Integrative gene-tissue microarray-based approach for identification of human disease biomarkers: application to amyotrophic lateral sclerosis
Integrative gene-tissue microarray-based approach for identification of human disease biomarkers: application to amyotrophic lateral sclerosis
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DOI：
10.1093/hmg/ddq232
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发表时间：
2010-08-15
影响因子：
3.5
通讯作者：
Karsten, Stanislav L.
中科院分区：
文献类型：
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作者：
Kudo, Lili C.;Parfenova, Liubov;Karsten, Stanislav L.
基因组学和蛋白质组学的进步使疾病相关基因和蛋白质的快速鉴定成为可能。挑战包括动物模型与人类疾病之间的生物学差异，DNA和蛋白质表达数据之间的高度不一致以及缺乏研究人类复杂疾病的实验模型。为了克服这些局限性，我们开发了一种综合方法，使用动物模型，死后人类材料和高通量微阵列方法相结合来鉴定肌萎缩性侧索硬化症（ALS）的新分子标记。我们使用激光捕获显微解剖结合微阵列来识别脊髓运动神经元或周围胶质细胞中发生的早期转录组变化。在症状前阶段采用转基因小鼠SOD1（G93A）和TAU（P301L）两种家族性运动神经元疾病模型。所鉴定的基因表达变化主要是模型特异性的。然而，两个模型中都有几个基因受到调控。在症状前SOD1（G93A）动物的外周血转录组中，使用定制设计的ALS微阵列检测鉴定基因作为临床生物标志物的相关性。为了确认所鉴定的基因与人类散发性ALS （SALS）的相关性，我们利用从人死后脊髓组织构建的组织微阵列，通过高通量免疫分析检测了选定的相应蛋白产物。通过这些实验鉴定的基因位于与家族性ALS/额颞叶痴呆相关的连锁区域，并在几个家庭中进行了测序。这项大规模的基因和蛋白表达研究指出了TAU-和sod1诱导的运动神经元变性的不同分子机制，发现了几种新的sals相关蛋白（CNGA3, CRB1, OTUB2, MMP14， SLK, DDX58, RSPO2）和推测的血液生物标志物，包括Nefh， Prph和Mgll。
Advances in genomics and proteomics permit rapid identification of disease-relevant genes and proteins. Challenges include biological differences between animal models and human diseases, high discordance between DNA and protein expression data and a lack of experimental models to study human complex diseases. To overcome some of these limitations, we developed an integrative approach using animal models, postmortem human material and a combination of high-throughput microarray methods to identify novel molecular markers of amyotrophic lateral sclerosis (ALS). We used laser capture microdissection coupled with microarrays to identify early transcriptome changes occurring in spinal cord motor neurons or surrounding glial cells. Two models of familial motor neuron disease, SOD1(G93A) and TAU(P301L), transgenic mice were used at the presymptomatic stage. Identified gene expression changes were predominantly model-specific. However, several genes were regulated in both models. The relevance of identified genes as clinical biomarkers was tested in the peripheral blood transcriptome of presymptomatic SOD1(G93A) animals using custom-designed ALS microarray. To confirm the relevance of identified genes in human sporadic ALS (SALS), selected corresponding protein products were examined by high-throughput immunoassays using tissue microarrays constructed from human postmortem spinal cord tissues. Genes that were identified by these experiments and located within a linkage region associated with familial ALS/frontotemporal dementia were sequenced in several families. This large-scale gene and protein expression study pointing to distinct molecular mechanisms of TAU- and SOD1-induced motor neuron degeneration identified several new SALS-relevant proteins (CNGA3, CRB1, OTUB2, MMP14, SLK, DDX58, RSPO2) and putative blood biomarkers, including Nefh, Prph and Mgll.