Discovery of Pancreatic Ductal Adenocarcinoma-Related Aberrant Glycosylations: A Multilateral Approach of Lectin Microarray-Based Tissue Glycomic Profiling With Public Transcriptomic Datasets
Discovery of Pancreatic Ductal Adenocarcinoma-Related Aberrant Glycosylations: A Multilateral Approach of Lectin Microarray-Based Tissue Glycomic Profiling With Public Transcriptomic Datasets
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DOI：
10.3389/fonc.2020.00338
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发表时间：
2020-03
影响因子：
4.7
通讯作者：
Takanori Wagatsuma;Chiaki Nagai-Okatani;A. Matsuda;Y. Masugi;M. Imaoka;K. Yamazaki;M. Sakamoto
中科院分区：
文献类型：
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作者：
Takanori Wagatsuma;Chiaki Nagai-Okatani;A. Matsuda;Y. Masugi;M. Imaoka;K. Yamazaki;M. Sakamoto
异常蛋白质糖基化是癌组织中最显著的特征之一，因此具有疾病相关糖基化改变的糖蛋白是开发生物标志物和治疗剂的迷人目标。为此，需要一种可靠的策略来分析癌症进展过程中特定糖蛋白上发生的糖基化改变。在这里，我们提出了一个双边的方法相结合的凝集素微阵列为基础的组织糖组学分析和数据库衍生的转录组数据集。首先，使用凝集素微阵列对来自胰腺导管腺癌（PDAC）的冷冻组织切片的非肿瘤和肿瘤区域的粗提取物进行差异糖组学分析。该分析揭示了PDAC样本中两个显著的组织糖组改变：唾液酸化聚糖和二等分N-乙酰葡糖胺增加以及ABO血型抗原减少。为了检查与这些糖组学改变相关的糖基化机制中的畸变，我们接下来分别采用了癌症基因组图谱和基因型组织表达项目提供的癌性和正常胰腺中基因表达谱的公共数据集。在该分析中，负责糖基化改变的糖基转移酶在癌组织中显示异常基因表达，与组织糖组学谱一致。然后通过在肿瘤和非肿瘤胰腺细胞中表达的膜N-糖蛋白basigin的差异聚糖谱来评价糖基转移酶表达和组织糖组学的相关改变。来自PDAC组织切片的非肿瘤和癌性区域的内源性basigin的聚焦差异糖组学分析表明，basigin的PDAC相关聚糖改变密切反映了组织糖组中疾病特异性改变的显著特征。总之，目前使用公共转录组数据集的多组学策略和使用少量临床样本的实验糖组学分析成功证明，basigin是一种代表性的N-糖蛋白，反映了PDAC相关的异常糖基化。这项研究表明，大的公共数据集，如糖基化相关基因的基因表达谱的高度敏感的组织糖组学分析结果的评估的有用性。该策略有望用于发现新的糖生物标志物和糖治疗靶点。
Aberrant protein glycosylation is one of the most notable features in cancerous tissues, and thereby glycoproteins with disease-relevant glycosylation alterations are fascinating targets for the development of biomarkers and therapeutic agents. For this purpose, a reliable strategy is needed for the analysis of glycosylation alterations occurring on specific glycoproteins during the progression of cancer. Here, we propose a bilateral approach combining lectin microarray-based tissue glycomic profiling and database-derived transcriptomic datasets. First, lectin microarray was used to perform differential glycomic profiling of crude extracts derived from non-tumor and tumor regions of frozen tissue sections from pancreatic ductal adenocarcinoma (PDAC). This analysis revealed two notable tissue glycome alterations in PDAC samples: increases in sialylated glycans and bisecting N-acetylglucosamine and a decrease in ABO blood group antigens. To examine aberrations in the glycosylation machinery related to these glycomic alterations, we next employed public datasets of gene expression profiles in cancerous and normal pancreases provided by The Cancer Genome Atlas and the Genotype-Tissue Expression projects, respectively. In this analysis, glycosyltransferases responsible for the glycosylation alterations showed aberrant gene expression in the cancerous tissues, consistent with the tissue glycomic profiles. The correlated alterations in glycosyltransferase expression and tissue glycomics were then evaluated by differential glycan profiling of a membrane N-glycoprotein, basigin, expressed in tumor and non-tumor pancreatic cells. The focused differential glycomic profiling for endogenous basigin derived from non-tumor and cancerous regions of PDAC tissue sections demonstrated that PDAC-relevant glycan alterations of basigin closely reflected the notable features in the disease-specific alterations in the tissue glycomes. In conclusion, the present multi-omics strategy using public transcriptomic datasets and experimental glycomic profiling using a tiny amount of clinical specimens successfully demonstrated that basigin is a representative N-glycoprotein that reflects PDAC-related aberrant glycosylations. This study indicates the usefulness of large public data sets such as the gene expression profiles of glycosylation-related genes for evaluation of the highly sensitive tissue glycomic profiling results. This strategy is expected to be useful for the discovery of novel glyco-biomarkers and glyco-therapeutic targets.