参考文献

TAR DNA-binding protein 43 (TDP-43) dysfunction is a hallmark of several neurodegenerative diseases,including frontotemporal dementia,amyotrophic lateral sclerosis,and Alzheimer’s disease. Although cryptic exon inclusion is a well-characterized consequence of TDP-43 loss of function,emerging evidence reveals broader roles in RNA metabolism,notably in the regulation of alternative polyadenylation (APA) of disease-relevant transcripts. In the present study,we examined 3′ untranslated region lengthening events in the brains of individuals with frontotemporal lobar degeneration with TDP-43 pathology (FTLD-TDP),focusing on the functional impact of APA dysregulation. To investigate whether TDP-43-mediated APA events occur in the postmortem brain,we measured the 3′ untranslated region length of the retromer component vacuolar protein sorting 35 (VPS35) and the ETS transcription factor (ELK1) in the frontal cortex of a large cohort of FTLD-TDP patients and of healthy controls,and evaluated if these APA events are associated with FTLD-TDP clinical characteristic,markers of TDP-43 pathology [e.g.,hyperphosphorylated TDP-43 and cryptic stathmin-2 RNA],or the expression of VPS35 and VPS29 proteins,the latter being essential to the retromer complex. We identified robust 3′ untranslated region lengthening of VPS35 and ELK1 in FTLD-TDP,which strongly associated with markers of TDP-43 pathology,and ELK1 APA also associated with an earlier age of disease onset. Functionally,VPS35 APA was associated with reduced VPS35 and VPS29 protein expression,and lower VPS35 levels were associated with increased hyperphosphorylated TDP-43 and cryptic stathmin-2 RNA. Together,these data implicate APA dysregulation as a critical downstream consequence of TDP-43 dysfunction and suggest that TDP-43 loss may contribute to retromer impairment through APA-mediated repression of retromer subunits. Recent work has shown that TDP-43 loss in frontotemporal dementia (FTD) induces changes in alternative polyadenylation,but the functional consequences of this are unclear. This study reports that 3′UTR lengthening of VPS35 in FTD patient brain samples correlates with reduced VPS35 and VPS29 protein levels,suggesting that TDP-43 loss induces retromer dysfunction. View Publication

Aberrant deposition of β‐amyloid (Aβ) and hyperphosphorylated tau,along with neuroinflammation,are key drivers of Alzheimer's disease (AD) pathology. Here,we identify ramalin,a natural antioxidant,as a promising therapeutic agent that alleviates AD pathology by modulating β‐site APP cleaving enzyme 1 (BACE1),histone deacetylase 6 (HDAC6),and the mitogen‐activated protein kinases (MAPK) pathway. Ramalin reduced BACE1 protein levels,independently of its transcription,translation,or enzymatic activity,an effect mediated by inhibition of HDAC6. Consistently,HDAC6 knockout similarly decreased BACE1 levels,highlighting HDAC6 as a key regulator of BACE1. Ramalin further suppressed neuroinflammatory responses by downregulating inducible nitric oxide synthase (iNOS) and the NLR family pyrin domain containing 3 (NLRP3) inflammasome. In AD mouse models,ramalin treatment significantly attenuated neuroinflammation,Aβ plaque burden,and tau hyperphosphorylation,while improving cognitive performance. Notably,ramalin reversed Aβ oligomer‐induced synaptic transmission impairment and restored synaptic vesicle recycling in hippocampal neurons. Transcriptomic analysis identified modulation of the MAPK pathway,with reduced phosphorylation of c‐Jun N‐terminal kinase (JNK) and extracellular signal‐regulated kinase (ERK) implicated in tau pathology. These findings establish ramalin as a disease‐modifying intervention that provides neuroprotection through concurrent regulation of BACE1,HDAC6,and MAPK signaling pathway. Collectively,our findings highlight ramalin as a compelling disease‐modifying candidate with the potential to drive a breakthrough approach targeting AD pathology. Ramalin alleviates Alzheimer's disease pathology by selectively inhibiting HDAC6,reducing BACE1 levels,and suppressing neuroinflammation through downregulation of the NLRP3 inflammasome and iNOS. It restores synaptic function impaired by Aβ toxicity and improves cognitive performance in AD mouse models,APP/PS1 and 3xTg‐AD. Additionally,ramalin modulates the MAPK signaling pathway,reducing tau phosphorylation by inhibiting JNK and ERK activation. View Publication

Many biological tissues,such as cardiac muscle,tendons,and the cornea,exhibit highly organized microstructural alignment that is critical for mechanical and physiological functions. Disruptions in this structural organization are commonly associated with pathological conditions such as fibrosis,infarction,and cancer. However,conventional histological imaging techniques rely on immunofluorescence or histochemical staining,and they evaluate tissue alignment via non-physical 2D gradient-based calculation,which is labor-intensive,antibody-dependent,and prone to variability. Here,we demonstrate label-free mid-infrared dichroism-sensitive photoacoustic microscopy (MIR-DS-PAM),an analytical imaging system for cardiac tissue assessments. By combining molecular specificity with polarization sensitivity,this method selectively visualizes protein-rich engineered heart tissue (EHT) and quantifies the extracellular matrix (ECM) alignment without any labeling. The extracted dichroism-sensitive parameters,such as the degree of dichroism and the orientation angle,enable histostructural evaluation of tissue integrity and reveal diagnostic cues in fibrotic EHT. This technique offers a label-free analytical tool for fibrosis research and tissue engineering applications. Mid-infrared dichroism-sensitive photoacoustic microscopy enables label-free,quantitative histostructural analysis by combining spectral specificity and polarization sensitivity to visualize protein-rich components and evaluate anisotropic tissue alignment. View Publication

BACKGROUND: The discovery of induced pluripotent stem cells revolutionized regenerative medicine,providing a source for generating induced pluripotent stem cell-derived mesenchymal stem cells (iMSCs). AIM: To evaluate and compare five iMSC differentiation protocols,assessing their efficiency,phenotypic characteristics,and functional properties relative to primary mesenchymal stem cells (MSCs). METHODS: Five iMSC differentiation protocols were assessed: SB431542-based differentiation (iMSC1,iMSC3),an iMatrix-free method (iMSC2),growth factor supplementation (iMSC4),and embryoid body formation with retinoic acid (EB-iMSC). iMSC identity was confirmed according to the International Society for Cell & Gene Therapy 2006 criteria,requiring expression of surface markers (CD105,CD73,CD90) and absence of pluripotency markers. Functional assays were conducted to evaluate differentiation potential (osteogenic and adipogenic),proliferation,mitochondrial function,reactive oxygen species,senescence,and migration. RESULTS: All iMSC types expressed MSC markers and lacked pluripotency markers. EB-iMSC and iMSC2 showed enhanced osteogenesis (runt-related transcription factor 2; P ≤ 0.01 and P ≤ 0.0001,respectively),while adipogenic potential was reduced in iMSC2 (Adipsin; P ≤ 0.01) and EB-iMSC (Adipsin and peroxisome proliferator-activated receptor gamma; P ≤ 0.0001 and P ≤ 0.01,respectively). Proliferation was comparable or superior to bone marrow MSCs,except in iMSC1,with iMSC4 showing the highest rate (MTT assay; P values ranged from 0.01 to 0.001). Despite reduced mitochondrial health in iMSC3 and iMSC4 (P ≤ 0.001),reactive oxygen species levels were lower in all iMSCs (P values ranged from 0.001 to 0.0001),and senescence was significantly reduced in all iMSCs with the exception of iMSC1 (P values ranged from 0.01 to 0.0001). Migration was most reduced in iMSC4 (P ≤ 0.001 at 24 hours and P ≤ 0.0001 at 48 hours). CONCLUSION: While all protocols generated functional iMSCs,variations in differentiation,proliferation,and function emphasize the impact of protocol selection. These findings contribute to optimizing iMSC generation for research and clinical applications. View Publication

Background: DDB1 and CUL4‐associated factor 7 (DCAF7) is a WD‐repeat adaptor that recruits substrates to the CUL4DDB1 ubiquitinligase complex,but its pan‐cancer relevance and mechanistic contribution to tumor progression remain unclear. Methods: Multi‐omics datasets (genomic,transcriptomic,epigenomic,proteomic and single‐cell) from 33 tumor types were integrated to define DCAF7 expression,regulation,and clinical significance. Somatic alterations and copy‐number variation were analysed across cohorts,and promoter methylation and RNA modification signatures were interrogated. Immune associations were assessed by computational deconvolution and checkpoint‐gene profiling. Pathway and network analyses were performed to infer DCAF7‐linked programmes. Mechanistic and functional validation was conducted in hepatocellular carcinoma (LIHC) cell lines (HepG2,Huh7) using DCAF7 perturbation and pharmacologic Wnt inhibition. Results: DCAF7 was overexpressed in most cancers,consistent with copy‐number gain,focal promoter hypomethylation and putative m6A‐linked post‐transcriptional regulation,whereas hypermethylation at two CpG loci predicted poor prognosis in LIHC. DCAF7 alterations,predominantly amplifications,were associated with shorter overall survival in LIHC and positively correlated with DCAF7 mRNA abundance across cohorts. Immunogenomic analyses linked high DCAF7 to CD4+ T‐cell enrichment,broad upregulation of checkpoint genes (PD‐1/PD‐L1,CTLA‐4,TIGIT),and increased tumour mutational burden,microsatellite instability and neoantigen load,suggesting an immune‐evasive phenotype. Network and enrichment analyses converged on canonical Wnt/β‐catenin,Hippo and cell‐cycle programs. In vitro,DCAF7 promoted LIHC cell proliferation and migration by stabilising β‐catenin via increased inhibitory Ser9 phosphorylation of GSK‐3β,thereby inducing c‐Myc and cyclin D1; DCAF7 knockdown or the Wnt inhibitor XAV939 attenuated these effects. Drug‐response modelling further predicted increased sensitivity of DCAF7‐high tumours to 17‐AAG,docetaxel and alsterpaullone. Conclusions: DCAF7 is frequently activated by genetic and epigenetic mechanisms across cancers,associates with an immunotherapy‐relevant tumour immune milieu,and drives Wnt/β‐catenindependent malignant phenotypes in LIHC. These findings support DCAF7 as a prognostic biomarker and a candidate therapeutic target,particularly for stratified intervention in LIHC. Key points: DCAF7 is up‐regulated in various tumours and correlates with poor prognosis,particularly in LIHC. High DCAF7 expression is linked to CD4+ T cell infiltration,up‐regulation of immune checkpoint genes and increased tumour mutational burden,suggesting a role in tumour immune escape. DCAF7 stabilises β‐catenin by enhancing GSK‐3β Ser9 phosphorylation,thereby driving c‐Myc/cyclin D1 expression and contributing to proliferation and migration in LIHC. DCAF7‐high tumours demonstrate therapeutic vulnerability to 17‐AAG,docetaxel and CDK/GSK‐3 inhibitor,revealing potential targeted treatment strategies. View Publication

The development of new alternative models is essential to overcome the limitations of traditional two-dimensional (2D) cell cultures and animal models. Three-dimensional (3D) models,such as organoids,better mimic the structural and functional complexity of mammalian organs,thereby reducing the ethical and economic issues related to animal experimentation. These systems provide more physiologically relevant environments,improving the accuracy of disease modeling and drug response prediction. In this context,we have developed mouse hepatic organoids from livers of adult BALB/c mice and characterized them by microscopy and transcriptional analysis. This model was applied to a robust and reproducible high-throughput screening (HTS) platform for testing cytotoxicity at the preclinical stage of drug discovery. In addition,mouse hepatic organoids were co-cultured with amastigotes of Leishmania donovani parasites to establish a model of host–parasite interaction,which was characterized by RNA-seq linked to differential expression analysis and cytokine production by the hepatic organoids. The findings provided in this work establish mouse hepatic organoids as an alternative model for drug discovery and pathogenesis studies. View Publication

Palmitic acid (PA) is the most common dietary saturated fatty acid,and is abundant in palm and cottonseed oil,butter,and cheese,whereas oleic acid (OA) is a monounsaturated omega-9 fatty acid found in olive oil. The differences in the cytotoxic and pro-inflammatory effects of PA and OA across endothelial cells (ECs) isolated from different vascular beds have not been investigated in detail. Here,we incubated primary human aortic valve (HAVEC),saphenous vein (HSaVEC),internal thoracic artery (HITAEC),and microvascular (HMVEC) ECs with albumin-bound PA or OA for 24 h and found that PA induced a considerable cytotoxic response,accompanied by an elevated expression of the genes encoding cell adhesion molecules (VCAM1,ICAM1,SELE,and SELP) and pro-inflammatory cytokines (MIF,PTX3,CSF2,CSF3,IL1A,IL6,CCL2,CCL5,CCL20,CSF2,CSF3,CXCL1,CXCL2,CXCL3,CXCL5,CXCL6,CXCL8,and CXCL10),followed by an increased release of interleukin-6 and interleukin-8. HAVEC and HSaVEC were more susceptible to PA,whereas OA had mild-to-moderate cytotoxic effects on HAVEC and HMVEC but did not induce generalized EC activation. Compared with other EC types,HITAEC was the most resistant to PA and OA treatment. Collectively,these results indicate considerable heterogeneity across the ECs of distinct origin in response to PA. View Publication

Multiple myeloma (MM) is a plasma cell malignancy that disrupts bone homeostasis by suppressing osteogenesis and promoting osteoclast activity. While most therapeutic interventions to date have focused on targeting tumor cells and reducing osteolysis,we investigate whether osteoinductive strategies can restore bone formation and counteract disease progression. Using a human bone marrow-like scaffold model that enables direct in vivo evaluation of tumor–stroma interactions and human bone formation,we demonstrate that MM-derived mesenchymal stromal cells (MSCs) retain osteogenic potential but are functionally suppressed by MM cells. Transcriptomic profiling of MM-primed MSCs revealed the downregulation of small leucine-rich proteoglycans (SLRPs),ASPN,OGN,and OMD,key mediators of bone morphogenetic protein (BMP) signaling,which governs osteoblast differentiation. Among the BMPs analyzed,BMP6 emerged as a potent inducer of osteogenesis and regulator of the expression of these SLRPs. Notably,BMP6 selectively promoted bone formation without enhancing osteoclastogenesis and attenuated inflammatory and tumor-supportive MSC phenotypes. BMP6 also directly inhibited MM cell proliferation and suppressed IL6-induced growth. These findings highlight BMP6 as a distinct multifunctional regulator warranting further investigation as a potential therapeutic approach,while establishing the humanized model as a valuable platform for dissecting tumor–bone interactions in MM. View Publication

CD8+ T cells are an important weapon in the therapeutic armamentarium against cancer. While CD8+CD103+ T cells with a tissue-resident memory T (TRM) cell phenotype are associated with favorable prognoses,the tumor microenvironment also contains dysfunctional exhausted T (TEX) cells that exhibit a variety of TRM-like features. Here we deconvolute TRM and TEX cells across human cancers,ascribing markers and gene signatures that distinguish these populations and enable their functional distinction. Although TRM cells have superior functionality and are associated with long-term survival post-tumor resection,they are not associated with responsiveness to immune checkpoint blockade. Tumor-associated TEX and TRM cells are clonally distinct,with the latter comprising tumor-independent bystanders and tumor-specific cells segregated from cognate antigen. Intratumoral TRM cells can be forced toward an exhausted fate when chronic antigen stimulation occurs,indicating that the presence or absence of continuous antigen exposure within the microenvironment is the key distinction between tumor-associated TEX and TRM populations. These results highlight unique functions for TRM and TEX cells in tumor control,underscoring the need for distinct strategies to harness these populations for cancer therapies. Here the authors show that tissue-resident memory and exhausted T cells in tumors are distinct populations that are shaped by relative presence or absence of TCR signals,suggesting that a tailored therapeutic strategy is needed to target each subset. View Publication

Propofol is a commonly used anesthetic for sedation during surgery. This drug is reported to exhibit nonanaesthetic immunomodulatory and anti-inflammatory effects. Herein,we investigated the impact of topical propofol delivery with the aim of mitigating psoriatic inflammation. The antipsoriatic potency of propofol was evaluated in a cell-based study in which keratinocytes,macrophages,and neutrophils were used as models. A significant reduction in the proinflammatory effectors interleukin (IL)-6,IL-8,and CXC motif chemokine ligand (CXCL)1 was found in activated keratinocytes (HaCaT) treated with propofol. This reduction could enable baseline control. Immunoblotting suggested that the antioxidant enzymes nuclear factor erythroid 2-related factor (Nrf)2 and heme oxygenase (HO)-1 were involved in the protective effect of propofol on keratinocyte stimulation. The increase in Nrf2 and HO-1 was mediated by kelch-like ECH-associated protein (KEAP)1 downregulation. Propofol presented scavenging activity and decreased 2,2-diphenyl-1-picrylhydrazyl (DPPH) by 47%. The downregulation of cytokines/chemokines in activated macrophages (differentiated THP-1) and mouse neutrophils was also found after propofol treatment. Macrophage migration triggered by the conditioned medium of activated keratinocytes could be blocked with the intervention of propofol. The absorption level of propofol (3 mM) into intact pig skin was 1.2 nmol/mg. Skin deposition was increased to 3.7 nmol/mg after SC lipid removal to mimic psoriasiform skin. In silico molecular docking demonstrated the facile interaction of propofol with ceramides in the stratum corneum (SC). The treatment of imiquimod (IMQ)-sensitized mice with topical propofol suppressed erythema,acanthosis,and macrophage/neutrophil infiltration. Propofol also dramatically decreased cytokine/chemokine levels and epidermal thickness in the lesion. In summary,propofol exhibits anti-inflammatory and antioxidant properties to treat psoriasiform lesions. Topical propofol delivery is useful as an ideal route to accomplish antipsoriatic therapy and avoid systemic effects. View Publication