Site-specific gene expression profiling as a novel strategy for unravelling keloid disease pathobiology.
Site-specific gene expression profiling as a novel strategy for unravelling keloid disease pathobiology.
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DOI：
10.1371/journal.pone.0172955
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发表时间：
2017
期刊：
影响因子：
3.7
通讯作者：
Bayat A
中科院分区：
文献类型：
--
作者：
Jumper N;Hodgkinson T;Paus R;Bayat A
瘢痕疙瘩病是一种纤维增生性皮肤肿瘤，其特征为异质性、过度胶原沉积和侵袭性局部浸润。缺乏有效的动物模型和对多种当前疗法的耐药性导致KD管理的临床结果不令人满意。为了从一个新的角度解决KD，我们首次应用了一种位点特异性原位显微切割和基因表达谱分析方法，通过结合激光捕获显微切割和转录组阵列。这里的目的是分析这种方法的效用相比，已建立的调查方法，包括全组织活检和单层细胞培养技术。本研究采用最先进的显微切割和基因表达谱分析技术，从无假设和特定区域的角度探讨KD。我们试图阐明特定瘢痕疙瘩病变部位之间的表达差异，并阐明显著失调的基因对瘢痕疙瘩病理生物学机制的潜在贡献，从而为未来对KD的探索性研究提供信息。在这里，我们强调了我们的原位显微切割策略在生成表达数据方面的优势，其灵敏度和准确性优于传统方法。这种方法学途径通过鉴定与上皮-间质转化、炎症和免疫调节有关的基因和上游调节因子，支持表皮在KD发病机制中的积极作用。我们描述了与TGF β介导的信号传导相关的对胶原沉积至关重要的真皮表达模式，这些模式以前在KD中未被检查过。此外，这项研究支持了先前提出的KD中存在癌症样干细胞群体，并探讨了基因失调对KD对常规治疗的抗性的可能贡献。通过这种创新的原位显微切割基因分析方法，我们提供了不同KD区域的更好定义的基因特征，从而解决KD异质性，通过转录指纹图谱促进与其他皮肤纤维化的鉴别诊断，并突出未来KD研究的关键领域。
Keloid disease (KD) is a fibroproliferative cutaneous tumour characterised by heterogeneity, excess collagen deposition and aggressive local invasion. Lack of a validated animal model and resistance to a multitude of current therapies has resulted in unsatisfactory clinical outcomes of KD management. In order to address KD from a new perspective, we applied for the first time a site-specific in situ microdissection and gene expression profiling approach, through combined laser capture microdissection and transcriptomic array. The aim here was to analyse the utility of this approach compared with established methods of investigation, including whole tissue biopsy and monolayer cell culture techniques. This study was designed to approach KD from a hypothesis-free and compartment-specific angle, using state-of-the-art microdissection and gene expression profiling technology. We sought to characterise expression differences between specific keloid lesional sites and elucidate potential contributions of significantly dysregulated genes to mechanisms underlying keloid pathobiology, thus informing future explorative research into KD. Here, we highlight the advantages of our in situ microdissection strategy in generating expression data with improved sensitivity and accuracy over traditional methods. This methodological approach supports an active role for the epidermis in the pathogenesis of KD through identification of genes and upstream regulators implicated in epithelial-mesenchymal transition, inflammation and immune modulation. We describe dermal expression patterns crucial to collagen deposition that are associated with TGFβ-mediated signalling, which have not previously been examined in KD. Additionally, this study supports the previously proposed presence of a cancer-like stem cell population in KD and explores the possible contribution of gene dysregulation to the resistance of KD to conventional therapy. Through this innovative in situ microdissection gene profiling approach, we provide better-defined gene signatures of distinct KD regions, thereby addressing KD heterogeneity, facilitating differential diagnosis with other cutaneous fibroses via transcriptional fingerprinting, and highlighting key areas for future KD research.