Tougan Takahiro, Onda Hiroaki, Okuzaki Daisuke, Kobayashi Shigeto, Hashimoto Hiroshi, Nojima Hiroshi
Department of Molecular Genetics, Osaka University, Osaka 565-0871, Japan.
DNA Res. 2008 Apr 30;15(2):103-14. doi: 10.1093/dnares/dsm035. Epub 2008 Feb 7.
DNA diagnostics are useful but are hampered by difficult ethical issues. Moreover, it cannot provide enough information on the environmental factors that are important for pathogenesis of certain diseases. However, this is not a problem for RNA diagnostics, which evaluate the expression of the gene in question. We here report a novel RNA diagnostics tool that can be employed with peripheral blood mononuclear cells (PBMCs). To establish this tool, we identified 290 genes that are highly expressed in normal PBMCs but not in TIG-1, a normal human fibroblast cell. These genes were entitled PREP after predominantly expressed in PBMC and included 50 uncharacterized genes. We then conducted PREP gene-focused microarray analysis on PBMCs from seven cases of Churg-Strauss syndrome (CSS), which is a small-vessel necrotizing vasculitis. We found that PREP135 (coactosin-like protein), PREP77 (prosaposin), PREP191 (cathepsin D), PREP234 (c-fgr), and PREP136 (lysozyme) were very highly up-regulated in all seven CSS patients. Another 28 genes were also up-regulated, albeit more moderately, and three were down-regulated in all CSS patients. The nature of these up- and down-regulated genes suggest that the immune systems of the patients are activated in response to invading microorganisms. These observations indicate that focused microarray analysis of PBMCs may be a practical, useful, and low-cost bedside diagnostics tool.
DNA诊断很有用,但受到棘手的伦理问题的阻碍。此外,它无法提供足够的关于某些疾病发病机制中重要的环境因素的信息。然而,这对RNA诊断来说不是问题,RNA诊断评估的是相关基因的表达。我们在此报告一种可用于外周血单个核细胞(PBMC)的新型RNA诊断工具。为建立此工具,我们鉴定出290个在正常PBMC中高表达但在正常人类成纤维细胞TIG-1中不表达的基因。这些基因在PBMC中主要表达后被命名为PREP,其中包括50个未表征的基因。然后,我们对7例变应性肉芽肿性血管炎(CSS,一种小血管坏死性血管炎)患者的PBMC进行了聚焦PREP基因的微阵列分析。我们发现PREP135(肌动蛋白结合蛋白样蛋白)、PREP77(prosaposin)、PREP191(组织蛋白酶D)、PREP234(c-fgr)和PREP136(溶菌酶)在所有7例CSS患者中均高度上调。另外28个基因也有上调,不过程度较轻,且有3个基因在所有CSS患者中下调。这些上调和下调基因的性质表明患者的免疫系统因入侵微生物而被激活。这些观察结果表明,对PBMC进行聚焦微阵列分析可能是一种实用、有用且低成本的床边诊断工具。