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

基本信息

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
项目类别：
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

项目摘要

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.

项目概要越来越明显的是，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。癌细胞通常具有通过减弱抗原性或主动解除免疫来适应逃避免疫监视的能力通过免疫检查点封锁激活。 (B) 化疗干预与治疗之间的联系免疫原性。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。如果没有功能性的免疫系统，细胞毒性小分子与健康细胞相比，证明对癌细胞的选择性降低。 (C) 高度异质性肿瘤在其原生环境中。肿瘤由多种肿瘤谱系的复杂混合物组成嵌入宿主组织微环境。肿瘤微环境的结构和复杂性直接影响治疗干预的效果。该提案开发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孔和生物测定多重分析与代谢组学相结合和化学信息学软件，从根本上加速生物活性化合物的发现，并专门解决在急性髓系白血病的发现背景下的上述考虑导致了抗癌药物的发现。此外，实验设计将为参考集的效果和机制提供新的见解。已知的合成和天然小分子，提供细胞对细胞毒性小分子反应的基础集用于评估发现工作中产生的先导化合物的分子。本提案的具体目标独立组织开发用于生物效应器发现的多路复用系统用于分析异质细胞混合物，以及通过多重免疫分析进行深度细胞反应分析细胞状态的标记。目标是为我们采用代谢组学的跨学科团队制定这一计划，天然产物化学生物学和发现（巴赫曼）、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）鉴定专门针对人类癌细胞的微生物代谢物从原发性肿瘤组织样本中提取来调节抗肿瘤免疫。 (2) 发现重塑代谢物免疫细胞群命运增强抗肿瘤免疫力，（3）确定深层单细胞代谢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反应要点相关性：拟议研究的成功完成与人类健康高度相关因为它将提供加速鉴定潜在抗癌天然产物的方法，已经并将继续对人类健康产生巨大影响。此外，多细胞的发现针对已知化合物家族的免疫肿瘤活性，新发现的化合物可能提供新的靶向治疗，具有更高的效率和更低的临床毒性。