Department of Otolaryngology, University of Adelaide, Adelaide, Australia.
Australian Cancer Research Foundation (ACRF) Genomics Facility, Centre for Cancer Biology, SA Pathology, Adelaide, Australia; School of Biological Sciences, University of Adelaide, Adelaide, Australia; School of Pharmacy and Medical Science, University of South Australia, Adelaide, Australia.
Rhinology. 2020 Jun 1;58(3):273-283. doi: 10.4193/Rhin19.403.
RNA sequencing (RNA-Seq) allows the characterization of a global transcriptomic signature in a least-biased fashion, but few studies have applied this method to investigate the pathophysiology of CRS.
We collected mucosal tissue samples from 6 CRS without nasal polyps (CRSsNP), 6 CRS with nasal polyps (CRSwNP), and 6 control patients. Additional matched polyp samples were collected from the 6 CRSwNP patients. RNA was extracted and sequenced on the Illumina HiSeq-2500. Differential gene expression and pathway analyses were performed.
CRSsNP showed evidence of upregulated interferon-mediated immunity, MHC-class-I mediated antigen presentation, CXCR3 binding, neutrophil chemotaxis and degranulation, and potential downregulation of genes related to cilia movement and production. CRSwNP polyp tissue showed upregulation of B-cell mediated immune responses, but reduced expression of genes related to epithelial morphogenesis and haemostasis. Polyps also showed a generalized reduction of positive gene regulation. The sinonasal transcriptomic signature was largely determined by tissue type (polyp versus mucosa) and disease phenotype, with minimal signal originating from the individual patient.
RNA-Seq is a useful tool to explore the complex pathophysiology of CRS. Our findings stress the importance of tissue selection in molecular research utilizing sinonasal tissue, and demonstrate the limitation of the sNP/wNP paradigm (and the importance of endotyping). On the other hand, classical CRSsNP/wNP disease phenotypes played some role in determining the global transcriptomic signature, and should not be hastily discarded. The value of RNA-Seq-described transcriptomic signatures in exploring endotypes is yet to be explored in future studies.
RNA 测序（RNA-Seq）允许以最少偏见的方式对全局转录组特征进行描述，但很少有研究应用这种方法来研究 CRS 的病理生理学。
我们从 6 名无鼻息肉的慢性鼻窦炎（CRSsNP）患者、6 名有鼻息肉的慢性鼻窦炎（CRSwNP）患者和 6 名对照患者中收集了粘膜组织样本。另外，从 6 名 CRSwNP 患者中收集了匹配的息肉样本。在 Illumina HiSeq-2500 上提取并测序 RNA。进行差异基因表达和通路分析。
CRSsNP 显示干扰素介导的免疫、MHC 类-I 介导的抗原呈递、CXCR3 结合、中性粒细胞趋化和脱颗粒的上调证据，以及与纤毛运动和产生相关的基因潜在下调。CRSwNP 息肉组织显示 B 细胞介导的免疫反应上调，但与上皮形态发生和止血相关的基因表达下调。息肉还表现出普遍的正向基因调控减少。鼻内转录组特征主要由组织类型（息肉与粘膜）和疾病表型决定，个体患者的信号很少。
RNA-Seq 是探索 CRS 复杂病理生理学的有用工具。我们的研究结果强调了在利用鼻内组织进行分子研究时选择组织的重要性，并证明了 sNP/wNP 范式的局限性（以及内型的重要性）。另一方面，经典的 CRSsNP/wNP 疾病表型在确定全局转录组特征方面发挥了一定作用，不应轻易摒弃。在未来的研究中，还需要探讨 RNA-Seq 描述的转录组特征在探索内型方面的价值。
