Proteomic profiling of human plasma extracellular vesicles identifies PF4 and C1R as novel biomarker in sarcopenia

IF 9.4 1区医学 Q1 GERIATRICS & GERONTOLOGY

Journal of Cachexia Sarcopenia and Muscle Pub Date : 2024-07-15 DOI:10.1002/jcsm.13539

Paula Aparicio, David Navarrete-Villanueva, Alba Gómez-Cabello, Tresa López-Royo, Enrique Santamaría, Joaquín Fernández-Irigoyen, Karina Ausín, Manuel Arruebo, Victor Sebastian, Germán Vicente-Rodríguez, Rosario Osta, Raquel Manzano

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引用次数: 0

Abstract

Background

Sarcopenia, the gradual and generalized loss of muscle mass and function with ageing, is one of the major health problems in older adults, given its high prevalence and substantial socioeconomic implications. Despite the extensive efforts to reach consensus on definition and diagnostic tests and cut-offs for sarcopenia, there is an urgent and unmet need for non-invasive, specific and sensitive biomarkers for the disease. Extracellular vesicles (EVs) are present in different biofluids including plasma, whose cargo reflects cellular physiology. This work analysed EV proteome in sarcopenia and robust patients in the search for differentially contained proteins that can be used to diagnose the disease.

Methods

Plasma small EVs (sEVs) from a total of 29 robust controls (aged 73.4 ± 5.6 years; 11 men and 18 women) and 49 sarcopenic patients (aged 82.3 ± 5.4 years; 15 men and 34 women) aged 65 years and older were isolated and their cargo was analysed by proteomics. Proteins whose concentration in sEVs was different between sarcopenic and robust patients were further validated using ELISA. The concentration of these candidates was correlated to the EWGSOP2 sarcopenia tests for low muscle strength and low physical performance, and receiver operating characteristic (ROC) curve analyses were carried out to evaluate their diagnostic power, sensitivity and specificity.

Results

Proteomic analysis identified 157 sEVs proteins in both sarcopenic and robust samples. Among them, 48 proteins had never been reported in the ExoCarta nor Vesiclepedia databases. Statistical analysis revealed eight proteins whose concentration was significantly different between groups: PF4 (log2 FC = 4.806), OIT3 (log2 FC = −1.161), MMRN1 (log2 FC = −1.982), MASP1 (log2 FC = −0.627), C1R (log2 FC = 1.830), SVEP1 (log2 FC = 1.295), VCAN (FC = 0.937) and SPTB (log2 FC = 1.243). Among them, platelet factor 4 (PF4) showed the lowest concentration while Complement C1r subcomponent (C1R) increased the most in sarcopenic patients, being these results confirmed by ELISA (P = 1.07E-09 and P = 0.001287, respectively). The concentrations of candidate proteins significantly correlated with EWGSOP2 tests currently used. ROC curve analysis showed an area under the curve of 0.8921 and 0.7476 for PF4 and C1R, respectively. Choosing the optimal for PF4, 80% sensitivity and 85.71% specificity was reached while the optimal cut-off value of C1R would allow sarcopenia diagnosis with 75% sensitivity and 66.67% specificity.

Conclusions

Our results support the determination of EV PF4 and C1R as plasma diagnostic biomarkers in sarcopenia and open the door to investigate the role of the content of these vesicles in the pathogeny of the disease.

Abstract Image

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人体血浆细胞外囊泡的蛋白质组分析发现 PF4 和 C1R 是肌少症的新型生物标记物。

背景：肌肉疏松症是一种随着年龄增长而逐渐和普遍出现的肌肉质量和功能丧失的疾病，是老年人的主要健康问题之一，因为其发病率高，对社会经济有重大影响。尽管人们为肌少症的定义、诊断测试和临界值达成了广泛共识，但对该疾病的非侵入性、特异性和敏感性生物标志物的需求仍十分迫切。细胞外囊泡 (EV) 存在于包括血浆在内的不同生物流体中，其载体反映了细胞的生理机能。这项研究分析了肌肉疏松症和健壮患者体内的EV蛋白质组，以寻找可用于诊断该疾病的不同蛋白质：方法：分离了29名壮年对照组患者（年龄为73.4 ± 5.6岁，男性11人，女性18人）和49名65岁及以上的肌肉疏松症患者（年龄为82.3 ± 5.4岁，男性15人，女性34人）的血浆小EVs（sEVs），并通过蛋白质组学分析了它们的载体。使用 ELISA 方法进一步验证了肌肉疏松患者和健壮患者 sEV 中浓度不同的蛋白质。这些候选蛋白的浓度与 EWGSOP2 肌肉疏松症测试（低肌力和低体能）相关，并进行了接收器操作特征曲线（ROC）分析，以评估其诊断能力、灵敏度和特异性：蛋白质组分析在肌少症和健壮样本中发现了 157 个 sEVs 蛋白质。其中有 48 个蛋白质从未在 ExoCarta 或 Vesiclepedia 数据库中出现过。统计分析显示，有 8 种蛋白质的浓度在不同组间存在显著差异：PF4（log2 FC = 4.806）、OIT3（log2 FC =-1.161）、MMRN1（log2 FC =-1.982）、MASP1（log2 FC =-0.627）、C1R（log2 FC = 1.830）、SVEP1（log2 FC = 1.295）、VCAN（FC = 0.937）和SPTB（log2 FC = 1.243）。其中，血小板因子 4（PF4）的浓度最低，而补体 C1r 亚组分（C1R）在肌无力患者中的浓度增加最多，ELISA 证实了这些结果（P = 1.07E-09 和 P = 0.001287）。候选蛋白质的浓度与目前使用的 EWGSOP2 检测方法有明显的相关性。ROC 曲线分析显示，PF4 和 C1R 的曲线下面积分别为 0.8921 和 0.7476。选择 PF4 的最佳值，灵敏度为 80%，特异性为 85.71%，而选择 C1R 的最佳截断值，则可诊断肌肉疏松症，灵敏度为 75%，特异性为 66.67%：我们的研究结果支持将 EV PF4 和 C1R 确定为肌肉疏松症的血浆诊断生物标志物，并为研究这些囊泡内容物在疾病病因中的作用打开了大门。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Journal of Cachexia Sarcopenia and Muscle MEDICINE, GENERAL & INTERNAL-

CiteScore

13.30

自引率

12.40%

发文量

234

审稿时长

16 weeks

期刊介绍： The Journal of Cachexia, Sarcopenia and Muscle is a peer-reviewed international journal dedicated to publishing materials related to cachexia and sarcopenia, as well as body composition and its physiological and pathophysiological changes across the lifespan and in response to various illnesses from all fields of life sciences. The journal aims to provide a reliable resource for professionals interested in related research or involved in the clinical care of affected patients, such as those suffering from AIDS, cancer, chronic heart failure, chronic lung disease, liver cirrhosis, chronic kidney failure, rheumatoid arthritis, or sepsis.