Topic 359: A Device Prototype for Isolation of Melanoma Exosomes for Diagnostics and Treatment Monitoring

主题 359：用于诊断和治疗监测的黑色素瘤外泌体分离设备原型

• 批准号: 10019069
• 负责人: Annette Marleau
• 金额: $ 186.06万
• 依托单位: AETHLON MEDICAL, INC.
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-09-12 至 2021-09-15
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10019069
• 关键词: Affinity; CSPG4 gene; DNA; Devices; Diagnostic; Fiber; Filtration; Laboratories; Malignant Neoplasms; MicroRNAs; Monitor; Monoclonal Antibodies; Non-Malignant; Performance; Population; Protein Analysis; Sampling; System; Testing; Tumor-Derived; Work; clinical care; clinical diagnostics; design; exosome; experimental study; liquid biopsy; melanoma; novel; oncology; prototype

The proposed work aims to develop a pre-commercial prototype of an exosome isolation device to be used in clinical diagnostic laboratories. We will advance the device to facilitate exosome isolation for downstream liquid biopsy tests requiring analysis of protein, DNA, or microRNA, which rely upon acquisition of intact and pure exosomes. The device strategy will be designed to isolate total exosomes and then to separate discrete populations of tumor-derived exosomes (TEX) and non-malignant exosomes (non-TEX). The device strategy will be advanced for melanoma exosome isolation in the proposed experiments but is expected to be applicable for other cancers. To achieve this study's objectives, we will develop and optimize a filtration system incorporating the GNA Hemopurifier for isolation of total exosomes from biofluids (Specific Aim 1). The capabilities of the GNA Hemopurifier for isolating melanoma TEX and non-TEX from diverse biofluids will be characterized by evaluating exosome depletion performance (Specific Aim 2). We will then construct a novel device, the CSPG4 cartridge, for the isolation of TEX from total exosomes, followed by testing the final workflow from the starting biofluid sample to the final TEX isolation (Specific Aim 3). Successful completion of this project will produce an exosome isolation system that will be applicable for high throughput exosome isolation and for TEX and non-TEX separation from diverse biofluids of varying complexity in a clinical care setting in oncology.

提出的工作旨在开发一种外泌体分离装置的预商业原型，用于临床诊断实验室。我们将推进该设备，以促进需要分析蛋白质、DNA或microRNA的下游液体活检测试的外泌体分离，这些测试依赖于获得完整和纯的外泌体。该设备策略将被设计用于分离总外泌体，然后分离肿瘤衍生外泌体（TEX）和非恶性外泌体（non-TEX）的离散种群。该设备策略将在拟议的实验中用于黑色素瘤外泌体的分离，但预计将适用于其他癌症。为了实现本研究的目标，我们将开发并优化一种过滤系统，该过滤系统包含GNA Hemopurifier，用于从生物体液中分离总外泌体（Specific Aim 1）。GNA血液净化器从多种生物体液中分离黑色素瘤TEX和非TEX的能力将通过评估外泌体消耗性能来表征（Specific Aim 2）。然后，我们将构建一种新型设备，CSPG4试剂盒，用于从总外泌体中分离TEX，随后测试从起始生物流体样品到最终TEX分离的最终工作流程（特定目标3）。该项目的成功完成将产生一种外泌体分离系统，该系统将适用于高通量外泌体分离，以及肿瘤临床护理环境中不同复杂性的各种生物体液中TEX和非TEX的分离。