Tumor models for the study of inflammation and oncogenesis

用于研究炎症和肿瘤发生的肿瘤模型

• 批准号: 8937889
• 负责人: Robert Wiltrout
• 金额: $ 39.11万
• 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/8937889
• 关键词: B-Lymphocytes; Bacterial Proteins; Binding; Blood; CD8B1 gene; Cancer Etiology; Carcinoma; Cells; Characteristics; Chronic; Code; Coupled; Cytokine Signaling; Data; Development; Engineering; Epidemiology; Fatty Liver; Frequencies; Gene Delivery; Gene Expression; Gene Silencing; Gene Targeting; Genes; Genetic; Green Fluorescent Proteins; Hepatic; Hepatocyte; Image; Immune; Immune response; Immune system; Immunodeficient Mouse; Incidence; Individual; Inflammation; Inflammation Mediators; Inflammatory; Lesion; Ligands; Liver; Liver Cell Adenoma; Liver diseases; Liver neoplasms; Luciferases; Malignant Neoplasms; Measures; Mediating; Modeling; Molecular; Monitor; Mouse Strains; Mus; Mutation; Neoplasm Metastasis; Obesity; Oncogenes; Oncogenic; Organ; Pathway interactions; Physiological; Play; Pre-Clinical Model; Predisposition; Primary carcinoma of the liver cells; Proto-Oncogene Proteins c-akt; Reagent; Reporter Genes; Role; Signal Transduction; Sleeping Beauty; Solid Neoplasm; Somatic Cell; Sorting - Cell Movement; Source; Study models; Surrogate Markers; T-Lymphocyte; TNF gene; TNFRSF1A gene; Tissues; Transgenic Mice; Tumor Promotion; Viral Proteins; beta catenin; cellular engineering; clinically relevant; cytokine; improved; in vivo; knockout gene; mouse model; non-alcoholic fatty liver; receptor; response; therapeutic development; tumor; tumor growth; tumor progression; tumorigenesis

Mouse tumor models are most useful when they reproduce as many known molecular lesions as possible, so as to most accurately portray the body's physiological response to tumorigenesis. With somatic cell engineering (Sleeping Beauty/SB), we can recapitulate multiple lesions simultaneously with ease and scale not possible by other means. Hepatocellular carcinoma is the third leading cause of cancer worldwide, with increasing incidence due to a number of epidemiologic and biologic reasons, such as obesity. Moreover, the liver is a frequent depot for metastases originating from solid tumors arising in other organs as well. We have developed oncogene-initiated liver tumors which recapitulate the development and progression of hepatocellular carcinoma (HCC) and hepatocellular adenoma (HCA). Our first Aim is to optimize and quantitate hepatic tumor development, using reporter genes. To date, we have reproducibly generated hepatocellular adenomas (HCA) by the co-delivery of SB-AKT and SB-beta-catenin (CAT), and hepatocellular (HCC) by the co-delivery of SB-MET and SB-CAT. In contrast, individual delivery these oncogenes does not induce tumor formation. We are using a co-delivered Gaussia luciferase transposon to track tumor growth from oncogene-initiated hepatocytes to overt tumor formation. Gaussia luciferase is secreted into the blood thereby allowing for a non-invasive, quantitative measure of gene delivery and tumor development. Additionally co-delivered transposons expressing red or green fluorescent protein will be used to tag oncogene-initiated hepatocytes using tumor imaging and flow cytometric sorting. Our second Aim is to define the immunological contributions to AKT+CAT and MET+CAT oncogenic pathways in vivo. Using RAG1-/- immunodeficient mice, we found that these mice were significantly protected from the development of AKT+CAT (HCA) liver tumors. This finding suggests that the host immune system collaborates and provides essential contributing factors towards the development of AKT+CAT initiated HCA. In contrast, RAG1-/- mice succumbed more rapidly to tumor development when MET+CAT was used, suggesting an important role for immune-mediated surveillance of MET+CAT initiated HCC. Our ongoing studies using T cell- and B cell-deficient mice suggest that AKT+CAT induced hepatosteatosis development and overall tumor growth is dependent on B cells, but not CD4+ or CD8+ T cells. B cell deficient mice also have a dramatic reduction in hepatic steatosis, a characteristic feature underlying non-alcoholic fatty liver disease and predisposition to liver tumor development. The tumor-promoting role of B cells is controversial, but consistent with a tumor-promoting role that has been proposed for these cells in some chronic inflammatory models. We hypothesize that tumors can develop when initiating mutations are coupled with inflammatory microenvironments that enhance tumor promotion. Therefore, in the third Aim of this project, we will identify those cytokines and inflammatory networks capable of modulating AKT+CAT or MET+CAT initiated tumors. Our results from studies using immunodeficient mice have identified a critical tumor promoting role for TNFR1 and LTBetaR signaling in the AKT+CAT model, since mice deficient in these receptors or their ligands have reduced tumor growth and improved survival. B cell associated LTBeta, in particular, may therefore play an indispensable role in AKT+CAT induced HCA and hepatic steatosis. We are monitoring tumor progression in wildtype and B cell deficient mice that have been treated with LTBetaR agonistic and antagonistic LTBetaR-binding reagents. The involvement of additional B cell-derived factors will also be investigated. Additional studies with MET+CAT will be completed to investigate oncogene specific inflammatory-mediators of tumor progression to carcinoma. Collectively, the approaches described in this project will provide us with accurate pre-clinical models for HCA and HCC and facilitate the development of therapeutics that more effectively target tumorigenesis.

当小鼠肿瘤模型尽可能多地复制已知的分子病变时，它们是最有用的，以便最准确地描绘身体对肿瘤发生的生理反应。通过体细胞工程（睡美人/SB），我们可以轻松地同时重现多个病变，并且通过其他方式无法实现规模化。肝细胞癌是世界范围内第三大癌症原因，由于许多流行病学和生物学原因（如肥胖），其发病率不断增加。此外，肝脏也是源自其他器官中产生的实体瘤的转移的常见仓库。我们已经开发了癌基因启动的肝脏肿瘤，其重演了肝细胞癌（HCC）和肝细胞腺瘤（HCA）的发展和进展。我们的第一个目标是优化和定量肝肿瘤的发展，使用报告基因。迄今为止，我们已经通过SB-AKT和SB-β-连环蛋白（CAT）的共同递送可重复地产生肝细胞腺瘤（HCA），并且通过SB-MET和SB-CAT的共同递送可重复地产生肝细胞腺瘤（HCC）。相反，单独递送这些癌基因不会诱导肿瘤形成。我们正在使用一种共递送的Gaussia荧光素酶转座子来跟踪肿瘤从癌基因启动的肝细胞到明显的肿瘤形成的生长。Gaussia荧光素酶分泌到血液中，从而允许基因递送和肿瘤发展的非侵入性定量测量。此外，表达红色或绿色荧光蛋白的共递送转座子将用于使用肿瘤成像和流式细胞术分选来标记癌基因启动的肝细胞。我们的第二个目的是确定免疫学对体内AKT+CAT和MET+CAT致癌途径的贡献。使用RAG 1-/-免疫缺陷小鼠，我们发现这些小鼠被显着保护免受AKT+CAT（HCA）肝肿瘤的发展。这一发现表明，宿主免疫系统协作并为AKT+CAT引发的HCA的发展提供了重要的促进因素。相比之下，当使用MET+CAT时，RAG 1-/-小鼠更快地死于肿瘤发展，这表明MET+CAT引发的HCC的免疫介导的监视的重要作用。我们正在进行的使用T细胞和B细胞缺陷小鼠的研究表明，AKT+CAT诱导的脂肪肝发展和整体肿瘤生长依赖于B细胞，而不是CD 4+或CD 8 + T细胞。B细胞缺陷小鼠的肝脂肪变性也显著减少，肝脂肪变性是非酒精性脂肪肝病的特征和肝肿瘤发展的易感性。B细胞的肿瘤促进作用是有争议的，但与在一些慢性炎症模型中提出的这些细胞的肿瘤促进作用一致。我们假设，当启动突变与增强肿瘤促进的炎症微环境相结合时，肿瘤可以发展。因此，在本项目的第三个目标中，我们将鉴定能够调节AKT+CAT或MET+CAT引发的肿瘤的那些细胞因子和炎症网络。我们使用免疫缺陷小鼠的研究结果已经确定了TNFR 1和LTBetaR信号在AKT+CAT模型中的关键肿瘤促进作用，因为这些受体或其配体缺陷的小鼠减少了肿瘤生长并提高了存活率。因此，B细胞相关的LT β可能在AKT+CAT诱导的HCA和肝脂肪变性中发挥不可或缺的作用。我们正在监测野生型和B细胞缺陷小鼠的肿瘤进展，这些小鼠已经用LTBetaR激动性和拮抗性LTBetaR结合试剂治疗。还将研究其他B细胞衍生因子的参与。将完成MET+CAT的其他研究，以研究肿瘤进展为癌症的癌基因特异性炎症介质。总的来说，本项目中描述的方法将为我们提供准确的HCA和HCC临床前模型，并促进更有效地靶向肿瘤发生的治疗方法的开发。