The estrogen receptor-positive, human epidermal growth factor receptor 2-negative metastatic breast cancer (MBC) treatment landscape continues to evolve with targeted therapy development. Estrogen receptor 1 (ESR1) testing is recommended at recurrence or each progression event to identify patients for effective therapies. However, there is limited information on testing and mutation patterns in the real world that can inform patient identification in the clinic. A retrospective observational study among patients with estrogen receptor-positive, human epidermal growth factor receptor 2-negative MBC who initiated first line (1L) therapy with aromatase inhibitors or selective estrogen receptor degraders ± CDK4/6 inhibitors from 1 January, 2020 to 31 March, 2025, using the Flatiron Health Research Database was conducted. ESR1 testing and mutation positivity rates at 1L to third line initiation, stratified by endocrine-resistant, 1L endocrine-sensitive, and endocrine-sensitive/de novo MBC were descriptively reported. Overall testing rates varied across subgroups, with one or more ESR1 test per patient in 61.2
Anbalcabtagene autoleucel (RIMQARTO®) is a CAR T therapy that includes CAR T cells with dual PD-1 and TIGIT downregulation to mitigate T cell exhaustion. It is being developed by Curocell Inc. for the treatment of diffuse large B-cell lymphoma, acute lymphoblastic leukaemia and autoimmune diseases, which received its first approval in April 2026 in South Korea. This article summarizes the milestones in the development of anbalcabtagene autoleucel leading to this first approval for the treatment of relapsed or refractory diffuse large B-cell lymphoma and primary mediastinal large B-cell lymphoma in adults following ≥ 2 lines of systemic therapy.
Molecular profiling is increasingly required for treatment selection in advanced biliary tract cancer, yet tumor tissue is often insufficient and plasma circulating tumor DNA has limited yield. Because bile directly contacts the biliary epithelium and is obtainable during routine biliary interventions, bile cell-free DNA (cfDNA) may provide a tumor-proximal molecular profiling source. We systematically reviewed and quantitatively synthesized bile cfDNA detection and profiling in biliary tract cancer. PubMed, EMBASE, and the Cochrane Library were searched from inception to 13 October, 2025 for studies that included at least ten patients with suspected or confirmed biliary tract cancer and extractable bile cfDNA outcomes. Pooled proportions were estimated using random-effects models with Freeman–Tukey double-arcsine transformation. Bile-versus-plasma detection was compared using pooled log risk ratios. Heterogeneity was quantified by I2, with a 95
Synaptic loss is a core pathological feature of neurodegenerative disorders and closely relates to cognitive and functional decline. Positron emission tomography (PET) targeting synaptic vesicle glycoprotein 2A (SV2A) has recently emerged as a promising tool for the indirect assessment of presynaptic alterations in the living human brain. This leading article discusses the evolving clinical landscape of SV2A PET across the neurodegenerative spectrum, emphasizing its translational trajectory and emerging applications. Current evidence spans Alzheimer's disease (AD), other dementias, movement disorders, Huntington's disease (HD), and amyotrophic lateral sclerosis (ALS). The literature is currently dominated by PET with [11C]UCB-J, while [18F]-labeled tracers, particularly [18F]SynVesT-1, are expanding clinical feasibility through longer half-life and broader distribution potential. Across disorders, PET consistently detected SV2A reductions that frequently correlated with cognition, disease severity, and complementary biomarkers, including amyloid, tau, glucose metabolism, and dopaminergic imaging. Although the field remains limited by small cohorts, heterogeneous quantification strategies, and incomplete longitudinal validation, SV2A PET is rapidly evolving into a promising translational tool for studying synaptopathies and monitoring disease progression.
Early recognition of sepsis in emergency departments (EDs) improves outcomes and reduces healthcare resource use. However, current clinical practice (CCP), largely based on the National Early Warning Score 2 (NEWS2) and clinical judgement, has limited diagnostic sensitivity at presentation. Consequently, some patients with early sepsis are not promptly identified, resulting in delayed treatment, disease progression, and increased use of high-cost care. Rapid point-of-care host-response diagnostics may complement clinical assessment by identifying patients at risk who do not initially meet conventional clinical thresholds. To estimate the cost-effectiveness of adding a rapid LPS-induced TNF-α release point-of-care assay (TARA) to current clinical practice, compared with procalcitonin (PCT) plus CCP and CCP alone, from the UK National Health Service (NHS) and Personal Social Services perspective, in line with NICE reference case methods. A decision-analytic model simulated diagnostic classification, timing of sepsis recognition (early, delayed, late), initiation of antibiotic treatment, hospital admission (general ward or intensive care unit [ICU]), discharge, and readmission among adults presenting to UK EDs with suspected sepsis without shock. Model inputs included diagnostic accuracy, clinical outcomes, health-related quality of life, and costs derived from published literature and UK-relevant sources. The base-case time horizon was lifetime, to fully capture the QALY consequences of prevented deaths and long-term sequelae (ESRD, amputation); a 6-month scenario analysis was also conducted. Deterministic and probabilistic sensitivity analyses assessed parameter uncertainty. In the deterministic base-case analysis, TARA plus CCP was associated with lower total costs and higher quality-adjusted life-years (QALYs) compared with both comparator strategies. Cost savings were approximately £370 per patient versus PCT plus CCP and £1,822 per patient versus CCP alone, with incremental QALY gains of 0.0834 and 0.0836, respectively. In the probabilistic analysis, mean cost savings were £391 versus PCT plus CCP and £1,835 versus CCP alone. Benefits were driven primarily by reductions in false-negative diagnoses, leading to earlier treatment, lower ICU use, and reduced downstream hospital costs. Integrating a rapid host-response point-of-care assay into ED sepsis pathways in the UK was associated with lower total costs (savings of approximately £370 per patient versus PCT plus CCP and £1,822 per patient versus CCP alone) and marginally higher quality-adjusted life-years (QALY gains of 0.0834 and 0.0836, respectively), driven by reductions in missed diagnoses and downstream use of high-cost care.
Non-invasive prenatal testing (NIPT) based on circulating cell-free DNA (cfDNA) has transformed prenatal screening for common aneuploidies, but its interpretation is less straightforward in pregnancies conceived with donor oocytes. In this setting, fetal chromosomal risk is largely donor-driven, whereas cfDNA interpretability is shaped mainly by the recipient and the placenta. Donor-oocyte pregnancies are enriched for advanced maternal age, obesity, hypertensive and immunologic complications, low-molecular-weight heparin exposure, multiple gestation, vanishing twin, and placental dysfunction – all factors that can destabilize the cfDNA signal. Fetal fraction should therefore be read not only as a technical quality metric but also as a biological marker of placental performance. Analytical platform behavior also matters; counting-based and single-nucleotide polymorphism-based approaches may behave differently in biologically complex samples, particularly for sex chromosome findings and in the setting of maternal confounding. In donor-oocyte pregnancies, a no-call result may be clinically informative rather than neutral. Pre-test counseling, explicit communication with the laboratory, availability of fetal-fraction reporting, and integration with first-trimester ultrasound are essential. Donor-oocyte pregnancy should be regarded as a biologically distinct model for cfDNA screening, one that exposes the limits of generic NIPT algorithms and argues for more tailored interpretation.
Endoplasmic reticulum (ER) is a multifunctional organelle essential for maintaining proteostasis, lipid and carbohydrate metabolism, and calcium homoestasis. Rapidly dividing cancer cells driven by oncogenes, elevated translational output, increased metabolic demands, and a hostile tissue microenvironment overwhelm the protein-folding machinery of ER, leading to massive accumulation of unfolded proteins within the ER's lumen leading to chronic ER stress. This activates the unfolded protein response (UPR), a conserved signaling network mediated by three principal sensors: protein kinase R-like endoplasmic reticulum kinase (PERK), inositol-requiring enzyme 1-alpha (IRE1α), and activating transcription factor 6 (ATF6), which functions to restore proteostasis or induce apoptosis under unresolved ER stress. Accumulating evidence indicates that malignant cells hijack the pro-adaptive function of the UPR pathway not only to thrive but also to promote cancer progression by invasion and metastasis. UPR activation modulates transcriptional and translational programs that contribute to angiogenesis, invasion, metastasis, immune escape, and chemoresistance. In this review, we dissect how cells balance this tightrope between adaptation and cell death in the context of cancer. We also explore how UPR signaling drives angiogenesis, metastasis, immune-evasion, and chemoresistance before finally discussing its therapeutic potential.
Neisseria gonorrhoeae is a global public health threat, exacerbated by the rapid dissemination of ceftriaxone-resistant strains harboring penA allele 60.001. Current diagnostics are limited by long turnaround times, reliance on thermal cycling equipment, and poor suitability for resource-limited settings. We developed an isothermal recombinase polymerase amplification assay coupled with a lateral flow strip (RPA-LFS) for separate identification of N. gonorrhoeae and the ceftriaxone resistance-associated SNP C932T (A311V) in penA 60.001 and limited closely related alleles. Species detection employed an RPA-Nfo probe targeting the conserved porA pseudogene. Resistance detection employed an allele-specific amplification refractory mutation system RPA (ARMS-RPA) strategy, incorporating artificial mismatches to specifically discriminate the C932T SNP. Assay performance was evaluated using genomic DNA of a panel of gonococcal and non-gonococcal strains and validated directly on unprocessed clinical urine samples. The porA RPA-LFS assay demonstrated a detection limit of 30 fg of genomic DNA with no cross-reactivity against closely related Neisseria species or other bacterial pathogens. The mismatch penA 60.001 RPA-LFS assay achieved a detection limit of 300 fg and exhibited absolute analytical specificity, with no amplification observed from wild-type or other mosaic penA alleles. Validation using 50 clinical urine samples yielded 100
Sudden cardiac death is the leading cause of death among athletes during exercise. These events are devastating for both the family and society, especially when they occur in children and young adults. Some of these unexpected deceases occur in athletes who carry genetic alterations that predispose them to malignant arrhythmias, and exercise is the main trigger for these lethal episodes. These genetic alterations are the origin of inherited arrhythmogenic syndromes, often latent, and the first clinical manifestation can be sudden death itself. It is also important to take into account the toxicology data, since high doses of certain substances, as well as mixtures of them, can also be significant triggers of malignant arrhythmias, especially in genetically predisposed hearts. Genetic studies that can provide a diagnosis can be performed on deceased individuals (molecular autopsy) or on their relatives, who may be carriers of the genetic defect and at risk of malignant arrhythmogenic events. Early identification of individuals at risk allows adoption of preventive measures helping to reduce risk of life-threatening arrhythmogenic episodes. Our study examines the main arrhythmogenic causes of sudden cardiac death in athletes during exercise, highlighting a multidisciplinary interpretation of forensic, clinical, genetic and molecular data to obtain a definite cause for the lethal episode.
Intrinsic and acquired resistance to temozolomide (TMZ), the standard chemotherapeutic agent for glioblastoma (GBM), is highly common and results in poor clinical outcomes. This review highlights the emerging dual role of Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR)-associated protein (Cas) technologies in addressing this challenge. First, it examines genome-wide CRISPR screens that revealed DNA damage repair networks, stress adaptations, stemness maintenance, and tumor heterogeneity as key drivers of resistance to TMZ. Second, it examines CRISPR/Cas-based strategies, including targeted gene disruption and epigenetic silencing of O6-methylguanine-DNA methyltransferase (MGMT), to restore TMZ sensitivity. Finally, it explores CRISPR/Cas-engineered brain tumor models. Alongside these approaches, CRISPR/Cas technologies highlight the value of decoding the multifactorial basis of TMZ resistance and guiding rational therapeutic strategies. Continued refinement of CRISPR/Cas tools may ultimately contribute to more effective treatments for GBM.
Marnetegragene autotemcel (KRESLADI™), an autologous stem cell-based gene therapy that adds functional copies of the ITGB2 gene to haematopoietic stem cells, was developed by Rocket Pharmaceuticals for the treatment of leukocyte adhesion deficiency-I (LAD-I). In March 2026, marnetegragene autotemcel received its first approval in the USA for the treatment of paediatric patients with severe LAD-I due to biallelic variants in ITGB2 without an available human leukocyte antigen (HLA)-matched sibling donor for allogeneic haematopoietic stem cell transplant. This article summarises the milestones in the development of marnetegragene autotemcel leading to this first approval for treatment of paediatric patients with severe LAD-I.
BACKGROUND AND OBJECTIVE:Circulating tumor DNA enables longitudinal monitoring in cancer, but current sequencing-based assays are limited by cost, turnaround time, performance in difficult genomic regions, and infrequent sampling. Universal Signal Encoding PCR (USE-PCR) is a highly multiplexed digital PCR chemistry that enables measurement of up to 32 tumor-informed targets per reaction. Here, we evaluated USE-PCR for dense longitudinal circulating tumor DNA monitoring in patients with metastatic melanoma undergoing immunotherapy. METHODS:Tumor and plasma sequencing identified subject-specific variants from nine subjects with melanoma, including variants in GC-rich and historically challenging loci such as the TERT promoter. Subject-specific USE-PCR panels were designed using an automated cloud workflow and applied to plasma cell-free DNA and matched peripheral blood mononuclear cell sample DNA, with up to 21 timepoints per subject, collected over a 2-year window. Variant allele fractions were quantified on a commercial digital PCR platform and interpreted with a peripheral blood mononuclear cell genomic DNA background. CONCLUSIONS:USE-PCR resolved variant-specific circulating tumor DNA trajectories across longitudinal plasma samples. Dense sampling revealed rapid and low-level circulating tumor DNA shifts, including subclonal variants below the 0.2% variant allele fraction that may not be captured through less frequent surveillance intervals. Panels supported the incorporation of new variants without compromising analytical performance.
Friedreich ataxia is a rare, autosomal recessive neurogenerative disorder caused by mutations to the frataxin (FXN) gene resulting in loss of functional FXN protein. Applications of cutting-edge genetic and cellular therapies expand therapeutic options for patients with rare, genetic diseases including Friedreich ataxia. Multiple investigational techniques and strategies seek to permanently alter the disease course in patients with Friedreich ataxia, although no product has established definitive benefit. This review catalogs both the history and ongoing efforts of genetic and cellular therapies applied to Friedreich ataxia and its disease models, including therapeutic efficacy and adverse effects. We list the key limitations and cautions of such therapies, chiefly those of potential FXN overexpression toxicity, critical therapeutic windows, and adverse effects of these therapies applicable to any disease target. As gene and cellular therapy continue to diversify in design and strategy, Friedreich ataxia patients will likely have multiple therapeutic options in the future from both investigational therapies described here and future ones yet to be optimized.
BackgroundColorectal cancer is one of the most common and deadly malignancies worldwide. Traditionally, all colon tumours have been treated as a single entity, but growing evidence shows that tumour behaviour differs markedly according to tumour sidedness. Right-sided and left-sided tumours exhibit distinct molecular profiles, prognoses and therapeutic responses. A liquid biopsy is a minimally invasive approach to monitor tumour dynamics in real time, offering valuable prognostic and predictive insights into metastatic colorectal cancer. However, despite increasing research on liquid biopsy-based biomarkers, the potential impact of tumour sidedness on their interpretation has not been systematically explored.MethodsWe evaluated circulating RAS mutational status, RAS mutant allele fraction and cell-free DNA concentration and fragmentation in plasma from 232 patients with metastatic colorectal cancer.ResultsOur findings reveal that tumour sidedness substantially modulates the prognostic value of these circulating biomarkers. In left-sided tumours, worse outcomes were associated with the presence of circulating RAS mutations, higher cell-free DNA concentrations and elevated carcinoembryonic antigen levels. In contrast, among right-sided tumours, only mutant allele fraction levels revealed significant prognostic relevance.ConclusionsOur results demonstrate that tumour sidedness critically influences liquid biopsy interpretation in metastatic colorectal cancer, underscoring the need to incorporate the primary tumour location into the clinical assessment of circulating biomarkers.
Tividenofusp alfa (tividenofusp alfa-eknm; AVLAYAH™) is an intravenous brain-penetrant enzyme replacement therapy being developed by Denali Therapeutics for the treatment of mucopolysaccharidosis type II (MPS II). Also known as Hunter syndrome, MPS II is a rare X-linked recessive lysosomal storage disease caused by a deficiency of the lysosomal enzyme iduronate-2-sulfatase resulting in accumulation of glycosaminoglycans, heparan sulphate and dermatan sulphate. Consisting of the iduronate-2-sulfatase enzyme fused to a transferrin receptor-binding Fc domain, tividenofusp alfa has been designed to enable distribution of exogenous iduronate-2-sulfatase into both the central nervous system and peripheral tissues. Based on a reduction in cerebrospinal fluid heparan sulphate levels in patients with MPS II treated with tividenofusp alfa in a phase I/II trial, in March 2026 tividenofusp alfa was approved in the USA, under accelerated approval, for the treatment of neurologic manifestations of MPS II when initiated in presymptomatic or symptomatic paediatric patients weighing ≥ 5 kg prior to advanced neurologic impairment. Continued approval for tividenofusp alfa may be contingent upon verification of clinical benefit in a confirmatory trial. Tividenofusp alfa has also been granted Priority Medicine designation by the European Medicines Agency. This article summarises the milestones in the development of tividenofusp alfa leading to this first approval for MPS II.
Cerebrotendinous xanthomatosis (CTX) is rare, autosomal recessive inborn error of metabolism caused by biallelic pathogenic variants in CYP27A1, which encodes sterile 27 hydroxylase, a key enzyme in bile acid biosynthesis. Enzyme deficiency results in reduced cholic and chenodeoxycholic acid synthesis with accumulation of cholestanol, bile acid intermediates, and bile alcohols, producing a progressive multisystem disorder characterized by chronic diarrhea, juvenile-onset cataracts, tendons xanthomas, and neurological dysfunction. Although CTX typically begins in childhood, diagnosis is frequently delayed until adulthood, limiting the benefit of effective disease modifying therapy with chenodeoxycholic acid. Since the identification of CYP27A1, more than 200 pathogenic variants have been reported, including canonical loss of function alleles and missense variants with variable residual enzyme activity. In this review, we summarize the medical genetics, population genetics and genotype phenotype relationships of CTX. We highlight insights from large population databases that refine global incidence estimates, revealing population-specific enrichment of pathogenic variants and persistent underdiagnosis. We further review functional and clinical data demonstrating that stratification of CYP27A1 variants by functional effect, complete loss of function versus hypomorph alleles, correlates with clinical severity and biochemical phenotype. Finally, we discuss emerging advances in biochemical testing and newborn screening strategies that offer the potential for presymptomatic diagnosis, enabling timely initiation of therapy and prevention of irreversible disease manifestations. Integrating molecular, functional, and population genetic data provides a framework for improved variant interpretation, earlier diagnosis, and optimized therapeutic intervention in CTX.
Activating ESR1 mutations are a major mechanism of resistance to aromatase inhibitors in hormone receptor-positive, HER2-negative metastatic breast cancer (mBC). International guidelines, including those from ASCO, NCCN, and ESMO, recommend liquid biopsy as the preferred approach for ESR1 mutation testing at progression on endocrine therapy, with digital PCR (dPCR) and next-generation sequencing (NGS) as the preferred analytical platforms. Although elacestrant was approved by the U.S. Food and Drug Administration together with Guardant360® Dx as its companion diagnostic, European regulatory frameworks allow the use of validated in-house assays for ESR1 testing, which are increasingly being implemented across clinical laboratories. To support the clinical implementation of ESR1 testing and improve analytical standardization in routine practice, we performed a European multicentre analytical verification study using dPCR- and NGS-based liquid biopsy workflows. Six referral institutions participated in this study. All laboratories verified dPCR workflows and four also verified NGS-based assays using standardized reference materials containing clinically relevant ESR1 mutations. Limit of detection (LoD) and limit of blank (LoB) were determined in each laboratory according to locally validated workflows following CLSI-based verification procedures. Analytical sensitivity and specificity were assessed across platforms, focusing on the two most frequently tested ESR1 hotspot mutations, p.Y537S and p.D538G. Total DNA input ranged from 10 to 30 ng per reaction for dPCR assays, while NGS input followed platform-specific requirements. LoD values ranged from 0.01
Ovarian cancer is a highly heterogeneous malignancy and remains a major cause of cancer-related mortality, mainly due to late diagnosis and frequent development of therapy resistance. Current diagnostic markers show limited sensitivity for early-stage disease, highlighting the need for novel biomarkers and improved understanding of tumor biology. Epigenetic deregulation is a hallmark of ovarian cancer, characterized by global DNA hypomethylation and focal hypermethylation of tumor suppressor genes. Among hypomethylated genomic regions, long interspersed nuclear elements-1 (LINE-1) represent a large fraction of the human genome and a widely used surrogate marker of global epigenetic instability. LINE-1 elements are normally epigenetically silenced. In ovarian cancer, loss of LINE-1 methylation leads to transcriptional reactivation, expression of ORF1p and ORF2p, and increased retrotransposition activity. These events contribute to genomic instability, DNA damage, insertional mutagenesis, tumor heterogeneity, and clonal evolution. LINE-1 deregulation is also associated with defects in DNA repair pathways, including homologous recombination deficiency, and may influence tumor progression and response to therapy. In addition, aberrant LINE-1 activity has been linked to activation of innate immune and interferon-related signaling pathways, supporting its biological relevance in cancer. Several studies indicate that LINE-1 hypomethylation is associated with aggressive tumor features and adverse clinical outcomes in ovarian cancer. Importantly, repetitive elements, including LINE-1, can be detected in circulating cell-free DNA (cfDNA), providing a technical basis for their evaluation as minimally invasive biomarkers. This review summarizes current knowledge on LINE-1 biology, epigenetic regulation, and functional consequences in ovarian cancer. In addition, we discuss the realistic potential of LINE-1 as a circulating epigenetic biomarker and therapeutic target.
The clinical success of cancer immunotherapy has highlighted the urgent need for biomarkers capable of capturing dynamic and functional anti-tumor immune activity. Granzyme B (GzmB), a serine protease released by activated cytotoxic CD8⁺ T lymphocytes and natural killer cells during target-cell killing, represents a proximal and mechanistically linked marker of immune effector function. Positron emission tomography (PET) tracers targeting GzmB enable non-invasive, whole-body assessment of cytotoxic activity in vivo, potentially overcoming the spatial and temporal limitations of tissue biopsy and conventional imaging. Preclinical investigations using ⁶⁸Ga- and 18F-labeled peptide-based tracers, including [⁶⁸Ga]Ga-NOTA-GZP and [18F]AlF-mNOTA-GZP, consistently demonstrate early discrimination between responders and non-responders to immune checkpoint blockade and combination therapies, often preceding measurable tumor shrinkage or metabolic changes on [18F]FDG PET. Emerging first-in-human studies further support the feasibility, safety, and predictive potential of GzmB PET across multiple tumor types. Despite promising results, several challenges remain, including optimal imaging timing, signal heterogeneity related to non-canonical GzmB sources, and the need for standardized quantitative thresholds and multicenter validation. Overall, GzmB PET represents a biologically grounded and translationally advanced approach for early response assessment, patient stratification, and potential theranostic applications in precision immuno-oncology.
Standard treatment for locally advanced rectal cancer involves neoadjuvant chemoradiation therapy (nCRT) followed by total mesorectal excision, but this approach carries significant morbidity and often results in incomplete resections owing to poor intraoperative tumor visualization. For patients with complete response to nCRT, a watch-and-wait (W W) strategy can spare surgery, but current imaging techniques inadequately identify complete responders, leading to regrowth in 30
