Comparative Proteomic and Metabolomic Analyses of Plasma Reveal the Novel Biomarker Panels for Thyroid Dysfunction

Abstract

Background: Thyroid dysfunction, including hypothyroidism (THO) and hyperthyroidism (THE), commonly arise from pathological processes in the thyroid gland. The current diagnosis of thyroid dysfunction varies because of the age and sex of the patients. The aim of this study was to explore novel candidate biomarker panels for hypothyroidism and hyperthyroidism screening with mass spectrometry and bioinformatics. Methods: Plasma samples were collected from 15 THE patients, 9 THO patients, and 15 healthy controls. Data Independent Acquisition(DIA)-based proteomic and untargeted metabolomic analyses were performed to identify novel biomarker panels for THO and THE patients. Finally, three candidate biomarkers were verified by ELISA in 34 samples. Results: A total of 2738 proteins and 6103 metabolites were identified, and 173 proteins and 2487 metabolites were found to be differentially expressed among the THE, THO and control groups. The results of the ensemble feature selection, K-means clustering and least absolute shrinkage and selection operator (LASSO) regression model showed that two proteins (C4-A and C3/C5 convertase) combined with two metabolites (L-arginine and L-proline), and proteins (APOL1 and ITIH4) combined with metabolites (cortisol, and cortisone) identified by plasma proteomics and metabolomics could help distinguish THO and THE patients from healthy controls, respectively. Conclusions: This study identified and verified two pairs of biomarker panels that can be used to distinguish THE and THO patients regardless of age and sex. Consequently, our findings represent a comprehensive analysis of thyroid dysfunction plasma, which is significant for clinical diagnosis.

Serum metabolome analysis in hyperthyroid cats before and after radioactive iodine therapy

“…This mirrors ndings in experimentally induced and naturally occurring human hyperthyroidism, where alterations in metabolomic signatures persist despite returning to the euthyroid state. [12][13][14][15] The metabolomic signature of hyperthyroid cats in our study corroborated altered cholesterol and steroid hormonogenesis observed in hyperthyroid humans. [16][17][18] Key components of the mevalonate pathway, the hydroxy fatty acids mevalonate and mevalonolactone, are essential for cholesterol biosynthesis.…”

Integrated proteomic and metabolomic analysis of plasma reveals regulatory pathways and key elements in thyroid cancer

“…The proteomic analysis of the plasma samples was performed, as described previously. 17 SDS free lysate (7 M urea, 2 M thiourea, 20 mM Tris–HCl, pH 8.0) was added to 100 μL of the plasma sample, and finally to make up a total volume of 1 mL; then the proteins were reduced with 10 mM DTT for 30 minutes at 37 °C and alkylated with 55 mM iodoacetamide for 30 minutes at room temperature. Using a C18 SPE (solid phase extraction) column, protein enrichment was conducted and the eluate was frozen and dried for further treatment.…”

“…The extraction of metabolites followed the following steps. 17 After being thawed slowly at 4 °C, each plasma sample (100 μL) was placed in 96-well plates. Then, 300 μL of extraction solution (methanol : ACN = 2 : 1, v : v, −20 °C precooling) and 20 μL of the internal standard were added to each sample.…”

LC-MS/MS based metabolomics and proteomics reveal candidate biomarkers and molecular mechanism of early IgA nephropathy

“…Samples of plasma were processed according to the previously described procedure with modifications [ 22 ]. Briefly, 100 μg of the protein was purified by methanol/chloroform precipitation.…”