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.

这项拟议的工作旨在开发一种用于临床诊断实验室的外显子分离设备的商业化前原型。我们将改进该设备，以促进下游液体活组织检查需要分析蛋白质、DNA或microRNA的外切体分离，这些检测依赖于获取完整和纯净的外切体。该设备的策略将被设计成分离总的外切体，然后分离肿瘤来源的外切体(TeX)和非恶性外切体(Non-TeX)的离散群体。在拟议的实验中，该设备的策略将用于黑色素瘤外显体的分离，但预计也将适用于其他癌症。为了实现这项研究的目标，我们将开发和优化一个包含GNA血液净化器的过滤系统，用于从生物液中分离总外切体(特定目标1)。GNA血液净化器从不同的生物体液中分离黑色素瘤TeX和非TeX的能力将通过评估外体消耗性能来表征(特定目标2)。然后，我们将构建一种新的设备，CSPG4盒，用于从总外体中分离TeX，然后测试从开始的生物流体样本到最终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体分离系统，该系统将适用于肿瘤学临床护理环境中高通量外体分离以及从不同复杂程度的不同生物液中分离特克斯和非特克斯。