Serum Detection of Medulloblastoma Metastasis

髓母细胞瘤（MB）是原始神经外胚层最常见的形式， 小儿中枢神经系统（CNS）肿瘤。MB的标准治疗包括手术 切除，然后化疗或放疗。癌细胞的扩散和随后的 转移是治疗失败的主要原因之一。MB转移的早期识别对于 及时正确的随访治疗，改善MB患者的预后。当前临床 检测MB转移的方法是：1）脊柱的磁共振成像（MRI），和2）细胞学 或通过腰椎穿刺收集的脑脊液（CSF）样本的流式细胞术分析 procedure.这两种方法对于转移的早期检测都不够敏感， 儿科患者。更重要的是，在资源匮乏的地区，这些服务不易获得。 如果没有早期发现，积极治疗儿科患者可能会导致过度治疗引起的CNS 损害因此，迫切需要用于检测稀有MB细胞的高灵敏度和稳健的方法 在CSF和血液中传播，特别是对于无法或有限获得成像和并发症的患者 医院程序。第一阶段STTR项目的目标是开发一种超灵敏的液体活检 技术捕获MB患者血液样本中的MB细胞， 培训和资源。虽然已经开发了各种循环癌细胞（CTC）检测方法， 当前技术的灵敏度和特异性受到检测之间的不希望的相互作用的限制 系统和血液样品中的非特异性生物材料和细胞，例如，蛋白质冠的形成， 将非癌细胞吸附在细胞捕获装置的表面上。这种生物污染效应不仅 减弱靶向配体的亲和力，导致生物标志物特异性的丧失，但也产生不期望的 背景，导致不准确的结果。因此，目前没有可用的CTC检测方法 检测血液中的MB扩散该STTR项目将通过几项专利解决这些问题 技术，包括：（1）抗生物污损涂层聚合物，用于减少蛋白质冠和非特异性细胞 摄取以改善生物标记物靶向和细胞捕获的特异性，和（2）高磁性磁性铁 氧化物纳米棒（IONR）用于有效的免疫磁性细胞捕获。该项目旨在：（1）开发和 优化作为细胞捕获剂的抗生物污染超磁IONR;（2）确定和优化 所开发的IONR用于捕获GFP的血液中的循环GFP标记的MB细胞的有效性 转基因MB小鼠;和（3）测试用于检测MB细胞的开发的免疫磁性捕获技术 临床确诊MB转移患者的血液样本中。如果成功， MB转移的早期检测将在第二阶段进一步开发，并将产品用于实践- 改变对MB患者临床管理的影响，特别是那些在低资源环境中的患者。

PROJECT SUMMARY Medulloblastoma (MB) is the most common form of primitive neuroectodermal tumor of the pediatric central nervous system (CNS). The standard treatment of MBs includes surgical resection, followed by chemo- or radiation treatments. Dissemination of cancer cells and subsequent metastasis is one of the major causes of treatment failure. Early identification of MB metastases is important in applying timely and proper follow-up treatment and improving outcome of MB patients. Current clinical methods for detection of MB metastases are: 1) magnetic resonance imaging (MRI) of spine, and 2) cytology or flow cytometry analysis of cerebrospinal fluid (CSF) samples collected through the lumbar puncture procedure. Neither method is sensitive enough for the early detection of metastases and well-tolerated by pediatric patients. More importantly, they are not easily accessible and available in low-resource areas. Without early detection, aggressively treating pediatric patients can lead to over-treatment caused CNS damage. Therefore, there is a critical need in highly sensitive and robust methods for detecting rare MB cells spread in CSF and blood, especially for patients having no or limited access to imaging and complicated hospital procedures. The goal of this phase I STTR project is to develop an ultra-sensitive liquid biopsy technology to capture MB cells in blood samples of MB patients, with the readout simply done with minimal training and resources. While various circulating cancer cells (CTCs) detection methods have been developed, sensitivity and specificity of current technologies are limited by the unwanted interactions between detecting systems and non-specific biomaterials and cells in blood samples, e.g., the formation of protein corona and adsorption of non-cancerous cells on the surface of cell capturing devices. Such biofouling effect not only attenuates the affinity of targeting ligands, causing loss of biomarker specificity, but also generates undesirable background, leading to inaccurate results. Consequently, there is no CTC detection method currently available to detect MB spread in the blood. This STTR project will address these problems with several patented technologies, including: (1) anti-biofouling coating polymer for reducing protein corona and non-specific cell uptake to improve biomarker targeting and specificity of cell capture, and (2) high magnetism magnetic iron oxide nanorods (IONRs) for efficient immuno-magnetic cell capturing. The project aims to: (1) develop and optimize an anti-biofouling ultra-magnetic IONR as a cell capturing agent; (2) determine and optimize the effectiveness of developed IONRs for capturing circulating GFP tagged MB cells in the blood of GFP transgenic MB mice; and (3) test the developed immuno-magnetic capturing technology for detecting MB cells in blood samples from patients with clinically confirmed MB metastases. If successful, proposed technology for early detection of MB metastasis will be further developed in the Phase II with a product to make a practice- changing impact on the clinical management of MB patients, especially those in low-resource settings.