New tools for tracking single cells in vivo

体内追踪单细胞的新工具

• 批准号: 10400200
• 负责人: Guillem Pratx
• 金额: $ 58.9万
• 依托单位: STANFORD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-09-01 至 2024-10-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10400200
• 关键词: 3-Dimensional; Algorithms; Animal Model; Basic Science; Biological; Biological Process; Biomedical Research; Cell Cycle Kinetics; Cell Therapy; Cell Transplantation; Cells; Circulation; Clinical; Clinical Research; Contrast Media; Development; Diabetes Mellitus; Disease; Embryonic Development; Engineering; Experimental Models; Felis catus; Genetic; Goals; Health; Heart Ventricle; Heart failure; Hematopoiesis; Human; Human body; Image; Imaging Device; Immunity; Immunotherapy; In Vitro; Individual; Injections; Investigation; Kinetics; Label; Left; Link; Lipids; Liver; Location; Measurement; Measures; Melanoma Cell; Metastatic Melanoma; Methods; Microfluidic Microchips; Microfluidics; Modeling; Molecular; Molecular Profiling; Mus; Neoplasm Metastasis; Organ; Outcome; Output; Pathology; Patients; Pattern; Phase; Phenotype; Physiological Processes; Population; Positioning Attribute; Positron-Emission Tomography; Pre-Clinical Model; Predictive Factor; Process; Proteins; Radioactivity; Radiolabeled; Reporter; Reproducibility; Role; Route; Signal Transduction; Silicon Dioxide; Surface; System; Technology; Testing; Time; Toxic effect; Tracer; Translating; Translational Research; Translations; Treatment Protocols; Validation; cell motility; computerized tools; design; human imaging; imaging modality; improved; in vivo; insight; migration; molecular imaging; molecular phenotype; nanoparticle; novel; novel strategies; novel therapeutics; pre-clinical; pre-clinical research; predicting response; programs; radiotracer; receptor expression; reconstruction; spatiotemporal; stem cell migration; stem cell therapy; survival prediction; tool; trafficking; treatment optimization

Abstract This project will develop a new pipeline for tracking the migration of single cells in vivo at the whole-body level. Cell migration is a crucial biological process involved in the pathology and treatment of some of the world’s most intractable diseases. Stem cell therapy and immunotherapy, for instance, are emerging as viable treatments for conditions previously thought incurable, such as heart failure and diabetes. Unfortunately, cell tracking methods remain inadequate to fully capitalize on these recent advances. Currently, cell tracking relies on imaging the distribution of a specific population of cell through a contrast agent, which is either directly affixed to the cells or targeted towards an engineered reporter protein. This approach precludes precise measurement of cell circulation kinetics or migration routes. Furthermore, due to efflux and non-specific retention, the distribution of the contrast agent does not necessarily match the underlying distribution of cells. In view of these challenges, we consider a novel approach that has the potential to revolutionize cell tracking. While current methods aim to track bulk populations of cells, we hypothesize that novel biological insight may be gained by tracking cells individually, in small numbers, with unprecedented temporal and spatial accuracy. We will pursue the development of CellGPS, a method capable of tracking the 3D position of individual cells continuously as these cells migrate through the body of a living subject. To accomplish this goal, we rely on a previously developed algorithm that can extract the position of a moving cell directly from the raw list-mode output of a positron emission tomography (PET) scanner. PET is the most sensitive imaging modality available for whole-body human imaging and, therefore, the ideal imaging modality for this project. Building on extensive preliminary studies, we plan to pursue the following four specific aims: (1) develop a rapid, safe and robust strategy for radiolabeling cells; (2) design and build a novel microfluidics pipeline to molecularly profile and isolate single cells for in vivo tracking; (3) evaluate single-cell tracking as a readout of cell dissemination in an experimental model of metastatic melanoma; and (4) explore translation of this technology to human imaging scanners. This project is expected to generate a positive impact for biomedical research both in the pre-clinical and clinical setting. For instance, single-cell tracking could be used to determine the spatiotemporal kinetics of cell migration during the earliest phase of the metastatic cascade. The method could also help determine the dynamic distribution of cells after transplantation for cell-based therapy, which could help predict response and optimize treatment regimen. This project will achieve critical milestones towards routine and reproducible tracking of single cells in vivo using PET.

抽象的 该项目将开发一种新的管道，用于在全身水平上跟踪体内单细胞的迁移。 细胞迁移是一个重要的生物过程，涉及世界上一些疾病的病理学和治疗。 例如，干细胞疗法和免疫疗法正在成为可行的治疗方法。 不幸的是，细胞治疗以前被认为无法治愈的疾病，例如心力衰竭和糖尿病。 目前，追踪方法仍不足以充分利用这些最新进展。 通过造影剂对特定细胞群的分布进行成像，该造影剂可以直接 固定在细胞上或针对工程报告蛋白，这种方法排除了精确性。 此外，由于外排和非特异性，细胞循环动力学或迁移路线的测量。 保留时，造影剂的分布不一定与细胞的底层分布相匹配。 鉴于这些挑战，我们考虑了一种有可能彻底改变细胞追踪的新方法。 虽然当前的方法旨在追踪大量细胞群，但我们认为新颖的生物学见解可能会 通过以前所未有的时间和空间精度单独跟踪少量细胞来获得。 我们将致力于开发 CellGPS，一种能够跟踪单个细胞 3D 位置的方法 当这些细胞在活体体内不断迁移时，我们依靠一种方法来实现这一目标。 先前开发的算法可以直接从原始列表模式中提取移动单元的位置 正电子发射断层扫描 (PET) 扫描仪的输出是最灵敏的成像方式。 用于全身人体成像，因此是该项目的理想成像方式。 初步研究后，我们计划追求以下四个具体目标：（1）开发快速、安全、稳健的 (2) 设计并构建新型微流体管道以进行分子分析和 分离单细胞进行体内追踪；（3）评估单细胞追踪作为细胞传播的读数 转移性黑色素瘤的实验模型；(4) 探索该技术在人体成像中的应用 该项目预计将对临床前的生物医学研究产生积极影响。 例如，单细胞追踪可用于确定时空动力学。 该方法还可以帮助确定转移级联最早阶段的细胞迁移。 细胞治疗移植后细胞的动态分布，有助于预测反应和 优化治疗方案将实现常规和可重复的关键里程碑。 使用 PET 追踪体内单细胞。