Urine Proteomics Profiling and Functional Characterization of Knee Osteoarthritis Using iTRAQ Technology

Abstract

Osteoarthritis (OA) is a degenerative chronic disease affecting the whole joint structures. With the increment in life expectancy and aging population, OA has become one of the largest socioeconomic burdens, associated with pain and loss of joint function. However, early laboratory tests of OA are still lacking. Therefore, new diagnostic tests for this disease are urgently needed. In this study, to gain an insight into the pathogenesis and the potential biomarkers of OA, we implemented a comparative urine proteomics study on OA patients and health people using iTRAQ-based mass spectrometry technology. Western blotting was used to validate the relative changes in urine protein levels for four of the identified proteins. We constructed a comprehensive urine proteome profile of the OA patients and identified 102 proteins differently changed in abundance. Forty-six proteins were upregulated and 56 proteins were significantly downregulated in OA patients. Furthermore, the proteins, COL-4, MMP9, adiponectin, and BBOX1 were validated through Western blots, which can serve as valuable candidate biomarkers and help to illustrate the pathogenesis of OA. These findings may provide clues for promising biomarkers for the early diagnosis and also offer a theoretical basis for the early treatment of OA.

Quantitative Mass Spectrometry-Based Proteomics for Biomarker Development in Ovarian Cancer

“…Stable isotope labeling uses the mass increase caused by the mass tags with incorporated stable isotopes to quantify peptides at the MS1 full scan (precursor ion) level or peptide fragments at the MS2 (production ion) scan level. There are several strategies to label peptides or proteins with stable isotopes: chemical labeling such as Isobaric Tags for Relative and Absolute Quantitation (iTRAQ) [ 97 , 100 , 101 , 102 ] and Tandem Mass Tags (TMT) [ 103 , 104 , 105 , 106 ], and metabolic labeling strategies such as Stable Isotope Labeling by Amino acids in Cell culture (SILAC) [ 107 , 108 , 109 , 110 ]. Data independent acquisition (DIA) methods can also be used for quantitative studies ( Figure 1 ).…”

Insights into the molecular landscape of osteoarthritis in human tissues

“…This resulted in the identification of 102 proteins that had significant differences in their abundances (46 upregulated and 56 downregulated in urine from osteoarthritis patients). Among these proteins, collagen type IV (COL-4), matrix metalloproteinase 9 (MMP9), adiponectin and gamma-butyrobetaine dioxygenase 1 (BBOX1) were highlighted as potential biomarkers for early diagnosis of osteoarthritis in urine [ 36 ].…”

Lifetime development changes in rats tracked by urinary proteome

“…More importantly, unlike blood, urine is not subjected to homeostatic control, and it accumulates small, sensitive, and early changes associated with systemic changes, some of which may be used as biomarkers1 (Gao, 2013). Urine proteomics has already been applied to various clinical studies (Rodríguez-Suárez et al, 2014; Zou and Sun, 2015), including studies on lung cancer (Chen et al, 2015; Wang et al, 2017; Zhang et al, 2015; Zhang et al, 2018), breast cancer (Beretov et al, 2015; Gajbhiye et al, 2016), bladder cancer (Duriez et al, 2017; Lei et al, 2013; Santoni et al, 2018), gastric cancer (Shimura et al, 2020), genitourinary cancer (Chen et al, 2019), and knee osteoarthritis (Xiao et al, 2019). Moreover, urine-filtered plasma proteins originate from distal organs, including the brain, etc.…”