Meso-plex miRNA and protein profiling for cancer diagnostics using chip-integrate

描述(由申请人提供)：随着对疾病发生和发展的复杂生物分子机制的日益清晰的了解，临床肿瘤学准备通过应用多种分析工具来改善诊断、预后和治疗能力，从而实现个性化医学的前景。然而，一般而言，缺乏适当的技术来支持临床上的多重分析--特别是在与疾病相关的miRNA和蛋白质面板的分析方面，尽管它们的效用已明确确立。癌症发生和发展的复杂和异常机制只能通过测量多个生物标记物的签名来揭示，并且在miRNA和蛋白质水平上，已经识别了大量可能的生物标记物，当与其他标记物组合一起考虑时，这些生物标记物显示出更高的预测价值。然而，许多发现技术并不适用于临床。对于miRNAs，qRT-PCR分析非常敏感、相对快速和成本效益高，但每次分析只能定量单个靶标的表达。相反，微阵列很容易实现多路传输，但速度很慢，成本也很高。因此，迫切需要中间复合体诊断能力，从而可以使用快速、成本效益高和高度可扩展的技术同时询问由10个miRNAs组成的聚焦面板。同样，在分析能力方面也存在着显著的差距，这限制了多重蛋白质组学向临床的转化。金标酶联免疫吸附试验(EL ISA)通常非常敏感、选择性和成本效益高，尽管大多数情况下是单链。蛋白质微阵列具有高度的多重性，但通常灵敏度低得多，选择性低，而且不能适应临床环境。新兴的多重分析方法提供了一些改进，但尚未发现广泛的临床实用价值。芯片集成的硅光子传感器阵列最近作为一种内在的可扩展和多路复用的生物分子分析技术而出现，这一应用旨在有力地验证这一强大的中神经丛癌症诊断技术。具有多达128个可唯一寻址的传感器元件的硅光子微环谐振器阵列先前已被用于以多路分析格式并从复杂的、临床相关的样本矩阵中定量检测核酸和蛋白质特征。重要的是，这项技术起源于成熟的半导体加工方法，传感器阵列芯片可以按比例制造，以实现低成本的分析(1美元/件)。此外，这种方法的分子一般性将被用来同时分析同一临床样本的microRNA(MiRNA)和蛋白质表达-一种转化能力。尽管适用于任何癌症，但致命性脑癌多形性胶质母细胞瘤将成为实验室和临床模型。胶质母细胞瘤已有成熟的miRNA和蛋白质生物标志物，因此建议的努力完全停留在技术验证上。

DESCRIPTION (provided by applicant): Armed with an increasingly clear picture of the complex biomolecular mechanisms of disease onset and progression, clinical oncology is poised to realize the promise of personalized medicine by applying multiplexed analytical tools for improved diagnostic, prognostic, and theranostic capabilities. However, there are, in general, a lack of suitable technologies to support multiplexed analyses in the clinic-particularly with respect to the analysis of disease-relevant miRNA and protein panels, despite their clearly established utility. Complex and aberrant mechanisms underlying cancer onset and progression can only be unraveled through the measurement of multiple biomarker signatures and at both the miRNA and protein level, a wealth of putative biomarkers have been identified that show enhanced predictive value when considered together with panels of other markers. However, many discovery technologies are not amenable to the clinic. For miRNAs, qRT-PCR assays are incredibly sensitive, relatively rapid, and cost effective, yet are only able to quantitte expression of a single target per assay. Conversely, microarrays are readily multiplexable, yet quite slow and expensive. Thus, there exists a pressing need for meso-plex diagnostic capabilities whereby focused panels of 10s of miRNAs can be simultaneously interrogated using rapid, cost effective, and highly scalable technologies. Again, there is a striking gap in analyticl capabilities that limit the translation of multiplexed proteomics into the clinic. The gold standar enzyme-linked immunosorbent assay (ELISA), is typically very sensitive, selective, and cost effective, though most often single-plex. Protein microarrays are highly multiplexable, but generally far less sensitive, less selective, and not amenable to the clinical setting. Emerging multiplexed analysis methodologies offer some improvements but have yet to find widespread clinical utility. Chip-integrated silicon photonic sensor arrays have recently emerged as an inherently scalable and multiplexable biomolecular analysis technology, and this application aims to robustly validate this powerful technology for meso-plex cancer diagnostics. Silicon photonic microring resonator arrays, having up to 128 uniquely address-able sensor elements, have been previously utilized to quantitatively detect nucleic acid and protein signatures in multiplexed assay formats and from within complex, clinically-relevant sample matrices. Importantly, the technology has its origin in well-established methods of semiconductor processing and sensor array chips can be scalably fabricated to allow low cost assays (<$1/measurement). Furthermore, the molecular generality of this methodology will be utilized to simultaneous profile micro-RNA (miRNA) and protein expression from the same clinical sample-a transformative capability. Although applicable to any cancer, the lethal brain cancer glioblastoma multiforme will be the laboratory and clinical model. Well-established miRNA and protein biomarkers exist for glioblastoma, thus keeping the proposed efforts entirely on technology validation.