Distinguishing Pancreatic Cancer from Benign Pancreatic Disease using Nanoparticle-based Biomarkers

项目摘要 该项目评估使用由肿瘤来源的外切体和其他细胞器携带的生物标记物的效果。 细胞外小泡在胰腺癌与胰腺良性病变鉴别诊断中的应用 疾病。我们将使用基于高导介电泳法的技术同时回收不同的 这些类型的癌症衍生纳米颗粒来自容量有限的患者血浆的单个样本。这将是 允许获取数量有限的早期癌症样本。这项研究将产生新的知识 支持开发基于纳米颗粒的早期和晚期胰腺癌生物标记物小组 并将支持将基于纳米颗粒的诊断引入临床环境的基本原理。 随着高分辨率CT和MRI的使用，胰腺囊肿的发现频率也在增加 影像增加，高达9.3%的患者有无症状的囊肿。一种有创的内窥镜超声 传统上，引导下细针吸取活检(EUS/FNA)用于确定该囊肿是否是胰腺导管。 腺癌(PDAC)。这会带来高昂的财务成本和相关的健康风险，每100名患者中就有1人会 发展成急性胰腺炎，其中十分之一的患者会死亡。目前60%-76%的患者接受了 EUS/FNA没有胰腺癌。这意味着临床上尚未满足的需要是开发血液检测来分层 胰腺囊肿患者分为高概率和低概率两类，其中 极有可能受益于EUS/FNA。目前，还没有基于血液的PDAC生物标记物存在。肿瘤- 衍生的纳米颗粒为潜在的PDAC相关生物标志物提供了一个新的来源。挑战在于，传统的 每种纳米颗粒类型的纳米颗粒回收方法要求等离子体体积太大而不能 由目前可用的PDAC患者血浆样本支持。我们的初步数据现在表明， 电导介电(DEP)技术可以回收足够量的纳米颗粒 用于检测癌前病变以及区分早期和晚期PDAC与对照组和 只需要30微米L的血浆。每个目标将把我们的DEP对单个生物标记物的验证转化为一个小组 这将在由PDAC患者和良性胰腺疾病患者组成的盲法队列研究中进行评估。 我们将目标1集中在肿瘤主动释放的颗粒上，目标2集中在不同的细胞器碎片上。 AIM 3评估了这些生物标志物，并从我们的初步数据中评估了5个，分别在阶段1和阶段2的PDAC样本上。 这项研究的意义在于，所产生的知识将导致潜在的验证小组 纳米颗粒相关生物标记物能够将PDAC与良性胰腺疾病区分开来。这个 这里提出的研究可能最终支持基于纳米颗粒的临床开发的理论基础 诊断血液测试，以确定将受益于EUS/FNA程序的胰腺囊肿患者。

Project Summary This project evaluates the efficacy of using biomarkers carried by tumor-derived exosomes and other organelle- derived extracellular vesicles to differentiate patients with pancreatic cancer from patients with benign pancreatic disease. We will use high conductance dielectrophoresis-based technology to simultaneously recover different types of these cancer-derived nanoparticles from individual samples of volume limited patient plasma. This will enable access to volume restricted early stage cancer samples. This research will generate new knowledge supporting the development of a nanoparticle-based biomarker panel for early and late stage pancreatic cancer detection and will support the rationale to bring nanoparticle-based diagnostics to the clinical setting. Pancreatic cysts are coincidently discovered with increasing frequency as the use of high resolution CT and MRI imaging increases, with up to 9.3% of patients having an asymptomatic cyst. An invasive endoscopic ultrasound guided fine needle aspiration biopsy (EUS/FNA) is traditionally used to determine if the cyst is pancreatic ductal adenocarcinoma (PDAC). This has a high financial cost and associated health risk where 1/100 patients will develop acute pancreatitis and 1/10 of these patients will die. Currently 60-76% of patients that undergo EUS/FNA do not have pancreatic cancer. This signifies the unmet clinical need to develop a blood test to stratify patients with pancreatic cysts into categories of high-probability and low-probability for having PDAC, where high-probability would benefit from the EUS/FNA. Currently, no blood based PDAC biomarkers exist. Tumor- derived nanoparticles offer a new source of potential PDAC related biomarkers. The challenge is that traditional nanoparticle recovery methods for each nanoparticle type require plasma volumes that are too large to be supported with currently available PDAC patient plasma samples. Our preliminary data now suggest that high conductance dielectrophoresis (DEP) techniques can recover sufficient amounts of nanoparticle derived biomarkers to detect precancerous lesions as well as differentiate early and late stage PDAC from controls and requires only 30 µl of plasma. Each Aim will translate our DEP validation of individual biomarkers into a panel that will be evaluated in a blinded cohort study consisting of patients with PDAC and benign pancreatic disease. We focus Aim 1 on particles actively released by tumors, and Aim 2 on different cellular organelle fragments. Aim 3 evaluates these biomarkers, and five from our preliminary data, on stage 1 and 2 PDAC samples. The significance of this research is that the knowledge generated will lead to potential validation of a panel of nanoparticle-associated biomarkers capable of differentiating PDAC from benign pancreatic disease. The research proposed here may ultimately support the rationale for the clinical development of a nanoparticle-based diagnostic blood test to identify patients with pancreatic cysts that would benefit from the EUS/FNA procedure.