. Liquid biopsy provides a powerful and rapidly evolving tool for the minimally invasive molecular assessment of CNS tumors. While CSF remains the optimal matrix for diagnosis and characterization, advances in ultrasensitive detection methods increasingly support the feasibility of plasma and urine for longitudinal follow-up. Integrating liquid biopsy with advanced imaging and tissue-based data will likely transform diagnostic accuracy, therapeutic decision-making, and real-time monitoring of CNS tumors.</p>","PeriodicalId":49797,"journal":{"name":"Molecular Diagnosis & Therapy","volume":" ","pages":""},"PeriodicalIF":4.4,"publicationDate":"2026-05-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147845287","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Therapeutic Strategies Targeting the Molecular Pathogenesis of Myotonic Dystrophy Type 1: Current Status and Future Directions.","authors":"Mohamed Chahine, Vamsi Krishna Murthy Ginjupalli, Dominic Jauvin, Mohamed Boutjdir","doi":"10.1007/s40291-026-00848-3","DOIUrl":"https://doi.org/10.1007/s40291-026-00848-3","url":null,"abstract":"<p><p>Myotonic dystrophy type 1 is the most prevalent adult-onset muscular dystrophy and is characterized by progressive muscle weakness, myotonia, cardiac conduction defects, endocrine dysfunction, and central nervous system involvement. Myotonic dystrophy type 1 is caused by an unstable CTG repeat expansion in the 3' untranslated region of the DMPK gene, which produces toxic CUG-expanded transcripts that sequester RNA-binding proteins such as Muscleblind-like, induce widespread alternative splicing defects, and drive an RNA gain-of-function mechanism rather than simple DMPK haploinsufficiency. Despite major advances in understanding the molecular pathogenesis of myotonic dystrophy type 1, there is still no approved cure or disease-modifying therapy. This review summarizes the molecular basis of myotonic dystrophy type 1 and provides an in-depth overview of emerging therapeutic strategies that directly target the underlying pathogenic cascade at the DNA and RNA levels. Gene therapy-based approaches, including CRISPR-mediated genome editing, aim to reduce or eliminate the expanded CTG repeats or expanded DMPK allele and its toxic transcripts. In parallel, a broad spectrum of RNA-directed interventions is being developed, encompassing antisense oligonucleotides, antibody-penetrating and cell-penetrating peptide-conjugated antisense oligonucleotides to enhance skeletal and cardiac muscle delivery, small interfering RNAs, and microRNA-based tools such as antagomiRs. Additional strategies exploit engineered RNA-binding proteins and peptide decoys to disrupt toxic ribonuclear aggregates, polyadenylation signal-driven premature transcriptional termination to selectively silence mutant DMPK, and small molecules that modulate RNA metabolism, dissolve CUG RNA foci, or correct downstream mis-splicing. By integrating data from preclinical models and ongoing clinical trials, including recent advances with muscle‑targeted antisense oligonucleotide conjugates and gene therapy, this review outlines the current status, strengths, and limitations of these mechanism-based therapies for myotonic dystrophy type 1. The discussion highlights key translational challenges such as efficient delivery to skeletal muscle, the heart, and brain, long-term safety, and robust pharmacodynamic biomarkers as well as opportunities for combination and next-generation approaches aimed at converting molecular correction into durable clinical benefit for patients with myotonic dystrophy type 1.</p>","PeriodicalId":49797,"journal":{"name":"Molecular Diagnosis & Therapy","volume":" ","pages":""},"PeriodicalIF":4.4,"publicationDate":"2026-04-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147718737","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Beyond Binary FISH Results: Targeted NGS Provides Deeper Characterization of MYC and Broader Genomic Alterations in DLBCL.","authors":"Mariana Bastos Oreiro, Sara Fernández de Córdoba Oñate, Ismael de la Iglesia San Sebastián, Francisco Díaz-Crespo, María Chicano Lavilla, Juan Carlos Triviño, Raquel Fernández González, Miguel López-Esteban, Marina Gómez-Llobel, Paula Fernández Caldas, Pablo Silva, Javier Menárguez, Ramón García Sanz, Ismael Buño, Carolina Martínez Laperche","doi":"10.1007/s40291-026-00846-5","DOIUrl":"https://doi.org/10.1007/s40291-026-00846-5","url":null,"abstract":"<p><strong>Introduction: </strong>Rearrangements involving MYC, BCL2, and BCL6 are central to the genetic classification and risk stratification of diffuse large B cell lymphoma (DLBCL). In routine practice, these alterations are primarily assessed by fluorescence in situ hybridization (FISH), which provides limited diagnostic information and lacks information on partner genes, breakpoint architecture, or clonal complexity.</p><p><strong>Methods: </strong>We investigated whether targeted next-generation sequencing (NGS) refines the interpretation of gene rearrangements in aggressive B cell lymphomas and alters their diagnostic classification. Tumor samples from 48 patients were analyzed using FISH, immunohistochemistry, and a custom hybrid-capture NGS panel enabling simultaneous detection of structural variants, copy number alterations, and somatic mutations.</p><p><strong>Results: </strong>Targeted NGS identified 59 fusion events in 37 patients, including 26 MYC rearrangements (MYC-Rs). A substantial proportion (58%) involved non-immunoglobulin partners, many of them previously unreported. NGS resolved inconclusive or discordant FISH results, uncovered cryptic rearrangements, and revealed genomic complexity. In several cases, these findings refined genetic classification, including the identification of additional double- and triple-hit lymphomas.</p><p><strong>Conclusions: </strong>These results demonstrate that a FISH-based assessment of MYC-, BCL2-, and BCL6-Rs is insufficient to capture the biological and diagnostic complexity of DLBCL. Targeted NGS provides structural and functional context that improves interpretation of rearrangements and supports a more refined genetic classification in routine practice.</p>","PeriodicalId":49797,"journal":{"name":"Molecular Diagnosis & Therapy","volume":" ","pages":""},"PeriodicalIF":4.4,"publicationDate":"2026-04-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147677986","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"From Gene to Hope: Rett Syndrome and the Rise of Molecular Therapies.","authors":"Yoann Leblay, Marie-Solenne Felix, Jean-Christophe Roux, Nicolas Panayotis","doi":"10.1007/s40291-026-00847-4","DOIUrl":"https://doi.org/10.1007/s40291-026-00847-4","url":null,"abstract":"<p><p>Rett syndrome is a rare X-linked neurodevelopmental disorder caused by mutations in MECP2, a gene critical for neuronal function, chromatin organization, and synaptic plasticity. After a period of apparently normal early development, individuals with Rett syndrome experience rapid regression followed by lifelong neurological impairment. Notably, preclinical studies have shown that restoration of MeCP2 expression can reverse established symptoms in adult mice, positioning Rett syndrome as a promising target for molecular therapies. However, MECP2 is extremely dosage sensitive and both insufficient and excessive expression are harmful, creating a narrow therapeutic window and a major challenge for treatment design. This review examines the evolving landscape of gene- and RNA-based therapies for Rett syndrome, with a focus on strategies that enable precise MeCP2 replacement, dosage control, and widespread central nervous system delivery. We discuss clinical-stage adeno-associated virus gene replacement programs, including TSHA-102 and NGN-401, highlighting their vector designs, regulatory elements, delivery approaches, and emerging clinical data. Advances in adeno-associated virus capsid engineering and vector optimization aimed at improving neuronal targeting while minimizing peripheral exposure and immune toxicity are also reviewed. Beyond gene supplementation, we explore approaches that restore endogenous MECP2 regulation, such as reactivation of the inactive X chromosome, as well as DNA and RNA editing strategies. Collectively, these advances reflect a shift toward precision-regulated therapies for Rett syndrome that may provide a model for treating other dosage-sensitive neurodevelopmental disorders.</p>","PeriodicalId":49797,"journal":{"name":"Molecular Diagnosis & Therapy","volume":" ","pages":""},"PeriodicalIF":4.4,"publicationDate":"2026-04-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147677968","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Neoantigen Targeting as a Novel Approach for Therapy-Resistant Tumors.","authors":"Masahiro Okada, Satoru Yamasaki, Go R Sato, Kanako Shimizu, Shin-Ichiro Fujii","doi":"10.1007/s40291-026-00842-9","DOIUrl":"https://doi.org/10.1007/s40291-026-00842-9","url":null,"abstract":"<p><p>Neoantigens have emerged as central targets in the development of individualized cancer vaccines, as they are recognized by the immune system as foreign and can elicit potent anti-tumor responses. Advances in computational algorithms and machine learning have improved the accuracy of neoantigen prediction, and clinical trials have reported promising results of vaccines incorporating validated neoantigens into peptide, RNA, or dendritic cell platforms. Despite this progress, the identification of immunologically effective neoantigens remains a complex multi-step process. Current attention is directed not only to conventional missense and indel mutations but also to non-coding RNA-derived neoantigens. A critical challenge is the establishment of reliable systems to verify whether candidate neoantigens can activate cytotoxic T lymphocytes. Moreover, neoantigen expression and immune responses are influenced by tumor-intrinsic and therapeutic factors. High programmed death-ligand 1 expression is known to suppress immune recognition, while ARID1A mutations, associated with tumor progression, can enhance neoantigen expression during chemoresistance, suggesting that drug resistance may be accompanied by new immunogenicity. This work provides an overview of current methodologies for neoantigen identification, advances in prediction and validation strategies, and the dynamic interplay between tumor-intrinsic and immune-related factors that regulate neoantigen expression. We also highlight recent clinical insights as well as novel analytical approaches and discuss challenges and future directions in this rapidly evolving field, emphasizing the potential of neoantigen-based therapies to transform cancer immunotherapy.</p>","PeriodicalId":49797,"journal":{"name":"Molecular Diagnosis & Therapy","volume":" ","pages":""},"PeriodicalIF":4.4,"publicationDate":"2026-03-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147582814","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"The Evolving Role of Genomic Technologies in Multiple Myeloma: Implications for Diagnosis, Risk Stratification and Resistance.","authors":"Klára Baďurová, Veronika Kapustová, Jana Kotulová, Vítězslav Brinsa, Lenka Piherová, Václav Janoušek, Michal Pohludka, Michal Richtář, Tomáš Jelínek, Roman Hájek, Zuzana Chyra, Tereza Ševčíková","doi":"10.1007/s40291-026-00836-7","DOIUrl":"https://doi.org/10.1007/s40291-026-00836-7","url":null,"abstract":"<p><p>Plasma cell disorders range from indolent precursors to aggressive malignancies, with multiple myeloma as the most common malignant form. Multiple myeloma biology is highly complex and driven by genomic heterogeneity. Although recent genomic advances offer increasingly precise treatment strategies, routinely used conventional diagnostic methods sometimes fail to reflect the evolution of up-to-date multiple myeloma risk stratification models, and the implementation of genomic discoveries into clinical guidelines is hindered. We reviewed the current state and emerging roles of genomic diagnostics in multiple myeloma and related conditions, emphasizing the clinical utility. We summarized clinical presentation and diagnostic criteria, mapped the mutational landscape across disease stages, and tracked the evolution of risk stratification from clinical staging to integrated genomic models. We also highlighted recent recommendations that incorporate a quantitative assessment of cytogenetic abnormalities and tumor suppressor gene mutations. Further, we compared routinely used and state-of-the-art technologies including fluorescence in situ hybridization, targeted gene panels, whole-exome and whole-genome sequencing, long-read platforms, optical genome mapping, and analyses of circulating tumor DNA, focusing on their strengths, limitations, and complementary roles in the baseline work-up, longitudinal monitoring, and detection of resistance. Last, we discussed practical barriers to implementation and proposed a framework for proactive integration of novel approaches into clinics. In summary, comprehensive genomic profiling is becoming central to modern multiple myeloma diagnostics and a key step for precise and personalized medicine that will, if integrated into routine care, enable a more precise risk assessment, earlier detection of relapse, and improved therapy selection.</p>","PeriodicalId":49797,"journal":{"name":"Molecular Diagnosis & Therapy","volume":" ","pages":""},"PeriodicalIF":4.4,"publicationDate":"2026-03-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147576342","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Exosomal Cargo-Derived Mediators of Ovarian Cancer Chemoresistance.","authors":"Szymon Rutecki, Krzysztof Książek, Justyna Mikuła-Pietrasik","doi":"10.1007/s40291-026-00844-7","DOIUrl":"https://doi.org/10.1007/s40291-026-00844-7","url":null,"abstract":"<p><p>Exosomes are small extracellular vesicles secreted by various cells. They play a vital role in intercellular communication due to their diverse molecular cargo. Recent advancements in biomedical research have enabled a more detailed characterization of exosomes and their significant role in cancer biology, particularly in understanding mechanisms that contribute to chemoresistance. This review focuses on the current understanding of exosomes in ovarian cancer, one of the deadliest gynecological malignancies, known for its high recurrence and treatment failure rates. Chemoresistance in ovarian cancer stems from several factors, including altered drug efflux, enhanced DNA repair mechanisms, changes in the tumor microenvironment, and modifications in signaling pathways. Emerging evidence suggests that exosomes facilitate these processes by transferring regulatory molecules such as proteins, microRNAs (miRNAs), and circular RNAs (circRNAs) between cells, which in turn modulate drug response and tumor progression. For instance, exosomal proteins such as DNA methyltransferase 1 (DNMT1) and circular forkhead box P1 (circFoxp1), along with miRNAs like miR-21-3p, miR-1246, and miR-6836, have been associated with promoting resistance to platinum- and taxane-based chemotherapies. Conversely, some exosomal miRNAs, including miR-30a-5p, may enhance drug sensitivity. Furthermore, circRNAs transported by exosomes, such as hsa_circ_0010467, circ-PIP5K1A, and circ_0025033, play a role in regulating key oncogenic pathways associated with chemoresistance. Overall, these findings highlight the multifaceted role of exosomes in ovarian cancer biology and underscore their potential as both biomarkers and therapeutic targets. A deeper understanding of how exosomes mediate molecular mechanisms may lead to novel strategies for overcoming chemoresistance and improving treatment outcomes for ovarian cancer patients.</p>","PeriodicalId":49797,"journal":{"name":"Molecular Diagnosis & Therapy","volume":" ","pages":""},"PeriodicalIF":4.4,"publicationDate":"2026-03-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147576301","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Etuvetidigene Autotemcel: First Approval.","authors":"Hannah A Blair","doi":"10.1007/s40291-026-00843-8","DOIUrl":"https://doi.org/10.1007/s40291-026-00843-8","url":null,"abstract":"<p><p>Etuvetidigene autotemcel (WASKYRA^™) is an autologous haematopoietic stem cell (HSC)-based gene therapy being developed by Fondazione Telethon for the treatment of Wiskott-Aldrich syndrome (WAS). In December 2025, etuvetidigene autotemcel was approved in the USA for the treatment of paediatric patients aged 6 months and older and adults with WAS who have a mutation in the WAS gene for whom HSC transplantation (HSCT) is appropriate and no suitable human leukocyte antigen (HLA)-matched related stem cell donor is available. Etuvetidigene autotemcel was approved in the EU for the same indication in January 2026. This article summarizes the milestones in the development of etuvetidigene autotemcel leading to this first approval for the treatment of WAS.</p>","PeriodicalId":49797,"journal":{"name":"Molecular Diagnosis & Therapy","volume":" ","pages":""},"PeriodicalIF":4.4,"publicationDate":"2026-03-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147516209","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"RNA Aptamers and Epitranscriptomics: Charting Unexplored Territories in RNA Biology.","authors":"Mehran Khorshid, Ehsan Pashay Ahi","doi":"10.1007/s40291-026-00841-w","DOIUrl":"https://doi.org/10.1007/s40291-026-00841-w","url":null,"abstract":"<p><p>The fields of RNA aptamers and epitranscriptomics have each emerged as powerful dimensions of RNA biology, yet their conceptual and mechanistic overlap remains largely unexplored. RNA aptamers as in vitro-selected single-stranded RNA oligonucleotides selected through in vitro evolution (Systematic Evolution of Ligands by Exponential Enrichment [SELEX]) and related methods are well known for their ability to fold into precise three-dimensional structures and bind specific targets with high affinity. These aptamers have been widely used in diagnostics, therapeutics, and molecular sensing. In parallel, the dynamic landscape of epitranscriptomic modifications, such as N⁶-methyladenosine (m⁶A), pseudouridine (Ψ), and 5-methylcytosine (m⁵C), has been recognized as a critical regulator of RNA structure, stability, and function. In this perspective article, we chart unexplored territories at the interface of these two mechanisms, focusing on how RNA modifications may influence aptamer folding or binding, and how aptamers could, in turn, be harnessed to detect or modulate the epitranscriptomic state. We also highlight opportunities for cross-disciplinary integration in synthetic biology, post-transcriptional regulation, and RNA-based therapeutics. While the direct regulatory connections between these domains remain to be fully elucidated, we propose their potential convergence as a fertile ground for innovation. The primary aim of this article is to stimulate discussion within and between these research communities and to attract attention toward a promising frontier for future investigations.</p>","PeriodicalId":49797,"journal":{"name":"Molecular Diagnosis & Therapy","volume":" ","pages":""},"PeriodicalIF":4.4,"publicationDate":"2026-03-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147469847","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Pre-analytical Best Practices for RNA Sequencing from Small Biopsies and Cytologic Specimens.","authors":"Gloria Hopkins Sura, Kelly B Engel, Sarah R Greytak, Sandra M Gaston, Kelsey Dillehay McKillip, Abhilasha Rao, Ping Guan, W Fraser Symmans, Rachael Clark, Maria Arcila, Sayak Ghatak, Karol Bomsztyk, Lokesh Agrawal","doi":"10.1007/s40291-026-00837-6","DOIUrl":"https://doi.org/10.1007/s40291-026-00837-6","url":null,"abstract":"<p><p>RNA sequencing is becoming increasingly common in precision oncology, in both clinical and research settings, for transcriptome profiling, gene-fusion detection, and biomarker discovery. RNA-sequencing analysis of specimens obtained via minimally invasive procedures such as small biopsy, fine needle aspiration, and exfoliation offers a powerful method for analyzing gene expression patterns and detecting RNA-level changes associated with cancers that are either difficult to collect or require longitudinal sampling. However, pre-analytical factors (e.g., details of specimen collection, processing, and storage workflow) influence not only RNA-sequencing success rates but also the quality and accuracy of sequencing results, which may affect patient care and research progress. Minimally invasive specimens are associated with a unique set of pre-analytical challenges owing to their small size, limited RNA yield, and distinct workflows. To address the need for evidence-based guidance, this review by a working group of National Cancer Institute grantees and intramural and extramural researchers identifies pre-analytical best practices for minimally invasive specimens destined for RNA-sequencing analysis, based on the available literature and their collective experience. Strategies for assessing specimen adequacy and RNA quality, maximizing tumor content, and minimizing specimen loss and RNA degradation due to pre-analytical handling are specified for small tissue and cytology specimens. By integrating current evidence and institutional insights, this review provides a practical framework for enhancing RNA-sequencing reliability and reproducibility in both clinical and research workflows.</p>","PeriodicalId":49797,"journal":{"name":"Molecular Diagnosis & Therapy","volume":" ","pages":""},"PeriodicalIF":4.4,"publicationDate":"2026-03-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147460703","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}