Control of Glioma Cell Invasion By Immunotherapy

通过免疫疗法控制胶质瘤细胞侵袭

• 批准号: 7286817
• 负责人: ELIZABETH W. NEWCOMB
• 金额: $ 18.46万
• 依托单位: NEW YORK UNIVERSITY SCHOOL OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-09-15 至 2009-05-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7286817
• 关键词: Animal Model; Animals; Awareness; Biological; Brain; Brain Neoplasms; Cancer Vaccines; Cell Line; Cells; Class; Clinical Protocols; Complementary DNA; Complex; Cranial Irradiation; Data; Development; Dose; Down-Regulation; Engineering; Exploratory/Developmental Grant; Extracellular Matrix; Frozen Sections; Gene Expression; Genes; Glioma; Goals; Green Fluorescent Proteins; Growth; Harvest; Human; Immune response; Immune system; Immunohistochemistry; Immunologic Surveillance; Immunotherapeutic agent; Immunotherapy; In Vitro; Inflammatory Response; Intracranial Neoplasms; Invaded; Invasive; Laboratories; Long-Term Survivors; Major Histocompatibility Complex; Major Histocompatibility Complex Gene; Mediating; Modeling; Molecular; Molecular Profiling; Mus; Neuraxis; Numbers; Patients; Peripheral; Process; Property; Protein Overexpression; Proteins; Radiation; Radiation therapy; Research Personnel; Residual Tumors; Retroviridae; Schedule; Site; Small Interfering RNA; Specimen; System; T-Lymphocyte; Technology; Testing; Tetanus Helper Peptide; Therapeutic Effect; Time; Transcriptional Activation; Up-Regulation; Vaccination; Week; X-Ray Computed Tomography; base; clinically relevant; day; immunogenic; immunogenicity; implantation; improved; in vivo; in vivo Model; interdisciplinary collaboration; irradiation; neoplastic cell; novel; novel therapeutics; programs; protein expression; small hairpin RNA; tumor

DESCRIPTION (provided by applicant): Presently, there is no known therapy that can target invading glioma tumor cells. In order to study the biological properties of invading glioma cells at the molecular level, we have developed an in vivo animal model using green fluorescent protein (GFP)-tagged murine GL261 glioma cells. This allows us to identify invading glioma cells into the brain adjacent to the main tumor mass using GFP immunohistochemistry. Microarray gene technology high-lighted differences in gene expression profiles between invading and non-invading human glioma cells identifying MHC class I molecules as a potential target for novel immunotherapeutic approaches. The GL261 glioma cell line is poorly immunogenic due to its low expression of MHC molecules. However, we have shown that low dose whole brain radiation therapy (WBRT) of an established GL261 tumor up-regulates MHC expression on invading GL261 glioma cells in vivo. In the present application we will continue to test the hypothesis that low dose WBRT to the GL261 intracranial tumor will up-regulate MHC expression providing a target(s) for a T-cell mediated antitumor immune response directed towards the invading cells and improve the efficacy of immunotherapy. In Aim 1 we will determine the optimal dose and time course to up-regulate MHC expression on invading glioma cells and optimal number of vaccinations to increase long-term survivals, respectively (1a); and to determine host's immune response elicited by the combination of WBRT and vaccination (V). The WBRT schedule of 4 Gy alone induces an inflammatory response associated with an influx of TILs at the tumor site. Brains will be characterized for TILs by FACS analysis and immunohistochemistry of frozen section at 1 week and 2 weeks following treatments. (1b). In Aim 2, we will determine the effects of MHC overexpression in GL261 tumor cells on their growth, invasion and immunogenicity in vitro and in vivo. GL261 tumor cells will be engineered to stably express an inducible construct (Tet-On) encoding the murine CIITA cDNA, a gene that regulates expression of the MHC complex. In Aim 3, we will determine whether up-regulation of MHC expression on invading GL261 tumor cells in vivo is required for the improved therapeutic effect of vaccination following WBRT. MHC expression will be inhibited using siRNA technology. The growing awareness that low dose irradiation can make tumors more amenable to recognition by the patients' immune system forms the basis of our rationale combining cancer vaccines with local irradiation of the brain tumor.

描述（由申请人提供）：目前，没有已知的疗法可以靶向入侵神经胶质瘤肿瘤细胞。为了研究分子水平入侵神经胶质瘤细胞的生物学特性，我们使用绿色荧光蛋白（GFP）标记的鼠类GL261胶质瘤细胞开发了一种体内动物模型。这使我们能够使用GFP免疫组织化学识别入侵的神经胶质瘤细胞与主要肿瘤质量相邻的大脑。微阵列基因技术在入侵和无侵蚀的人神经胶质瘤细胞之间的基因表达谱差异差异，将MHC I类分子鉴定为新型免疫治疗方法的潜在靶标。 GL261神经胶质瘤细胞系由于MHC分子的低表达而具有不良免疫原性。但是，我们已经表明，已建立的GL261肿瘤的低剂量全脑放射治疗（WBRT）上调了在体内入侵GL261神经胶质瘤细胞上MHC的表达。在本应用中，我们将继续检验以下假设：GL261颅内肿瘤的低剂量WBRT将上调MHC表达，为T细胞介导的针对入侵细胞的T细胞介导的抗肿瘤免疫反应提供靶标，并提高免疫疗法的效率。在AIM 1中，我们将确定在入侵神经胶质瘤细胞上上调MHC表达的最佳剂量和时间过程，并分别增加长期生存的疫苗接种数量（1a）；并确定WBRT和疫苗接种（V）的组合引起的宿主的免疫反应。仅4 Gy的WBRT时间表就会引起与肿瘤部位涌入TIL有关的炎症反应。在治疗后的1周和2周，将通过FACS分析和冻结截面的免疫组织化学来表征大脑。 （1b）。在AIM 2中，我们将确定MHC过表达在GL261肿瘤细胞中对其生长，侵袭和体内免疫原性的影响。 GL261肿瘤细胞将被设计为稳定地表达编码鼠CIITA cDNA的诱导构建体（TET-ON），该基因调节MHC复合物的表达。在AIM 3中，我们将确定在WBRT后疫苗接种的治疗效应改善所必需的MHC表达是否在入侵GL261肿瘤细胞上的上调。 MHC表达将使用siRNA技术抑制。人们对低剂量照射的越来越多的意识会使肿瘤更容易受到患者的免疫系统的识别，这是我们将癌症疫苗与脑肿瘤局部辐射相结合的基本原理的基础。