P0356QUANTITATIVE ASSESSMENT OF GALNAC NUMBER IN THE HINGE REGION OF IGA1 IN IGA NEPHROPATHY WITH CRESCENTS FORMING

Abstract Background and Aims Aberrant IgA1 O-linked glycosylation of IgA1 hinge region (HR) has been shown playing an important role in the pathogenesis of IgA nephropathy (IgAN). Serum levels of galactose-deficient IgA1 (Gd-IgA1) was associated with the development and progression in IgAN, however variations in the composition of IgA1 HR glycoforms are unknown. In this study we aim to quantitative assessment of GalNAc number in the hinge region of IgA1 in IgAN with crescents forming using mass spectrometry. Method Plasma polymeric IgA1 (pIgA1) was purified from a discovery cohort including crescentic IgAN (n=11) non-crescentic IgAN (n=10) and healthy controls (HC, n=10) and a validation cohort (crescentic IgAN, n=10; non-crescentic IgAN, n=10 and healthy controls, n=11). After denaturation, reduction, alkylation and trypsin digestion, liquid chromatography-high-resolution mass spectrometry (LC-MS) was used to analyse the IgA1 HR glycans. The intensity of the identified IgA1 O-glycopeptide was calculated and expressed as the relative abundance for each glycopeptide. The molecular weight (MW) of intact O-glycopeptide was calculated as the formula following: MW = HR + x GalNAc + y Gal + z NeuAc + H+ (x, y and z represent the number of GalNAc, galactose and NeuAc which bind to one HR in IgA1 O-linked glycopeptides respectively) Glycopeptide content ratio = (glycopeptide peak intensity/ total glycopeptide intensity) ×100%. Results In the discovery cohort population, the level of Gd-IgA1 was highest in patients with crescentic IgAN, middle in patients with non-crescentic IgAN and lowest in healthy controls (347.69±57.89 U/ml vs 336.32±38.44 U/ml vs 330.14±33.22 U/ml), albeit that didn’t reach the statistical significance. There were significantly difference in GalNAc number of IgA1 HR among patients with IgAN and healthy controls. Overall the numbers of GalNAc bound to one HR was much lower in the patients than healthy controls. As shown in the Figure 1, the proportion of GalNAc 3, defined as O-glycans that were bound to one HR at 3 sites, were highest in patients with crescentic IgAN, then the non-crescentic IgAN and lowest in the healthy controls (9.92%±3.37% vs 6.65%±1.53% vs 4.05%±1.24%; p-value for the trend<0.001). Similar results were observed in GalNAc 4 (45.91%±4.75% vs 41.13%±2.95% vs 40.98%±2.95%; P=0.004. However regarding the GalNAc 5 and GalNAc 6, crescentic IgAN was lowest, then non-crescentic IgAN and highest in the healthy control (GalNAc 5: 45.17%±5.46% vs 46.90%±2.78% vs 48.05%±3.02%, P=0.001 GalNAc 6:1.62%±1.60% vs 3.95%±1.92% vs 5.39%±2.38%; P<0.001). These results were consistent in the validation cohort (Figure 1). Conclusion GalNAc numbers in IgA1 HR was lower in patients with IgAN especially in crescentic IgAN. These results suggest that the O-glycans of IgA1 were associated with the severe phenotype in IgA nephropathy.

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AB0156 NMR-BASED MUSCLE METABOLOMICS IN INFLAMMATORY MYOSITIS- UNDERSTANDING CHANGES IN SERUM AS A REFLECTION OF THE MUSCLE

Annals of the Rheumatic Diseases ◽

10.1136/annrheumdis-2020-eular.316 ◽

2020 ◽

Vol 79 (Suppl 1) ◽

pp. 1378.2-1379

Author(s):

L. Gupta ◽

U. Kumar ◽

A. Anuja ◽

P. Sharma ◽

A. Guleria ◽

...

Keyword(s):

Statistical Significance ◽

Muscle Disease ◽

P Value ◽

Healthy Controls ◽

Inflammatory Myositis ◽

Activity Assessment ◽

Muscle Tissues ◽

Low Levels ◽

Serum Metabolomics ◽

Research Grant

Background:We have previously found promise in NMR as a tool to distinguish sera of active from inactive inflammatory myositis (IIM)1. To understand the changes previously found in sera and urine we studied muscle tissue of patients with myositis.Objectives:To identify differences in metabolome on inflamed muscle tissues of patients with active myositis from that of healthy controls and infectious polymyositis.Methods:Muscle (n=17) from patients classifiable as myositis by the ACR-EULAR criteria [34 years (23.5 - 50.5 IQR), M/F 1:3] were compared with healthy controls [n=11, age= 44 (35-50) years, M/F-1:1]. Two disease controls with infectious polymyositis were also compared. Findings were applied to muscle biopsy tissues of two patients with established myositis and superadded infections (HBV, Histoplasmosis) to assess discriminatory potential.Metabolic profiles were obtained at 800 MHZ NMR spectrometer and compared using multivariate partial least-squares discriminant analysis (PLS-DA). The discriminatory metabolites were identified based on variable importance in projection (VIP) statistics and further evaluated for statistical significance (p-value <0.05). Paired T tests, ANNOVA and correlation of individual metabolites were done after normalizing for formate.Results:Metabolomics profiles in IIM were distinct from healthy controls (Fig. 1A).Of the various discriminatory metabolites (Fig. 1B), Succinate had the highest discriminatory potential (AUC 0.8, P=0.01) followed by citrate, glycine, glycerol, glucose, creatine and lactate. (Fig. 1C) Both glucose and creatine were decreased in IIM (Fig. 1D,E) and this was uniform across all types of IIM. However, glycine levels differed across different myositis subsets supporting the fact that they might differ in pathogenesis. (Fig. 1E) Amongst various serum biomarkers of muscle disease and damage, serum Aspartate Transaminase correlated with glutamate (r=0.6, p=0.01), and serum creatinine correlated negatively with glycerol (r-0.8, p=0.04),Biopsies of infectious polymyositis suggested difference in spectra from IIM (Fig. 2A). Trends were observed towards lower succinate and higher citrate levels suggesting metabolomics could possibly be useful to differentiate the two. Muscle of both patients with IIM with superadded infectious polymyositis also exhibited low succinate and elevated citrate.Conclusion:Muscle metabolomics of active myositis is distinctive. Amino acids and creatine are lower in diseases muscle suggesting active breakdown and loss, in turn explaining previous findings of low levels in serum in active disease. Certain metabolite composition differ in different types of myositis supporting different pathogenesis.Infectious polymyositis might exhibit different metabolome from IIM with potential as a biomarker though this needs to be confirmed in larger numbers.Disclosures:Funded by APLAR research grant 2017 awarded to Dr Latika Gupta.References:[1]Gupta L, Kumar D, Gulerai A, Kumar U, Misra R. “NMR-Based Serum, Urine and Muscle Metabolomics in Inflammatory Myositis for Diagnosis and Activity Assessment: Serum Metabolomics Can Differentiate Active from Inactive Myositis” Oral presentation at the ACR, Atlanta 2019.Acknowledgments:MSA and metabolomics supported by APLAR research grant.Disclosure of Interests:None declared

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Untargeted LC-HRMS-based metabolomics to identify novel biomarkers of metastatic colorectal cancer

Scientific Reports ◽

10.1038/s41598-019-55952-8 ◽

2019 ◽

Vol 9 (1) ◽

Cited By ~ 7

Author(s):

Ariadna Martín-Blázquez ◽

Caridad Díaz ◽

Encarnación González-Flores ◽

Daniel Franco-Rivas ◽

Cristina Jiménez-Luna ◽

...

Keyword(s):

Colorectal Cancer ◽

Mass Spectrometry ◽

Liquid Chromatography ◽

High Resolution ◽

Metastatic Colorectal Cancer ◽

High Resolution Mass Spectrometry ◽

Healthy Controls ◽

High Resolution Mass ◽

Novel Biomarkers ◽

Resolution Mass

AbstractColorectal cancer is one of the main causes of cancer death worldwide, and novel biomarkers are urgently needed for its early diagnosis and treatment. The utilization of metabolomics to identify and quantify metabolites in body fluids may allow the detection of changes in their concentrations that could serve as diagnostic markers for colorectal cancer and may also represent new therapeutic targets. Metabolomics generates a pathophysiological ‘fingerprint’ that is unique to each individual. The purpose of our study was to identify a differential metabolomic signature for metastatic colorectal cancer. Serum samples from 60 healthy controls and 65 patients with metastatic colorectal cancer were studied by liquid chromatography coupled to high-resolution mass spectrometry in an untargeted metabolomic approach. Multivariate analysis revealed a separation between patients with metastatic colorectal cancer and healthy controls, who significantly differed in serum concentrations of one endocannabinoid, two glycerophospholipids, and two sphingolipids. These findings demonstrate that metabolomics using liquid-chromatography coupled to high-resolution mass spectrometry offers a potent diagnostic tool for metastatic colorectal cancer.

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AB1242 A NOVEL BIOMARKER OF MMP-CLEAVED PROLARGIN IS ELEVATED IN PATIENTS WITH PSORIATIC ARTHRITIS COMPARED TO OTHER FIBRO-INFLAMMATORY DISEASES

Annals of the Rheumatic Diseases ◽

10.1136/annrheumdis-2020-eular.981 ◽

2020 ◽

Vol 79 (Suppl 1) ◽

pp. 1912-1913

Author(s):

D. Sinkeviciute ◽

A. Schlemmer ◽

E. Berg Schmidt ◽

A. C. Bay-Jensen ◽

M. Karsdal ◽

...

Keyword(s):

Psoriatic Arthritis ◽

Disease Activity ◽

Validation Cohort ◽

Full Time ◽

Healthy Controls ◽

Healthy Individuals ◽

Discovery Cohort ◽

Low Disease Activity ◽

Technical Validation ◽

Full Time Employee

Background:Psoriatic Arthritis (PsA) is a chronic inflammatory disease, characterized by involvement of skin, axial and peripheral skeleton. Prolargin is a class II small leucine-rich proteoglycan found to be expressed in connective tissues of patients with PsA, and previously suggested to be remodelled upon treatment. Fragments of prolargin could quantify tissue turnover in individuals with PsA and reflect pathological tissue changes in these patients.Objectives:This study aimed at developing an immunoassay targeting a neo-epitope of prolargin cleaved by matrix metalloproteinases (MMPs), named PROM; and measure PROM levels in serum from two cohorts of patients affected by PsA and healthy controls.Methods:Development of a novel immunoassay targeting a specific MMP-generated neo-epitope fragment of prolargin (PROM) together with technical validation was performed, and then evaluated in serum from two independent cohorts. The technical validation included inter- and intra-variation, linearity, spiking recovery, stability and specificity. Specificity was tested using an elongated peptide, a truncated peptide and a non-sense peptide. The Discovery Cohort consists of 13 healthy individuals and 11 PsA patients, mean age 58, 60.3% female and 100% caucasian. The Validation Cohort included 35 healthy individuals and 112 PsA patients with low disease activity included in a 24-week randomized, double-blind, placebo-controlled trial of 3g n-3 polyunsaturated fatty acids (PUFA), a cohort of patients diagnosed with PsA by the CASPAR criteria. These patients had a mean age of 50.8, 57.8 % female and 100 % caucasian. Clinical variables and serum samples were collected at baseline and after 24 weeks of follow-up. An unpaired t-test was used for evaluation of healthy individuals and patients affected by PsA, while a paired t-test was used for evaluation of treatment at baseline and after 24 weeks.Results:A technically robust and specific assay was developed. The inter- and intra-assay variation of PROM was determined as 14% and 4 % respectively. PROM showed a good dilution recovery, spiking recovery, and storage /freeze-thaw stability (All, 100%±20%). PROM showed to be specific towards the targeted sequence, and did not show any reactivity towards the truncated peptide, elongated peptide or non-sense peptide. In the Discovery Cohort, serum levels of PROM were increased in patients with PsA compared to healthy individuals (p=0.032, Figure 1A). This increase was confirmed by the Validation Cohort, where PsA patients were significantly increased compared to healthy individuals at baseline (p=0.002, Figure 1B). After 24 weeks, the levels of PROM were unchanged in the n-3 PUFA treated group.Figure 1.Conclusion:The novel biomarker PROM, reflecting connective tissue remodeling, is elevated in PsA patients compared to healthy controls in two independent cohorts. No significant association was found for PROM in a low disease activity group of PsA patients treated with n-3 PUFA.References:NoneAcknowledgments:We thank the Innovation Foundation and Danish Research Foundation for providing funding for this study.Disclosure of Interests:Dovile Sinkeviciute Grant/research support from: Industrial PhD Student, Employee of: Industrial PhD Student, Annette Schlemmer: None declared, Erik Berg Schmidt: None declared, Anne-Christine Bay-Jensen Shareholder of: Nordic Bioscience A/S, Employee of: Full time employee at Nordic Bioscience A/S., Morten Karsdal Shareholder of: Nordic Bioscience A/S., Employee of: Full time employee at Nordic Bioscience A/S., Jeppe Hagstrup Christensen: None declared, Salome Kristensen: None declared, Signe Holm Nielsen Employee of: Full time employee at Nordic Bioscience

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Poly-IgA Complexes and Disease Severity in IgA Nephropathy

Clinical Journal of the American Society of Nephrology ◽

10.2215/cjn.01300121 ◽

2021 ◽

pp. CJN.01300121

Author(s):

Xue Zhang ◽

Jicheng Lv ◽

Pan Liu ◽

Xinfang Xie ◽

Manliu Wang ◽

...

Keyword(s):

Mass Spectrometry ◽

Iga Nephropathy ◽

Immune Complex ◽

Healthy Controls ◽

Peking University ◽

Lower Egfr ◽

One Step ◽

Treatment Responses ◽

Severity Of The Disease ◽

Immune Complex Formation

Background and objectives: Poly-IgA immune complex formation and glomerular deposition play a key role in IgA nephropathy. Our study sought to develop a new methodology for one-step serological detection of poly-IgA levels. Design, setting, participants, and measurements: A novel ELISA method using recombinant CD89 as 'capturing' probe was established for detecting poly-IgA immune complex in the plasma. We applied semiquantitative measurements of these poly-IgA indices in patients recruited at Peking University First Hospital with IgA nephropathy or other kidney disease types, as compared to healthy controls. The longitudinal trend of the poly-IgA index, together with the association with pathological parameters and treatment responses were evaluated. Finally, we analyzed the molecular composition of poly-IgA complexes in patients by mass spectrometry. Results: Recombinant CD89-mounted ELISA plates specifically captured plasma poly-IgA. The levels of poly-IgA immune complex (26.7, IQR 17.1-42.6 units/ml) in IgA nephropathy were significantly higher than those in healthy (15.5, IQR 10.7- 20.0 units/ml; P<0.001), or non-IgA nephropathy disease controls (14.8, IQR 10.5-21.9 units/ml; P<0.001). Higher levels of poly-IgA immune complex were associated with lower eGFR and worse kidney outcome. Accuracy parameters and concordant statistics showed good discrimination between IgA nephropathy and healthy controls based on poly-IgA index levels (AUC, 0.78; 95% CI, 0.72-0.83; P<0.001), significantly outperforming galactose deficient-IgA1 levels (AUC, 0.70; P=0.05). Corticosteroid and immunosuppressant treatments lowered poly-IgA indices. Following a recombinant CD89-directed workflow in conjunction with mass spectrometry, we also analyzed the molecular compositions of IgA immune complex in IgA nephropathy patients. Conclusions: Higher level of recombinant CD89-bound poly-IgA immune complex was associated with the severity of the disease, as well as treatment response to steroids and immunosuppressants.

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Toward Noninvasive Diagnosis of IgA Nephropathy: A Pilot Urinary Metabolomic and Proteomic Study

Disease Markers ◽

10.1155/2016/3650909 ◽

2016 ◽

Vol 2016 ◽

pp. 1-9 ◽

Cited By ~ 11

Author(s):

Michaela Neprasova ◽

Dita Maixnerova ◽

Jan Novak ◽

Colin Reily ◽

Bruce A. Julian ◽

...

Keyword(s):

Mass Spectrometry ◽

Renal Biopsy ◽

Iga Nephropathy ◽

Invasive Procedure ◽

Pilot Project ◽

Urine Samples ◽

Renal Diseases ◽

Single Individual ◽

Healthy Controls ◽

Spot Urine

IgA nephropathy is diagnosed by renal biopsy, an invasive procedure with a risk of significant complications. Noninvasive approaches are needed for possible diagnostic purposes and especially for monitoring disease activity or responses to treatment. In this pilot project, we assessed the utility of urine samples as source of biomarkers of IgA nephropathy. We used spot urine specimens from 19 healthy controls, 11 patients with IgA nephropathy, and 8 renal-disease controls collected on day of renal biopsy. Urine samples were analyzed using untargeted metabolomic and targeted proteomic analyses by several experimental techniques: liquid chromatography coupled with mass spectrometry, immunomagnetic isolation of target proteins coupled with quantitation by mass spectrometry, and protein arrays. No single individual biomarker completely differentiated the three groups. Therefore, we tested the utility of several markers combined in a panel. Discriminant analysis revealed that combination of seven markers, three metabolites (dodecanal, 8-hydroxyguanosine, and leukotriene C4), three proteins (α1-antitrypsin, IgA-uromodulin complex, and galactose-deficient IgA1), and heparan sulfate, differentiated patients with IgA nephropathy from patients with other renal diseases and healthy controls. Future studies are needed to validate these preliminary findings and to determine the power of these urinary markers for assessment of responses to therapy.

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Identification of feature autophagy-related genes in patients with acute myocardial infarction based on bioinformatics analyses

Bioscience Reports ◽

10.1042/bsr20200790 ◽

2020 ◽

Vol 40 (7) ◽

Author(s):

Yajuan Du ◽

Enfa Zhao ◽

Yushun Zhang

Keyword(s):

Myocardial Infarction ◽

Acute Myocardial Infarction ◽

Cellular Response ◽

Recurrent Events ◽

Validation Cohort ◽

Recursive Feature Elimination ◽

Support Vector ◽

Svm Classifier ◽

Healthy Controls ◽

Discovery Cohort

Abstract Objective: To identify feature autophagy-related genes (ARGs) in patients with acute myocardial infarction (AMI) and further investigate their value in the diagnosis of AMI. Methods: Gene microarray expression data of AMI peripheral blood samples were downloaded from the GSE66360 dataset. The data were randomly classified into a discovery cohort (21 AMI patients and 22 healthy controls) and a validation cohort (28 AMI patients and 28 healthy controls). Differentially expressed ARGs between patients with AMI and healthy controls in the discovery cohort were identified using a statistical software package. Feature ARGs were screened based on support vector machine-recursive feature elimination (SVM-RFE), and an SVM classifier was constructed. Receiver operating characteristic (ROC) analysis was used to investigate the predictive value of the classifier, which was further verified in an independent external cohort. Results: A total of seven genes were identified based on SVM-RFE. The SVM classifier had an excellent discrimination ability in both the discovery cohort (area under the curve [AUC] = 0.968) and the validation cohort (AUC = 0.992), which was further confirmed in the GSE48060 dataset (AUC = 0.963). Furthermore, the SVM classifier showed outstanding discrimination between AMI patients with and without recurrent events in the independent external cohort (AUC = 0.992). The identified genes are mainly involved in the cellular response to autophagy, macroautophagy, apoptosis, and the FoxO signaling pathway. Conclusion: Our study identified feature ARGs and indicated their potential roles in AMI diagnosis to improve our understanding of the molecular mechanism underlying the occurrence of AMI.

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Breaking the cycle

Neurology Neuroimmunology & Neuroinflammation ◽

10.1212/nxi.0000000000000562 ◽

2019 ◽

Vol 6 (3) ◽

pp. e562 ◽

Cited By ~ 4

Author(s):

Arie R. Gafson ◽

Constantinos Savva ◽

Tom Thorne ◽

Mark David ◽

Maria Gomez-Romero ◽

...

Keyword(s):

Adverse Events ◽

Time Course ◽

Validation Cohort ◽

Tca Cycle ◽

Therapeutic Effects ◽

Healthy Controls ◽

Discovery Cohort ◽

Cross Sectional ◽

Abdominal Symptoms ◽

Tca Cycle Intermediates

ObjectiveTo infer molecular effectors of therapeutic effects and adverse events for dimethyl fumarate (DMF) in patients with relapsing-remitting MS (RRMS) using untargeted plasma metabolomics.MethodsPlasma from 27 patients with RRMS was collected at baseline and 6 weeks after initiating DMF. Patients were separated into discovery (n = 15) and validation cohorts (n = 12). Ten healthy controls were also recruited. Metabolomic profiling using ultra-high-performance liquid chromatography mass spectrometry (UPLC-MS) was performed on the discovery cohort and healthy controls at Metabolon Inc (Durham, NC). UPLC-MS was performed on the validation cohort at the National Phenome Centre (London, UK). Plasma neurofilament concentration (pNfL) was assayed using the Simoa platform (Quanterix, Lexington, MA). Time course and cross-sectional analyses were performed to identify pharmacodynamic changes in the metabolome secondary to DMF and relate these to adverse events.ResultsIn the discovery cohort, tricarboxylic acid (TCA) cycle intermediates fumarate and succinate, and TCA cycle metabolites succinyl-carnitine and methyl succinyl-carnitine increased 6 weeks following treatment (q < 0.05). Methyl succinyl-carnitine increased in the validation cohort (q < 0.05). These changes were not observed in the control population. Increased succinyl-carnitine and methyl succinyl-carnitine were associated with adverse events from DMF (flushing and abdominal symptoms). pNfL concentration was higher in patients with RRMS than in controls and reduced over 15 months of treatment.ConclusionTCA cycle intermediates and metabolites are increased in patients with RRMS treated with DMF. The results suggest reversal of flux through the succinate dehydrogenase complex. The contribution of succinyl-carnitine ester agonism at hydroxycarboxylic acid receptor 2 to both therapeutic effects and adverse events requires investigation.

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Serum metabolic fingerprinting of psoriasis and psoriatic arthritis patients using solid-phase microextraction—liquid chromatography—high-resolution mass spectrometry

Metabolomics ◽

10.1007/s11306-021-01805-3 ◽

2021 ◽

Vol 17 (7) ◽

Author(s):

Nikita Looby ◽

Anna Roszkowska ◽

Nathaly Reyes-Garcés ◽

Miao Yu ◽

Tomasz Bączek ◽

...

Keyword(s):

Mass Spectrometry ◽

Liquid Chromatography ◽

Fatty Acid ◽

Psoriatic Arthritis ◽

Solid Phase Microextraction ◽

High Resolution Mass Spectrometry ◽

Solid Phase ◽

Healthy Controls ◽

Serum Samples ◽

Resolution Mass

Abstract Introduction Psoriatic arthritis (PsA), an inflammatory arthritis that develops in individuals with psoriasis, is associated with reduced quality of life. Identifying biomarkers associated with development of PsA as well as with PsA disease activity may help management of psoriatic disease. Objectives To use metabolomic fingerprinting to determine potential candidate markers of disease conversion (psoriasis to PsA) and/or PsA activity. Methods A novel sample preparation protocol based on solid-phase microextraction (SPME) was used to prepare serum samples obtained from: (1) individuals with psoriasis, some of whom develop psoriatic arthritis (n = 20); (2) individuals with varying PsA activity (mild, moderate, severe; n = 10 each) and (3) healthy controls (n = 10). Metabolomic fingerprinting of the obtained extracts was performed using reversed-phase liquid chromatography coupled to high resolution mass spectrometry. Results Psoriasis patients who developed PsA had similar metabolomic profiles to patients with mild PsA and were also indistinguishable from patients with psoriasis who did not develop PsA. Elevated levels of selected long-chain fatty acids (e.g., 3-hydroxytetradecanedioic acid) that are associated with dysregulation of fatty acid metabolism, were observed in patients with severe PsA. In addition, 1,11-undecanedicarboxylic acid—an unusual fatty acid associated with peroxisomal disorders—was also identified as a classifier in PsA patients vs. healthy individuals. Furthermore, a number of different eicosanoids with either pro- or anti-inflammatory properties were detected solely in serum samples of patients with moderate and severe PsA. Conclusion A global metabolomics approach was employed to analyze the serum metabolome of patients with psoriasis, PsA, and healthy controls in order to examine potential differences in the biochemical profiles at a metabolite level. A closer examination of circulating metabolites may potentially provide markers of PsA activity.

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Serum miRNAs Are Pharmacodynamic Biomarkers Associated With Therapeutic Response in Pediatric Inflammatory Bowel Disease

Inflammatory Bowel Diseases ◽

10.1093/ibd/izaa209 ◽

2020 ◽

Vol 26 (10) ◽

pp. 1597-1606 ◽

Cited By ~ 2

Author(s):

Suruchi K Batra ◽

Christopher R Heier ◽

Lina Diaz-Calderon ◽

Christopher B Tully ◽

Alyson A Fiorillo ◽

...

Keyword(s):

Inflammatory Bowel Disease ◽

Clinical Response ◽

Therapeutic Response ◽

Validation Cohort ◽

Activity Index ◽

Healthy Controls ◽

Discovery Cohort ◽

Serum Mirnas ◽

Tnf Α ◽

Inflammatory Bowel

Abstract Background We sought to identify microRNAs (miRNAs) associated with response to anti-TNF-α or glucocorticoids in children with inflammatory bowel disease (IBD) to generate candidate pharmacodynamic and monitoring biomarkers. Methods Clinical response was assessed by Pediatric Crohn’s Disease Activity Index and Pediatric Ulcerative Colitis Activity Index. Quantitative real-time polymerase chain reaction via Taqman Low-Density Array cards were used to identify miRNAs in a discovery cohort of responders (n = 11) and nonresponders (n = 8). Seven serum miRNAs associated with clinical response to treatment, along with 4 previously identified (miR-146a, miR-146b, miR-320a, miR-486), were selected for further study. Candidates were assessed in a validation cohort of serum samples from IBD patients pre- and post-treatment and from healthy controls. Expression of miRNA was also analyzed in inflamed mucosal biopsies from IBD patients and non-IBD controls. Results Discovery cohort analysis identified 7 miRNAs associated with therapeutic response: 5 that decreased (miR-126, miR-454, miR-26b, miR-26a, let-7c) and 2 that increased (miR-636, miR-193b). In the validation cohort, 7 of 11 candidate miRNAs changed in the same direction with response to anti-TNF-α therapies, glucocorticoids, or both. In mucosal biopsies, 7 out of 11 miRNAs were significantly increased in IBD vs healthy controls. Conclusions Five candidate miRNAs associated with clinical response and mucosal inflammation in pediatric IBD patients were identified (miR-126, let-7c, miR-146a, miR-146b, and miR-320a). These miRNAs may be further developed as pharmacodynamic and response monitoring biomarkers for use in clinical care and trials.

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Genetic variations in miRNA binding site of TPST1 and ZG16B associated with prognosis for patients with colorectal cancer.

Journal of Clinical Oncology ◽

10.1200/jco.2013.31.15_suppl.3553 ◽

2013 ◽

Vol 31 (15_suppl) ◽

pp. 3553-3553 ◽

Cited By ~ 2

Author(s):

Byung Woog Kang ◽

Soo Jung Lee ◽

Yoo Jin Lee ◽

Jong Gwang Kim ◽

Yee Soo Chae ◽

...

Keyword(s):

Colorectal Cancer ◽

Target Genes ◽

Validation Cohort ◽

Disease Free Survival ◽

Stage Iv ◽

In Silico Analysis ◽

Recessive Model ◽

P Value ◽

Nucleotide Polymorphisms ◽

Discovery Cohort

3553 Background: MicroRNAs (miRNA) may play important roles in tumorigenesis by regulating the expressions of proto-oncogenes or tumor suppressor genes. Single nucleotide polymorphisms (SNP) located in the 3’-UTR of miRNA target genes might affect miRNA-mediated gene regulation, thereby contributing to the susceptibility or prognosis of cancer. Accordingly, the present study analyzed SNPs located in the putative miRNA-binding sites of the 3’-UTR of various genes and their impact on the prognosis for patients with colorectal cancer by altering miRNA binding efficiency. Methods: Eight hundred and thirty-one consecutive patients (discovery cohort, n=309; validation cohort, n=522) with curatively resected colorectal adenocarcinoma were enrolled in the present study. The genomic DNA was extracted from fresh colorectal tissue. One hundred fifty-seven SNPs were selected in Silico analysis for the current study, which was based on several miRNA database and HapMap database. The SNP genotyping was performed using the Sequenom MassARRAY. Results: The median age of all patients was 65 years, and 468 (56.3%) patients had colon cancer, while 363 (45.1%) patients had rectal cancer. The pathologic stages after the surgical resection were as follows: stage I (n=150, 18.1%), stage II (n=332, 40.0%), stage III (n=333, 40.1%), and stage IV (n=16, 1.9%). In the discovery cohort, 19 SNPs were identified as possible prognostic biomarkers in a multivariate survival analysis [disease-free survival (DFS) and/or overall survival (OS)] adjusted for age, preoperative CEA level, and pathologic stage. In the validation cohort, the TPST1 rs3757417T>G and ZG16B rs12373A>C were significantly associated with prognosis as same direction in the discovery cohort (discovery + validation cohort; TPST1 rs3757417T>G, DFS, p value=0.0007, OS, p value=0.0091 in recessive model; ZG16B rs12373A>C, DFS, p value=<0.0001, OS, p value=0.0009 in dominant model). Conclusions: The current study provides evidence that the rs3757417T>G and rs12373A>C polymorphism in the 3’-UTR of TPST1 and ZG16B, respectively, are possible prognostic biomarker for patients with colorectal cancer.

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