A novel preclinical model of spontaneous metastasis

一种新型的自发转移临床前模型

• 批准号: 7966147
• 负责人: GLENN T. MERLINO
• 金额: $ 46.15万
• 依托单位: DIVISION OF BASIC SCIENCES - NCI
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/7966147
• 关键词: Adjuvant Chemotherapy; Adjuvant Therapy; Angiogenesis Inhibitors; Animal Model; Antineoplastic Agents; Biological Assay; Biological Markers; Bioluminescence; C57BL/6 Mouse; Cancer cell line; Cell Culture Techniques; Cisplatin; Clinic; Clinical; Clinical Trials; Collaborations; Complex; Data; Development; Diagnosis; Disease; Disease Progression; Distant; Drug Delivery Systems; Drug Monitoring; Excision; Exhibits; Goals; Growth; Human; Immune; Immune system; Immunocompetent; Immunocompromised Host; In Vitro; Kinetics; Label; Laboratories; Lesion; Lewis Lung Carcinoma; Luciferases; Lung; Malignant Epithelial Cell; Malignant Neoplasms; Malignant neoplasm of lung; Modeling; Monitor; Mus; Neoplasm Metastasis; Neoplasm Transplantation; Non-Small-Cell Lung Carcinoma; Operative Surgical Procedures; Organ; Paclitaxel; Patients; Pharmaceutical Preparations; Plastics; Pre-Clinical Model; Primary Neoplasm; Proteins; Reaction Time; Recurrence; Reporter; Reporting; Resected; Science; Signal Transduction; Site; Staging; Subfamily lentivirinae; System; Technology; Testing; Therapeutic; Time; Tissues; Transplantation; Treatment Efficacy; base; cancer therapy; chemotherapeutic agent; chemotherapy; clinically relevant; clinically significant; design; experience; host neoplasm interaction; in vivo; meetings; melanoma; mouse model; neoplastic cell; novel; novel strategies; pre-clinical; prevent; programs; rapid growth; response; subcutaneous; success; tool; tumor; tumor growth; tumor progression

The major goal of this Sub-Project is to develop and exploit a tractable, preclinical mouse model of metastatic disease that recapitulates the clinical progression of surgical removal of primary tumor, treatment with adjuvant therapy and clinical recurrence at distant sites. We see a translational opportunity to devise a novel strategy to prevent the clinical recurrence of metastatic disease. The model will be designed to incorporate normal immune system function to maintain an appropriate tumor microenvironment and provide superior tumor-host interactions, an approach that is significantly more likely to yield biologically and clinically relevant data. Since metastasis often targets internal organs and the timing of its recurrence may vary greatly, the preclinical model will allow for the non-invasive, long-term monitoring of disease progression within the immunocompetent mouse. To meet these criteria, we have developed a syngeneic immunocompetent mouse model of Lewis Lung Carcinoma (LLC) using tissue that was never adapted to cell culture and labeled using a high-titer lentivirus encoding a luciferase/GFP fusion reporter protein, developed by this laboratory in collaboration with the Advanced Technology Program. Our decision to employ the NSCLC mouse LLC model is well aligned with the new NCI Lung Cancer Program, and provides a robust, well-characterized system to validate our preclinical approach. LLC exhibits highly favorable growth kinetics and virtually 100% take rate of lung metastases in C57BL/6 mice, and was instrumental in the successful development of early conventional chemotherapies that are still used in the clinical today. LLC has been subcloned in vitro and many sublines have been established since being discovered in 1952; however, for this preclinical model we felt it was essential to use an in vivo-maintained LLC stock that has never been adapted to cell culture, an approach that has been reported to yield more reliable preclinical data. In full collaboration with Dr. Melinda Hollingshead (DTP, NCI), we have now established a stably labeled bank of highly metastatic LLC tissue (LLC-Luc/GFP) through multiple rounds of subcutaneous transplantation, resection, and selection of Luc/GFP+ metastatic clones in syngeneic C57BL/6 mice. Our preclinical approach will be to subcutaneously inoculate Luc/GFP-labeled LLC cells into albino C57BL/6 mice and resect consistently at a selected tumor size, at which time drug treatment can be initiated. Recurrence of clinically significant LLC macrometastases is evident in the lungs as Luc/GFP+ lesions within a week, and an entire drug study can be completed in less than one month. The BL signal allows the monitoring of disease after resection of the primary tumor, the progression of metastatic growth over time, and the response of the tumor to therapy; moreover, the BL intensity is well correlated with the pathological score of the metastatic disease. We have found that this resection/recurrence metastasis model fulfills multiple requirements for a tractable and robust syngeneic preclinical model of advanced stage disease, one that we anticipate may more accurately predict therapeutic response. To date we have treated mice in a setting akin to post-surgical first-line adjuvant chemotherapy using cisplatin, paclitaxel and/or antiangiogenic agents. As in the clinic these drugs were found to be most effective against progression in combination. However, the response of metastases to agents could not be predicted from, and often opposed, their effects on subcutaneous tumors. Moreover, time to macrometastasis onset, rather than growth, correlated with both mouse survival and treatment efficacy.

该亚项目的主要目的是开发和利用一种可进行的临床前小鼠转移性疾病模型，该模型概括了手术切除原发性肿瘤的临床进展，辅助治疗和远处部位的临床复发治疗。我们看到了一个翻译的机会，可以制定一种新的策略，以防止转移性疾病的临床复发。该模型将旨在结合正常的免疫系统功能，以维持适当的肿瘤微环境并提供优质的肿瘤宿主相互作用，这种方法更有可能在生物学和临床上相关的数据产生。由于转移通常靶向内部器官，并且其复发的时间可能有很大变化，因此临床前模型将允许对免疫能力小鼠内疾病进展的无创，长期监测。为了符合这些标准，我们开发了一种使用从未适应细胞培养的组织的Lewis Lung Carcinoma（LLC）的合成性免疫能力的小鼠模型，并使用编码高细胞慢病毒编码荧光素酶/GFP Fusion Fusion Reporter蛋白质，该蛋白质由此实验室合作开发。我们采用NSCLC Mouse LLC模型的决定与新的NCI肺癌计划非常一致，并提供了一个健壮，良好的系统来验证我们的临床前方法。 LLC在C57BL/6小鼠中表现出高度有利的生长动力学和几乎100％的肺转移率，并且在当今仍在临床中仍然使用的早期常规化学疗法的成功开发中发挥了作用。 LLC在体外已被亚克隆，自1952年被发现以来已经建立了许多下议院。但是，对于这种临床前模型，我们认为使用从未适用于细胞培养的体内维护限制性有限责任公司的股票至关重要，这种方法已据报道产生更可靠的临床前数据。在与Melinda Hollingshead博士（DTP，NCI）的完全合作中，我们现在已经建立了稳定的高度转移性LLC组织（LLC-LUC/GFP）的标签库，通过皮下移植，切除，LUC/GFP+ LUC/GFP+转移性CLONES在Syngenec C57bl/6 mice中的选择。我们的临床前方法是皮下接种LUC/GFP标记的LLC细胞进入白化病C57BL/6小鼠，并在选定的肿瘤大小下始终如一地切除，在此期间可以启动药物治疗。在肺部的LUC/GFP+病变中，临床上显着的LLC大量重温的复发是明显的，并且整个药物研究都可以在不到一个月的时间内完成。 BL信号允许切除原发性肿瘤后监测疾病，随着时间的流逝，转移性生长的进展以及肿瘤对治疗的反应；此外，BL强度与转移性疾病的病理评分很好地相关。我们发现，这种切除/复发转移模型满足了晚期疾病的可触犯和稳健的促临床前模型的多种要求，我们预计这种临床疾病可能会更准确地预测治疗反应。迄今为止，我们已经使用顺铂，紫杉醇和/或抗血管生成剂进行了类似于手术后一线辅助化疗的情况。与诊所一样，发现这些药物最有效地抵抗了组合的进展。 但是，转移对剂的反应不能从它们对皮下肿瘤的影响中预测，并且通常反对。此外，大量开始出现而不是生长的时间与小鼠的存活和治疗效果相关。