Dataset Information

Heterogeneity of proteome dynamics between connective tissue phases of adult tendon

ABSTRACT: Turnover of matrix proteins is essential to maintain tissue homeostasis, and dysregulation of protein turnover has been implicated in a variety of diseases. However, proteome dynamics in the musculoskeletal system, particularly tendon, remain relatively unexplored. The aim of this study was therefore to calculate the turnover rate of individual proteins within tendon and its constituent components. We hypothesised that turnover of proteins in the interfascicular matrix would be greater than in the fascicles. Achilles tendons were harvested from rats fed heavy labelled water over a period of 127 days. Proteins were extracted from one Achilles tendon, while laser capture microdissection was used to isolate regions of interfascicular and fascicular matrix from the other Achilles prior to protein extraction. Samples were analysed using tandem mass spectrometry and the rate of label incorporation used to calculate turnover rate of individual proteins. Results demonstrate complex proteome dynamics within tendon, with differences in protein turnover rates approaching 1000-fold. Collagen turnover was relatively slow, with much more rapid turnover of proteoglycans and glycoproteins. Data support the hypothesis, demonstrating overall faster protein turnover within the interfascicular matrix compared to the fascicles. More rapid turnover of the interfascicular matrix may be a mechanism to repair microdamage to this region which is exposed to high levels of shear during locomotion. The techniques used here provide a powerful approach to interrogate alterations in protein turnover in the musculoskeletal system with ageing and/or disease which are likely to influence injury risk.

INSTRUMENT(S): Q Exactive

ORGANISM(S): Rattus Norvegicus (rat)

TISSUE(S): Tendon

SUBMITTER: Chavaunne Thorpe

LAB HEAD: Chavaunne Thorpe

PROVIDER: PXD015928 | Pride | 2020-07-08

REPOSITORIES: Pride

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Heterogeneity of proteome dynamics between connective tissue phases of adult tendon.

Choi Howard H Simpson Deborah D Wang Ding D Prescott Mark M Pitsillides Andrew A AA Dudhia Jayesh J Clegg Peter D PD Ping Peipei P Thorpe Chavaunne T CT

eLife 20200512

Maintenance of connective tissue integrity is fundamental to sustain function, requiring protein turnover to repair damaged tissue. However, connective tissue proteome dynamics remain largely undefined, as do differences in turnover rates of individual proteins in the collagen and glycoprotein phases of connective tissue extracellular matrix (ECM). Here, we investigate proteome dynamics in the collagen and glycoprotein phases of connective tissues by exploiting the spatially distinct fascicular ...[more]

PMID: 32393437

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Project description:Healing process after connective tissues (CT) injury is complex but important as CT are critical for human moving, and patient outcome of CT injured patients is protracted with high individual variation. Specific markers which could be used for healing prognosis will be great help to monitor patient outcome, and furtherly improve target treatment and patient recovery. Although several common factors like age, gender and body mass index (BMI) were reported to predict ATR healing outcome [references], more specific markers which can be used as predictors of patient outcome after ATR are still lacking. Several biomarkers have been reported to associate with tendon healing, and indicated their relationship with patient outcome(1-4). These results highlight the potential of protein expression detection and analysis for tendon study in general and healing outcome prognosis in particular. Collagen type I (Col1a1) is the main component of Achilles tendon and plays vital role in tendon healing via involving in collagen fibrils production among tendon fibroblasts(5). Higher production of Col1a1 is associated with faster tendon repair(6), previous study on animal model reported the increased Col1a1 synthesis can exert a positive effect on tendon healing[?]. Thus, functional proteins with high impact on Col1a1 can be used not only to improve tendon healing, also can be as targets for healing improvement and outcome prediction. The proteome file of Achilles tendon which is important to increase understanding of tendon healing and help to identify accurate outcome-related biomarkers is however lagged behind. The identification of proteomic profile of Achilles tendon is very much limited in number of proteins and sample size(7-9), and is even few on human. Recent improvement in proteomic assay based on mass spectrometry (MS) made it possible to assemble the proteomic landscape of human tissues(10, 11) with high quality and sensitivity. To characterize the proteomic components in human Achilles tendon, we utilized biopsies from ATR patients with both good and poor 1-year healing outcome. A validated platform including quantitative MS and clinical database was used to identify proteomic landscape and potential bio-predictors of clinical outcome. Our study may provide a more comprehensive landscape of proteins for human Achilles tendon, as well as specific biomarkers which can be used for long term outcome prognosis after ATR.

Dataset Information

Heterogeneity of proteome dynamics between connective tissue phases of adult tendon

Publications

Heterogeneity of proteome dynamics between connective tissue phases of adult tendon.

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