Phosphoproteome and Biological Evidence Revealed Abnormal Calcium Homeostasis in Keloid Fibroblasts and Induction of Aberrant Platelet Aggregation
Phosphoproteome and Biological Evidence Revealed Abnormal Calcium Homeostasis in Keloid Fibroblasts and Induction of Aberrant Platelet Aggregation
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DOI：
10.1021/acs.jproteome.0c00984
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发表时间：
2021-03-12
影响因子：
4.4
通讯作者：
Wang, Tong
中科院分区：
文献类型：
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作者：
Yan, Ziqi;Zhang, Wanling;Wang, Tong
瘢痕疙瘩是一种良性肿瘤，其特征在于持续的炎症、增加的成纤维细胞增殖和伤口中胶原蛋白的异常沉积。瘢痕疙瘩的病因尚不清楚。在这里，我们探讨了磷酸化信号的变化，人类瘢痕疙瘩成纤维细胞通过磷酸化蛋白质组质谱分析。我们发现比较磷酸化蛋白质组学可以在统计学上区分瘢痕疙瘩和对照成纤维细胞。差异表达的磷蛋白可以预测已知的瘢痕疙瘩相关上游调节因子的激活，包括转化生长因子β 1、白细胞介素（IL）-4和IL-5。通过多种生物信息学分析，磷酸化FLNA、TLN 1和VCL在钙稳态和血小板聚集方面显著富集。我们从生物学上证实了瘢痕疙瘩成纤维细胞在离子霉素刺激后比对照成纤维细胞具有更高水平的Ca 2+内流。通过共培养分析，我们发现人瘢痕疙瘩成纤维细胞可以直接促进血小板聚集。如PhosphoPath和基因集富集分析所示，pFLNA集中为与瘢痕疙瘩表型相关的最高磷蛋白。我们验证了pFLNA在瘢痕疙瘩成纤维细胞和瘢痕疙瘩组织切片中都上调，暗示了其生物标志物的潜力。总之，我们首次报道了瘢痕疙瘩成纤维细胞的磷酸化蛋白质组，揭示了瘢痕疙瘩成纤维细胞具有异常的钙稳态，并可直接诱导血小板聚集。
Keloid is a benign tumor characterized by persistent inflammation, increased fibroblast proliferation, and abnormal deposition of collagen in the wound. The etiology of keloid is unclear. Here, we explored the phospho-signaling changes in human keloid fibroblasts via phosphoproteome mass spectrometry analysis. We found that comparative phosphoproteomics could statistically distinguish keloid from control fibroblasts. Differentially expressed phosphoproteins could predict the activation of known keloid-relevant upstream regulators including transforming growth factor-beta 1, interleukin (IL)-4, and IL-5. With multiple bioinformatics analyses, phosphorylated FLNA, TLN1, and VCL were significantly enriched in terms of calcium homeostasis and platelet aggregation. We biologically verified that keloid fibroblasts had a higher level of Ca2+ influx than the control fibroblasts upon ionomycin stimulation. Via co-cultivation analysis, we found that human keloid fibroblasts could directly promote platelet aggregation. As suggested by PhosphoPath and gene set enrichment analysis, pFLNA was centered as the top phosphoproteins associated with keloid phenotypes. We validated that pFLNA was upregulated both in keloid fibroblasts and keloid tissue section, implicating its biomarker potential. In conclusion, we reported the first phosphoproteome on keloid fibroblasts, based on which we revealed that keloid fibroblasts had aberrant calcium homeostasis and could directly induce platelet aggregation.