Biotechnology Resource Center of Biomodular Multi scale Systems CBM2 for Precision Molecular Diagnostics

 描述（由申请人提供）：来自血液的循环标志物代表了一种令人兴奋的体外诊断方案，因为固定这些标志物的微创性质和血液中发现的大量标志物类型，如生物细胞、无细胞分子（蛋白质和无细胞DNA）和囊泡（纳米组装体，如外来体）。不幸的是，这些血液传播的标志物中的许多尚未在临床实践中有效地用于管理具有挑战性的疾病，例如癌症、传染病和中风。这种缺陷主要是由于以下事实引起的：疾病相关的血液标志物在混合人群中占绝大多数，由于缺乏用于其分离的有效平台和可以确定其分子生物学特性的系统，使得它们难以被发现和分析。 它们可能隐藏的结构变化。为了满足这一迫切需求，提出了一个新的生物技术资源中心（CBM 2），该中心由一个高度成就和多学科的团队组成，将产生用于从全血中选择循环标志物和处理疾病特异性分子特征的创新系统。设想的系统利用多个长度尺度（mm到nm）来影响系统提供的独特处理能力。该系统将处理全血（=1 mL），并将临床相关标记物浓缩至nL体积（>106富集因子），并使用固相连接酶检测反应（spLDR）从DNA和RNA分子中搜索各种序列变异，该反应在使用基于复制的技术在一个步骤中制造的数百万个聚合物柱上进行。 spLDR产品被电动地扫入纳米飞行管中，其识别基于分子依赖的电泳迁移率;单分子处理将使用纳米飞行管进行，检测以非光学方式进行。该系统将提供从单次抽血中选择所有临床相关标志物（细胞、无细胞DNA和外泌体）的能力，并以全自动方式保护这些标志物的相关信息，以允许将平台过渡到临床实践中。该研究将促进广泛的基础设施已经到位和积极的合作和服务项目中心组合。将通过由第二次建立信任措施和成员广泛的网络提供的引人注目的讲习班，促进向生物医学界传播新技术和提供培训。

 DESCRIPTION (provided by applicant): Circulating markers from blood represents an exciting in vitro diagnostic scenario because of the minimally invasive nature of securing these markers and the plethora of marker types found in blood, such as biological cells, cell-free molecules (proteins and cell-free DNA) and vesicles (nanometer assemblies such as exosomes). Unfortunately, many of these blood-borne markers have not been effectively utilized in clinical practice to manage challenging diseases such as cancer, infectious diseases and stroke to name a few. This deficiency has arisen primarily from the fact that disease-associated blood markers are a vast minority in a mixed population making them difficult to find and analyze due to the lack of efficient platforms for their isolation and systems that can determine the molecular structural variations they may harbor. To address this pressing need, a new Biotechnology Resource Center is proposed (CBM2), which consists of a highly accomplished and multidisciplinary team that will generate innovative systems for the selection of circulating markers from whole blood and process disease-specific molecular signatures. The envisioned system takes advantage of multiple length scales (mm-to-nm) to affect unique processing capabilities offered by the system. The system will process whole blood (=1 mL) and concentrate clinically relevant markers to nL volumes (>106 enrichment factor) and search for a variety of sequence variations from both DNA and RNA molecules using a solid-phase ligase detection reaction (spLDR) carried out on millions of polymer pillars fabricated in a single step using replication-based technologies. spLDR products are electrokinetically swept into nanometer flight tubes with their identification based on molecular-dependent electrophoretic mobilities; single-molecule processing will be carried out using nanometer flight tubes with detection performed non-optically. The system will provide the ability to select all clinically relevant markers (cells, cell-free DNA and exosomes) from a single blood draw and secure pertinent information from those markers in a fully automated fashion to allow transitioning the platform into clinical practice. The research will be facilitated by extensive infrastructure alreay in place and an aggressive Collaborative and Service Project Center portfolio. Novel technology dissemination and training to the biomedical community will be facilitated through compelling workshops offered by CBM2 and the members' extensive networks.