Label-free imaging of CAR T cell metabolism

CAR T 细胞代谢的无标记成像

基本信息

批准号：
10751581
负责人：
Christian Capitini
金额：
$ 66.2万
依托单位：
MORGRIDGE INSTITUTE FOR RESEARCH, INC.
依托单位国家：
美国
项目类别：
财政年份：
2023
资助国家：
美国
起止时间：
2023-08-01 至 2028-07-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10751581
关键词：
Antitumor Response Autologous B-Cell Lymphomas Benchmarking Biological Assay Biological Markers CAR T cell therapy CD19 gene Cancer Patient Cell physiology Cells Cellular Metabolic Process Clinical Clinical Trials Combined Modality Therapy Cytolysis Dasatinib Data Disease remission FDA approved Flow Cytometry Frequencies Genetic Engineering Goals Human Hypoxia Image Imaging technology Immunofluorescence Immunologic In Vitro Infiltration Infusion procedures Intervention Knowledge Label Malignant Neoplasms Measurement Measures Memory Metabolic Methods Modeling Monitor Neuroblastoma Optics Patient Selection Patients Phenotype Process Production Resolution Resources Sampling Solid Neoplasm Source Specificity Spleen T cell response T cell therapy T memory cell T-Lymphocyte Technology Technology Assessment Testing Time Xenograft procedure alternative treatment cancer cell cancer therapy cellular imaging chimeric antigen receptor chimeric antigen receptor T cells cytokine exhaust exhaustion fitness fluorescence imaging improved in vivo manufacture metabolic imaging mouse model neoplastic cell patient screening predictive modeling process optimization response response biomarker single cell technology stem cells therapy development tumor

项目摘要

PROJECT SUMMARY / ABSTRACT The goal of this proposal is to develop non-invasive single-cell technologies to improve the potency of T cell therapies against cancer. The first 6 chimeric antigen receptor (CAR) T cell therapies were recently approved and >800 CAR and T cell therapies are in clinical trials. However, barriers remain in achieving durable remissions (>1 year) for ~50% of patients who receive CAR T cell therapy. Due to the rapid development of these therapies and a great need for process optimization, we focus on improving three translational roadblocks to effective CAR T cell therapy: (1) screening patients whose T cells are unfit for CAR T cell manufacturing, (2) optimizing in vitro CAR T cell production for higher potency, and (3) identifying metabolic features of potent CAR T cells in vivo. CAR T cell therapy could be improved by enriching for naïve and stem cell memory (SCM) T cells in starting materials and final products. Deficiencies in naïve and SCM T cells occurs in ~50% of untreated cancer patients, and manufacturing autologous CAR T cell products from these sources has been unsuccessful. Even if SCM T cells can be isolated, after CAR incorporation, the expansion process typically diminishes potency through T cell exhaustion. After infusion, the presence of memory-like phenotypes in vivo correlate with better responses. To date, there are no robust, non-destructive technologies to monitor CAR T cell manufacturing to optimize production and assess potency in vivo at a single-cell level. These issues limit the impact of CAR T cell therapy. Current approaches to measure T cell function are labor-intensive, destructive, or lack single-cell resolution, which limits the frequency or specificity of these measurements. For CAR T cell therapy to realize its clinical potential, new methods are needed to monitor T cells for optimal potency throughout manufacturing and post- infusion. Changes in cell metabolism provide an attractive yet under-explored assay to track T cell potency. Previous studies, including our own, show that T cells undergo drastic metabolic changes with activation, and that naïve, exhausted, and memory-like T cells have distinct metabolic features. Our preliminary data shows that non-invasive single-cell imaging of the fluorescence intensity and lifetime of NAD(P)H and FAD (optical metabolic imaging, or OMI) can predict CAR T cell manufacturing conditions that produce a more vs. less potent anti-tumor response in vivo. Given these metabolic features of CAR T cell potency, we propose to determine whether label-free OMI of T cell autofluorescence and multivariate models can identify patient T cell fitness, optimal in vitro expansion conditions, and in vivo cell biomarkers of potent and persistent CAR T cell response. Overall, these technologies will streamline processes and interventions for consistently potent T cell therapy and increase our knowledge of CAR T cell metabolism in vitro and in vivo.

项目摘要 /摘要该提案的目的是开发非侵入性的单细胞技术来提高T细胞的效力针对癌症的疗法。最近批准了前6个嵌合抗原受体（CAR）T细胞疗法 > 800辆汽车和T细胞疗法正在临床试验中。但是，障碍仍然在实现持久的减免方面（> 1年）接受约50％的接受CAR T细胞疗法的患者。由于这些疗法的快速发展并且非常需要流程优化，我们专注于改善三个翻译的障碍到有效的汽车 T细胞疗法：（1）筛查T细胞不适合CAR T细胞制造的患者，（2）优化体外 CAR T细胞的产生具有更高的效力，（3）鉴定体内有效的CAR T细胞的代谢特征。可以通过富集幼稚和干细胞记忆（SCM）T细胞来改善CAR T细胞疗法。材料和最终产品。幼稚和SCM T细胞的缺陷发生在约50％的未经治疗的癌症患者中，这些来源的制造自动汽车T细胞产品没有成功。即使SCM T 可以隔离细胞，在合并汽车后，扩展过程通常会降低通过T细胞的效力精疲力尽。输注后，体内内存样表型的存在与更好的反应相关。到日期，没有强大的，无损的技术来监视CAR T细胞制造以优化单细胞水平的体内生产和评估效力。这些问题限制了汽车T细胞疗法的影响。当前测量T细胞功能的方法是劳动密集型，破坏性或缺乏单细胞分辨率，这限制了这些测量值的频率或特异性。为了使汽车T细胞疗法实现其临床潜在的，需要新的方法来监测T细胞以在整个制造业和之后的整个制造业中获得最佳效力输液。细胞代谢的变化为跟踪T细胞效力提供了一种有吸引力但探索的测定不足的测定。先前的研究，包括我们自己的研究，表明T细胞会随着激活而发生急剧的代谢变化，并且那个天真，疲惫和记忆状的T细胞具有独特的代谢特征。我们的初步数据表明 NAD（P）H和FAD（光学）的荧光强度和寿命的非侵入性单细胞成像代谢成像或OMI）可以预测产生更少而更少的汽车T细胞制造条件体内有效的抗肿瘤反应。鉴于CAR T细胞效力的这些代谢特征，我们建议确定T细胞自动荧光和多元模型的无标签OMI可以识别患者T细胞适应性，最佳体外膨胀条件以及潜力和持续性CAR T细胞的体内生物标志物回复。总体而言，这些技术将简化一贯潜在T细胞的过程和干预措施治疗并增加我们对体外和体内汽车T细胞代谢的了解。