Zhongjun Zhou

INTRODUCTION: Pancreatitis and pancreatic cancer lead to excess mortality in the general population. Excessive intrapancreatic fat deposition (IPFD) is a common driver of diseases of the exocrine pancreas according to the PANDORA hypothesis. However, the relationship of IPFD with mortality has never been investigated. We aimed to explore the association of IPFD with mortality. METHODS: Participants from the UK Biobank were divided into 2 cohorts based on the availability of IPFD quantified using MRI. The Kaplan-Meier survival analysis and multivariable Cox-proportional hazard model were used. Genome-wide association study on IPFD and Mendelian randomization analysis were performed. An IPFD-linked polygenic risk score (PRS) was applied in the MRI-naïve cohort. RESULTS: A total of 55,058 participants were analyzed, 695 (1.26%) of whom died during a median follow-up of 4.9 years. Excessive baseline IPFD was significantly associated with an increased risk of all-cause mortality (hazard ratio = 1.081, P < 0.05) and mortality from vascular diseases (hazard ratio = 1.247, P < 0.001). Genome-wide association study identified 38 significant IPFD-associated single nucleotide polymorphisms. The PRS derived from these single nucleotide polymorphism showed significant associations with all-cause mortality and mortality from vascular diseases. In the MRI-naïve cohort of 354,761 participants, consistent results were obtained using the PRS as a genetic proxy for IPFD. DISCUSSION: Excessive IPFD is associated with elevated risk of future mortality especially from vascular diseases in the general population.

NLRP3-associated autoinflammatory diseases (NLRP3-AIDs) are rare autoinflammatory disorders caused by uncontrolled inflammasome activation. While IL-1β blockade is first-line therapy, many patients respond inadequately, highlighting a need for alternative strategies. Transcriptomic analysis was performed on immune cells from a patient with an NLRP3 L573W mutation. Functional validation of CD177 as a downstream effector of NLRP3 activation was conducted. A novel NLRP3 L573W knock-in mouse model was established. Correlation between CD177 expression, disease severity, neutrophilia, and tissue damage was assessed. Therapeutic efficacy of siRNA-mediated CD177 silencing was evaluated and compared with IL-1β blockade. CD177, a neutrophil-specific protein, was significantly upregulated in NLRP3-mutant cells and confirmed as a direct downstream effector of NLRP3 activation. The NLRP3 L573W knock-in mouse recapitulated human disease heterogeneity, from mild self-limited inflammation to severe multi-organ pathology. CD177 expression correlated with disease severity, neutrophilia, and tissue damage. siRNA-mediated CD177 silencing attenuated systemic inflammation, reduced neutrophil infiltration and cytokine levels (IL-1β, IL-6, TNFα), and ameliorated multi-organ damage, with effects comparable to or exceeding those of IL-1β blockade. CD177 is a non-canonical amplifier of NLRP3-driven inflammation. Targeting CD177 represents a superior therapeutic strategy for NLRP3-AIDs, including IL-1β-refractory cases.

The global surge in the population of people 60 years and older, including that in China, challenges healthcare systems with rising age‐related diseases. To address this demographic change, the Aging Biomarker Consortium (ABC) has launched the X-Age Project to develop a comprehensive aging evaluation system tailored to the Chinese population. Our goal is to identify robust biomarkers and construct composite aging clocks that capture biological age, defined as an individual’s physiological and molecular state, across diverse Chinese cohorts. This Perspective outlines the core objectives, methodological framework and key deliverables of the X-Age Project, including cohort recruitment, standardized sample collection, multimodal data acquisition and clock model development. By integrating interdisciplinary expertise, we aim to provide a practical and scalable platform for understanding aging complexity and heterogeneity, early detection of accelerated aging and evaluation of aging interventions.

Cellular senescence is the major hallmark and therapeutic target of aging and age-related diseases. The role of ALKBH5, one of the main m6A demethylases, in cellular senescence emerges however remains contentious. Herein, we show the reversible ALKBH5 aggregation in cytoplasm promotes cellular senescence. Mechanically, ALKBH5 aggregation causes cytosolic retention, resulting in the m6A dysregulation and m6A hypermethylation of Cdk2, which promotes Cdk2 RNA instability to drive senescence. In addition, m6A imbalance aggravates ALKBH5 cytosolic aggregation in a feedback loop. We further demonstrate that ALKBH5 nuclear translocation required the formation of ALKBH5 droplet phase via binding Nucleoporin p62 (Nup62), while the aggregation of ALKBH5 traps with Nup62 in the cytoplasm. Reduced Nup62 prevents ALKBH5 nuclear entry leading to cellular senescence. Importantly, administration of m6A labeled RNA efficiently reverses ALKBH5 cytosolic aggregates and restores its nuclear entry to alleviate cellular senescence. Forced nuclear entry by NLS-ALKBH5 can prevent senescence in vitro and in vivo. Taken together, these findings unravel a novel paradigm for m6A epigenetic regulation in cellular senescence and offer promising therapeutic targets and strategies for the intervention of aging and age-associated diseases.

Single-cell profiling protein-chromatin interactions is often constrained by complex workflows, high cost, or dependence on specialized equipment. We present indexed tagmentation-based single-cell CUT&Tag-sequencing (IT-scC&T-seq), a modular, plate-based strategy using three-round combinatorial barcoding. IT-scC&T-seq robustly profiles histone modifications and transcription factors with high specificity and throughput, supporting simultaneous analysis of multiple samples and epitopes. Notably, it enables sensitive single-cell mapping of lamina-associated domains, low-abundance chromatin features previously difficult to resolve. Applied to adult mouse mammary gland, the method reveals cell-type-specific chromatin landscapes and lineage-regulatory dynamics. Together, IT-scC&T-seq provides a scalable, cost-effective, and broadly accessible approach for high-resolution chromatin profiling.

Single-cell ATAC-seq (scATAC-seq) enables high-resolution mapping of chromatin accessibility but is often limited by throughput, cost, and equipment requirements. Here, we present indexed Tn5 tagmentation-based scATAC-seq (IT-scATAC-seq), a semi-automated, cost-effective, and scalable approach that leverages indexed Tn5 transposomes and a three-round barcoding strategy. This workflow prepares libraries for up to 10,000 cells in a single day, reduces the per-cell cost to approximately $0.01, and maintains high data quality. Comprehensive benchmarking demonstrates that IT-scATAC-seq achieves robust library complexity, high signal specificity, and improved cost-efficiency compared to existing methods. We apply IT-scATAC-seq to mouse embryonic stem cells, capturing chromatin remodelling during early differentiation, and to human peripheral blood mononuclear cells, resolving cell-type–specific regulatory programs. Here, we show that IT-scATAC-seq provides a robust and efficient approach for high-resolution single-cell epigenomic investigations, balancing scalability, data quality, and accessibility.

Single-cell ATAC-seq (scATAC-seq) allows for detailed mapping of chromatin accessibility but often faces challenges related to throughput, cost, and equipment demands. In this study, we introduce indexed Tn5 tagmentation-based scATAC-seq (IT-scATAC-seq), a semi-automated, economical, and scalable method that utilizes indexed Tn5 transposomes along with a three-round barcoding strategy. This workflow can prepare libraries for up to 10,000 cells in just one day, lowers the cost per cell to around $0.01, and preserves strong data resolution.

ISGylation is the post-translational modification of protein substrates covalently conjugated with mediated by the ubiquitin-like protein, interferon-stimulated gene 15 (ISG15). Although initially linked to antiviral immunity, recent evidence highlights important roles for ISGylation in various biological processes, such as maintaining genomic stability, promoting tumourigenesis, and being involved in other pathological conditions. In this review, we examine the molecular mechanisms underlying ISGylation, its interplay with other post-translational modifications, and its involvement in diverse biological and pathological processes. We propose future research directions to advance the field and discuss how ISGylation might be harnessed to ensure human health, particularly genome instability-associated diseases.