Objective:Multisystem proteinopathy (MSP) is a pleiotropic group of disorders initially presenting as inclusion body myopathy (IBM), amyotrophic lateral sclerosis (ALS), frontotemporal dementia (FTD), and/or Paget disease of bone (PDB). Additional genes including MATR3, OPTN, and ANXA11, have recently been implicated in MSP-like disorders, further expanding the genetic spectrum. This research aims to study the genetic and clinical characteristics of MSP and related disorders in a large Chinese cohort. Methods:Twenty-nine patients were identified in 953 patients diagnosed with ALS, IBM, or dementia at Huashan Hospital between 2000 and 2024. Variants in MSP-related genes were detected using next-generation sequencing and confirmed by Sanger sequencing. Clinical, pathological, imaging, and electromyography data were collected and analyzed. Results:A total of 29 patients (3.0%) were identified as carrying MSP-related gene variants. Most patients were male (72.4%), with disease onset predominantly in the third to fifth decades of life. The majority of patients (21/29) presented with a single clinical phenotype. ALS was the most common phenotype (20/29), followed by IBM (10/29), FTD (7/29), and PDB (1/29). The most frequent variants were in ANXA11 (34.5%) and VCP (20.7%), followed by OPTN (17.2%), SQSTM1 (10.3%), MATR3 (10.3%), and HNRNPA1 (6.9%). All patients with VCP variants presented with initial lower limb involvement, whereas those carrying ANXA11 or OPTN variants predominantly showed upper limb or bulbar onset. Patients harboring OPTN variants had a later age at onset compared with those carrying VCP or MATR3 variants. Patients with ALS-onset exhibited faster progression compared with those with myopathy-onset, even when harboring identical variants. Conclusion:This study broadens the clinical and genetic landscape of MSP and related disorders in a Chinese cohort. These results emphasize the clinical utility of next-generation sequencing for improving diagnostic accuracy in patients with unexplained neuromuscular or cognitive presentations, especially in the presence of multisystem involvement.

Background Repetitive head impacts in former contact sport athletes are associated with cognitive impairment, accelerated cerebral atrophy and risk of neurodegenerative disease. Epigenetic clocks derived from age-associated DNA methylation (DNAm) profiles may capture accelerated biological ageing in neurodegenerative conditions; however, their application in former athletes remains unexplored. Here we aim to explore the application of epigenetic clocks as a measure of accelerated biological ageing in a cohort of former athletes. Methods In 126 former athletes (96% male; mean age: 54.5±14.4 years; mean concussions: 6.8±6.7), we examined associations of brain volumes, plasma neurofilament light levels and cognitive/behavioural scores with DNAmAge-acceleration, AgeAccelResidual and DNAmFitAge-acceleration. Results We only found an association between the number of concussions and DNAmFitAge-acceleration (p=0.003, B=0.46, R²=0.063), indicating that every two additional concussions were associated with a 5-year increase in DNAmFitAge-acceleration. There was also a trend towards an association between years of play and DNAmAge-acceleration in older athletes. Conclusions These preliminary findings suggest that specific epigenetic clock measures may serve as early markers of biological ageing related to repetitive head impacts.

Amyotrophic lateral sclerosis (ALS) remains a major therapeutic challenge, with immune dysregulation increasingly recognized as a critical driver of disease progression. Despite extensive mechanistic research, no immunotherapeutic approach has achieved consistent disease-modifying effects, raising questions about whether this translational gap reflects biological complexity or structural misalignment within the research ecosystem. To characterize the intellectual evolution of ALS immunotherapeutics research, identify immune targets with translational potential, and evaluate collaboration patterns that may influence translational efficiency, we performed a bibliometric analysis of 2,256 publications indexed in Web of Science and Scopus using network-based approaches including co-citation clustering, keyword co-occurrence, and citation burst detection implemented in CiteSpace, VOSviewer, and R-Bibliometrix. Publication output increased 8.4-fold over the study period, delineating three developmental phases. Thematic analyses revealed a shift from early emphasis on microglial biology and SOD1-based models toward recent focus areas including the gut-brain axis, C9orf72-associated immune dysregulation, and advanced immunomodulatory strategies. Collaboration networks remain predominantly regional despite strong contributions from the United States, Europe, and Asia, with limited integration between mechanistic research groups and clinical trial consortia. Among immune-directed therapeutic strategies, regulatory T cell modulation and microglial-targeted approaches exhibit the highest translational readiness. These findings suggest that the lack of effective ALS immunotherapeutics reflects not only biological complexity but also structural and strategic misalignment within the research ecosystem. This bibliometric analysis provides a systems-level framework to guide more integrated translational strategies in ALS immunotherapeutics development.

Spatial architecture of cell types and gene expression is the foundation of cell-cell interactions, biological function and disease pathology, but is not well investigated in human cortex, which are highly related to neurodegenerative diseases, such as Alzheimer's disease (AD) and Amyotrophic Lateral Sclerosis (ALS). Recent studies indicated single nucleus transcriptomic features of excitatory neuron vulnerability in AD entorhinal cortex, and motor neuron vulnerability in ALS motor cortex. However, it remains unclear what is the brain regional vulnerability of AD or ALS associated genes. We developed an entropy-weighted differential gene expression matrix-based tool (SpatialE) to identify the spatial enrichment of gene sets in spatial transcriptomics (ST). We benchmarked SpatialE against another enrichment tool (Multimodal Intersection Analysis, MIA) using ST data from human and mouse brain tissues. To investigate regional vulnerability, we analyzed three human motor cortex and two dorsolateral prefrontal cortex (DLPFC) tissues for spatial enrichment analyses of AD and ALS associated genes. We also used Cell2location to estimate the abundance of cell types in AD and ALS related cortex layers. SpatialE showed more accurate and specific spatial enrichment of regional cell type markers than MIA in both mouse brain and human DLPFC. Spatial transcriptomic analyses of human motor cortex and dorsolateral prefrontal cortex showed heterogenous cell types and spatial gene expression profiles. We found that the expression of AD associated genes ( n  = 139, n  = 162) that were indicated from two AD genome-wide association studies are significantly enriched in layer 1 (L1) motor cortex (P_bonferroni<0.01 for 3 ST data), but not in DLPFC. Cell type deconvolution analysis identified abundant expression of astrocytes in human L1 motor cortex. Additionally, the expression of 260 manually curated ALS-associated genes are significantly enriched in layer 5 (L5) motor cortex and DLPFC (P_bonferroni<0.03 for 5 ST data). Cell type deconvolution analysis revealed an abundant expression of upper motor neurons and L5 excitatory neurons in human L5 motor cortex. We developed a novel computational tool (SpatialE) to characterize spatial enrichment expression of a gene set in spatial transcriptomics data. Spatial enrichment analyses identified regional brain vulnerability of AD and ALS associated genes in human brain.

Amyotrophic lateral sclerosis (ALS) is a severe motor neuron disease, with most sporadic cases lacking clear genetic causes. Abnormal pre-mRNA splicing is a fundamental mechanism in neurodegenerative diseases. For example, TAR DNA-binding protein 43 (TDP-43) loss of function causes widespread RNA mis-splicing events in ALS. Additionally, splicing mutations are major contributors to neurological disorders. However, the role of intronic variants driving RNA mis-splicing in ALS remains poorly understood.To address this, we developed Spliformer to predict RNA splicing. Spliformer is a transformer-based deep learning model trained and tested on splicing events from the GENCODE database, in addition to RNA-sequencing data from blood and CNS tissues. We benchmarked Spliformer against SpliceAI and Pangolin using testing datasets and paired whole-genome sequencing with RNA-sequencing data. We also developed the Spliformer-motif model to identify splicing regulatory motifs. We analysed the Clinvar dataset to identify the link of splicing variants with disease pathogenicity. Additionally, we analysed whole-genome sequencing data of ALS patients and controls to identify common intronic splicing variants linked to ALS risk or disease phenotypes. We also profiled rare intronic splicing variants in ALS patients to identify known or novel ALS-associated genes. Minigene assays were used to validate candidate splicing variants. Finally, we measured spine density in neurons with a specific gene knockdown or those expressing a TDP-43 disease-causing mutant.Spliformer accurately predicts the possibilities of a nucleotide within a pre-mRNA sequence being a splice donor, acceptor or neither. Spliformer outperformed SpliceAI and Pangolin in both speed and accuracy in tested splicing events and/or paired whole-genome sequencing/RNA-sequencing data. Spliformer-motif successfully identified canonical and novel splicing regulatory motifs. In the Clinvar dataset, splicing variants are highly related to disease pathogenicity. Genome-wide analyses of common intronic splicing variants nominated one variant linked to ALS progression. Deep learning analyses of whole-genome sequencing data from 1370 ALS patients revealed rare splicing variants in reported ALS genes (such as PTPRN2 and CFAP410, validated through minigene assays and RNA sequencing) and TDP-43 loss-of-function-related RNA mis-splicing genes (such as PTPRD). Further genetic analysis and minigene assays nominated PCP4 and TMEM63A as ALS-associated genes. Functional assays demonstrated that PCP4 is crucial for maintaining spine density and can rescue spine loss in neurons expressing a disease-causing TDP-43 mutant. In summary, we developed Spliformer and Spliformer-motif, which accurately predict and interpret pre-mRNA splicing. Our findings highlight an intronic genetic mechanism driving RNA mis-splicing in ALS and nominate PCP4 as an ALS-associated gene. Tang et al. present a new deep learning tool, Spliformer, that accurately predicts RNA splicing. Genome-wide analyses using this tool identified intronic variants driving RNA mis-splicing in amyotrophic lateral sclerosis, while additional genetic and functional analyses pinpointed PCP4 as an ALS-associated gene.

The long-term consequences of repetitive head impacts in contact sports include cognitive deficits, accelerated brain atrophy, and neurodegenerative diseases. Currently, no studies of former athletes have addressed the connection between brain aging and biological aging, which can be assessed using age-related DNA methylation (DNAm) profiles. The most studied epigenetic clock is DNAm-age, which is providing consistent results across tissues (e.g., blood and brain). It includes several measures like DNAm-age acceleration (DNAmAA), the difference between DNAm-age and chronological age, and DNAmAA-residual (DNAmAAr), independent of chronological age. In a cohort of retired athletes, we explored the link between measures of brain age and epigenetic age, including the recently developed DNAmFit-age reported to be younger in physically fit individuals. We investigated 126 former contact sports athletes (mean age: 54.5±14.4; 96% male; mean concussion number: 6.8±6.7). Longitudinal assessments were available for 21 athletes (2–3 time points over 1–10 years). Bisulfite-converted blood DNA was analyzed using the Infinium MethylationEPIC chip, and the DNAm data were submitted to the Horvath calculator ( https://dnamage.clockfoundation.org/ ) to obtain DNAmFit-age, DNAm-age, and DNAmAAr. T1-weighted MRIs were processed using the CAT12-Toolbox. Regional gray matter volume was examined in relation to DNAmAA, DNAmAAr and DNAmFit-age acceleration (DNAmFitAA). Consistent with previous findings, chronological age was significantly associated with brain volumes and cortical thickness. DNAmAA, DNAmAAr, and DNAmFitAA remained stable over a period of up to 10 years, and none of these measures were associated with brain volumes or cortical thickness. Notably, multivariate linear regression analysis revealed a significant positive association between DNAmFitAA and the number of concussions ( p  = 0.0027, B=0.46, R 2 =0.063), indicating that every additional two concussions correspond to a 5-year increase in DNAmFitAA. In former athletes, we confirmed that chronological age is associated with cerebral atrophy and identified an association between increased DNAmFitAA and a higher number of concussions. This finding requires validation in independent studies, along with an assessment of the relationship between DNAmFitAA and neurodegeneration. Our analysis did not reveal a link of the examined epigenetic clocks with brain volumes and cortical thickness. Exploration of other epigenetic clocks may provide new insights into the mechanisms underlying brain aging.

The mislocalization of RNA-binding proteins (RBPs) from nucleus to cytoplasm and the formation of aggregates are hallmarks of neurodegeneration. Amyotrophic lateral sclerosis (ALS) disease-causing mutations in the fused in sarcoma (FUS) gene, encoding an RNA-binding protein, cluster at the C-terminal proline/tyrosine-nuclear localization signal (PY-NLS) domain, which is crucial for mediating nucleus-cytoplasm translocation by binding to Transportin-1. However, the mechanisms underlying heterogeneous protein mislocalization and age at onset (AAO) of ALS cases carrying FUS PY-NLS mutations remain unclear. Here, we screened FUS mutations in 416 ALS patients, and identified 12 patients carrying four FUS mutations at the p.R521 locus of PY-NLS domain (p.R521P, p.R521C, p.R521G, p.R521H), exhibiting highly variable AAO (20–56 years). AlphaFold-2 predicted protein structures classified FUS p.R521 mutants into alpha-helix containing (p.R521C, p.R521H) and alpha-helix disrupted (p.R521P, p.R521G) subgroups. Isothermal titration calorimetry experiment showed that the FUS alpha-helix disrupted subgroup had a reduced binding affinity with transportin-1, which is essential for mediating the nucleus-cytoplasm translocation. Furthermore, immunofluorescence in HEK-293 T and SH-SY5Y cells revealed more protein mislocalization in the FUS alpha-helix disrupted subgroup compared to the alpha-helix containing subgroup. FUS mislocalization status is also significantly associated with ALS AAO. Finally, the alpha-helix structure based FUS-ALS subgroups exhibited significantly different AAO (P = 0.036) in our cohort, but not in a Chinese cohort including published dataset. In summary, we showed highly diverse phenotypes in ALS patients with FUS R521 mutants, and implicated a link between genetic mutation related C-terminal structure with the status of FUS protein mislocalization.

Background Amyotrophic lateral sclerosis (ALS) is a severe motor neuron disease, with highly diverse survival time. However, genetic and epigenetic factors influencing ALS survival across diverse populations remain unclear. Methods We performed whole-genome sequencing (WGS) and DNA methylome array in blood DNA of patients with ALS. For survival analysis, we used Cox proportional hazards model for genetic variants, DNA methylation (DNAm) of CpG sites or CpG-SNPs in Chinese and Canadian cohorts, followed by meta-analysis. We performed pathway enrichment analysis for candidate genes inferred from DNAm events associated with survival. In paired genome and methylome data, we analysed the effect of the candidate CpG-SNP genotypes on DNAm status. Findings Genome-wide cross-population meta-analysis of common variants in 511 patients with ALS showed a suggestive association of CAV1/CAV2 rs117002347 genotypes with survival. Epigenome-wide cross-population meta-analysis in 459 patients revealed that ALS survival was significantly linked to DNAm of 88 CpGs on 40 genes, and highlighted the AMPK and cytoskeleton pathways. Epigenome-wide cross-population meta-analysis of CpG-SNPs in 459 patients identified 8 loci on 4 genes, including BAG6 (cg27014438/rs28732154), which was further validated in another 204 patients with ALS. Moreover, analysis of paired genome/epigenome data (n = 454) indicated that BAG6 rs28732154 genotypes may modulate cg27014438 methylation, which is also a cis-eQTM of BAG6 expression in blood. Interpretation Our study identified BAG6 cg27014438 methylation as a potential epigenetic modifier of ALS survival. BAG6 cg27014438 methylation is modulated by rs28732154 genotypes, and linked to BAG6 expression. Our findings extended our understanding of epigenetic modifiers in ALS survival. Funding This work was supported by the National Natural Science Foundation of China (82071430, 82371878) (MZ), Shanghai Municipal Natural Science Foundation General Program (22ZR1466400) (MZ), the Fundamental Research Funds for the Central Universities (MZ), the G. Harry Sheppard Memorial Research Fund, and Canadian Consortium on Neurodegeneration in Aging (ER).

MEF2D fusions are found in a special subtype of B-cell precursor acute lymphoblastic leukemia (BCP-ALL) with poor prognosis. In this study, we conducted high-throughput drug screenings using cell line and ex vivo cell model harboring, respectively, MEF2D::HNRNPUL1(MH) and MEF2D::BCL9(MB), the two major MEF2D fusions. We identified CUDC-907 as a highly potent dual-target inhibitor of PI3K/HDAC, demonstrating remarkable efficacy in inducing robust lethality while maintaining selectivity for MEF2D fusion-expressing cells. CUDC-907 effectively induced apoptosis and promoted the down-regulation of pre-BCR signaling. We discovered that the hyperactivation of the PI3K-AKT signaling pathway, HDAC9, and BCL2 contributed to the sustained state of MEF2D fusion (+) BCP-ALL. Importantly, CUDC-907 exerted dual regulatory function by targeting the integrative pathways of MEF2D fusions. It suppressed the PI3K-CREB pathway and fusion gene expression, while simultaneously inhibited transcriptional activity regulated by the MEF2D fusion-HDAC axis. CUDC-907 demonstrated remarkable efficacy in patient samples carrying distinct MEF2D fusion variants in vitro. Furthermore, this compound’s effectiveness and safety were confirmed in both MH/NRASG12D BCP-ALL mouse model and MB patient-derived xenograft (PDX) model, outperforming conventional therapies. These results support the therapeutic potential of dual-pathway inhibition in MEF2D fusion (+) BCP-ALL and suggest CUDC-907 as a promising candidate for precision treatment in fusion-driven leukemias with similar molecular dependencies.

Precise regulation of pre-mRNA splicing underpins molecular diversity and is linked to aging and disease. Genetic variants are key drivers of RNA mis-splicing, yet how they induce tissue-specific splicing remains largely unclear. Here, we introduce Spliformer-v2, a deep learning model based on SegmentNT architecture to predict multi-tissue RNA splicing. Spliformer-v2 is trained on paired genome/transcriptome data from 18 human tissues, including 12 central nervous system and 6 peripheral tissues. Spliformer-v2 accurately predicts the effects of heterozygous and homozygous variants on splicing, outperforming existing models (such as SpliceTransformer) across tissues for both regression evaluations (R2: 0.83-0.89 vs 0.10-0.41) and classification evaluations (AUPRC: 0.90-0.95 vs 0.45-0.61). Analysis of ClinVar dataset indicated the link of tissues-specific splicing variants with disease pathogenicity. Analysis of genome-wide association study hits from neurological diseases (such as Amyotrophic Lateral Sclerosis, Alzheimer’s disease and Parkinson’s disease) identified tissue-specific splicing variants in genes like CREB3, SCFD1, MAPT and TOMM40 , suggesting potential splicing mechanisms in disease. In summary, Spliformer-v2 is a powerful tool for predicting RNA splicing across the most comprehensive set of human brain and spinal cord tissues to date. This advancement enhances our understanding of tissue-specific splicing variants and their roles in neurological and other complex diseases, thereby advancing the discovery of novel drug targets and biomarkers in precision medicine. ### Competing Interest Statement The authors have declared no competing interest. National Natural Science Foundation of China, 82371878 Fundamental Research Funds for the Central Universities

Amyotrophic lateral sclerosis (ALS) is a severe motor neuron disease with uncertain genetic predisposition in most sporadic cases. The spatial architecture of cell types and gene expression are the basis of cell-cell interactions, biological function and disease pathology, but are not well investigated in the human motor cortex, a key ALS-relevant brain region. Recent studies indicated single nucleus transcriptomic features of motor neuron vulnerability in ALS motor cortex. However, the brain regional vulnerability of ALS-associated genes and the genetic link between region-specific genes and ALS risk remain largely unclear.Here, we developed an entropy-weighted differential gene expression matrix-based tool (SpatialE) to identify the spatial enrichment of gene sets in spatial transcriptomics. We benchmarked SpatialE against another enrichment tool (multimodal intersection analysis) using spatial transcriptomics data from both human and mouse brain tissues. To investigate regional vulnerability, we analysed three human motor cortex and two dorsolateral prefrontal cortex tissues for spatial enrichment of ALS-associated genes. We also used Cell2location to estimate the abundance of cell types in ALS-related cortex layers. To dissect the link of regionally expressed genes and ALS risk, we performed burden analyses of rare loss-of-function variants detected by whole-genome sequencing in ALS patients and controls, then analysed differential gene expression in the TargetALS RNA-sequencing dataset.SpatialE showed more accurate and specific spatial enrichment of regional cell type markers than multimodal intersection analysis in both mouse brain and human dorsolateral prefrontal cortex. Spatial transcriptomic analyses of human motor cortex showed heterogeneous cell types and spatial gene expression profiles. We found that 260 manually curated ALS-associated genes are significantly enriched in layer 5 of the motor cortex, with abundant expression of upper motor neurons and layer 5 excitatory neurons. Burden analyses of rare loss-of-function variants in Layer 5-associated genes nominated NOMO1 as a novel ALS-associated gene in a combined sample set of 6814 ALS patients and 3324 controls (P = 0.029). Gene expression analyses in CNS tissues revealed downregulation of NOMO1 in ALS, which is consistent with a loss-of-function disease mechanism. In conclusion, our integrated spatial transcriptomics and genomic analyses identified regional brain vulnerability in ALS and the association of a layer 5 gene (NOMO1) with ALS risk. Using spatial transcriptomics and single-nucleus RNA sequencing, Guo et al. show that ALS-associated gene expression is enriched in layer 5 of the motor cortex. Genomic analyses revealed a novel association between a layer 5 gene called NOMO1 and ALS risk. Understanding region-specific vulnerability in ALS will aid precision medicine approaches.

The hyper-unstable Chr9p21 locus, harbouring the interferon gene cluster, oncogenes and C9orf72, is linked to multiple diseases. C9orf72 (GGGGCC)n expansions (C9orf72Exp) are associated with incompletely penetrant amyotrophic lateral sclerosis, frontotemporal dementia and autoimmune disorders. C9orf72Exp patients display hyperactive cGAS-STING-linked interferon immune and DNA damage responses, but the source of immunostimulatory or damaged DNA is unknown. Here, we show C9orf72Exp in pre-symptomatic and amyotrophic lateral sclerosis-frontotemporal dementia patient cells and brains cause the folate-sensitive chromosomal fragile site, FRA9A. FRA9A centers on >33 kb of C9orf72 as highly compacted chromatin embedded in an 8.2 Mb fragility zone spanning 9p21, encompassing 46 genes, making FRA9A one of the largest fragile sites. C9orf72Exp cells show chromosomal instability, heightened global- and Chr9p-enriched sister-chromatid exchanges, truncated-Chr9s, acentric-Chr9s and Chr9-containing micronuclei, providing endogenous sources of damaged and immunostimulatory DNA. Cells from one C9orf72Exp patient contained a highly rearranged FRA9A-expressing Chr9 with Chr9-wide dysregulated gene expression. Somatic C9orf72Exp repeat instability and chromosomal fragility are sensitive to folate deficiency. Age-dependent repeat instability, chromosomal fragility and chromosomal instability can be transferred to CNS and peripheral tissues of transgenic C9orf72Exp mice, implicating C9orf72Exp as the source. Our results highlight unappreciated effects of C9orf72 expansions that trigger vitamin-sensitive chromosome fragility, adding structural variations to the disease-enriched 9p21 locus, and likely elsewhere.

Objective: Genetic mutations of fused in sarcoma (FUS) causing amyotrophic lateral sclerosis (ALS) may disrupt mRNA splicing events. For example, the FUS c.1394-2delA variant was reported in two western ALS patients, but its molecular mechanism is unclear. In this study, we aim to investigate FUS splice site mutations in Chinese ALS patients. Methods: Sanger sequencing was used to identify FUS splicing mutations in Chinese ALS patients. We combined a deep learning tool (SpliceAI), RNA sequencing, and RT-PCR/RT-qPCR to analyze the effect of FUS c.1394-2delA mutation on RNA splicing and expression. AlphaFold was used to predict the protein structure of mutant FUS. In transfected cell lines, we used immunofluorescence to assess cytoplasmicmislocalization of mutant FUS protein. Results: We identified a de novo FUS splice acceptor site mutation (c.1394-2delA, p. Gly466Valfs*14) in one Chinese sporadic ALS patient, which is linked to exon 14 skipping, and upregulated total FUS mRNA expression. The FUS splice site mutation was predicted to be translated into a truncated protein product at Cterminal. In vitro studies revealed that the FUS mutation increased cytoplasmic mislocalization in both HEK293T and SH-SY5Y cells. Conclusions: We identified a de novo FUS splicing mutation (c.1394-2delA, p. Gly466Valfs*14) in 1 out of 233 Chinese ALS patients. It caused abnormal RNA splicing, upregulated gene expression, truncated FUS translation, and cytosolic mislocalization. Our findings suggested that FUS splice site mutation is rare in Chinese ALS patients and extended our knowledge of molecular mechanisms of the FUS c.1394-2delA mutation.

Background: Genetic and epigenetic modifiers of age at onset of Parkinson’s disease (PD) are largely unknown. It remains unclear whether DNA methylation (DNAm) age acceleration is linked to age at onset in PD patients of different ethnicities with a similar genetic background. We aim to characterize the clinical, genomic and epigenomic features of three pairs of Chinese monozygotic twins discordant for PD onset by up to 10 years. Methods: We conducted whole genome sequencing, multiplex ligation-dependent probe amplification and genome-wide DNAm array to evaluate the three pairs of Chinese monozygotic twins discordant for age at onset of PD (families A–C). Results: We identified two heterozygous PRKN mutations (exon 2–4 deletion and p.Met1Thr) in PD affected members of one family. Somatic mutation analyses of investigated families did not reveal any variants that could explain the phenotypic discordance in the twin pairs. Of note, our epigenetic study revealed that the twins with earlier-onset had a trend of faster DNAm age acceleration than the later-onset/asymptomatic twins, but without statistical significance. Conclusion: The link between DNAm age acceleration and PD onset in Chinese patients should be interpreted with cautious, and need to be further verified in an extended PD cohort with similar genetic background.

[目的]分析男科门诊早泄(PE)患者的临床特征及影响因素.[方法]收集2020年1月至2022年5月医院男科门诊20～60岁男性患者,详细记录一般人口学资料、病史、性生活史等,对符合要求的患者行早泄诊断量表(PEDT)、自我评估阴道内射精潜伏期(IELT)、国际勃起功能问卷-5(IIEF-5)、勃起硬度评分(EHS)、广泛性焦虑障碍量表(GAD-7)和健康问卷抑郁量表等量表评估.采用多元线性回归及Logistic回归分析PE的相关因素及独立危险因子.[结果]共纳入男科门诊患者973例,其中PE组445例(31.12±6.72)岁,非PE组528例(32.78±7.95)岁.两组间年龄、PEDT、IELT、IIEF-5、EHS、GAD-7及PHQ-9的差异均存在统计学意义(P<0.05).回归分析校正后,PEDT与年龄(b=-0.11,P=0.001)、IIEF-5(b=-0.17,P<0.001)及PHQ-9(b=0.19,P<0.001)存在显著相关性.多因素校正后,年龄≤30岁、勃起功能障碍及抑郁状态三个变量有统计学意义(P<0.01),其OR(95％CI)值分别为1.63(1.23,2.16)、2.05(1.45,2.92)、1.90(1.37,2.65).[结论]PE与年龄、勃起功能评分、焦虑及抑郁评分等因素密切相关.年龄≤30岁、勃起功能障碍及抑郁状态是PE的独立危险因子.

Parkinson’s disease is a clinically and genetically heterogeneous movement disorder with highly variable age-at-onset. DNA methylation (DNAm) age is an epigenetic clock that could reflect biological aging. Studies of DNAm-age acceleration (difference between DNAm-age and chronological age) are pertinent to neurodegenerative diseases (e.g., Parkinson’s disease), for which aging is the strongest risk-factor. We assessed DNAm-age in idiopathic Parkinson’s disease (n=96) and a longitudinal LRRK2 cohort at four time-points over a 3-year period (n=220), including manifesting (n=91) and non-manifesting (n=129) G2019S-carriers. A highly variable age-at-onset was observed in both the idiopathic cohort (26-77 years) and manifesting G2019S-carriers (39-79 years). Increased DNAm-age acceleration was significantly associated with younger onset in idiopathic and LRRK2 -related Parkinson’s disease, suggesting that every 5-year increase in DNAm-age acceleration is linked to about 6-year earlier onset. At an individual level, DNAm-age acceleration remained steady over a 3-year period for most G2019S-carriers, indicating that it might serve as a stable biomarker of biological aging. Future studies should evaluate the stability of DNAm-age acceleration over longer time-periods, especially for phenoconverters from non-manifesting to manifesting subjects. In conclusion, DNAm-age acceleration is linked to disease onset, and could be used in disease-modifying clinical trials of prodromal Parkinson’s disease.

Objective:To explore the correlation between post-transplant non-HLA antibodies and humoral rejection(HR)after kidney transplantation(KT).Methods:A retrospective study was conducted for KT recipients with non-HLA antibody level detected from September 2019 to January 2021.The recipients with biopsy confirmed HR and donor-specific HLA antibodies negative or feeble positive at the time of HR were designated as HR group while recipients with stable renal allograft function from 2 weeks post-KT to the time of detecting non-HLA antibody as stable group.The levels of HLA antibody, MHC classⅠchain-related gene A(MICA)antibody and 32 non-HLA antibodies were tested by Luminex single antigen bead and the levels of angiotensin Ⅱ type 1 receptor(AT1R)antibody quantified by enzyme-linked immunosorbent assay (ELISA). Inter-group differences in positive rate of non-HLA antibodies and number of positive non-HLA antibodies were analyzed.Results:Twenty-four recipients had positive non-HLA antibodies while the remainders had no positive non-HLA antibodies.Three HR recipients were positive for actin antibody, collagen Ⅲ antibody, glutathione S-transferase theta-1 antibody or IFN-γ antibody respectively.However, all four non-HLA antibodies of stable recipients were negative.There was significant inter-group difference( P=0.017). Four HR recipients were positive for collagenⅡantibody while only 1 stable recipient was positive for collagenⅡantibody.The positive rate of collagenⅡ antibody was significantly higher in HR recipients than that in stable recipients( P=0.023). HR recipients had an average of 2.36 positive non-HLA antibodies while stable recipients had an average of 0.90.There was significant inter-group difference ( P=0.008). Conclusions:A high level of non-HLA antibodies may elevate the risk of HR after KT.

Spinocerebellar ataxia type 3 (SCA3), also known as Machado Joseph disease (MJD), is a common dominantly inherited ataxia, and has heterogeneous clinical features and variable age of onset, ranging from 10 to 78 years. Repeats variability of ATXN3, HTT, ATN1 and ATXN2 can explain partially but not fully SCA3 age of onset heterogeneity. Aging is a reported modifier of SCA3 severity and closely linked to DNA methylation (DNAm). DNAm age acceleration was associated with disease risk and/or variable disease phenotypes in several repeat associated neurodegenerative diseases (such as Huntington's disease and Amyotrophic lateral sclerosis). To understand if DNAm age acceleration is associated with SCA3 age of onset, we performed a genome-wide DNAm study of a Chinese SCA3 family with variable age of onset and clinical presentations. All patients showed unsteady gait, deterioration of extremities coordination, speech (dysarthria) and swallowing problems (dysphagia, choking on eating and/or drinking) and oculomotor abnormalities, with variable age of onset ranging from 27 to 52 years. We found that DNAm age acceleration is associated with age of onset (p-value = 0.0023, B = -1.26), suggesting that every 5 year increase in DNAm-age acceleration is corresponding to a 6.3 year earlier disease onset. This association remains significant after the adjustment to ATXN3 CAG repeats (adjusted p-value = 0.037, adjusted B = -1.0). In an independent SCA3 cohort (n = 40), we also observed the association between DNAm age acceleration and age of onset (adjusted p-value = 0.007, adjusted B = -0.69). Of note, we found no significant association between DNAm of single-CpG locus and/or CpG-SNPs and SCA3 age of onset in the current family or the SCA3 cohort. Our findings suggested that DNAm age acceleration might be a SCA3 age of onset modifier, and encourage further investigations in extended SCA3 cohorts to clarify the role of epigenetic aging in modifying disease onset.

目的 探索供体来源的细胞游离DNA(dd-cfDNA)在肾移植术后移植肾缺血再灌注损伤中的应用价值.方法 前瞻性的纳入2020年3月至5月于上海交通大学医学院附属仁济医院肾移植中心接受同种异体肾移植术的40例受试者,在术后第1、3、7天采集患者外周血,利用二代测序的方法对患者外周血中的dd-cfDNA％进行检测,同时检测患者相应时间点的肌酐.结果 本研究所有患者肾移植术后肾功能恢复顺利,无移植物功能延迟恢复及排斥反应的发生.整体分析发现:肌酐测量值、dd-cfDNA％在3个时间点的整体比较,差异有高度统计学意义(P<0.01).进一步两两比较,组内任意2个时间点的肌酐测量值、dd-cfDNA％浓度比较,差异均有高度统计学意义(P<0.01).术后第3天的肌酐测量值比术后第1天降低245.65μmol/L(95％置信区间:172.70～318.60μmol/L);术后第7天的肌酐测量值较术后第3天进一步降低195.95μmol/L(95％置信区间:153.02～238.88μmol/L).术后第3天的dd-cfDNA％浓度比术后第1天降低8.97％(95％置信区间:7.71％～10.24％);术后第7天的dd-cfDNA％浓度较术后第3天进一步降低1.63％(95％置信区间:1.32％～1.94％).结论 dd-cfDNA在肾移植术后移植肾缺血再灌注的不同时间段有差异性表达,对移植肾缺血再灌注损伤程度的评估具有一定价值.

目的 建立大鼠神经损伤勃起功能障碍(CNI-ED)动物模型,通过在阴茎海绵体注射脂肪来源间充质干细胞(ASCs),探究ASCs对双侧海绵体神经损伤后勃起功能障碍的治疗效果和作用机制.方法 实验动物分为3组,每组8只,分别为年龄匹配组(AMC组,假手术治疗),对照组(双侧海绵体损伤+海绵体注射PBS,PBS组),实验组(双侧海绵体损伤+海绵体注射ASCs,ASCs组).手术后4周和12周,检测阴茎海绵体测压(ICP/MAP比值)和组织学结果.结果 手术4周后,对照组大鼠的ICP/MAP比值明显低于AMC组(p＜ 0.001),而实验组的ICP/MAP比值较对照组有显著提高(p＜ 0.05);12周后,对照组和实验组大鼠ICP/MAP比值仍低于AMC组(p＜ 0.001),但实验组显著高于对照组(p＜ 0.05),提示ASCs治疗能够促进大鼠的勃起功能恢复.Masson染色结果表明,术后4周和12周对照组平滑肌/胶原纤维比值较AMC组明显降低(p＜ 0.001),实验组平滑肌/胶原纤维比值高于对照组(p＜ 0.05).免疫荧光结果显示,术后4周和12周AMC组大鼠nNOS数量显著高于对照组和实验组(p＜ 0.001),而实验组显著高于对照组(p＜0.05).12周后实验组的nNOS数量比4周有显著提高(p＜ 0.05).结论 实验组在功能学(ICP/MAP比值)和形态学(平滑肌胶原纤维比值)及海绵体nNOS恢复上均优于对照组,证实海绵体注射ASCs能够促进CNI-ED大鼠勃起功能恢复,主要机制是促进海绵体神经再生和抗海绵体纤维化.