Project description:Here, three colorectal tumors were subjected to anti-ORF1p LFQ, IP-MS: (A) Krukenberg Carcinoma, Ovary; (B) Metastatic Rectal Adenocarcinoma, Liver; (C) Adenocarcinoma, Colon. These were accompanied by matched normal IP controls and/or mouse IgG IP controls. The objective is to map LINE-1 RNP interactions in cancer.
Project description:<p><strong>BACKGROUND:</strong> Novel biomarkers are urgently needed to distinguish between benign and malignant thyroid nodules and detect thyroid cancer in the early stage. The associations between serum IgG N-glycosylation and thyroid cancer risk have been revealed. We aimed to explore the potential of IgG N-glycan traits as biomarkers in the differential diagnosis of thyroid cancer.</p><p><strong>METHODS:</strong> Plasma IgG N-glycome analysis was applied to a discovery cohort followed by independent validation. IgG N-glycan profiles were obtained using a robust quantitative strategy based on matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. IgG N-glycans were relatively quantified, and classification performance was evaluated based on directly detected and derived glycan traits.</p><p><strong>RESULTS: </strong>Four directly detected glycans were significantly changed in thyroid cancer patients compared to that in non-cancer controls. Derived glycan traits and a classification glycol-panel were generated based on the directly detected glycan traits. In the discovery cohort, derived trait BN (bisecting type neutral N-glycans) and the glyco-panel showed potential in distinguishing between thyroid cancer and non-cancer controls with AUCs of 0.920 and 0.917, respectively. The diagnostic potential was further validated. Derived trait BN and the glycol-panel displayed “accurate” performance (AUC>0.8) in discriminating thyroid cancer from benign thyroid nodules and healthy controls in the validation cohort. Meanwhile, derived trait BN and the glycol-panel also showed diagnostic potential in detecting early-stage thyroid cancer.</p><p><strong>CONCLUSIONS:</strong> IgG N-glycome analysis revealed N-glycomic differences that allow classification of thyroid cancer from non-cancer controls. Our results suggested that derived trait BN and the classification glyco-panel rather than single N-glycans may serve as candidate biomarkers for further validation.</p>
Project description:We aimed to characterize changes within the SUMOylation pathway that are driven by the tRNA-like molecule mascRNA. For the experiments otulined below, we used 293FT cells (a derivative of HEK293T cells).First, we identified the SAE2-associating proteins. Sample IDs: 92072 NRS control, 92073 antiSAE2 IP. Secondly, we identified proteins pulled-down by biotinylated mascRNA and mascRNA-3'CCA, using scrambled mascRNA oligo as a control. Sample IDs: 92199: scrambled mascRNA-Biotin control; 92200: mascRNA3'biotinTEG; 92201: mascRNA-CCA3'biotinTEGThirdly, we verified the specificity of SUMO1 IPs, by comparing two beads elution approaches: 1) by SUMO1 competitive peptide elution (processed gel pieces). Sample IDS: 99699-99704: Proteomic analysis of SDS PAGE gel pieces for SUMO1 IP and control IgG IP. (Details below) 2) urea eluates from the SUMO1-IPs. Sample IDs: 99705 control Rabbit IgG IP; 99706 SUMO1 IP Lastly, we used urea eluates from SUMO1-IPs from cells transfected with 5’P-mascRNA vs. 5’Pscrambled RNA. Sample IDs: 100479, 100480, 100481: Triplicates (1A, 1B, 1C) of control IgG IP from cells treated with scrambled mascRNA oligo (control IP)100482, 100483, 100484: Triplicates (3A, 3B, 3C) of control IgG IP from cells treated with mascRNA oligo (control IP)100485, 100486, 100487: Triplicates (2A, 2B, 2C) of SUMO1 IP from cells treated with scrambled mascRNA oligo (control transfection)100488, 100489, 100490: Triplicates (4A, 4B, 4C) of SUMO1 IP from cells treated with mascRNA oligo Sample details can be found in the file named Experimental_Design_Proteomics_ELIAPI_2024.txt below.
Project description:Chronic inflammatory and immune dysregulation are critical drivers in the development and progression of chronic obstructive pulmonary disease (COPD). Posttranslational modifications, such as glycosylation of Immunoglobulin G (IgG), modulates systemic inflammatory homeostasis. This study aims to profile plasma IgG glycopeptides (IgGPs) in COPD patients to uncover new insights into its pathogenesis and to identify novel biomarkers. Plasma IgG N-glycopeptides from 90 COPD patients, 45 clinical defined early COPD (CECOPD) patients and 90 healthy individuals were analyzed using an integrated platform that combines Fe3O4@PDA@DETA nanospheres enrichment with high-resolution mass spectrometry measurement. Correlations between IgG N-glycoforms and clinical parameters were assessed to explore underlying mechanisms of COPD progression. Disease-specific IgGPs were identified in both ECOPD and COPD cohorts. Notably, IgG glyco-pattern, rather than IgG levels, changed with disease progression. Early COPD patients showed decreased bisection and increased site-specific afucosylated galactosylation and fucosylation of IgG, indicating an anti-inflammatory state. In contrast, COPD patients gave increased inflammation, characterized by reduced galactosylation and sialylation. Interestingly, a subset of healthy controls displayed IgGPs patterns similar to early COPD, possibly reflecting the impact of substantial smoking exposure and associated immune responses. These findings suggest that plasma IgG glycosylation could serve as a potential biomarker for early COPD diagnosis, providing valuable insights into immune system changes during disease progression.
Project description:The antiphospholipid syndrome (APS) is an autoimmune disease in which antiphospholipid antibodies (aPL) cause vascular thrombosis (VT+) and/or pregnancy morbidity (PM+). The experiment aimed at determining whether specific aPL are associated with thrombotic or obstetric manifestations. In the present study we carried out microarray analysis comparing patterns of mRNA expression in monocytes from a healthy volunteer exposed to IgG from patients with a history of vascular thrombosis (VT+/PM-) or pregnancy morbidity (VT-/PM+) or to IgG from healthy controls (HC).
Project description:While antibodies in bullous pemphigoid (BP) are known to activate innate immune response, their direct effect on keratinocytes and the overall contribution of keratinocytes to disease pathogenesis is largely unknown. We sought to understand the mechanism of keratinocyte response to autoantibodies in BP. We affinity purified IgG from patients with a known diagnosis of bullous pemphigoid or healthy controls, and performed bulk RNA-seq. We identified numerous differentially expressed genes including upregulation of CXCL16, IL-24, and TGFB1 in BP-IgG treated keratinocytes
Project description:While antibodies in bullous pemphigoid (BP) are known to activate innate immune response, their direct effect on keratinocytes and the overall contribution of keratinocytes to disease pathogenesis is largely unknown. We sought to understand the mechanism of keratinocyte response to autoantibodies in BP. We affinity purified IgG from patients with a known diagnosis of bullous pemphigoid or healthy controls, and performed bulk RNA-seq. We identified numerous differentially expressed genes including upregulation of CXCL16, IL-24, and TGFB1 in BP-IgG treated keratinocytes
Project description:NOD.B10Sn-H2b/J mice are a mouse model of primary Sjogren's syndrome. We compared IgM and IgG autoantibody levels from NOD.B10Sn-H2b/J females with clinical disease to those of healthy controls.