Detection of Melanoma by Canine Olfactory Receptors.

描述（申请人提供）：黑色素瘤是最严重的皮肤癌，早期诊断是对抗它的最好方法。区分良性色素性病变和早期黑色素瘤可能很困难。几个世纪以来，恶臭一直被用作疾病的症状。在检测阈值和检测癌症的能力方面，犬科动物的极端嗅觉能力的证据已经被记录在案。该项目的目标是直接证明犬嗅觉受体对黑色素瘤肿瘤标志物（挥发性化合物）的反应。核心假设是黑色素瘤组织含有独特的挥发物--可以被嗅觉受体识别的生物标志物。将通过气相色谱法和质谱法分析来自人类活检样本的黑色素瘤挥发物。该提案的主要创新之处在于为基于代谢改变的挥发性副产物的黑色素瘤临床诊断奠定基础。两侧对称的嗅上皮将被分成亚段。将含有从上皮左侧分离的受体的膜片注射到非洲爪蟾卵母细胞中，并通过高通量电生理学针对分离的黑素瘤生物标志物进行测试。将使用从右侧平行子部分分离的mRNA的微阵列分析来确定犬嗅觉受体的遗传鉴定。在鉴定之后，候选犬嗅觉受体将在异源表达系统中进行初步表征。识别黑色素瘤标志物的特定犬嗅觉受体的鉴定将使我们能够追求长期目标：开发一种安全和非侵入性的诊断工具-用于早期检测黑色素瘤的生物传感器。公共卫生相关性：黑色素瘤是最严重的皮肤癌，早期诊断是最好的策略，以打击它。该项目的目标是确定犬嗅觉受体识别挥发性化合物-生物标志物目前是人类黑色素瘤，以开发诊断筛选测定-生物传感器，其早期检测。

DESCRIPTION (provided by applicant): Melanoma is the most serious form of skin cancer and early diagnosis is the best way to combat it. Distinguishing benign pigmented lesions from early melanomas can be difficult. Smell or odor has been used as a symptom of disease for centuries. The evidence for the extreme olfactory ability of canines, both in terms of detection threshold and in terms of ability to detect cancer has been documented. The goal of this project is to directly demonstrate that canine olfactory receptors respond to melanoma tumor markers (volatile compounds). The central hypothesis is that melanoma tissue contains unique volatiles - biomarkers that can be recognized by olfactory receptors. Melanoma volatiles from human biopsy samples will be profiled by gas chromatography and mass spectrometry. The major innovation of this proposal is to lay foundation for the clinical diagnosis of melanoma based on volatile by-products of altered metabolism. The bilaterally symmetrical olfactory epithelium will be divided into subsections. Membrane sheets containing receptors isolated from the left side of the epithelium will be injected into Xenopus oocytes and tested by high throughput electrophysiology against isolated melanoma biomarkers. The genetic identify of the canine olfactory receptors will be determined using microarray analysis of mRNA isolated from the parallel subsections on the right side. After identification, candidate canine olfactory receptors will be pharmacologically characterized in heterologous expression system. Identification of the specific canine olfactory receptors that recognize melanoma markers will enable pursuit of our long-term objective: the development of a safe and non-invasive diagnostic tool - a biosensor for the early detection of melanoma. PUBLIC HEALTH RELEVANCE: Melanoma is the most serious form of skin cancer, and early diagnosis is the best strategy to combat it. The goal of this project is to identify canine olfactory receptors that recognize volatile compounds - biomarkers present is human melanoma in order to develop diagnostic screen assay - a biosensor, for its early detection.