Using [6-13C,6-15N3]-L-Arginine for the molecular imaging of in vivo tumor arginase flux, and towards understanding the role of arginase isoforms in cancer metabolism.

使用 [6-13C,6-15N3]-L-精氨酸对体内肿瘤精氨酸酶通量进行分子成像，并了解精氨酸酶亚型在癌症代谢中的作用。

• 批准号: 10082445
• 负责人: Andrew Cho
• 金额: $ 3.21万
• 依托单位: WEILL MEDICAL COLL OF CORNELL UNIV
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-01-22 至 2021-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10082445
• 关键词: 4T1; Aftercare; Arginine; Biological Assay; Breast Cancer cell line; Cancer Biology; Cancer Cell Growth; Cancer Patient; Cell Proliferation; Cells; Characteristics; Clinical; Coculture Techniques; Complex; Cytoplasm; Development; Dichloromethylene Diphosphonate; Disease; Disease Progression; Enzymes; Fluorescence-Activated Cell Sorting; Future; Goals; Histopathologic Grade; Human; Immunocompetent; Immunosuppression; Immunotherapy; In Vitro; Individual; Infiltration; Isotopes; Knowledge; Laboratories; Lead; Liposomes; Magnetic Resonance Imaging; Malignant Neoplasms; Mammals; Measurement; Measures; Metabolic; Metabolism; Methods; Mitochondria; Modality; Modeling; Monitor; Mus; Neoplasm Metastasis; Patient Care; Patient-Focused Outcomes; Patients; Pharmaceutical Preparations; Phase; Physicians; Plasma; Population; Process; Prognosis; Prognostic Marker; Protein Isoforms; Proteins; Research; Research Proposals; Research Training; Role; Scientist; Selection for Treatments; Testing; Treatment Efficacy; Tumor-associated macrophages; Variant; Western Blotting; Work; arginase; base; cancer cell; cancer therapy; cell type; clinically relevant; density; design; experimental study; field study; human disease; imaging modality; improved; in vivo; in vivo Model; inhibitor/antagonist; insight; knock-down; liquid chromatography mass spectrometry; macrophage; metabolic phenotype; metabolomics; molecular imaging; novel therapeutics; overexpression; patient stratification; prognostic; prognostic significance; skills; small hairpin RNA; stem; targeted treatment; tool; tumor; tumor growth; tumor initiation; tumor metabolism; tumor microenvironment; tumor progression; uptake

Project Summary & Abstract The various cellular components of the tumor microenvironment undergo metabolic changes to support tumor growth and metastasis. Cancer metabolism is a clinically relevant and exciting field of study, as it provides additional mechanistic insight into the onset and progression of this disease. An improved understanding of this highly complex process can lead to the development of novel therapies, and characterization of a tumor's metabolic phenotype may provide additional criteria for determining prognosis and therapy selection. Towards the latter, the field of hyperpolarized MRI has emerged with the goal of establishing an imaging modality that can non-invasively measure enzymatic flux in vivo. One way in which cancer metabolism is altered involves arginine utilization, stemming from the overexpression of various arginase isoforms in cancer cells and tumor associated macrophages (TAMs), which is thought to promote cellular proliferation and immunosuppression. Elevated arginase activity in the plasma of cancer patients is associated with increasingly aggressive histological grading, and non-invasive quantification of intratumoral arginase activity with hyperpolarized MRI may be an improved prognostic metric. Furthermore, with the rising population of TAM-depleting immunotherapies, differences in arginase flux before and after the initiation of TAM-depleting therapies may correlate to changes in TAM infiltration and provide insight into therapeutic efficacy, which is another potential application of this modality. In addition, two arginase isoforms exist (arginase-1, A1, and -2, A2) which differ in cell-type-specific expression and subcellular localization. Cytosolic A1 is favorably expressed in TAMs, whereas mitochondrial A2 is expressed to some degree across most cell types, including cancer cells. The population and concentrations of downstream enzymes also differs between the cytoplasm and mitochondrion, leading to the hypothesis that A1 and A2 have different, cell-type-specific, pro-tumor functions in TAMs and cancer cells. The individual cell-type-specific contributions of A1 and A2 to cellular metabolism and proliferation have yet to be studied in the setting of cancer. With therapies that target arginine metabolism currently in Phase I and II clinical trails, this knowledge will support the development of the future iterations of this class of therapy. I have optimized the synthesis of [6-13C,6-15N3]-L-arginine as a dual purpose probe for 1) use as a hyperpolarized MRI probe for in vivo arginase activity measurements, and 2) LC/MS-based isotopic tracing metabolomics experiments to test the working hypothesis. Information gained from this project may yield new tools and metrics for patient stratification, and will add to the general understanding of cancer metabolism and its role in cancer cell proliferation, collectively contributing towards the improvement of patient care and providing additional mechanistic insight into this disease. In addition, the skills and knowledge gained from this research proposal and training plan will prepare me to achieve my goal of becoming a physician scientist.

项目概要和摘要 肿瘤微环境的各种细胞成分经历代谢变化，以支持肿瘤微环境的形成。 肿瘤生长和转移。癌症代谢是一个临床相关和令人兴奋的研究领域，因为它 为这种疾病的发病和进展提供了额外的机制见解。一种改进 了解这个高度复杂的过程可以导致新疗法的发展， 肿瘤代谢表型的特征可以为确定预后提供额外的标准 和治疗选择。对于后者，超极化MRI领域已经出现，其目标是 建立可以非侵入性地测量体内酶通量的成像模式。 改变癌症代谢的一种方式涉及精氨酸的利用，其来源于精氨酸。 在癌细胞和肿瘤相关巨噬细胞（TAM）中， 被认为促进细胞增殖和免疫抑制。血浆中的脱氢酶活性升高 癌症患者与越来越积极的组织学分级和非侵入性定量相关 超极化MRI检测肿瘤内端粒酶活性可能是一种改善的预后指标。此外，委员会认为， 随着TAM消耗免疫疗法的人群不断增加， 启动TAM消耗疗法可能与TAM浸润的变化相关，并提供有关以下方面的见解 治疗效果，这是该模式的另一个潜在应用。 此外，存在两种在细胞类型特异性上不同的脱氢酶同种型（脱氢酶-1，A1和-2，A2）。 表达和亚细胞定位。胞浆A1在TAM中有利地表达，而线粒体A1在TAM中有利地表达。 A2在大多数细胞类型中都有一定程度的表达，包括癌细胞。人口和 下游酶的浓度在细胞质和细胞质之间也不同，导致 假设A1和A2在TAM和癌症中具有不同的细胞类型特异性促肿瘤功能 细胞A1和A2对细胞代谢和增殖的个体细胞类型特异性贡献具有以下特征： 还有待于在癌症的背景下进行研究。针对精氨酸代谢的治疗目前处于I期 和II临床试验，这些知识将支持这类治疗的未来迭代的发展。 我已经优化了[6- 13 C，6- 15 N3]-L-精氨酸的合成，作为一种双重用途的探针，1）用作 超极化MRI探针用于体内脱氢酶活性测量，和2）基于LC/MS的同位素示踪 代谢组学实验来验证工作假设。从这个项目中获得的信息可能会产生新的 用于患者分层的工具和指标，并将增加对癌症代谢的一般理解， 它在癌细胞增殖中的作用，共同促进改善患者护理， 提供了对这种疾病的额外的机制性见解。此外，从中获得的技能和知识 我的研究计划和培训计划将帮助我实现成为一名医生科学家的目标。