Characterization of Biological Nanoparticle Subsets

生物纳米颗粒子集的表征

• 批准号: 9343961
• 负责人: Jennifer Jones
• 金额: $ 53.74万
• 依托单位: DIVISION OF BASIC SCIENCES - NCI
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/9343961
• 关键词: Antigen-Presenting Cells; B-Lymphocytes; Biological; Biological Markers; Biology; Biopsy; Blood; CD44 gene; Calibration; Cell Line; Cells; Characteristics; Clinical; Cooperative Research and Development Agreement; Data; Detection; Diagnostic Neoplasm Staging; Dipeptidyl-Peptidase IV; Epitopes; FOLH1 gene; Flow Cytometry; Fluorescence; Goals; Hypoxia; Immune; Immune response; Immunobiology; Immunology; Immunotherapy; Individual; Label; Leukocytes; Malignant Neoplasms; Malignant neoplasm of prostate; Mentors; Methods; MicroRNAs; Molecular; Monitor; Monoclonal Antibodies; Organism; PDCD1LG1 gene; Plasma; Population; Prostate; Proteins; Protocols documentation; RNA; Radiation; Reporting; Research; Resolution; Sorting - Cell Movement; Source; T-Lymphocyte; TACSTD1 gene; TGFB1 gene; Technology; Testing; Tumor Biology; Tumor stage; Tumor-Derived; United States National Institutes of Health; Vaccines; Vesicle; Work; base; burden of illness; cancer stem cell; cancer therapy; exosome; extracellular vesicles; high reward; high risk; improved; instrumentation; liquid biopsy; member; nanoparticle; neoplastic cell; novel marker; oncology; outcome forecast; particle; personalized medicine; programs; pyrrolidin-3-yl-methanesulfonic acid; receptor; response; single molecule; stressor; therapeutic target; treatment response; tumor; tumor progression

In the two years since the installation of the Vaccine Branch Astrios-EQ (installed mid-2014, in conjunction with a CRADA that we initiated with Beckman-Coulter in 2013), we developed nanoFACS protocols for fluorescence detection and calibration, for counting, and for high-throughput sorting, to sort EV subsets from plasma and other biofluids by nanoFACS. Using multiple distinct vesicle populations, we find that nanoFACS sorting is uniquely able to produce high fidelity EV subsets (95-97% pure), with functional biological cargo (RNA and protein). During the past 9-10 months, our focus has been exclusively on application of this method for clinically important studies, for the study of cancer disease burden, metastatic potential, and responses to treatment. Our NanoFACS approach that we developed as part of this ACI program is the only method currently able to analyze and preparatively sort functional EVs based on single EV characteristics, rather than bulk attributes. Furthermore, we have devised a new class of labels (Molecular NanoTags) that can be used with nanoFACS, not only to improve the detection of EV epitopes, but also to enable single molecule counting by flow cytometry. The specialized instrumentation and expertise assembled for this nanoFACS program does not exist anywhere else in the world, is a distinctive asset for NCI. We now have a unique opportunity to leverage this nanoFACS program in the Clinical Center to demonstrate the potential impact of this approach for oncology, immunology, and personalized medicine. Toward the goal of utilizing nanoFACS in the study of tumor-derived EVs as a liquid biopsy, we have tested and confirmed nanoFACS resolution of cancer and cancer stem-cell markers: PSMA, CD147, CD44, and CD26. During the past 6 months, we also found that immune cell markers such as MHC-II and PD-L1 identify distinct EV subsets. We predict that the co-receptors and miRNA that are expressed on or contained as cargo of MHC-II EV subsets will correlate with immune responses- of the cells or the organism that produced those EVs. We have validated PD-L1 detection on cell line-derived EVs and are beginning to study detection of PD-L1 on biofluid-derived EVs. We are working with James Gulley and Tim Greten to determine whether circulating PD-L1 levels on tumor-derived EVs or antigen presenting cell-derived EVs correlate with tumor biopsy levels of PD-L1 and responses to PD-L1/PD-1-directed therapies. Similarly, we are working now with members of the Senior Prostate Group to determine whether tumor-derived EVs with PSMA, EpCAM, and CD147 correlate with prostate cancer disease burden, prognosis, and responses to treatment.

自从我们在2013年与Beckman-Coulter启动的Crada一起安装了疫苗分支ARTARIOS-EQ以来的两年中，我们开发了用于计数的荧光检测和校准的纳米型协议，用于计数，用于高通量分类，以从plasma和其他plassm和其他plusofs和其他luofluids中进行分类。使用多个不同的囊泡种群，我们发现纳米型分类具有独特的能力，能够产生高富达EV子集（95-97％纯），具有功能性生物货物（RNA和蛋白质）。在过去的9-10个月中，我们的重点一直放在该方法中用于临床重要的研究，用于研究癌症疾病负担，转移性潜力和对治疗的反应。我们作为此ACI程序的一部分开发的纳米方法是目前能够基于单个EV特征而不是批量属性来分析和制备功能性EV的唯一方法。此外，我们设计了一类新的标签（分子纳米塔），可与纳米量一起使用，不仅可以改善对EV表位的检测，而且还可以通过流式细胞仪进行单分子计数。为该纳米素计划组装的专业仪器和专业知识在世界其他任何地方都不存在，这是NCI的独特资产。现在，我们有一个独特的机会来利用临床中心的该纳米量计划来证明这种方法对肿瘤学，免疫学和个性化医学的潜在影响。为了在肿瘤来源的EV研究中利用纳米量作为液体活检，我们已经测试并确认了癌症和癌症干细胞标记的纳米含量分辨率：PSMA，CD147，CD44和CD26。在过去的6个月中，我们还发现免疫细胞标记（例如MHC-II和PD-L1）鉴定了不同的EV子集。我们预测，以MHC-II EV子集的货物表达或包含的共受体和miRNA将与产生这些EVS的细胞或生物体的免疫反应相关。我们已经验证了细胞系衍生EV的PD-L1检测，并开始研究生物流体衍生的EV的PD-L1检测。我们正在与James Gulley和Tim Greten合作，以确定在肿瘤衍生的电动汽车或抗原上循环的PD-L1水平是否与PD-L1的肿瘤活检水平以及对PD-L1/PD-L1/PD-1导向疗法的反应有关。同样，我们现在正在与高级前列腺组成员合作，以确定肿瘤衍生的EV是否与PSMA，EPCAM和CD147与前列腺癌疾病负担，预后和对治疗的反应有关。