Fluorescence molecular tomography to study T cell infiltration into tumors

荧光分子断层扫描研究 T 细胞浸润肿瘤

• 批准号: 8902076
• 负责人: VICTOR H ENGELHARD
• 金额: $ 16.88万
• 依托单位: UNIVERSITY OF VIRGINIA
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-08-01 至 2017-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8902076
• 关键词: Adoptive Transfer; Affinity; Animals; Antigens; Behavior monitoring; Biological Markers; Biology; Blood Vessels; CD8B1 gene; Cancer Patient; Cell surface; Cells; Cellular Immunology; Collaborations; Color; Consensus; Data; Disease; Elements; Epitopes; Evaluation; Extravasation; Flow Cytometry; Future; Generations; Genetic; Histology; Homing; Imaging Techniques; Imaging technology; Immune; Immunosuppression; Immunotherapy; In Situ; In Vitro; Infiltration; Inflammation; Integrins; Knock-out; Ligands; Location; Lymphoid; Measurement; Microscopy; Modeling; Monitor; Mus; Normal tissue morphology; Organ; Ovalbumin; Patients; Pattern; Peripheral; Phage Display; Population; Process; Prognostic Marker; Property; Regulatory T-Lymphocyte; Research Personnel; Role; Staging; System; T-Cell Receptor; T-Lymphocyte; Technical Expertise; Techniques; Technology; Time; Tissues; Transgenic Organisms; Tumor-Associated Vasculature; Vaccination; Work; base; clinically relevant; design; expectation; fluorescence molecular tomography; improved; in vivo; melanoma; migration; neoplastic cell; non-invasive imaging; public health relevance; receptor; receptor expression; research study; single cell analysis; success; therapeutic effectiveness; trafficking; tumor; tumor immunology; tumor microenvironment; whole animal imaging

DESCRIPTION (provided by applicant): The presence of CD8 T cell (TCD8) in tumors is a positive indicator of patient survival. It has also become clear that patients who respond clinically to new generation immunotherapies are those in which an immunological tumor infiltrate is already evident prior to treatment. On the other hand, the fraction of adoptively transferred tumor-specific effectors that infiltrate tumors is surprisingly small, necessitating in vitro expansion of large numbers of T cells in conjunction with lymphodepletion to support further expansion in vivo. Thus, limited infiltration of T cells is a major barrier to tumor contro over and above the hurdle of immunosuppression. This is an application to understand the basis for this limited infiltration and seek ways to overcome it. Homing receptors (HR) on T cells and their corresponding vascular ligands are essential for TCD8 effectors to infiltrate peripheral tissues. However, we lack a full understanding of which HR control migration into tumors, how this is influenced by expression of the corresponding ligands on tumor vasculature, and the extent to which these same HR also enable infiltration into normal tissues, potentially in competition with tumor infiltration. In addition, the retention of TCD8 effectors after extravasatin may be determined by their ability to recognize Ag on tumor cells or intratumoral Ag presenting cells, or by the action of retention integrins. Most studies of intratumoral T cells have relied on flow cytometry, histology, and blockade or knockout approaches to investigate some of these issues at single time points. Here, we propose to use a new imaging technique, Fluorescence Molecular Tomography (FMT), to longitudinally monitor TCD8 in tumor bearing animals in real time. FMT has been used to visualize TCD8 in vivo in only a single preliminary study. Thus, some aspects of this application are designed to optimize the use of FMT, with the expectation that it will become widely used. The more fundamental aspects of the application will provide greater understanding of the trafficking, distribution and dynamics of T cell entry and exit into tumors and other peripheral tissues; the importance of homing receptors (HR) on T cells, their corresponding vascular ligands, and antigen in promoting tumor entry and retention; and the role of the tumor microenvironment in controlling these processes. This application is a collaboration between two investigators with complementary intellectual expertise in tumor immunology and tumor vascular biology and technical expertise in cellular immunology / single cell analysis by multi-color flow cytometry and advanced whole animal imaging techniques. The specific aims of this application are: 1) To establish a system for long-term measurements of T cell distribution and trafficking using FMT; 2) To investigate the distribution and dynamics of TCD8 effectors in tumor bearing mice; and 3) To establish the role of inflammation, endogenous TCD8 effectors, and Treg in controlling TCD8 infiltration into tumors and the properties of the tumor- associated vasculature. We expect the results from the work proposed in this application will support a future multi-investigator R01 or P01 project application in which such comprehensive studies will be undertaken.

描述（由申请人提供）：肿瘤中CD8 T细胞（TCD8）的存在是患者存活的积极指标。同样，对新一代免疫疗法的临床反应的患者是免疫学肿瘤浸润之前已经明显的患者。另一方面，浸润肿瘤的传递转移的肿瘤特异性效应子的比例非常小，需要 大量T细胞与淋巴结序列的体外扩展，以支持体内进一步膨胀。因此，T细胞的有限浸润是在免疫抑制障碍之上和之上肿瘤防治的主要障碍。这是了解这种有限渗透的基础并寻求克服它的方法的应用。 T细胞及其相应的血管配体上的归位受体（HR）对于TCD8效应子浸润周围组织至关重要。但是，我们对哪些人力资源控制迁移到肿瘤的迁移中缺乏完全的理解，这如何受到肿瘤脉管系统中相应配体的表达的影响，以及这些相同的HR也使肿瘤浸润的竞争可能在正常组织中渗透到正常组织中。另外，胎盘素外TCD8效应子的保留可以取决于其对肿瘤细胞或肿瘤内Ag呈现细胞的能力，或者通过保留整合素的作用。大多数肿瘤内T细胞的研究都依赖于 流式细胞仪，组织学和封锁或敲除方法在单个时间点研究其中一些问题。在这里，我们建议使用一种新的成像技术，即荧光分子断层扫描（FMT），以实时监测肿瘤轴承动物的TCD8。 FMT仅在一项初步研究中用于可视化体内TCD8。因此，本应用程序的某些方面旨在优化FMT的使用，并期望它将被广泛使用。应用程序的更基本方面将提供对T细胞进入的运输，分布和动力学的更多了解，并退出肿瘤和其他周围组织。 HOHON受体（HR）对T细胞的重要性，它们相应的血管配体以及抗原在促进肿瘤进入和保留方面的重要性；以及肿瘤微环境在控制这些过程中的作用。该应用是两位研究人员通过多色流式细胞术和先进的全动物成像技术在肿瘤免疫学和肿瘤血管生物学方面具有互补智力专业知识和技术专业知识的合作。本应用的具体目的是：1）建立使用FMT长期测量T细胞分布和运输的系统； 2）研究肿瘤轴承小鼠TCD8效应子的分布和动力学； 3）为了确定炎症，内源性TCD8效应子和Treg在控制TCD8浸润到肿瘤中的作用和肿瘤相关脉管系统的特性。我们预计，本申请中提出的工作的结果将支持将来的多启示剂R01或P01项目应用程序，其中将进行此类全面研究。