In vivo real-time detection of circulating melanoma cells

体内循环黑色素瘤细胞实时检测

• 批准号: 7657021
• 负责人: Vladimir P Zharov
• 金额: $ 27.73万
• 依托单位: UNIV OF ARKANSAS FOR MED SCIS
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-03-19 至 2014-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7657021
• 关键词: African American; Animal Model; Biological Assay; Blood; Blood Circulation; Blood flow; Blood specimen; Caucasians; Caucasoid Race; Cells; Cessation of life; Characteristics; Control Groups; Data; Detection; Development; Diagnostic Neoplasm Staging; Disease; Fiber; Flow Cytometry; Goals; Human; Human Cell Line; In Vitro; Individual; Injection of therapeutic agent; Label; Lasers; Malignant Neoplasms; Melanoma Cell; Metastatic Melanoma; Methods; Micrometastasis; Monitor; Mus; Neoplasm Metastasis; Noise; Painless; Patients; Physiologic pulse; Public Health; Signal Transduction; Skin Pigmentation; Skin Tissue; Spectrum Analysis; Staging; Tail; Techniques; Time; Translating; Treatment Efficacy; Tumor stage; Veins; absorption; base; cancer cell; cancer recurrence; healthy volunteer; in vivo; innovation; melanoma; mouse model; neoplastic cell; public health relevance

DESCRIPTION (provided by applicant): Death from cancer, especially melanoma, is most often related to metastasis. As the quantitative detection of rare cancer cells in the blood circulation appears to be an early marker of metastatic development, cancer recurrence, and therapeutic efficacy, and as assays for detecting metastatic cells are currently only in vitro techniques that are inadequate to these tasks, our proposal has as its ultimate goal the development of photoacoustic (PA) flow cytometry (FC) in vivo, a new method for ultrasensitive real-time, label-free noninvasive quantitative detection of circulating melanoma cells in blood flow. We will pursue this goal through accomplishment of the following Specific Aims: Aim 1. Develop an advanced PA flow cytometer (PAFC) and verify its parameters in vitro. A new, advanced PA flow cytometer will be developed that uses a near- infrared (IR) laser with a high pulse repetition rate. Its main characteristics, especially its sensitivity threshold, will be evaluated in vitro with the use of static and flowing melanoma cells. Aim 2. Estimate the capability of PAFC to detect single melanoma cells in real time in vivo in an animal model. The absorption contrast of melanoma cells in relation to blood and skin tissues will be determined by PA spectroscopy in vivo in a mouse model at different laser wavelengths. The capability of PAFC for real-time, label-free detection of melanoma cells will be verified by the injection of cells into the tail veins of mice. Aim 3. Ascertain the capability of PAFC to monitor circulating metastatic melanoma cells at different stages of tumor development in an animal model. Metastatic tumor cells will be detected in real time with the PA technique in blood circulation in an animal model at different stages of tumor development, with available conventional assays serving as independent controls. Aim 4. Translate PAFC to human application for quantitative monitoring of CTCs at different stages of melanoma development. The capability of a painless, non-invasive, label-free, fiber- based PAFC for quantitative detection of circulating melanoma cells in vivo in humans will be assessed in four stages: (1) ex vivo PA study of blood samples from healthy donors spiked with melanoma human cell lines; (2) ex vivo PA study of blood samples from melanoma patients verified with conventional assays; (3) in vivo study of healthy individuals as a control group with different skin pigmentation; and 4) melanoma patients at different stages of disease. In the course of this study, we will obtain statistically significant data that will demonstrate this innovative technique's unprecedented capability for quantitatively monitoring circulating melanoma cells in vivo without the need for labeling. The benefits to the public health of achieving this goal extend to routinely monitoring circulating tumor cells as early marker for the micrometastasis development and cancer recurrence in vivo in melanoma patients, as well as to evaluating the efficacy of therapy. PUBLIC HEALTH RELEVANCE: The capability of a painless, noninvasive, label-free, fiber-based photoacoustic flow cytometry (PAFC) for selective, time-resolved detection of PA signals from different vessels will be assessed in (1) Caucasian and African American healthy volunteers and (2) melanoma patients at different stages of disease, in whom circulating metastatic melanoma cells will be quantitatively determined. In the course of this study, we will obtain statistically significant data that will demonstrate this innovative technique's unprecedented capability for quantitatively monitoring circulating melanoma cells in vivo without the need for labeling. The benefits to the public health of achieving this goal extend to the routine monitoring of circulating cells as early markers of micrometastatic development and cancer recurrence in vivo in melanoma patients, as well as to evaluating the efficacy of therapy.

描述（由申请人提供）：癌症的死亡，尤其是黑色素瘤，通常与转移有关。 As the quantitative detection of rare cancer cells in the blood circulation appears to be an early marker of metastatic development, cancer recurrence, and therapeutic efficacy, and as assays for detecting metastatic cells are currently only in vitro techniques that are inadequate to these tasks, our proposal has as its ultimate goal the development of photoacoustic (PA) flow cytometry (FC) in vivo, a new method for ultrasensitive实时的无标记的无创定量检测血流中循环黑色素瘤细胞。我们将通过实现以下特定目的来实现这一目标：目标1。开发高级PA流式细胞仪（PAFC）并在体外验证其参数。将开发出一种新的，先进的PA流式细胞仪，该流式细胞仪使用具有较高脉冲重复速率的近红外（IR）激光器。它的主要特征，尤其是其敏感性阈值，将通过使用静态和流动的黑色素瘤细胞在体外进行评估。目标2。估计PAFC在动物模型中实时检测单个黑色素瘤细胞的能力。黑色素瘤细胞相对于血液和皮肤组织的吸收对比度将通过在不同激光波长下的小鼠模型中的体内PA光谱来确定。 PAFC在实时，无标记的黑色素瘤细胞中的能力将通过将细胞注射到小鼠的尾静脉中来验证。目标3。确定在动物模型中肿瘤发育的不同阶段，PAFC监测循环转移性黑色素瘤细胞的能力。在肿瘤发育的不同阶段，在动物模型的血液循环中，将实时检测转移性肿瘤细胞，并具有可用的常规分析作为独立对照。 AIM 4。将PAFC转化为人类黑色素瘤发育不同阶段中CTC的定量监测。将在人类体内进行定量检测体内循环黑色素瘤细胞的定量检测无痛的，无标记的，无标签的，无标签的PAFC的能力：（1）在体内对来自黑色素瘤人类瘤人类细胞系的健康捐赠者的血液样本研究； （2）在体内对经常测定的黑色素瘤患者的血液样本研究； （3）在体内研究健康个体作为具有不同皮肤色素沉着的对照组； 4）处于疾病不同阶段的黑色素瘤患者。在这项研究的过程中，我们将获得具有统计学意义的数据，这些数据将证明这种创新技术在体内定量监测循环黑色素瘤细胞而无需标记的循环黑色素瘤细胞。实现这一目标的公共卫生的好处扩展到常规监测循环肿瘤细胞，作为黑色素瘤患者体内微阶段发育和癌症复发的早期标志物，以及评估治疗的疗效。公共卫生相关性：无痛，无创，无标签，基于纤维的光声流式细胞仪（PAFC）的能力，用于选择性，时间分解的检测来自不同血管的PA信号的能力，将在（1）青年人和非裔美国人健康的志愿者和（2）个疾病中，在不同阶段的疾病中，将在（1）产生循环范围内得到循环的临床群体。在这项研究的过程中，我们将获得具有统计学意义的数据，这些数据将证明这种创新技术在体内定量监测循环黑色素瘤细胞而无需标记的循环黑色素瘤细胞。实现这一目标的公共卫生的好处扩展到了对循环细胞的常规监测，作为黑色素瘤患者体内微转移性发育和癌症复发的早期标志，以及评估治疗的疗效。