Clarifying the role of chr11q13.3 and the related genes in this amplicon in HNSCC

IP-mass analysis in Flag-FADD-overexpressed 293T cells.

BackgroundType 1 diabetes (T1D) is an autoimmune disease characterized by progressive β-cell destruction, yet current risk stratification tools, which rely mainly on genetic susceptibility and autoantibody profiles, remain insufficient for accurately predicting disease progression. We aimed to characterize macrophage-related inflammatory transcriptional activity in T1D and to develop peripheral blood–based biomarkers for diagnosis and risk stratification.MethodsWe integrated bulk RNA-seq, single-cell RNA-seq, and spatial transcriptomic data from human islets with public and in-house peripheral blood transcriptomic datasets. Macrophage heterogeneity and remodeling trajectories were analyzed in the islet microenvironment, and machine learning was used to derive tissue- and blood-based proinflammatory macrophage-related genes (PMRG). Diagnostic and prognostic models were then constructed and validated in peripheral blood cohorts, including a longitudinal islet autoimmunity (IA) cohort. SHAP analysis was applied to improve model interpretability. Independent PBMC RT-qPCR and mouse pancreatic immunofluorescence were performed to validate selected PMRG-related genes.ResultsT1D islets showed marked immune remodeling with myeloid enrichment and five distinct macrophage subtypes. Pseudotime analysis identified a pro-inflammatory macrophage trajectory and 265 PMRGs, from which a 9-gene islet-derived PMRG (iPMRG) was obtained. Spatial transcriptomics supported the association of iPMRG-high macrophage signals with disrupted β-cell regions, and CellChat analysis inferred altered inflammatory communication programs. In peripheral blood mononuclear cells (PBMCs), the iPMRG-based diagnostic classifier distinguished T1D from healthy controls with an optimism-corrected AUC of 0.736. For prognosis, a 15-gene prognostic PMRGs was used to construct a risk score that, when integrated with clinical variables, predicted progression from IA to clinical T1D with time-dependent AUCs of 0.825, 0.814, and 0.860 at 12, 36, and 60 months, respectively. SHAP analysis identified the PMRG risk score as the dominant predictor and highlighted six core driver genes (PID1, TFPI2, SERPINB2, SOX4, DUSP2, and MT1X). The computational findings were further supported by independent validation in PBMCs and mouse pancreatic tissues.ConclusionsOur study highlights the heterogeneous and dynamic nature of macrophage remodeling in the T1D islet microenvironment, which is translated into accessible peripheral blood signatures. The resulting diagnostic and prognostic models provide an interpretable framework for T1D risk stratification and may support future strategies for earlier detection and precision prevention.

A proportion of patients with new-onset diabetes share similar symptoms with type 1 diabetes (T1D) patients but they are negative for islet antigen-specific autoantibodies. This study was to develop an islet antigen-specific CD8+ T-cell assay to provide autoimmune evidence regarding these “suspected” T1D patients. HLA-A*0201 individuals with autoAbs+ T1D, autoAbs− suspected T1D, and type 2 diabetes, along with HLA-A*0201 healthy controls were recruited. Using interferon-γ enzyme-linked immunospot assays, the percentages of participants in each group with various islet antigen-specific CD8+ T cells were determined. Sixteen out of the 28 islet antigen-specific epitopes tested were T1D specific, meaning that there was a significantly (P < 0.05) greater epitope positivity rate in the autoAbs+ T1D cohort than in the healthy controls. Using a cutoff value of two positive epitopes, the 16-epitope panel led to a sensitivity of 75.0

Abnormal hematopoiesis is inherently linked to metabolic reprogramming. Protein phosphatase 2A (PP2A), a master regulator of hematopoietic homeostasis, has been implicated in multiple hematological disorders. However, the precise mechanisms by which PP2A coordinates metabolic networks to govern hematopoietic fate decisions remain poorly defined. Herein, we identify lactate as a critical mediator of myeloid-biased differentiation triggered by PP2A inactivation. Genetic ablation of PPP2CA, the catalytic subunit of PP2A, results in aberrant myeloid proliferation and lymphoid depletion. Transcriptomic profiling reveals that Ppp2ca deficiency alters the expression of transcriptional regulators governing hematopoietic lineage commitment and energy metabolism. Metabolomic analyses further demonstrate enhanced lactate metabolism in Ppp2ca-deficient hematopoietic progenitors. Importantly, either haploinsufficiency or pharmacological inhibition of lactate dehydrogenase A (LDHA) in vivo effectively reverses the abnormal hematopoiesis induced by Ppp2ca deficiency. Mechanistically, Ppp2ca deletion directly promotes the transcriptional initiation of glycolytic genes (e.g., Ldha) via RNA polymerase II (Pol II). This leads to heightened lactylation of histone deacetylases (HDAC) at specific residues - lysine 412 in HDAC1 and lysine 451 in HDAC2 - impairing the assembly of the HDAC1/2/SIN3A co-repressor complex on chromatin, enhancing histone acetylation, and ultimately dysregulating hematopoietic gene expression. Collectively, our work establishes the "PP2A-lactate- HDAC lactylation" axis as a pivotal regulator of hematopoiesis and identifies LDHA as a promising therapeutic target for PP2A-associated hematological disorders.

Acute myeloid leukemia (AML), which is most common in adults, is a challenging hematological malignancy. The occurrence and the progression of AML are often accompanied by various gene fusions and/or mutations. Herein, we report the first case of a TOR1AIP2::ETV6 fusion transcript with a translocation of t(1;12)(q25;p13) in AML progressed from myelodysplastic syndrome (MDS) combined with an FLT3-ITD (internal tandem duplication) mutation. Further studies should focus on the biological functions of these novel chimeric products in disease onset and progression, as well as their potential as monitoring markers in disease regression.

Objective: Genome-wide association studies (GWAS) have identified that 6p22.2 region is associated with type 1 diabetes (T1D) risk in the Chinese Han population. This study aims to reveal associations between this risk region and T1D subgroups and related clinical features, and further identify causal variant(s) and target gene(s) in this region. Methods: 2608 T1D and 4814 healthy controls were recruited from East, Central, and South China. Baseline data and genotyping for rs4320356 were collected. The most likely causal variant and gene were identified by bioinformatics analysis, dual-luciferase reporter assays, expression quantitative trait loci (eQTL), and functional annotation of the non-coding region within the 6p22.2 region. Results: The leading variant rs4320356 in the 6p22.2 region was associated with T1D risk in the Chinese and Europeans. However, this variant was not significantly associated with islet function or autoimmunity. In silico analysis suggested rs9379874 was the most potential causal variant for T1D risk among thymus, spleen, and T cells, overlapping with the enhancer-related histone mark in multiple T cell subsets. Dual luciferase reporter assay and eQTL showed that the T allele of rs9379874 increased BTN3A1 expression by binding to FOXA1. Public single-cell RNA sequencing analysis indicated that BTN3A1 was related to T-cell activation, ATP metabolism, and cytokine metabolism pathways, which might contribute to T1D development. Conclusion: This study indicates that a functional variant rs9379874 regulates BTN3A1 expression, expanding the genomic landscape of T1D risk and offering a potential target for developing novel therapies.

Context Single positive islet autoantibodies (IAbs), sometimes detected in healthy individuals and patients with low-risk of developing type 1 diabetes (T1D), are considered to be irrelevant to the development of diabetes, making it difficult to diagnose and classify adult-onset diabetes. Objective To determine the significance and clinical value of IAbs in T1D diagnosis in the low-prevalence population, and to explore whether an electrochemiluminescence IAb detection assay can improve the clinical utility of IAbs in the immunodiagnosis of T1D in the low-prevalence population. Methods A total of 633 newly diagnosed patients with adult-onset diabetes (>= 18 years old) were divided into 2 groups according to their clinical phenotypes: 575 patients with age at diagnosis >= 35 years and body mass index (BMI) >= 24 kg/m(2) were considered a low-prevalence population (population with a low prevalence of T1D) and the other 58 patients were considered a high-prevalence population. All the samples from 633 participants were tested with IAbs using standard radiobinding assays (RBAs) and electrochemiluminescence (ECL) assays in parallel. Results Compared with the high-prevalence population, fewer positive IAbs (94/575, 16.3% vs 28/58, 48.3%) were detected in the low-prevalence population, and more of whom (69/94, 73.4% vs 9/28, 32.2%) were positive for a single IAb, with glutamate decarboxylase antibodies being the most prevalent single IAb. Single-IAb detection in the low-prevalence population did not always suggest the T1D phenotype. Combined detection of IAbs by RBA and ECL assay had a significant clinical utility to distinguish autoimmune diabetes in the low-prevalence population with low BMI, poor beta-cell function at the diagnosis, and an accelerated decline in beta-cell function during the follow-up. Conclusion Combined autoantibody detection by RBA and ECL assays improved differentiating autoimmune from nonautoimmune diabetes in the low-prevalence population.

In this research, lipid microbubbles (MBs) were prepared first, and then Tat peptide, carboxylated heparin, amino biotin, and amino folic acid were successively synthesized. After a certain amount of paclitaxel was added, paclitaxel nanoparticles (NPs) with a double-ligand were obtained through reaction. The lipid MBs prepared above were added to streptavidin. After washing and purification, the lipid MBs and double-ligand paclitaxel NPs were added. After incubation on ice, the lipid MB-double-ligand paclitaxel NP complex was obtained. In addition to the physical characterization of the materials, human breast cancer cells MDA-MB-231 and lung cancer cells A549 were first utilized to test the biological properties of the NP complex In Vitro and then utilized to study the effects of gastric cancer (GC) cells. The results revealed that the lipid MBs were uniformly distributed and did not aggregate. The concentration of the NP complex reached 7.75±0.93×10 8 NPs/mL, and the particle size was 2.23±0.68 μ m. At various radiation intensities, blue fluorescently stained MDA-MB-231 cells and A549 cells showed greener fluorescently labeled double-ligand paclitaxel NPs around and inside the nucleus of Hoechst 33342. According to the prepared products and byproducts, they were grouped to compare different prepared products. The fluorescence uptake of the two cells at 4 h was the highest under the condition of the NP complex combined with ultrasonic radiation, and the destruction of cancer cells (MDA-MB-231 and A549) was the strongest under the condition of the NP complex combined with ultrasonic radiation. In GC cells, NP complexes inhibited cell migration and invasion relative to the other groups ( P <0.05), the level of Bax protein increased ( P <0.05), while that of Bcl-2, pSTAT3/STAT3, and phosphorylation of NF-kappa B (PNF- κ B)/NF- κ B protein were markedly decreased ( P <0.05).

This study aims to investigate the correlations between islet function/ insulin resistance and serum lipid levels, as well as to assess whether the strength of such correlations is affected by the GCKR rs1260326 variant in healthy and T2D individuals. We performed an oral glucose tolerance test (OGTT) on 4889 middle-aged adults, including 3135 healthy and 1754 T2D individuals from the REACTION population study in the Nanjing region. We also measured their serum lipid levels and genotyped for rs1260326. We found that serum high-density lipoprotein (HDL) cholesterol and triglyceride (TG) levels were independently correlated with indexes of islet function (HOMA-β and IGI [insulinogenic index]) and insulin resistance (HOMO-IR and ISIMatsuda) in both healthy and T2D individuals. The correlations were significantly decreased in T2D individuals, with significant heterogeneities compared to healthy controls (I2 > 75%, Phet < 0.05). Although no correlation was observed between serum total cholesterol (TC) level and islet function/ insulin resistance in healthy controls, significant correlations were found in T2D individuals, with significant heterogeneity to healthy controls in the correlation with ISIMatsuda(I2 = 85.3%, Phet = 0.009). Furthermore, we found significant interactions of the GCKR rs1260326 variant for the correlations between serum HDL cholesterol and HOMA-β/ISIMatsuda in T2D subjects (P = 0.015 and 0.038, respectively). These findings illustrate that distinct correlations between serum lipid levels and islet function/ insulin resistance occurred in T2D subjects compared to healthy individuals. Common gene variants, such as rs1260326, might interact substantially when studied in specific populations, especially T2D disease status.

Objective:To examine the distribution and effects of the subclass of insulin antibodies on glucose control and side events in patients with type 2 diabetes treated with premixed insulin analog.Methods:A total of 516 patients treated with premixed insulin analog were sequentially enrolled from the First Affiliated Hospital of Nanjing Medical University from June 2016 to August 2020. Subclass-specific insulin antibodies (IAs) (IgG1-4, IgA, IgD, IgE, and IgM) were detected in IA-positive patients by electrochemiluminescence. We analyzed glucose control, serum insulin, and insulin-related events between IA-positive and IA-negative groups, as well as among patients with different IA subclasses.Results:Overall, 98 of 516 subjects (19.0%) were positive for total IAs after premixed insulin analog therapy; of these participants, 92 had subclass IAs, and IgG-IA was the predominant subclass, followed by IgE-IA. IAs were associated with serum total insulin increase and local injection-site reactions but not glycemic control and hypoglycemia. In the subgroup analysis in patients with IA-positive, the IgE-IA and IA subclass numbers were more associated with increased serum total insulin levels. Additionally, IgE-IA might be correlated more strongly with local responses and weakly with hypoglycemia, while IgM-IA might be correlated more strongly with hypoglycemia.Conclusion:We concluded that IAs or IA subclasses might be associated with unfavorable events in patients receiving premixed insulin analog therapy, which can be used as an adjunctive monitoring indicator in clinical insulin trials.

ObjectivesN6-methyladenosine (m6A) is essential in the regulation of the immune system, but the role that its single nucleotide polymorphisms (SNPs) play in the pathogenesis of type 1 diabetes (T1D) remains unknown. This study demonstrated the association between genetic variants in m6A regulators and T1D risk based on a case-control study in a Chinese population.MethodsThe tagging SNPs in m6A regulators were genotyped in 1005 autoantibody-positive patients with T1D and 1257 controls using the Illumina Human OmniZhongHua-8 platform. Islet-specific autoantibodies were examined by radioimmunoprecipitation in all the patients. The mixed-meal glucose tolerance test was performed on 355 newly diagnosed patients to evaluate their residual islet function. The functional annotations for the identified SNPs were performed in silico. Using 102 samples from a whole-genome expression microarray, key signaling pathways associated with m6A regulators in T1D were comprehendingly evaluated.ResultsUnder the additive model, we observed three tag SNPs in the noncoding region of the PRRC2A (rs2260051, rs3130623) and YTHDC2 (rs1862315) gene are associated with T1D risk. Although no association was found between these SNPs and islet function, patients carrying risk variants had a higher positive rate for ZnT8A, GADA, and IA-2A. Further analyses showed that rs2260051[T] was associated with increased expression of PRRC2A mRNA (P = 7.0E-13), and PRRC2A mRNA was significantly higher in peripheral blood mononuclear cell samples from patients with T1D compared to normal samples (P = 0.022). Enrichment analyses indicated that increased PRRC2A expression engages in the most significant hallmarks of cytokine-cytokine receptor interaction, cell adhesion and chemotaxis, and neurotransmitter regulation pathways. The potential role of increased PRRC2A in disrupting immune homeostasis is through the PI3K/AKT pathway and neuro-immune interactions.ConclusionThis study found intronic variants in PRRC2A and YTHDC2 associated with T1D risk in a Chinese Han population. PRRC2A rs2260051[T] may be implicated in unbalanced immune homeostasis by affecting the expression of PRRC2A mRNA. These findings enriched our understanding of m6A regulators and their intronic SNPs that underlie the pathogenesis of T1D.

AbstractBackgroundAcetylcholine (ACh) and norepinephrine (NE) are representative neurotransmitters of parasympathetic and sympathetic nerves, respectively, that antagonize each other to coregulate internal body functions. This also includes the control of different kinds of hormone secretion from pancreatic islets. However, the molecular mechanisms have not been fully elucidated, and whether innervation in islets is abnormal in diabetes mellitus also remains unclear.Methods and resultsImmunofluorescence colocalization and islet perfusion were performed and the results demonstrated that ACh/NE and their receptors were highly expressed in islet and rapidly regulated different hormones secretion. Phosphorylation is considered an important posttranslational modification in islet innervation and it was identified by quantitative proteomic and phosphoproteomic analyses in this study. The phosphorylated islet proteins were found involved in many biological and pathological processes, such as synaptic signalling transduction, calcium channel opening and insulin signalling pathway. Then, the kinases were predicted by motif analysis and further screened and verified by kinase‐specific siRNAs in different islet cell lines (αTC1‐6, Min6 and TGP52). After functional verification, Ksr2 and Pkacb were considered the key kinases of ACh and NE in insulin secretion, and Cadps, Mlxipl and Pdcd4 were the substrates of these kinases measured by immunofluorescence co‐staining. Then, the decreased expression of receptors, kinases and substrates of ACh and NE were found in diabetic mice and the aberrant rhythm in insulin secretion could be improved by combined interventions on key receptors (M3 (pilocarpine) or α2a (guanfacine)) and kinases (Ksr2 or Pkacb).ConclusionsAbnormal innervation was closely associated with the degree of islet dysfunction in diabetic mice and the aberrant rhythm in insulin secretion could be ameliorated significantly after intervention with key receptors and kinases in the early stage of diabetes mellitus, which may provide a promising therapeutic strategy for diabetes mellitus in the future.

T2D and T1D are phenotypically heterogeneous. This study aims to reveal the relationship between the common SLC30A8 rs13266634 variant and subgroups of T2D and T1D and their clinical characteristics. We included 3158 OGTT-based healthy controls, unrelated 1754 T2D, and 1675 autoantibody-positive T1D individuals. The associations between rs13266634 and subtypes of T2D, T1D, autoantibody status and glycemic-related quantitative traits were performed by binary logistic regression analysis under the additive model and multiple linear regression with appropriate adjustment. We found that the T allele of rs13266634 was protectively associated with lean (OR = 0.810, P = 6.91E–04) but not obese T2D with considerable heterogeneity (P = 0.018). This allele also conferred significant protection with T1D of single (OR = 0.847, P = 9.76E–03), but not multi autoantibodies with substantial heterogeneity (P = 0.005). This variant significantly affected OGTT-related insulin release in lean (P = 2.66E–03, 3.88E–03 for CIR and DI, respectively) but not obese healthy individuals. Furthermore, rs13266634 T allele correlated with the risk of ZnT8A (OR = 1.440, P = 3.31E–05) and IA-2A (OR = 1.219, P = 1.32E–03) positivity, with more effect size in children/adolescents compared with adult-onset T1D subtypes. These suggested that the SLC30A8 rs13266634 variant might be put into genetic risk scores to assess the risk of the subtypes of T1D and T2D and their related clinical features.

Circulating CD25hi B cells, a subset of regulatory B cells in humans, are closely related to inflammation and autoimmune diseases. This study is aimed at investigating the alternation of CD25hi Bregs and their correlation with CD4 effector and regulatory T cells in T1D individuals. We included 68 autoantibody-positive T1D and 68 age-matched healthy individuals with peripheral blood mononuclear cells (PBMCs) and assessed them with CD25hi Bregs and CD4 effector or regulatory T cells by flow cytometry. Here, we demonstrate that the frequency of CD25hi Bregs was significantly decreased in T1D subjects (P = 0.0016), but they were not affected by disease status (age at T1D diagnosis or duration) or T1D risk loci (rs2104286 or rs12251307) in IL2RA (all P > 0.05). Moreover, higher IgD (P = 0.043) and lower CD27 (P = 0.0003) expression was observed in CD25hi Bregs of T1D individuals, but not the expression of IgM, CD24, or CD38 (all P > 0.05). Although there was no correlation between CD25hi Bregs and CD4 effector T cell subsets in either T1D or healthy individuals (all P > 0.05), we found a positive correlation between CD25hi Bregs and CD4 Tregs in healthy controls (Sp. r = 0.3544, P = 0.0249), which disappeared in T1D subjects (Sp. r = 0.137, P = 0.401). In conclusion, our results suggest that decreased CD25hi Bregs and alternation of their phenotypes are features of T1D regardless of disease duration and T1D genetic risk loci, and an impaired balance between CD25hi Bregs and CD4 Tregs might contribute to the pathogenesis of T1D.

The critical role of IL-10-producing B cells (B10 cells) with a unique CD1d(hi) CD5(+) phenotype in suppressing autoimmune responses and relieving inflammation has been demonstrated in several models of autoimmune diseases. However, the regulatory role of B10 cells in T cell-mediated autoimmune responses during the natural history of type 1 diabetes is unclear. In this study, we used the NOD mouse model of autoimmune diabetes to clarify the changes and potential mechanisms of B10 cells for disease. Compared with B10 cells present in the 4-wk-old normoglycemic NOD mice, the frequency of B10 cells was increased in the insulitis and diabetic NOD mice, with the highest proportion in the insulitis NOD mice. The changes in the relative number of B10 cells were most pronounced in the pancreas-draining lymph nodes. The pathogenic T cells, including Th1 and Th17 cells, remarkably increased. The assays in vitro showed that B10 cells in the NOD mice did not inhibit the proliferation of CD4(+) CD25(-) T cells. They also had no regulatory effect on IFN-gamma and IL-4 secretion or on Foxp3 expression of T cells. B10 cells suppressed T cell-mediated autoimmune responses via an IL-10-dependent pathway. In contrast, B10 cells in the NOD mice exhibited a significant reduction in IL-10 production. In summary, a defect in the number and function of B10 cells may participate in the development and progression of type 1 diabetes.

Objective:To explore whether palmitic acid (PA) can induce ferroptosis in pancreatic beta-cell line MIN6 cells.Methods:Pancreatic beta-cell line MIN6 cells were divided into normal group, PA group (1.0 mmol/L), PA+apoptosis inhibitor (Z-VAD-FMK) group (5 μmol/L), PA+ ferrostain-1(Fer-1) group (5 μmol/L), PA+ necrostatin-1(Nec-1) group (5 μmol/L). Each group was set three parallels. Cell viability was detected by CCK8 method. Cellular ultrastructure was assessed with electron microscopy. FerroOrange was used to detect intracellular Fe 2+. The lipid peroxidation assay kit was used to detect the concentration of malondialdehyde (MDA). Flow cytometry was used to detect intracellular reactive oxygen species (ROS) levels. Real-time quantitative polymerase chain reaction was used to detect the mRNA levels of caspase3, receptor-interacting protein kinase 3 ( RIPK3), acyl-CoA synthetase long-chain family member 4 ( ACSL4), prostaglandin endoperoxidase synthase 2 ( Ptgs2), arachidonate lipoxygenase 15 ( ALOX15), ferritin heavy chain ( FTH), ferritin light chain ( FTL), and glutathione peroxidase 4 ( GPX4). Western blotting was used to detect the proteins levels of cleaved caspase3, RIPK3, ACSL4, ALOX15 and Ptgs2. The t-test was used to compare two groups. Results:Compared with the PA group, cell viability of MIN6 was higher in PA+Z-VAD-FMK group, PA+Fer-1 group or PA+Nec-1 group ( P<0.01). MIN6 cells, which were treated with PA to exhibit the characteristic morphologic features, were associated with apoptosis, necrosis and ferroptosis under electron microscope. Compared with the control group, in PA group, the levels of the cellular intracellular Fe 2+, MDA and total lipid ROS were increased. The mRNA relative expression levels of caspase3, RIPK3, ACSL4, Ptgs2 and ALOX15 were increased. The mRNA relative expression levels of FTH and GPX4 were decreased. The protein relative expression levels of cleaved caspase3, RIPK3, ACSL4, ALOX15 and Ptgs2 were increased (all P<0.01). Compared with the PA group, in PA+Fer-1 group, the levels of cellular intracellular Fe 2+, MDA and total lipid ROS were decreased. The mRNA relative expression levels of ACSL4, Ptgs2 and ALOX15 were decreased. The mRNA relative expression levels of FTH and GPX4 were increased. The protein relative expression levels of ACSL4, ALOX15 and Ptgs2 were decreased (all P<0.01). Conclusion:PA could induce ferroptosis in pancreatic beta-cell line MIN6 cells.

Colorectal cancer is one of the most commonly occurring cancers worldwide. Although clinical reports have indicated the anticancer effects of Chinese herbal medicine, the multiple underlying molecular and biochemical mechanisms of action remain to be fully characterized. Chinese medicine (CM) monomers, which are the active components of CM, serve as the material basis of the functional mechanisms of CM. The aim of this review is to summarize the current experimental evidence from in vitro, in vivo, and clinical studies for the effects of CM monomers in colorectal cancer prevention and treatment, providing some useful references for future research.