Continuous, Non-Invasive Optical Monitoring of Circulating Tumor Cell-Mediated Metastasis in Awake Mice

连续、非侵入性光学监测清醒小鼠循环肿瘤细胞介导的转移

• 批准号: 10387600
• 负责人: Mark Jonathan Niedre
• 金额: $ 44.12万
• 依托单位: NORTHEASTERN UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-03-03 至 2027-02-28
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10387600
• 关键词: Adjuvant; Affect; Algorithms; Animal Model; Animals; Anti-Inflammatory Agents; Biology; Blood; Blood Circulation; Blood Vessels; Blood Volume; Blood specimen; Brain; Cancer Model; Cell Count; Cell Survival; Cerebrospinal Fluid; Cessation of life; Childhood Malignant Brain Tumor; Childhood Medulloblastomas; Clinical; Complex; Data; Data Collection; Data Set; Detection; Development; Diffuse; Disease; Distant; Dose; Fiber; Flow Cytometry; Fluorescence; Future; Goals; Hematogenous; Hour; In Vitro; Individual; Inflammatory Response; Lasers; Lead; Light; Lighting; Longitudinal Studies; Malignant Neoplasms; Measures; Mediating; Methods; Modeling; Monitor; Multiple Myeloma; Mus; Neoplasm Circulating Cells; Neoplasm Metastasis; Optics; Organ; Pharmaceutical Preparations; Pilot Projects; Primary Neoplasm; Process; Proteins; Radiation; Radiation therapy; Research; Sampling; Scanning; Signal Transduction; Site; Surface; System; Techniques; Technology; Testing; Tissues; Transgenic Mice; Transgenic Organisms; Treatment Protocols; Vertebral column; Xenograft Model; Xenograft procedure; anticancer treatment; awake; cancer therapy; design; detection sensitivity; improved; in vivo; insight; instrument; liquid biopsy; medulloblastoma; member; mouse model; non-invasive monitor; optical fiber; peripheral blood; pre-clinical; radiation response; response; signal processing; tool; tumor

Project Summary Hematogenous metastasis is responsible for a large majority of cancer-related deaths, where circulating tumor cells (CTCs) shed from the primary tumor into the peripheral blood (PB). A small number of CTCs may form secondary sites, which are extremely difficult to control clinically. Most methods for studying CTCs rely on drawing and analyzing fractionally small PB blood samples (“liquid biopsy”). Although CTCs and multicellular CTC clusters (CTCCs) have been studied for decades, little is known about their “dynamics” in vivo (transient changes in their numbers in PB), and how these may affect metastasis development and response to anti-cancer treatment. For example, it is know that radiation therapy may encourage metastatic dissemination of cancer, yet the mechanisms for this are still poorly understood. Our team recently developed a new method for in vivo enumeration of CTCs in small animals called “diffuse in vivo flow cytometry” (DiFC). DiFC uses diffuse light to detect fluorescent-protein expressing CTCs in large, deeply-seated blood vessels. DiFC can sample approximately 100 microliters of blood per minute, permitting detection of fewer than 1 CTC per mL of PB, and sampling of the entire peripheral blood volume in minutes. We previously used DiFC to study rare CTC and CTCC dissemination in mouse xenograft models. DiFC revealed that CTC numbers are highly dynamic and may change by an order-of-magnitude or more over 24 hour periods. These changes are largely missed by CTC enumeration methods that involve infrequent blood draws. The goal of this project is to build a “wearable” tethered w-DiFC instrument that will allow continuous, non- invasive monitoring of CTC numbers over extended periods in mice. The w-DiFC optical probe and signal processing design will permit data collection in freely-moving mice in ambient lighting conditions. We will first use w-DiFC to study CTC dynamics during disease development in an orthotopic xenograft and transgenic mouse model of metastasis. We will use also w-DiFC to measure continuous CTC dynamics after radiation therapy in a medulloblastoma (MB) mouse model. MB is a common form of childhood brain cancer that aggressively metastasizes to the leptomeningeal surfaces of the brain and spine via the PB. There is significant evidence that radiation may exacerbate metastasis by triggering mobilization of CTCs into the blood. We expect that the ability of w-DiFC to measure CTCs over short-, medium-, and long-term timescales will provide unique insights into this process. We will also use w-DiFC to study the use of anti-inflammatory drugs to block the pro-metastatic effect. Hence, the studies proposed here could ultimately lead to better understanding of metastasis and improved treatment protocols for childhood MB. We anticipate that the unique technologies that will be developed here will have broad application to other cancers and anti-cancer therapies in the future.

项目概要 血行转移是造成绝大多数癌症相关死亡的原因，其中循环肿瘤 细胞 (CTC) 从原发肿瘤脱落到外周血 (PB) 中，可能会形成少量 CTC。 大多数研究 CTC 的方法都依赖于次要位点。 抽取并分析少量 PB 血液样本（“液体活检”）。 CTC 簇（CTCC）已经被研究了几十年，但人们对它们在体内的“动态”（瞬态）知之甚少。 PB 中数量的变化，以及这些变化如何影响转移发展和抗癌反应 例如，众所周知，放射治疗可能会促进癌症的转移扩散。 其机制仍知之甚少。 我们的团队最近开发了一种小动物体内 CTC 计数的新方法，称为“弥漫性 体内流式细胞术”（DiFC）使用漫射光来检测大的、表达荧光蛋白的 CTC。 DiFC 每分钟可采集约 100 微升的血液，允许进行。 每毫升 PB 检测不到 1 个 CTC，并在几分钟内完成整个外周血容量的采样。 先前使用 DiFC 研究小鼠异种移植模型中罕见的 CTC 和 CTCC 传播。 CTC 数字高度动态，可能在 24 小时内发生一个数量级或更多的变化。 涉及不频繁抽血的 CTC 计数方法在很大程度上遗漏了这些变化。 该项目的目标是构建一种“可穿戴”系留 w-DiFC 仪器，该仪器将允许连续、非 w-DiFC 光学探针和信号对小鼠体内 CTC 数量进行长期侵入性监测。 处理设计将允许在环境照明条件下自由移动的小鼠中收集数据。 使用 w-DiFC 研究原位异种移植和转基因小鼠疾病发展过程中的 CTC 动态 转移模型。 我们还将使用 w-DiFC 测量髓母细胞瘤放射治疗后的连续 CTC 动态 (MB) 小鼠模型。MB 是儿童脑癌的一种常见形式，可侵袭性转移至大脑。 有重要证据表明辐射可能会通过 PB 影响大脑和脊柱的软脑膜表面。 通过触发 CTC 动员到血液中来加剧转移，我们预计 w-DiFC 具有以下能力： 在短期、中期和长期时间范围内衡量 CTC 将为这一过程提供独特的见解。 还将使用w-DiFC来研究使用抗炎药物来阻断促转移作用。 这里提出的研究最终可能会导致更好地了解转移并改善治疗 我们预计这里将开发的独特技术将具有儿童MB的协议。 未来将广泛应用于其他癌症和抗癌治疗。