The proteomic landscape of extracellular vesicles derived from human intervertebral disc cells

IF 3.4 3区 医学 Q1 ORTHOPEDICS JOR Spine Pub Date : 2024-11-05 DOI:10.1002/jsp2.70007
Li Li, Hadil Al-Jallad, Aiwei Sun, Miltiadis Georgiopoulos, Rakan Bokhari, Jean Ouellet, Peter Jarzem, Hosni Cherif, Lisbet Haglund
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Abstract

Background

Extracellular vesicles (EVs) function as biomarkers and are crucial in cell communication and regulation, with therapeutic potential for intervertebral disc (IVD)-related low back pain (LBP). EV cargo is often affected by tissue health, which may affect the therapeutic potential. There is currently limited knowledge of how the cargo of IVD cell-derived EVs varies with tissue health and how differences in proteomic profile affect the predicted biological functions.

Methods

Our study purified EVs from human IVD cell conditioned media by size-exclusion chromatography. Nanoparticle tracking analysis was conducted to measure EV size and concentration. Transmission electron microscopy and Western blot were performed to examine EV structure and markers. Tandem mass tag-mass spectrometry was conducted to determine protein cargo.

Results

Most EVs were exosomes and intermediate microvesicles with an increasing amount linked to disease progression. Of the proteins detected, 88.6% were shared across the non-degenerate, mildly-degenerate, and degenerate samples. GO and KEGG analyses revealed that cargo from the mildly-degenerate samples was the most distinct, with the proteins in high abundance strongly associated with extracellular matrix (ECM) organization and structure. Shared proteins, highly expressed in the non-degenerate and degenerate samples, showed strong associations with cell adhesion, ECM–receptor interaction, and vesicle-mediated transport, respectively.

Conclusions

Our findings indicate that EVs from IVD cells from tissue with different degrees of degeneration share a majority of the cargo proteins. However, the level of expression differs with degeneration grade. Cargo from the mildly-degenerate samples exhibits the most differences. A better understanding of changes in EV cargo in the degenerative process may provide novel information related to molecular mechanisms underlying IVD degeneration and suggest new potential treatment modalities for IVD-related LBP.

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从人类椎间盘细胞中提取的细胞外囊泡的蛋白质组图谱。
背景:细胞外囊泡(EVs)具有生物标志物的功能,在细胞通讯和调节中起着关键作用,对与椎间盘(IVD)相关的腰背痛(LBP)具有治疗潜力。EV 货物通常会受到组织健康状况的影响,这可能会影响治疗潜力。目前,人们对 IVD 细胞衍生 EVs 的载体如何随组织健康状况而变化,以及蛋白质组谱的差异如何影响预测的生物功能了解有限:我们的研究通过大小排阻色谱法从人类 IVD 细胞条件培养基中纯化了 EVs。我们采用纳米粒子追踪分析来测量EV的大小和浓度。透射电子显微镜和 Western 印迹检查了 EV 的结构和标记。采用串联质量标签质谱法确定蛋白质货物:结果:大多数EV是外泌体和中间微囊泡,其数量的增加与疾病进展有关。在检测到的蛋白质中,88.6%是非变性、轻度变性和变性样本共有的。GO和KEGG分析显示,轻度变性样本中的货物最为独特,高丰度蛋白质与细胞外基质(ECM)的组织和结构密切相关。在非变性和变性样本中高表达的共有蛋白质分别与细胞粘附、ECM-受体相互作用和囊泡介导的运输密切相关:我们的研究结果表明,来自不同变性程度组织的 IVD 细胞的 EVs 共享大多数载货蛋白。结论:我们的研究结果表明,来自不同退化程度组织的 IVD 细胞的 EVs 分享了大部分载体蛋白。来自轻度变性样本的货物差异最大。更好地了解变性过程中EV载体的变化可能会提供与IVD变性的分子机制有关的新信息,并为IVD相关的腰椎间盘突出症提供新的潜在治疗方法。
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来源期刊
JOR Spine
JOR Spine ORTHOPEDICS-
CiteScore
6.40
自引率
18.90%
发文量
42
审稿时长
10 weeks
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