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

 描述(申请人提供)：血液循环标记物代表了一种令人兴奋的体外诊断方案，因为保护这些标记物的微创性质，以及血液中发现的过多的标记物类型，如生物细胞、无细胞分子(蛋白质和无细胞DNA)和囊泡(纳米组件，如外切体)。不幸的是，许多这些血液传播的标记物并没有被有效地用于临床实践，以管理具有挑战性的疾病，如癌症、传染病和中风等。这一缺陷主要是因为在混合人群中，与疾病相关的血液标志物是绝大多数，由于缺乏有效的分离平台和能够确定分子的系统，因此很难找到和分析它们 它们可能蕴藏着结构性的变化。为了满足这一迫切需求，建议成立一个新的生物技术资源中心(CBM2)，该中心由一支高度成功的多学科团队组成，将产生从全血中选择循环标志物的创新系统，并处理疾病特定的分子签名。设想的系统利用多个长度尺度(毫米到纳米)来影响由该系统提供的独特处理能力。该系统将处理全血(=1毫升)，并将临床相关标记浓缩到NL体积(&gt；106浓缩因子)，并使用固相连接酶检测反应(SpLDR)搜索DNA和RNA分子的各种序列变异，该反应采用基于复制的技术在一步制造的数百万个聚合物柱上进行。SPLDR产品被电动扫描到纳米飞行管中，基于分子相关的电泳迁移率进行识别；单分子加工将使用纳米飞行管进行，并进行非光学检测。该系统将提供从一次抽血中选择所有临床相关标记物(细胞、无细胞DNA和外切体)的能力，并以全自动方式从这些标记物中获得相关信息，从而使该平台能够过渡到临床实践。广泛的基础设施和积极的协作和服务项目中心产品组合将促进这项研究。将通过CBM2及其成员广泛的网络提供引人注目的讲习班，为生物医学界传播和培训新技术提供便利。

 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.