ULTRASENSITIVE NANOLASERS FOR EPIGENETICS INVESTIGATIONS

摘要：本项目的目标是开发一种新的无标记传感模式，能够检测和定量DNA甲基化。该传感器平台基于稀土掺杂的光学腔，形成纳米激光器，并结合基于受体的表面功能化。这种光学传感器可以在光学或电学领域进行检测，将被开发来检测和量化DNA序列中多个位置的CpG甲基化，而不使用PCR。RASS F1a和BRCA1中多个CpG甲基化位点的检测将展示其在卵巢癌早期检测和监测中的应用。外周血和肿瘤组织中的表观遗传标志物的分析在癌症的早期发现、预后和治疗方面显示出了希望。然而，目前的检测技术缺乏敏感性、特异性和重复性，阻碍了对这些标志物的分析。这项拟议的研究将开发一种全新的传感器平台，具有前所未有的检测表观遗传学变化的能力，并将其应用于卵巢癌的早期检测和监测，以及研究癌症进展的基本潜在机制。目前使用单一血清蛋白生物标记物筛查卵巢癌的技术还不成功。然而，越来越多的证据表明，使用多个标记物将对筛查和监测治疗效果有帮助。PI之前的纳米传感器研究已经导致了基于光学共振腔的无标记、单蛋白质分子传感技术的发展，该腔在缓冲液和血清中都起作用。有人建议，纳米激光传感器将用于检测血清DNA的表观遗传变化，为利用蛋白质和表观遗传变化检测和监测卵巢癌提供一个通用平台。

Abstract: The objective of this project is to develop a new label-free sensing modality capable of detecting and quantifying DNA methylation. The sensor platform is based on a rare- earth doped optical cavity, which forms a nanolaser, in combination with a receptor- based surface functionalization. This optical sensor, which can perform detection in either the optical or electrical domain, will be developed to detect and to quantify CpG methylation at multiple sites in DNA sequences without the use of PCR. Its application to early ovarian cancer detection and monitoring will be demonstrated with the detection of multiple CpG methylation sites in RASS F1A and BRCA1. Analysis of epigenetic markers in peripheral blood and tumor tissue has shown promise in the early detection, prognosis, and treatment of cancer. However, the analysis of these markers is hampered by the lack of sensitivity, specificity, and reproducibility of current detection techniques. This proposed research will develop a completely novel sensor platform with unprecedented capabilities for detecting epigenetic changes, and applies it to early ovarian cancer detection and monitoring, and to studying the fundamental underlying mechanisms of cancer progression. Current techniques to screen for ovarian cancer using single serum protein biomarkers have not been successful. However, there is growing evidence that use of multiple markers will have utility for screening and monitoring therapeutic efficacy. Previous nanosensor research by the PI has resulted in the development of label-free, single protein molecule sensing techniques based on optical resonant cavities which function in both buffer and serum. It is proposed that the nanolaser sensor will be useful in detecting epigenetic changes in serum DNA, providing a universal platform for detecting and monitoring ovarian cancer utilizing both protein and epigenetic changes.