Biomarkers for haemic neoplasia in the blue mussel, mytilus trossulus

本研究的总体目标是开发贻贝血液瘤样疾病（HN）的生物标志物。这种疾病在不列颠哥伦比亚省海岸的土生螺（Mytilus trossulus）中有很高的流行率。虽然这种疾病可能是由病毒引起的，但有一些迹象表明，在污染物存在的情况下，个人对这种疾病的易感性会加剧。贻贝被广泛用作生物指示生物，并被评估为大温哥华地区（GVRD）接受污水处理厂环境监测计划的潜在监测工具。血液肿瘤是被评估的各种终点之一。我们的目标是开发高通量技术，如基因表达分析，以监测这种疾病。我们的假设是白血病贻贝血细胞（LMH）的细胞凋亡过程受损。我们建议通过将LMH置于各种细胞凋亡诱导剂中来验证这一假设。了解LMH中哪些通路失调将有助于确定这种疾病的潜在生物标志物。我们已经在另一项研究中监测促凋亡基因p53和p63/p73。为了扩大生物标记基因的数量，作为本提案的第二个目标，我们的目标是对抗凋亡基因MDM2进行测序，并制定定量聚合酶链反应方案，以便MDM2基因在LMH中的表达也可以被监测。监测多种生物标志物，包括特定途径的主要和拮抗剂，可以提供有关该途径潜在失调的更多信息。

The overall objective of this research is to develop biomarkers for a leukemia-like disease in mussels called haemic neoplasia (HN). This disease has a high prevalence in Mytilus trossulus, the species native to the British Columbia coast. Although the disease likely has a virus aetiology there are some indications that susceptibility of individuals to the disease is exacerbated in the presence of pollutants. The mussel is used widely as a bioindicator organism, and is being evaluated as a potential monitoring tool for the Greater Vancouver Regional District (GVRD) receiving environment-monitoring program for one wastewater treatment plant. Haemic neoplasia is one of the various endpoints being evaluated. Our goal is to develop high throughput techniques such as gene expression analysis for monitoring of this disease. Our hypothesis is that apoptosis processes are impaired in leukemic mussel haemocytes (LMH). We propose to test this hypothesis by subjecting LMH to various apoptosis inducing agents. Knowing which pathways are dysregulated in LMH will help identify potential biomarkers for this disease. We are already monitoring the pro-apoptotic genes p53 and p63/p73 in a separate study. To extend the number of biomarker genes, and as a second objective of this proposal, we aim to sequence MDM2, an anti-apoptotic gene, and develop a quantitative polymerase chain reaction protocol so that MDM2 gene expression in LMH can be monitored also. Monitoring multiple biomarkers that include both protagonists and antagonists of a particular pathway gives more information about potential dysregulation of that pathway.