Inhibiting glioma dispersal using non-toxic, natural products

使用无毒的天然产品抑制神经胶质瘤的扩散

基本信息

批准号：
8449571
负责人：
John S Kuo
金额：
$ 18.46万
依托单位：
UNIVERSITY OF WISCONSIN-MADISON
依托单位国家：
美国
项目类别：
财政年份：
2012
资助国家：
美国
起止时间：
2012-04-01 至 2014-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8449571
关键词：
Adverse effects Apoptosis Astrocytes Attenuated Basement membrane Biological Assay Biological Factors Blood - brain barrier anatomy Brain Brain Neoplasms Cancer Cell Growth Cell Line Cell model Cells Clinic Complement Data Development Diagnosis Dominant-Negative Mutation Dose Drug Formulations Foundations Future Genetic Genetic Variation Glioblastoma Glioma Goals Histology Human Immigration Immunoblotting Immunohistochemistry Implant In Vitro Infiltration Inflammation Intervention Knock-out Magnetic Resonance Imaging Malignant Glioma Malignant Neoplasms Malignant neoplasm of brain Mediating Modeling Mus NIH Program Announcements Neuroglia Oncogenic Operative Surgical Procedures Patients Plants Pre-Clinical Model Primary Brain Neoplasms Property Proto-Oncogene Proteins c-akt Quality of life Radiation Radio Radiosurgery Recurrence Resveratrol Role Serum Signal Transduction Surgically-Created Resection Cavity Testing Time Transgenic Mice Translating Treatment Efficacy Treatment Protocols Tumor Cell Invasion angiogenesis attenuation brain tissue cancer care cancer cell cancer therapy cancer type cell growth chemotherapy follow-up glioma cell line improved in vitro Assay in vivo inhibitor/antagonist matrigel melanoma mouse model neoplastic cell novel prevent protective effect research clinical testing response standard care therapy resistant tumor

项目摘要

DESCRIPTION (provided by applicant): The long-term objective of this R21 proposal is developing the non-toxic plant product resveratrol as an agent to specifically inhibit glioma cell dispersal, for future use alongside current surgical, radiation, and chemotherapeutic strategies. Malignant gliomas such as glioblastoma multiforme (GBM) are the most common and deadly primary brain neoplasms, and even with aggressive interventions average survival remains less than 2 years after diagnosis. Extensive dispersal of therapy-resistant glioma cells into surrounding brain prevents efficient treatment, resulting in tumor recurrence immediately adjacent to the resection cavity in greater than 90% of cases. Therefore, new treatments or approaches specifically inhibiting invasive properties of glioma cells are highly desirable, and ideal candidates in this respect would be non-toxic so that the toxic load of the treatment regimen remains manageable. The natural product resveratrol satisfies these needs, causing virtually no side effects when administered in vivo and demonstrating wide-ranging anti-oncogenic effects, including invasion inhibition, on diverse cancer types. Our collaborators have demonstrated resveratrol's ability to inhibit infiltration of highly aggressive melanoma in a mouse model by disrupting AKT signaling. In brain tumors, oncogenic AKT is critical to continued cell growth, survival, and invasion. Additionally, we have developed novel resveratrol formulations that can increase serum levels of unmodified resveratrol 10-fold and can be delivered intratumorally as well as orally. We hypothesize that resveratrol can inhibit dispersal of highly aggressive glioma cells by disrupting AKT. To test this hypothesis, we will use our novel glioma initiating cell (GIC) model, established through sphere culture of 5 independent human GBMs. These GICs are highly invasive when implanted into a mouse orthotopic model, unlike historical glioma cell lines, and our multiple independent lines better represent the genetic diversity of human GBM compared to single transgenic mouse models. Our preliminary data already demonstrate in vitro resveratrol-mediated inhibition of GIC invasion in a basement membrane assay. In specific aim 1, we will determine the ability of resveratrol to inhibit glioma invasion ad dispersal of our multiple genetically diverse GIC lines, testing novel formulations and delivery of resveratrol as well. We will use basement membrane invasion assays in vitro and our orthotopic mouse model in vivo to test resveratrol's anti-invasive properties across physiologically relevant doses. In specific aim 2, we will elucidate the role of resveratrol-modulated AKT on glioma infiltration. We will first analyze changes in activated and total AKT upon resveratrol administration in vitro and in vivo. Secondly, we will use over-expression and dominant-negative (knock-out) AKT strategies in our GIC lines and analyze effects on resveratrol responsiveness. We believe that successful completion of these aims will lay the foundation for follow-up clinical testing of the non-toxic natural agent, resveratrol, combined with current GBM therapies to improve patient survival and quality of life by inhibiting tumor invasion into functional brain.

描述（由申请人提供）：该R21提案的长期目标是开发无毒植物产品白藜芦醇作为特定抑制神经胶质瘤细胞分散剂的药物，供将来与当前的外科手术，放射线和化学治疗策略一起使用。恶性神经胶质瘤（例如胶质母细胞瘤多形（GBM））是最常见和致命的原发性脑肿瘤，即使有侵略性干预措施，平均生存率仍不在诊断后不到2年。在大脑周围的大脑中，广泛的耐药性神经胶质瘤细胞可阻止有效的治疗，从而导致肿瘤复发在大于90％的病例中与切除腔相邻。因此，非常需要抑制神经胶质瘤细胞的侵入性特性的新治疗方法或方法是非常需要的，并且在这方面的理想候选物将无毒，因此治疗方案的毒性负荷仍然可以控制。天然产品白藜芦醇满足了这些需求，在体内给药时几乎没有副作用，并证明了对多种癌症类型的较广泛的抗结合作用，包括侵袭抑制。我们的合作者证明了白藜芦醇抑制小鼠高度侵略性黑色素瘤的能力通过破坏AKT信号传导模型。在脑肿瘤中，致癌AKT对于持续的细胞生长，生存和侵袭至关重要。此外，我们开发了新型的白藜芦醇制剂，可以提高未修饰的白藜芦醇10倍的血清水平，并且可以在肿瘤内和口服交付。我们假设白藜芦醇可以通过破坏AKT来抑制高度攻击性神经胶质瘤细胞的分散。为了检验这一假设，我们将使用我们的新型神经胶质瘤启动细胞（GIC）模型，该模型是通过5种独立人类GBM的球培养而建立的。与历史神经胶质瘤细胞系不同，这些GIC在植入小鼠原位模型中时具有高度侵入性，而与单个转基因小鼠模型相比，我们的多个独立线更好地代表了人类GBM的遗传多样性。我们的初步数据已经证明了体外白藜芦醇介导的基底膜测定中GIC侵袭的抑制作用。在特定的目标1中，我们将确定白藜芦醇抑制神经胶质瘤侵袭AD的能力，我们多种遗传多样的GIC线，测试新的配方和递送白藜芦醇也是如此。我们将在体内使用基底膜侵袭测定和我们的原位小鼠模型来测试白藜芦醇跨生理相关剂量的抗侵入性。在特定的目标2中，我们将阐明白藜芦醇调节Akt在神经胶质瘤浸润中的作用。我们将首先分析白藜芦醇在体外和体内给药后激活和总AKT的变化。其次，我们将在GIC线中使用过表达和主导性（淘汰）AKT策略，并分析对白藜芦醇反应性的影响。我们认为，这些目标的成功完成将为无毒天然剂，白藜芦醇加上当前的GBM疗法的后续临床测试奠定基础，以通过抑制肿瘤侵入功能性大脑来改善患者的生存和生活质量。