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.

摘要 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学过程，涉及世界上一些恶性肿瘤的病理学和治疗。 大多数疑难杂症。例如，干细胞疗法和免疫疗法正在成为可行的 治疗以前认为无法治愈的疾病，如心力衰竭和糖尿病。不幸的是，细胞 追踪方法仍不足以充分利用这些最新进展。目前，细胞跟踪依赖于 通过造影剂对特定细胞群的分布进行成像， 附着于细胞或靶向工程化的报道蛋白。这种方法排除了精确的 细胞循环动力学或迁移途径的测量。此外，由于外排和非特异性 由于保留，造影剂的分布不一定匹配细胞的潜在分布。 鉴于这些挑战，我们考虑一种新的方法，有可能彻底改变细胞跟踪。 虽然目前的方法旨在跟踪大量细胞群体，但我们假设新的生物学见解可能 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。 我们将继续开发CellGPS，这是一种能够跟踪单个细胞3D位置的方法 当这些细胞迁移通过活体时，为了实现这一目标，我们依靠 一种以前开发的算法，可以直接从原始列表模式中提取移动单元格的位置 正电子发射断层摄影（PET）扫描仪的输出。PET是最敏感的成像方式 用于全身人体成像，因此是该项目的理想成像模式。建立在广泛的 在初步研究中，我们计划追求以下四个具体目标：（1）开发一种快速，安全和强大的 放射性标记细胞的策略;（2）设计和构建一种新的微流体管道， 分离单细胞用于体内追踪;（3）评价单细胞追踪作为细胞散布在 转移性黑色素瘤的实验模型;以及（4）探索将该技术转化为人类成像 扫描仪该项目预计将对生物医学研究产生积极影响，无论是在临床前， 临床设置。例如，单细胞跟踪可以用于确定细胞的时空动力学。 在转移级联的最早阶段期间的细胞迁移。该方法还可以帮助确定 移植后细胞的动态分布，用于基于细胞的治疗，这可以帮助预测反应， 优化治疗方案。该项目将实现对常规和可重复的关键里程碑 使用PET在体内追踪单细胞。