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Fluorescence Molecular In Vivo Liquid Biopsy of Circulating Tumor Cells

循环肿瘤细胞的荧光分子体内液体活检

基本信息

批准号：
10322183
负责人：
Mark Jonathan Niedre
金额：
$ 17.98万
依托单位：
NORTHEASTERN UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-01-01 至 2023-12-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10322183
关键词：
Affinity Animals Arteries Back Basic Cancer Research Basic Science Biological Blood Blood Cells Blood Circulation Blood Vessels Blood Volume Blood specimen Cancer Model Cavia Cell Count Cells Cessation of life Clinical Clinical Management Clinical Research Clinical Trials Communities Contrast Media Detection Development Diagnosis Diffuse Disease Progression Distant Early Diagnosis Epithelial Cells Evaluation Excision Exhibits Flow Cytometry Fluorescence Forearm Goals Human Individual Injectable Label Laboratories Light Limb structure Longitudinal Studies Malignant Neoplasms Malignant neoplasm of liver Malignant neoplasm of lung Malignant neoplasm of ovary Mediating Methods Modeling Molecular Molecular Target Mus Neoplasm Circulating Cells Neoplasm Metastasis Optical Readers Optics Organ Pathway interactions Patients Phase III Clinical Trials Property Reader Recurrence Sampling Secondary to Skin Specificity Staging System Technology Testing Time Tissues Tracer Translations Universities Variant Wrist arm attenuation cancer surgery cancer type design diagnostic tool epithelial to mesenchymal transition experience first-in-human fluorescence-guided surgery folate-binding protein high reward high risk in vitro Model in vivo instrument interest light scattering liquid biopsy neoplastic cell new technology novel diagnostics novel therapeutics peripheral blood phantom model scale up small molecule tool treatment response uptake

项目摘要

Project Summary The goal of this project is to develop new technology to detect circulating tumor cells (CTCs) directly in the bloodstream without having to draw blood samples. Metastasis is responsible for the majority of cancer-related deaths, and is often mediated by dissemination of CTCs via the vasculature. CTCs are therefore of great interest clinically and in basic cancer research. Nearly all methods for the study of CTCs rely on drawing and analyzing small (7.5 mL) blood samples, which is broadly known as ‘liquid biopsy’. However, due to the rarity of CTCs estimation of CTC numbers from small blood samples is extremely inaccurate, and rare cells may escape detection entirely. Liquid biopsy is also insensitive to natural changes in CTC numbers that occur over short time periods. Our team recently developed new technology, Diffuse in vivo Flow Cytometry (DiFC), to detect rare, fluorescently-labeled CTCs directly in vivo in small animals. DiFC uses diffuse light to sample large circulating blood volumes in bulk tissue. The main advantage of DiFC is therefore sensitivity: we showed that DiFC can accurately detect fewer than 1 CTC per mL of PB. Because it does not require blood draws, DiFC also allows longitudinal studies in individual animals. DiFC is also readily scalable to larger limbs, species, and potentially even to humans. However, use of DiFC in humans would require bright and specific fluorescent labeling of target CTCs in vivo. Fortunately, there has been major technical and regulatory progress in injectable molecular tracer technology for fluorescence guided surgery of cancer. Of particular interest for this proposal, OTL38 is a near-infrared (NIR) small-molecule folate-receptor (FR)-targeted probe that is in phase-III clinical trials for ovarian and liver cancer. OTL38 has high specificity and affinity for CTCs in blood, with low non-specific uptake by other blood cells or vessel walls. In addition, because NIR light experiences minimal light attenuation through biological tissue, OTL38 is suitable for detection from deeper-seated blood vessels. The goal of this project is to develop and validate the enabling technology for ‘fluorescence molecular in vivo liquid biopsy of CTCs’. In Aim 1, we will build an H-DiFC system for use in the human wrist or forearm, where arterial flow rates are 100s of mL per minute. In Aim 2, we will validate labeling of FR+ CTCs with OTL38, and detectability with H-DiFC in an arm-mimicking flow phantom model in vitro. We will quantify specificity, brightness, and external detectability up to 4 mm deep in tissue. In Aim 3, we will test OTL38 and H-DiFC in a mouse metastasis model and in hairless guinea pigs (which have similar optical properties to human skin) in vivo. If successful, H-DiFC would allow sensitive and accurate enumeration of CTCs continuously without drawing blood samples. This would represent a completely new diagnostic tool for staging, managing, and studying metastasis for a broad range of malignancies. Moreover, because OTL38 (and other fluorescent tracers) are already in advanced clinical trials, there would be a rapid pathway to first-in-human testing.

项目摘要该项目的目标是开发新技术，直接在肿瘤细胞中检测循环肿瘤细胞（CTC）。而不需要抽取血液样本。转移是导致大多数癌症相关的死亡，并且通常是由CTCs通过血管系统的传播介导的。因此，CTC引起了极大的兴趣。临床和基础癌症研究。几乎所有的CTC研究方法都依赖于绘制和分析小的（7.5 mL）血液样本，广泛称为“液体活检”。然而，由于CTC的稀有性，从少量血液样本中估计CTC数量是非常不准确的，并且稀有细胞可能会逃脱检测完全是液体活检对短时间内发生的CTC数量的自然变化也不敏感。我们的团队最近开发了一种新技术，弥散体内流式细胞术（DiFC），用于检测罕见的，荧光标记的CTC直接在小动物体内。DiFC使用漫射光采样大循环大量组织中的血量。因此，DiFC的主要优点是灵敏度：我们表明，DiFC可以准确检测不到1 CTC/mL PB。因为它不需要抽血，DiFC还允许对个体动物的纵向研究。DiFC也很容易扩展到更大的肢体，物种，甚至对人类来说然而，在人类中使用DiFC将需要靶CTC的明亮和特异性荧光标记， vivo.幸运的是，在可注射分子示踪技术方面已经有了重大的技术和监管进展用于癌症的荧光引导手术。对于该提议特别感兴趣的是，OTL 38是一种近红外（NIR）小分子叶酸受体（FR）靶向探针，该探针正处于卵巢癌和肝癌的III期临床试验中。 0 TL 38对血液中的CTC具有高特异性和亲和力，具有低的被其他血细胞或淋巴细胞的非特异性摄取。血管壁此外，因为NIR光通过生物组织经历最小的光衰减， OTL 38适用于从更深的血管进行检测。本项目的目标是开发和验证“荧光分子在体”的使能技术 CTC液体活检。在目标1中，我们将构建用于人类手腕或前臂的H-DiFC系统，其中动脉流速为每分钟100毫升。在目标2中，我们将验证使用OTL 38对FR+ CTCs的标记，并且 H-DiFC在体外手臂模拟流动体模模型中的可检测性。我们将量化特异性，亮度和外部可探测性高达4毫米深的组织。在目标3中，我们将测试OTL 38和H-DiFC，小鼠转移模型和无毛豚鼠（与人类皮肤具有相似的光学特性）体内。如果成功，H-DiFC将允许连续灵敏和准确地计数CTC，而无需绘图。血液样本这将代表一种全新的诊断工具，用于分期、管理和研究广泛的恶性肿瘤的转移。此外，因为OTL 38（和其他荧光示踪剂）是已经在高级临床试验中，将有一个快速的途径进行首次人体试验。