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.

项目概要该项目的目标是开发新技术来直接检测循环肿瘤细胞（CTC）无需抽取血液样本即可进行血流转移是大多数癌症相关的原因。死亡，并且通常是通过脉管系统传播 CTC 来介导的，因此引起了极大的兴趣。在临床和基础癌症研究中，几乎所有 CTC 研究方法都依赖于绘制和分析。小量（7.5 mL）血液样本，通常被称为“液体活检”，但是，由于 CTC 的稀有性。从小量血液样本中估算 CTC 数量极其不准确，稀有细胞可能会逃脱检测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新技术，即弥散体内流式细胞术 (DiFC)，用于检测罕见、荧光标记的 CTC 直接在小动物体内使用漫射光对大型循环进行采样。因此，DiFC 的主要优点是敏感性：我们证明 DiFC 可以。准确检测每毫升 PB 中少于 1 个 CTC 由于不需要抽血，DiFC 还允许。对个体动物的纵向研究也很容易扩展到更大的肢体、物种和潜在的。甚至对人类。然而，在人类中使用 DiFC 需要对目标 CTC 进行明亮且特异的荧光标记。幸运的是，注射分子示踪剂技术在技术和监管方面取得了重大进展。对于荧光引导癌症手术，OTL38 是一种近红外 (NIR) 技术。小分子叶酸受体 (FR) 靶向探针，正在进行卵巢癌和肝癌的 III 期临床试验。 OTL38 对血液中的 CTC 具有高特异性和亲和力，其他血细胞或其他血细胞的非特异性摄取较低此外，由于近红外光穿过生物组织的光衰减最小， OTL38适合检测较深层的血管。该项目的目标是开发和验证“体内荧光分子”的使能技术在目标 1 中，我们将构建一个用于人类手腕或前臂的 H-DiFC 系统。动脉流速为每分钟 100 毫升在目标 2 中，我们将使用 OTL38 验证 FR+ CTC 的标记，并且 H-DiFC 在体外模拟手臂流动体模模型中的可检测性我们将量化特异性，在 Aim 3 中，我们将在组织中测试 OTL38 和 H-DiFC。小鼠转移模型和无毛豚鼠（具有与人类皮肤相似的光学特性）体内。如果成功，H-DiFC 将允许连续灵敏、准确地计数 CTC，而无需绘图这将代表一种用于分期、管理和研究的全新诊断工具。此外，由于 OTL38（和其他荧光示踪剂）是多种恶性肿瘤的转移。已经进入高级临床试验，将有一条快速途径进行首次人体测试。

项目成果

期刊论文数量（2）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Near-infrared diffuse in vivo flow cytometry.

近红外漫射体内流式细胞术。

DOI：
发表时间：
2022-09
期刊：
影响因子：
3.5
作者：
Pace, Joshua;Ivich, Fernando;Marple, Eric;Niedre, Mark
通讯作者：
Niedre, Mark

In Vivo Labeling and Detection of Circulating Tumor Cells in Mice Using OTL38.

使用 OTL38 体内标记和检测小鼠循环肿瘤细胞。

DOI：
10.1007/s11307-024-01914-0
发表时间：
2024-04-09
期刊：
Molecular imaging and biology
影响因子：
3.1
作者：
Joshua Pace;Jane Lee;Madduri Srinivasarao;Shivakrishna Kallepu;Philip S. Low;M. Niedre
通讯作者：
M. Niedre