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

• 批准号: 10112518
• 负责人: Mark Jonathan Niedre
• 金额: $ 20.95万
• 依托单位: NORTHEASTERN UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-01-01 至 2022-12-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10112518
• 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; 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

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 该项目的目的是开发新技术，以直接检测循环肿瘤细胞（CTC） 血液不必抽血。转移负责大多数与癌症有关的 死亡，通常是通过脉管系统传播CTC来介导的。因此，CTC引起了极大的兴趣 临床和基本癌症研究。几乎所有用于研究CTC的方法都依赖于绘图和分析 小（7.5 mL）血液样本，通常称为“液体活检”。但是，由于CTC的稀有性 小血样的CTC数量的估计极为不准确，稀有细胞可能逃脱检测 完全。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，散射体体流式细胞仪（DIFC），以检测稀有的， 荧光标记的CTC直接在小动物中体内。 DIFC使用漫射光来取样大型循环 散装组织中的血量。因此，DIFC的主要优点是灵敏度：我们表明DIFC可以 准确地检测到每毫升Pb少于1 CTC。因为它不需要抽血，因此DIFC也允许 单个动物的纵向研究。 DIFC也很容易扩展到较大的四肢，物种和潜在 甚至对人类。 但是，在人类中使用DIFC将需要目标CTC的明亮而特定的荧光标记 体内。幸运的是，可注射的分子示踪剂技术取得了重大的技术和法规进展 用于荧光引导性癌症手术。该提案特别有趣，OTL38是一个近红外（NIR） 小分子叶酸纤维受体（FR）靶向的探针，该探针正在III期临床试验中进行卵巢癌和肝癌。 OTL38对血液中的CTC具有很高的特异性和亲和力，其他血细胞或其他血细胞的非特异性摄取或 船墙。此外，由于NIR光经历了通过生物组织的最小衰减，所以 OTL38适用于从较深的血管中检测。 该项目的目的是开发和验证体内荧光分子的能力技术 CTCS的液体活检。在AIM 1中，我们将构建一个H-DIFC系统，用于在人的腕部或森林中使用 动脉流速为每分钟100毫升。在AIM 2中，我们将用OTL38验证FR+ CTC的标记，并且 在体外模仿手臂的流动幻影模型中使用H-DIFC可检测性。我们将量化特异性， 亮度和外部可检测性在组织中深4毫米。在AIM 3中，我们将在A中测试OTL38和H-DIFC 在体内，小鼠转移模型和无毛豚鼠（具有与人皮相似的光学特性）。 如果成功，H-DIFC将不断允许CTC的敏感且准确的枚举，而无需绘图 血液样本。这将代表用于分期，管理和学习的全新诊断工具 多种恶性肿瘤的转移。而且，因为OTL38（和其他荧光示踪剂）是 已经在高级临床试验中，将有一个人类测试的快速途径。