RECIPROCAL FEEDBACK MECHANISMS OF GLIOBLASTOMA AND NEURONAL NETWORK HYPEREXCITABILITY

胶质母细胞瘤与神经网络过度兴奋的交互反馈机制

• 批准号: 10629813
• 负责人: Jochen Meyer
• 金额: $ 37.93万
• 依托单位: BAYLOR COLLEGE OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-03-14 至 2028-02-29
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10629813
• 关键词: AMPA Receptors; Acceleration; Acute; Address; Affect; Area; Astrocytes; Behavior monitoring; Behavioral; Brain; CRISPR/Cas technology; Calcium; Cell Death; Cell Proliferation; Cells; Chronic; Clinical Research; Communication; Coupled; Coupling; Cystine; Deceleration; Disease; Disease Progression; Distant; Electroencephalography; Epilepsy; Epileptogenesis; Evolution; Feedback; Floods; GPC3 gene; Genes; Genetic; Genetic Models; Genetic study; Glioblastoma; Glutamate Receptor; Glutamates; Glypican; Goals; Growth; Homeostasis; Hyperactivity; Image; Immunocompetent; Impairment; Inflammation; Interneurons; Intervention; Invaded; Link; Malignant - descriptor; Measures; Mediating; Methods; Modeling; Monitor; Motion; Mus; N-Methyl-D-Aspartate Receptors; N-Methylaspartate; NF1 gene; Nature; Neuroepithelial, Perineurial, and Schwann Cell Neoplasm; Neuroglia; Neurologic; Neurons; Pathway interactions; Patients; Pattern; Phase; Photons; Population; Process; Proliferating; Protein Secretion; Publishing; Quality of life; Reporting; Resolution; Risk Reduction; Runaway; Seizures; Signal Pathway; Source; Symptoms; Synapses; TP53 gene; Techniques; Testing; Therapeutic; Time; Tissues; Tumor Cell Invasion; Tumor Tissue; Variant; antiporter; awake; cell motility; comorbidity; curative treatments; density; designer receptors exclusively activated by designer drugs; effective intervention; effective therapy; experience; experimental study; gain of function; genetic variant; glutamatergic signaling; human disease; imaging study; improved; in vivo; in vivo imaging; indexing; insight; islet; mRNA Expression; model organism; mortality; neoplastic cell; nervous system disorder; neural; neural network; neuronal tumor; neuroregulation; novel; pre-clinical research; synaptic inhibition; synaptogenesis; tumor; tumor growth; tumor progression; tumor xenograft; two-photon

Glioblastoma (GBM) and tumor-related epilepsy (TRE) are intimately linked and devastating neurological disorders lacking effective therapies despite decades of promising pre-clinical and clinical research. TRE is reported in 40-60% of all GBM patients, often as the presenting symptom, and therefore new treatments would be highly significant, not only for slowing tumor progression but also improving seizure-free quality of life. In recent years, multiple lines of evidence have shown that malignant GBM cells can utilize numerous pathways of interaction to drive peritumoral neural tissue into hyperactive states, thereby facilitating their own proliferation and setting in motion a vicious feedback loop resulting in runaway disease progression in most patients. Specifically, excess glutamate seems to be involved in many cellular neuron-glia interactions including, increased synaptogenesis and hyperexcitability, and we now have evidence for specific genes expressed in our genetic murine tumor model that drive malignant processes. However, we do not know how these processes unfold over time in a native, immunocompetent mammalian tumor model, and how potential therapeutic windows could be exploited to intervene and slow tumor growth. I propose to dissect the mechanisms of tumor-induced glutamate dysregulation and, vice versa, tumor regulation by neural activity using, for the first time, chronic in vivo cellular and macroscale imaging in a CRISPR/Cas9 genetic model of GBM. We recently published the first widefield-calcium imaging study of this genetic GBM model and characterized spatial and temporal profiles of seizures and spreading depolarization waves. We will use these techniques, as well as cellular resolution 2-photon imaging, simultaneous EEG and behavioral monitoring to address the following questions:1) How does GBM cause the degradation of glutamate homeostasis and calcium activity over time? Using chronic widefield imaging of genetically expressed glutamate and calcium activity indicators, we will follow neural activity and malignant glia invasion in vivo to determine spatial and temporal dynamics of hyperexcitability and GBM growth. 2) How do these dysregulation dynamics change when we perturb the genetic tumor driver composition? We will add genes encoding glypicans 3 and 6, recently identified as synaptogenic proteins secreted by astrocytes, to our CRISPR/Cas9 construct. 3) How do different methods of controlling neural hyperexcitability affect GBM growth? We will determine which targets are more amenable to effective intervention than others by studying the effect of NMDA- and AMPA receptor blockers over time in vivo. In addition, we will assess the contribution of non-synaptic glutamate release using mice with a genetic deletion of the xCT cystine-glutamate astrocytic antiporter. Finally, we will directly silence local peritumoral neurons using inhibitory DREADD constructs and measure subsequent GBM deceleration.

胶质母细胞瘤(Gbm)和肿瘤相关癫痫(Tre)是密切相关的和破坏性的神经系统疾病。 尽管数十年的临床前和临床研究前景看好，但仍缺乏有效的治疗方法。特雷是 报告在所有GBM患者中有40%-60%，通常作为主要症状，因此新的治疗方法将 具有非常重要的意义，不仅对于减缓肿瘤的进展，而且还可以提高无癫痫发作的生活质量。在……里面 近年来，多种证据表明恶性肾小球系膜细胞可以利用多种途径。 相互作用，以驱动瘤周神经组织进入过度活跃状态，从而促进其自身 扩散和启动恶性反馈循环，在大多数情况下导致失控的疾病进展 病人。具体地说，过量的谷氨酸似乎参与了许多细胞神经元-神经胶质细胞的相互作用。 包括，增加突触发生和过度兴奋，我们现在有证据表明特定的基因 在我们的基因小鼠肿瘤模型中表达，这种基因驱动了恶性过程。然而，我们不知道如何 这些过程随着时间的推移在天然的、具有免疫功能的哺乳动物肿瘤模型中展开，以及 治疗窗口可以被用来干预和减缓肿瘤的生长。我提议解剖一下 肿瘤引起的谷氨酸调节失调的机制，反之亦然，神经活动对肿瘤的调节 首次在CRISPR/Cas9遗传模型中使用慢性活体细胞和大尺度成像 GBM。我们最近发表了对这种遗传性GBM模型的第一个宽领域钙成像研究，并 描述了癫痫发作和传播的除极化波的空间和时间分布。我们将使用这些 技术，以及细胞分辨率双光子成像，同步脑电和行为监测，以 解决以下问题：1)GBM如何引起谷氨酸动态平衡和 随着时间的推移，钙的活动？使用基因表达的谷氨酸和钙的长期广域成像 活动指标，我们将跟踪神经活动和恶性胶质细胞在体内的侵袭来确定空间和 过度兴奋和基底膜生长的时间动力学。2)这些失调的动态如何改变 当我们扰乱基因肿瘤驱动成分的时候？我们将添加编码龟头3和6的基因， 最近被鉴定为星形胶质细胞分泌的突触生成蛋白，与我们的CRISPR/Cas9构建有关。3)怎么做 控制神经过度兴奋的不同方法会影响基底膜的生长吗？我们将确定哪些目标 通过研究NMDA-和AMPA受体的作用，比其他人更容易受到有效的干预 随着时间的推移，阻滞剂在体内。此外，我们将评估非突触谷氨酸释放的贡献 具有XCT半胱氨酸-谷氨酸星形细胞逆向转运蛋白基因缺失的小鼠。最后，我们将直接沉默 使用抑制性DREADD结构的局部瘤周神经元，并测量随后的GBM减速。