MOLECULAR DISSECTION OF SEIZURE MICROENVIRONMENT IN MALIGNANT GLIOMA

恶性胶质瘤癫痫微环境的分子解剖

基本信息

批准号：
10062889
负责人：
Benjamin Deneen
金额：
$ 63.21万
依托单位：
BAYLOR COLLEGE OF MEDICINE
依托单位国家：
美国
项目类别：
财政年份：
2017
资助国家：
美国
起止时间：
2017-12-15 至 2022-11-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10062889
关键词：
Adult Adult Glioma Anaplastic astrocytoma Appearance Astrocytes Benchmarking Biological Markers Brain Cell Lineage Cells Cessation of life Childhood Glioma Chlorides Clinical Clustered Regularly Interspaced Short Palindromic Repeats Coculture Techniques Cognitive Data Deltastab Detection Deterioration Development Developmental Biology Dissection Early Diagnosis Electroencephalography Electrophysiology (science)Electroporation Engineering Epilepsy Epileptogenesis Evolution Excitatory Synapse Functional disorder Gene Deletion Generations Genes Genetic Glioblastoma Glioma Glutamates Goals Growth Hippocampus (Brain)Homeostasis Human Hyperactivity Image Impairment In Vitro Intractable Epilepsy Knowledge Label Lead Life Location Longevity Malignant - descriptor Malignant Glioma Malignant Neoplasms Malignant neoplasm of brain Medical Modeling Molecular Molecular Profiling Monitor Mouse Strains Mus Mutation Nature Neuroglia Neurologic Deficit Neurons Neurosciences Pathogenicity Pathologic Patients Personal Satisfaction Physiology Population Prognosis Property Proteins Reproducibility Resistance Role Seizures Slice Specific qualifier value Subgroup Synapses Testing Time Tissues Tumor Burden Tumor Cell Invasion Tumor Expansion Tumor stage Validation Wild Type Mouse analog comorbidity density gamma-Aminobutyric Acid genetic signature genomic biomarker gray matter host neoplasm interaction in utero in vivo molecular dynamics molecular marker mouse model mutant neoplastic cell neural network neuronal circuitry neuronal excitability neuroregulation next generation novel pre-clinical prevent relating to nervous system reuptake synaptic inhibition synaptogenesis tau Proteins transcriptome sequencing transcriptomics tumor tumor growth tumor microenvironment tumor progression

项目摘要

Glioblastoma cells trigger pharmacoresistant seizures that may promote tumor growth and diminish the quality of remaining life. To define the relationship between growth of glial tumors and their neuronal microenvironment, and to identify genomic biomarkers and mechanisms that may point to better prognosis and treatment of drug resistant epilepsy in brain cancer, we are analyzing a new generation of genetically defined CRISPR/in utero electroporation inborn glioblastoma (GBM) tumor models engineered in mice. The molecular pathophysiology of glioblastoma cells and surrounding neurons and untransformed astrocytes will be compared at serial stages of tumor development in three genetic mouse strains: wild type, seizure prone, and seizure resistant. Preliminary data reveal that epileptiform EEG spiking is a very early and reliable preclinical signature of GBM expansion preceding other neurological deficits in these mice, followed by rapidly progressive seizures and death within weeks. Transcriptomic analysis of cortical astrocytes reveals the expansion of a subgroup enriched in pro-synaptogenic genes that may drive hyperexcitability, a novel mechanism of epileptogenesis. In Specific Aim 1 we will systematically define the earliest appearance of cortical hyperexcitability in wild type mice with a prototypical GBM and correlate its progression with in vivo and neuropathological imaging of invasive tumor cell location, in vitro electrophysiology, and molecular markers of key epilepsy pathogenic cascades in peritumoral neurons, including impaired glutamate reuptake, altered GABA gated-chloride gradients, and synaptic densities. In Specific Aim 2 we will correlate these findings with detailed FACS-sorted transcriptomic profiles of both transformed and wild type astrocytes in the peritumoral region to test the novel hypothesis that peritumoral hyperexcitability is driven in part by astrocytic subtypes that disrupt synaptic E/I homeostasis. In Specific Aim 3, we will use this benchmark approach in WT brain to compare growth, electrophysiological and molecular pathological profiles of the same tumor generated in a hyperexcitable brain bearing a single gene deletion (Kcna1) that dramatically lowers the threshold for seizures and shortens lifespan, and in a monogenic deletion strain (MapT/tau) that raises cortical seizure threshold and prolongs life, in order to examine the contribution of host neuronal excitability to tumor expansion. Our approach sets the stage to broadly explore the developmental biology of personalized tumor/host interactions in mice engineered with novel human tumor mutations in specified glial cell lineages.

胶质母细胞瘤细胞引发耐药性癫痫发作，可能促进肿瘤生长并降低质量的剩余寿命。定义胶质瘤生长与其神经元之间的关系微环境，并识别基因组生物标志物和机制，可能表明更好的预后和脑癌耐药性癫痫的治疗，我们正在分析新一代基因定义的 CRISPR/子宫内电穿孔小鼠先天性胶质母细胞瘤 (GBM) 肿瘤模型。分子胶质母细胞瘤细胞和周围神经元以及未转化的星形胶质细胞的病理生理学将比较了三种基因小鼠品系的肿瘤发展的连续阶段：野生型、易癫痫发作型和抗癫痫发作。初步数据显示癫痫样脑电图尖峰是一种非常早期且可靠的临床前研究 GBM 扩张的特征先于这些小鼠的其他神经功能缺陷，然后迅速进行性癫痫发作并在几周内死亡。皮质星形胶质细胞的转录组分析揭示了富含促突触基因的亚群的扩展可能会导致过度兴奋，这是一种新的癫痫发生机制。在具体目标 1 中，我们将系统地定义最早出现的具有典型 GBM 的野生型小鼠的皮质过度兴奋性，并将其进展与体内和侵袭性肿瘤细胞定位的神经病理学成像、体外电生理学和分子标记瘤周神经元中关键的癫痫致病级联，包括谷氨酸再摄取受损，改变 GABA 门控氯化物梯度和突触密度。在具体目标 2 中，我们将把这些发现与瘤周转化型星形胶质细胞和野生型星形胶质细胞的详细 FACS 分类转录组图谱区域来测试新的假设，即瘤周过度兴奋部分是由星形胶质细胞亚型驱动的，破坏突触 E/I 稳态。在具体目标 3 中，我们将在 WT 大脑中使用这种基准方法来比较在同一肿瘤中产生的相同肿瘤的生长、电生理学和分子病理学特征过度兴奋的大脑带有单基因缺失（Kcna1），可显着降低癫痫发作的阈值并缩短寿命，并且在单基因缺失菌株（MapT/tau）中，会提高皮质癫痫阈值并延长寿命，以检查宿主神经元兴奋性对肿瘤扩张的贡献。我们的该方法为广泛探索个性化肿瘤/宿主相互作用的发育生物学奠定了基础在特定神经胶质细胞谱系中携带新型人类肿瘤突变的小鼠中。