Reverse Phase Protein Array Reveals Correlation of Retinoic Acid Metabolism With Cardiomyopathy in Friedreich's Ataxia
| العنوان: | Reverse Phase Protein Array Reveals Correlation of Retinoic Acid Metabolism With Cardiomyopathy in Friedreich's Ataxia |
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| المؤلفون: | Lauren A. Hauser, Marek Napierala, Amanda Clark, Jennifer M. Farmer, Kimal Rajapakshe, David A. Lynch, Peining Xu, Clementina Mesaros, Dean P. Edwards, Yu-Yun Chen, Ian A. Blair, Shixia Huang, Cristian Coarfa, Jill S. Napierala |
| المصدر: | Molecular & Cellular Proteomics : MCP |
| بيانات النشر: | American Society for Biochemistry and Molecular Biology, 2021. |
| سنة النشر: | 2021 |
| مصطلحات موضوعية: | Male, Proteomics, Retinoic acid, Biochemistry, Analytical Chemistry, chemistry.chemical_compound, reverse phase protein array, Iron-Binding Proteins, Gene expression, RA, retinoic acid, Cells, Cultured, ROL, retinol, 0303 health sciences, frataxin, biology, LDH, lactate dehydrogenase, 030302 biochemistry & molecular biology, PDT, population doubling time, Reverse phase protein lysate microarray, Middle Aged, DE, differentially expressed, Aldehyde Oxidoreductases, H2O2, hydrogen peroxide, CTRL, control, Biomarker (medicine), biomarker, Female, medicine.symptom, Cardiomyopathies, Adult, IRB, Institutional Review Board, Ataxia, Adolescent, retinoid metabolism, ALDH1A3, aldehyde dehydrogenase family 1 member A3, Protein Array Analysis, UAB, University of Alabama at Birmingham, GAA, guanine–adenine–adenine, CHOP, Children's Hospital of Philadelphia, 03 medical and health sciences, Retinoids, Young Adult, FRDA, Friedreich's ataxia, medicine, Humans, RAL, retinal, Molecular Biology, 030304 developmental biology, AOX1, aldehyde oxidase 1, Aged, RPPA, reverse phase protein array, HL, hearing loss, Research, Friedreich's ataxia, FARS, Functional Assessment Rating Scale, FC, fold change, Fibroblasts, Molecular biology, Fold change, CMP, cardiomyopathy, FXN, frataxin, chemistry, ISTD, internal standard, Friedreich Ataxia, BCM, Baylor College of Medicine, Frataxin, biology.protein, integrins, Biomarkers |
| الوصف: | Identifying biomarkers is important for assessment of disease progression, prediction of symptom development, and determination of treatment effectiveness. While unbiased analyses of differential gene expression using next-generation sequencing methods are now routinely conducted, proteomics studies are more challenging because of traditional methods predominantly being low throughput and offering a limited dynamic range for simultaneous detection of hundreds of proteins that drastically differ in their intracellular abundance. We utilized a sensitive and high-throughput proteomic technique, reverse phase protein array (RPPA), to attain protein expression profiles of primary fibroblasts obtained from patients with Friedreich's ataxia (FRDA) and unaffected controls (CTRLs). The RPPA was designed to detect 217 proteins or phosphorylated proteins by individual antibody, and the specificity of each antibody was validated prior to the experiment. Among 62 fibroblast samples (44 FRDA and 18 CTRLs) analyzed, 30 proteins/phosphoproteins were significantly changed in FRDA fibroblasts compared with CTRL cells (p < 0.05), mostly representing signaling molecules and metabolic enzymes. As expected, frataxin was significantly downregulated in FRDA samples, thus serving as an internal CTRL for assay integrity. Extensive bioinformatics analyses were conducted to correlate differentially expressed proteins with critical disease parameters (e.g., selected symptoms, age of onset, guanine–adenine–adenine sizes, frataxin levels, and Functional Assessment Rating Scale scores). Members of the integrin family of proteins specifically associated with hearing loss in FRDA. Also, RPPA data, combined with results of transcriptome profiling, uncovered defects in the retinoic acid metabolism pathway in FRDA samples. Moreover, expression of aldehyde dehydrogenase family 1 member A3 differed significantly between cardiomyopathy-positive and cardiomyopathy-negative FRDA cohorts, demonstrating that metabolites such as retinol, retinal, or retinoic acid could become potential predictive biomarkers of cardiac presentation in FRDA. Graphical abstract Highlights • RPPA analyzing Friedreich's ataxia cells reveals changed expression of 30 proteins. • Integrin expression correlates with hearing loss in Friedreich's ataxia samples. • Retinol levels are lower in serum of patients with Friedreich's ataxia than controls. In Brief We conducted a sensitive and high-throughput reverse phase protein array to attain protein expression profiles of primary fibroblasts from patients with Friedreich's ataxia (FRDA) and unaffected controls using a pool of 217 validated antibodies. Our extensive bioinformatics analyses correlated differentially expressed (DE) proteins with critical disease parameters. Expression levels of several integrin proteins specifically associated with hearing loss in FRDA. Also, reverse phase protein array data integrated with transcriptome data uncovered defects in retinoic acid metabolism in FRDA samples. |
| اللغة: | English |
| تدمد: | 1535-9484 1535-9476 |
| الوصول الحر: | https://explore.openaire.eu/search/publication?articleId=doi_dedup___::239d5431dbcbfd7df7c1a470624aee8dTest http://europepmc.org/articles/PMC8214145Test |
| حقوق: | OPEN |
| رقم الانضمام: | edsair.doi.dedup.....239d5431dbcbfd7df7c1a470624aee8d |
| قاعدة البيانات: | OpenAIRE |
| تدمد: | 15359484 15359476 |
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