Tethered Cationic Lipoplex Nanoparticle Assay for Liver Cancer Detection and Surv

描述（由申请人提供）：肝癌是全球癌症死亡的主要原因之一，近年来在美国增长迅速。一个对病人友好的早期发现和监测方法将大大降低这种严重疾病的死亡率。依靠检测患者血液样品中的循环肿瘤细胞（CTC）或包封在外泌体中的细胞外RNA的“液体活检”具有实现这一目标的巨大潜力。但现有的检测方法还远远不够完美，因为CTC很罕见（每1 mL人血液约5-100个），而且外来体非常小（直径<100 nm）。此外，现有的方法都不能同时捕获和检测循环外泌体和CTC。我们最近开发了一种新的和低成本的方法“拴系阳离子脂质体纳米颗粒（TCLN）生物芯片”，这是第一次，可以实现这一目标。在TCLN中，分子信标（MB）被预先装载在脂质体纳米颗粒中，以捕获循环外泌体并检测来自患者血液或流体样品的封装的细胞外RNA，而无需额外的复杂性。在不稀释样品的情况下原位检测靶RNA导致 - 现有方法（例如qRT-PCR）无法实现的非常高的检测灵敏度。相同的生物芯片还可以通过lipoplex内化检测捕获的CTC中的细胞内生物标志物。使用miR-21和miR-181 b作为生物标志物的MYC转基因小鼠模型和HCC患者血液样本的初步数据非常有希望。TCLN可以扩展到多重阵列设计，以允许检测一组靶RNA。在这项提案中，我们计划评估其用于肝癌早期检测和监测的可行性。我们的主要目的是（1）比较来自不同疾病阶段的MYC转基因和miR-122 KO小鼠模型的肿瘤组织和血液样品中作为生物标志物的一组靶RNA，以评估使用TCLN测定进行早期癌症检测的可行性，和（2）鉴定肝癌中的靶microRNA/mRNA生物标志物，并使用来自健康人的血液样品进行初步研究。定义的肝癌患者组，以评估TCLN检测在临床上用于癌症的相关性。

DESCRIPTION (provided by applicant): Liver cancer is one of the leading causes of cancer deaths worldwide with fast growing rate in the US in recent years. A patient-friendly early detection and surveillance method would substantially reduce the mortality in this serious disease. 'Liquid biopsy' relying on detection of either circulating tumor cells (CTCs) or extracellular RNAs encapsulated in exosomes in patient blood samples has great potential to achieve this goal. But existing detection methods are far from perfection because CTCs are rare (~5-100 per 1 mL human blood) and exosomes are very small (<100 nm in diameter). Furthermore, none of the existing methods are able to simultaneously capture and detect both circulating exosomes and CTCs. We recently developed a novel and low-cost method 'Tethered Cationic Lipoplex Nanoparticle (TCLN) biochip' that, for the first time, may achieve this goal. In TCLN, molecular beacons (MBs) are pre-loaded in liposome nanoparticles to capture circulating exosomes and detect encapsulated extracellular RNAs from the patient blood or fluid sample without extra complicity. The in situ detection of target RNAs without diluting the sample leads to very high detection sensitivity not achievable by existing methods, e.g. qRT-PCR. The same biochip can also detect intracellular biomarkers in the captured CTCs by lipoplex internalization. Preliminary data in a MYC transgenic mouse model and HCC patient blood samples using miR-21 and miR-181b as biomarkers are very promising. TCLN can be extended to a multiplexing array design to allow the detection of a panel of target RNAs. In this proposal, we plan to evaluate its feasibility for liver cancer early detection and surveillance. Our primary objectives are (1) to compare a panel of target RNAs as biomarkers in tumor tissue and blood samples from MYC transgenic and miR-122 KO mouse models at different disease stages to evaluate the feasibility of using TCLN assay for early cancer detection, and (2) to identify target microRNA/mRNA biomarkers in liver cancer and to conduct a pilot study using blood samples from well-defined liver cancer patient groups to evaluate the relevancy of TCLN assay for cancer use in clinic.