Single Cell Methods for Bioeffector Discovery and Analysis

用于生物效应器发现和分析的单细胞方法

• 批准号: 10329957
• 负责人: BRIAN O BACHMANN
• 金额: $ 57.15万
• 依托单位: VANDERBILT UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-12-18 至 2023-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10329957
• 关键词: Actinobacteria class; Acute Myelocytic Leukemia; Address; Affect; Antigens; Antineoplastic Agents; Bacteria; Benchmarking; Biological Assay; Biology; Biopsy; Cancer Biology; Cancerous; Cell Death; Cells; Chemicals; Clinical; Complex; Complex Mixtures; Computer software; Cytometry; Data Set; Diagnostic; Disease; Disease remission; Ecosystem; Environment; Evaluation; Evolution; Excision; Experimental Designs; Family; Health; Human; Immune; Immune response; Immune system; Immunoassay; Immunologic Surveillance; Immunooncology; Immunotherapeutic agent; Immunotherapy; Impairment; In Vitro; Individual; Intervention; Investigation; Lead; Malignant Neoplasms; Measures; Methodology; Methods; Molecular; Molecular Profiling; Multidimensional NMR Techniques; Myxococcales; Natural Products; Nature; Non-Malignant; Patients; Pattern; Population; Primary Neoplasm; Process; Repression; Research; Resolution; Sampling; Scientist; Signal Pathway; Signal Transduction; Source; Stromal Cells; Structure; System; Systems Analysis; T-Lymphocyte; Therapeutic Intervention; Tissue Sample; Tissues; Toxic effect; Treatment Efficacy; Tumor Antigens; Tumor Immunity; Tumor-infiltrating immune cells; Work; adaptive immune response; anti-cancer; base; cancer cell; cancer therapy; cancer type; cell preparation; cell type; cheminformatics; chemotherapy; cytotoxic; high dimensionality; immune activation; immune checkpoint blockade; immunogenic cell death; immunogenicity; insight; interest; metabolome; metabolomics; microbial; neoplastic cell; new therapeutic target; novel; novel strategies; novel therapeutics; patient response; patient subsets; precision medicine; recruit; response; small molecule; translational applications; translational potential; tumor; tumor microenvironment

PROJECT SUMMARY It is becoming increasingly apparent that discovering new therapeutics for the treatment of cancer must involve a consideration of: (A) The interplay between the host immune system and tumors. Cancer cells often have adapted the ability to evade immune surveillance, either by muted antigenicity or via actively disarming immune activation via immune checkpoint blockade. (B) The connection between chemotherapeutic interventions and immunogenicity. It has become apparent that the efficacy of many traditional chemotherapies is dependent upon enhancing the immunogenicity of cancer cells. Without a functional immune system, cytotoxic small molecules demonstrate decrease selectivity for cancer cells versus healthy ones. (C) The highly heterogeneous nature of tumors in their native environments. Tumors are comprised of a complex mixture of multiple tumor lineages embedded host tissue microenvironments. The structure and complexity of the tumor microenvironment has a direct bearing on the efficacy of therapeutic interventions. This proposal develops a new methodology for natural product discovery using biopsied human tumors and tumor infiltrating immune cells. Multiplexed activity metabolomics (MAM) merges flow cytometric microtiter well and bioassay multiplexing with metabolomics and cheminformatics software to radically accelerate bioactive compound discovery, and specifically addresses the above considerations in the context of the discovery of acute myeloid leukemia anticancer lead discovery. Additionally, experimental designs will provide new insights into the effect and mechanism of a reference set of known synthetic and natural small molecules, providing a basis set of cellular responses to cytotoxic small molecules for the evaluation of lead compounds generated during discovery efforts. Specific aims of this proposal are organized independently to develop a multiplexing system for bioeffector discovery, a multiplexing system for analysis of heterogeneous cell mixtures, and a deep cell response profiling via multiplexed immunoassay of markers of cell status. Aims circumscribe this plan for our cross-disciplinary team employing metabolomics, natural product chemical biology, and discovery (Bachmann), cytometry and cancer biology (Irish), and clinical cancer biology (Ferrell). We aim to: (1) Identify microbial metabolites that specifically target human cancer cells from primary tumor tissue samples to modulate anti-tumor immunity. (2) Discover metabolites that remodel immune cell population fates to enhance anti-tumor immunity, (3) Determine deep single cell metabolite responses of malignant and tumor-associated immune cells using known, clinically active molecules as reference points Relevance: This successful completion of the proposed research is highly relevant to human health because it will provide methods to accelerate the identification of potential anticancer natural products, which have had and continue to have a large impact on human health. Furthermore, the discovery of the multi-cell targeting immuno-oncological activity of known compound families, and newly discovered compounds may provide new targeted therapeutics, with greater efficiency and reduced clinical toxicity.

项目摘要 越来越明显的是，发现治疗癌症的新疗法必须涉及 考虑：（A）宿主免疫系统和肿瘤之间的相互作用。癌细胞通常具有 适应逃避免疫监视的能力，无论是通过减弱抗原性或通过主动解除免疫 通过免疫检查点阻断激活。(B)化疗干预与 免疫原性很明显，许多传统化疗的疗效取决于 增强癌细胞的免疫原性。如果没有正常的免疫系统，细胞毒性小分子 显示对癌细胞的选择性相对于健康细胞降低。(C)的高度异质性 肿瘤在它们的自然环境中。肿瘤由多种肿瘤谱系的复杂混合物组成 包埋宿主组织微环境。肿瘤微环境的结构和复杂性与肿瘤的发生发展密切相关。 直接影响治疗干预的效果。该提案提出了一种新的方法， 使用活组织检查的人类肿瘤和肿瘤浸润免疫细胞发现天然产物。复用 活性代谢组学（MAM）将流式细胞仪微量滴定和生物测定多路复用与代谢组学相结合 和化学信息学软件，从根本上加快生物活性化合物的发现，并具体解决 上述考虑在发现急性髓系白血病的背景下率先发现抗癌药物。 此外，实验设计将提供新的见解的影响和机制的参考集， 已知的合成的和天然的小分子，提供了细胞对细胞毒性小分子的细胞应答的基础集合。 用于评估发现过程中产生的先导化合物的分子。本提案的具体目标 是独立组织的，以开发一个用于生物效应器发现的多路复用系统， 用于异质细胞混合物的分析，以及通过多重免疫测定进行的深层细胞反应分析， 细胞状态的标志物。我们的跨学科团队采用代谢组学， 天然产物化学生物学和发现（Bachmann），细胞计数和癌症生物学（爱尔兰），以及临床 癌症生物学（费雷尔）。我们的目标是：（1）识别特异性靶向人类癌细胞的微生物代谢产物 以调节抗肿瘤免疫。(2)发现代谢物，重塑 免疫细胞群体命运增强抗肿瘤免疫，（3）确定深层单细胞代谢产物 使用已知的临床活性分子作为参考的恶性和肿瘤相关免疫细胞的应答 相关性：成功完成拟议的研究与人类健康高度相关 因为它将提供加速识别潜在抗癌天然产品的方法， 已经并将继续对人类健康产生重大影响。此外，多细胞的发现 已知化合物家族的靶向免疫肿瘤学活性，新发现的化合物可 提供新的靶向治疗，具有更高的效率和降低的临床毒性。