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.

拟议的工作旨在开发外泌体隔离装置的商业前原型，以用于临床诊断实验室。我们将推进该装置，以促进外泌体分离，以进行下游液体活检测试，需要分析蛋白质，DNA或microRNA，这些测试依赖于依赖完整和纯外泌体的获取。该装置策略将设计为隔离总外泌体，然后分离肿瘤来源的外泌体（TEX）和非恶性外泌体（非TEX）的离散群体。在拟议的实验中，将采用用于黑色素瘤外泌体隔离的设备策略，但预计将适用于其他癌症。为了实现这项研究的目标，我们将开发并优化融合GNA肢体疗法的过滤系统，以从生物流体中隔离总外泌体（特定的AIM 1）。通过评估外泌体耗竭性能，可以表征GNA肢体临床素从不同生物流体中分离出黑色素瘤和非TEXX的能力（特定的AIM 2）。然后，我们将构建一种新型设备CSPG4墨盒，以从总外泌体中隔离Tex，然后测试从起始生物流体样品到最终Tex隔离的最终工作流程（特定的AIM 3）。该项目的成功完成将产生外部隔离系统，该系统适用于高吞吐量外泌体隔离，以及在肿瘤学临床护理环境中不同复杂性的Tex和非Tex分离。