Imaging Apoptosis for Chemotherapy Responses

化疗反应的细胞凋亡成像

• 批准号: 6778584
• 负责人: ANNA E LOKSHIN
• 金额: $ 16.34万
• 依托单位: UNIVERSITY OF PITTSBURGH AT PITTSBURGH
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-05-05 至 2006-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6778584
• 关键词: antineoplastics; apoptosis; athymic mouse; bioimaging /biomedical imaging; breast neoplasms; cell line; disease /disorder model; doxorubicin; drug administration rate /duration; drug screening /evaluation; fluorescent dye /probe; immunocytochemistry; laboratory mouse; melanoma; method development; neoplasm /cancer chemotherapy; neoplastic growth; pharmacokinetics; technetium; toxicology

DESCRIPTION (provided by applicant): Recent studies indicate that therapeutic efficacy of anti-cancer therapy could be correlated with early tumor apoptotic responses. Therefore, evaluation of early tumor apoptosis could be instrumental in prediction of therapy outcome. In vivo imaging of the whole tumor may represent a sensitive and powerful approach, as this method does not involve invasive techniques and provides complete information about apoptosis in the entire tumor. The long-term goal of this proposal is to develop a novel method for sensitive and specific apoptosis imaging for prediction of early tumor apoptotic response to chemotherapy. The main objectives of this proposal are (i) to use a fluorescent dye, TO-PRO-3, for optical imaging of tumor apoptosis in vivo; and (ii) to apply this technique for prediction of drug efficacy. We selected to use TO-PRO-3 because it is permeable only to dead and dying cells and becomes strongly fluorescent only upon binding the DNA of these cells, thus minimizing the background from non-specific absorption to tissues. In addition, its spectral characteristics make it highly suitable for optical imaging. The following Specific Aims are proposed: 1. To develop and optimize optical imaging of tumor apoptosis with TO-PRO-3 in mouse xenografi model of human melanoma. Nude mouse xenograft model of MCF-7 breast carcinoma will be utilized. We will evaluate possible toxicity of TO-PRO-3 and determine dose and time schedules of TO-PRO-3 for the most sensitive detection of tumor apoptosis. Furthermore, we will perform immunohistochemical analysis of drug-treated tumors in order to correlate the in vivo and ex vivo imaging results. Finally, we will confirm the advantage of TO-PRO-3 over 99mTc-labeled annexin V methods for in vivo imaging of tumor drug responses. 2. To correlate the level of drug-induced apoptosis with therapeutic drug responses in melanoma and breast cancer models. In this Specific Aim, we will evaluate drug-induced tumor apoptosis by optical imaging after multiple drug treatments and its correlation with tumor growth inhibition. This will allow determination of its predictive value for disease outcome. Nude mouse xenografts of MCF-7 tumor cell lines will be utilized and the effects of doxorubicin will be evaluated. The proposed study would result in the development of the novel highly sensitive and specific technique for early prediction of tumor drug response. This will allow rapid individual assessment and optimization of treatment of cancer patients while minimizing the unnecessary drug toxicity.

描述（由申请人提供）： 最近的研究表明，抗癌疗法的疗效可能与早期肿瘤凋亡反应相关。因此，早期肿瘤细胞凋亡的评估可能有助于预测治疗结果。整个肿瘤的体内成像可能代表一种敏感且强大的方法，因为该方法不涉及侵入性技术并提供有关整个肿瘤细胞凋亡的完整信息。该提案的长期目标是开发一种灵敏且特异的细胞凋亡成像新方法，以预测早期肿瘤细胞凋亡对化疗的反应。该提案的主要目标是（i）使用荧光染料 TO-PRO-3 对体内肿瘤细胞凋亡进行光学成像； (ii) 应用该技术来预测药物功效。我们选择使用 TO-PRO-3，因为它仅对死亡和垂死的细胞具有渗透性，并且仅在结合这些细胞的 DNA 时才会发出强烈的荧光，从而最大限度地减少组织非特异性吸收的背景。此外，其光谱特性使其非常适合光学成像。提出以下具体目标： 1. 在人类黑色素瘤的小鼠异种移植模型中开发和优化 TO-PRO-3 肿瘤细胞凋亡的光学成像。将使用MCF-7乳腺癌的裸鼠异种移植模型。我们将评估 TO-PRO-3 可能的毒性，并确定 TO-PRO-3 的剂量和时间安排，以最灵敏地检测肿瘤细胞凋亡。此外，我们将对药物治疗的肿瘤进行免疫组织化学分析，以便关联体内和离体成像结果。最后，我们将确认 TO-PRO-3 相对于 99mTc 标记的膜联蛋白 V 方法在肿瘤药物反应体内成像方面的优势。 2. 将黑色素瘤和乳腺癌模型中药物诱导的细胞凋亡水平与治疗药物反应相关联。在这个具体目标中，我们将通过光学成像评估多次药物治疗后药物诱导的肿瘤细胞凋亡及其与肿瘤生长抑制的相关性。这将有助于确定其对疾病结果的预测价值。将利用 MCF-7 肿瘤细胞系的裸鼠异种移植物并评估阿霉素的作用。拟议的研究将导致开发用于早期预测肿瘤药物反应的新型高灵敏度和特异性技术。这将允许快速个体评估和优化癌症患者的治疗，同时最大限度地减少不必要的药物毒性。