New tools for tracking single cells in vivo

体内追踪单细胞的新工具

• 批准号: 10248540
• 负责人: Guillem Pratx
• 金额: $ 62.01万
• 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/10248540
• 关键词: 3-Dimensional; Algorithms; Animal Model; Basic Science; Biological; Biological Process; Biomedical Research; Blood Circulation; Cell Cycle Kinetics; Cell Therapy; Cell Transplantation; Cells; 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 该项目将开发一条新的管道，用于跟踪整个体内体内单个细胞的迁移。 细胞迁移是世界上一些的病理学和治疗涉及的至关重要的生物学过程 最棘手的疾病。例如，干细胞疗法和免疫疗法已成为可行的 治疗以前认为无法治愈的疾病，例如心力衰竭和糖尿病。不幸的是，牢房 跟踪方法仍然不足以完全利用这些最新进展。目前，细胞跟踪依赖 通过对比剂成像特定细胞群的分布，这要么是直接的 固定在细胞上或针对工程的报告基因蛋白。这种方法排除了精度 细胞循环动力学或迁移途径的测量。此外，由于排出和非特异性 保留，对比剂的分布不一定与细胞的潜在分布相匹配。 鉴于这些挑战，我们考虑了一种新颖的方法，该方法有可能改变细胞跟踪。 虽然目前的方法旨在跟踪细胞的大容量种群，但我们假设新型的生物学见解可能 通过以少量的暂时和空间精度单独跟踪细胞来获得。 我们将追求CellGP的开发，该方法能够跟踪单个单元的3D位置 当这些细胞通过活物体的身体迁移时，连续不断。为了实现这一目标，我们依靠 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格的位置 正电子发射断层扫描（PET）扫描仪的输出。宠物是可用的最敏感的成像方式 用于全身人类成像，因此，是该项目的理想成像方式。建立广泛的建设 初步研究，我们计划追求以下四个具体目标：（1）发展快速，安全和强大 放射标记细胞的策略； （2）设计和建造一种新型的微流体管道，以分子剖面和 分离单个细胞用于体内跟踪； （3）评估单细胞跟踪作为在一个细胞传播中的读数 转移性黑色素瘤的实验模型； （4）探索该技术向人类成像的翻译 扫描仪。预计该项目将对临床前的生物医学研究产生积极的影响 和临床环境。例如，单细胞跟踪可用于确定时空动力学 在转移级联的最早阶段的细胞迁移。该方法还可以帮助确定 移植后细胞的动态分布进行基于细胞的治疗，这可以帮助预测反应和 优化治疗方案。该项目将实现至关重要的里程碑，以实现常规和可重现 使用PET在体内跟踪单细胞。