Analysis of Cell Sense Labeled Human Cells

描述（由申请人提供）：细胞疗法在治疗和减轻与衰弱性人类疾病相关的症状方面具有巨大潜力，如癌症、心血管疾病和神经系统疾病。目前，没有一种通用的方法来跟踪细胞产品在给药后的存在，导致这些疗法延迟到达临床。无创性地可视化细胞在全身的转移过程，将使临床医生能够更好地理解为什么某些细胞移植是成功的，而另一些则不是。此外，确定转移细胞的生物分布对于克服政府对这些疗法的监管障碍至关重要。在这项研究中，我们提出表征一种新的荧光化学细胞跟踪试剂最近创建用于MRI，称为细胞感。该产品是含氟纳米颗粒的乳液，用于标记培养中的细胞，并使转移的细胞在体内使用19F MRI具有很高的特异性。细胞感觉已经在小鼠细胞中进行了广泛的测试，并使用体内啮齿动物模型进行了成像。它尚未在临床相关的人类细胞上进行测试。在本提案中，我们将研究各种主要的人类免疫细胞类型，包括树突状细胞，自然杀伤细胞和T细胞，目前正在免疫治疗试验中使用的细胞感标记特征。此外，干细胞，包括人类神经祖细胞（hNPC）和CD34+细胞类型，将被研究。本研究有两个具体目的：(1)开发离体细胞标记方案，并在治疗相关的人类细胞中检测细胞感觉标记剂量；(2)确定荧光化学标记是否对人体细胞活力或正常功能有不利影响。为了实现这些目标，我们将采用广泛的体外试验来评估细胞负荷、细胞毒性、增殖、表型和功能。结合这些结果，我们将能够为目前用于人体临床试验的各种细胞类型设计最佳的标记方案。通过建立Cell Sense对多种毒性有限的人类细胞进行标记的能力，我们将在向临床级产品迈进的道路上取得一个关键的里程碑，该产品可用于跟踪当前移植细胞的治疗用途。我们相信Cell Sense的潜在影响是巨大的，它消除了临床细胞疗法开发者目前面临的许多科学和监管障碍。公共卫生相关性：了解与治疗相关的细胞类型在过继性转移或移植后的行为可能有助于我们开发新的治疗方法。本项目研究了使用新型荧光化学MRI试剂对标记的移植细胞进行无创成像和检测。这项研究的结果将使MRI细胞跟踪更接近临床应用，并将为研究人员和医生提供有选择地跟踪标记细胞的机会。

DESCRIPTION (provided by applicant): Cellular therapeutics represent great potential for curing and alleviating symptoms associated with debilitating human diseases, such as cancer, cardiovascular disease, and neurological disorders. Currently, no versatile methodology for tracking cellular products following administration exists, leading to delays in these therapies reaching the clinic. The ability to visualize the trafficking of transferred cells throughout the body non-invasively will give clinicians the ability to better understand why certain cellular transplants are successful and others are not. In addition, determining the biodistribution of transferred cells is of paramount importance to overcome government regulatory hurdles for these therapies. In this study, we propose characterizing a novel fluorochemical cell tracking reagent recently created for use with MRI, called Cell Sense. This product is an emulsion of fluorine-containing nanoparticles that is used to label cells in culture and enables transferred cells to be visualized with great specificity in vivo using 19F MRI. Cell Sense has been tested extensively in mouse cells and imaged using in vivo rodent models. It has not yet been tested on clinically relevant human cells. In this proposal, we will examine the Cell Sense labeling characteristics in a variety of primary human immune cell types, including dendritic cells, natural killer cells, and T cells, that are currently being used in immunotherapeutic trials. Also, stem cells, including human neural progenitor cells (hNPC) and CD34+ cell types, will be studied. This proposal has two Specific Aims: (1) to develop ex vivo cell labeling protocols and examine the Cell Sense labeling dose in therapeutically relevant human cells; and (2) to determine whether the fluorochemical labeling has any detrimental impact on human cell viability or normal function. To achieve these Aims, we will employ an extensive battery of in vitro assays to assess cell loading, cytotoxicity, proliferation, phenotype, and function. Combining these results, we will be able to devise optimal labeling protocols for a diverse range of cell types that are currently being used for human clinical trials. By establishing Cell Sense's ability to label a wide variety of human cells with limited toxicity, we will have achieved a critical milestone in moving closer toward a clinical-grade product that can be used to track current transplanted cells for therapeutic use. We believe that the potential impact of Cell Sense is significant, by removing many of the scientific and regulatory hurdles that developers of cellular therapeutics for the clinic currently face. PUBLIC HEALTH RELEVANCE: Understanding how therapeutically relevant cell types behave following adoptive transfer or transplantation may help us develop novel therapies. This project examines the use of novel fluorochemical MRI reagents to image and detect labeled, transplanted cells non-invasively. The results of this study will move MRI cell tracking closer to clinical adoption and will provide researchers and physicians the opportunity to selectively follow labeled cells following administration.