Characterization of circadian rhythms in Glioblastoma multiforme and investigation of chronotherapy as a novel therapy to prolong patient survival

项目摘要 多形性胶质母细胞瘤(GBM)是最常见的成人原发脑肿瘤，发病15个月。 采用目前标准的护理治疗，包括替莫唑胺化疗的预后 (Temodar，TMZ)。计时疗法，一种在治疗疾病时考虑一天中的时间的做法，已经被证明。 以改善几种癌症的预后，如结直肠癌和急性淋巴细胞性白血病，但已经 从未应用于GBM。我们之前在培养的小鼠间充质GBM细胞系上的工作显示 DNA双链断裂、凋亡途径激活和细胞的时间依赖性最大值 死亡与BMal1和PER2两个核心时钟基因的表达高峰和谷值相对应。我们的 初步数据显示，颅内基底细胞瘤在体内的基因表达具有昼夜节律 这可以在整个疾病发展过程中进行长期监测。最值得注意的是，我们的试点数据显示，肿瘤 基因表达的节律与宿主的节律一致，导致肿瘤基因表达的反转模式 在反转的明/暗周期和在恒定的黑暗中与宿主对准的自由运行模式中。这 该提案将评估这样一种假设，即GBM肿瘤在体内有昼夜节律，并伴随着宿主 这些日常节律可以通过时间疗法来提高存活率。要解决这个问题 假设，这一建议将进一步评估基底膜肿瘤是否与体内宿主的日常节律一致作为以下因素的函数 通过在节律性肿瘤中植入心律失常肿瘤来改变光时间表和昼夜节律基因 小鼠和心律失常小鼠中的节律性肿瘤，并测量肿瘤生长率的变化以及 宿主和肿瘤之间的同步性，根据昼夜节律基因(目标1)。此外，这项建议 将确定肿瘤细胞死亡和宿主存活的差异取决于TMZ剂量的一天 测量TMZ对宿主和肿瘤中基因表达的日节律的影响，以及 TMZ对肿瘤缩小和血液毒性严重程度的影响(目标2)。这项建议将会有所改善 20年来首次通过最大限度地破坏肿瘤同时最大限度地减少肿瘤损害的GBM患者的结局 根据患者的性别和理想的治疗时间进行个性化治疗的副作用。它还将作为 重要的下一步是实现高通量筛查，以获得治疗疾病的最佳时间 胶质母细胞瘤。

Project Summary Glioblastoma multiforme (GBM) is the most common primary adult brain tumor diagnosed, with a 15-month prognosis using current standard of care treatment which includes chemotherapy with Temozolomide (Temodar, TMZ). Chronotherapy, the practice of considering time of day in treating a disease, has been shown to improve outcome in several cancers such as colorectal cancer and acute lymphoblastic leukemia, but has never been applied to GBM. Our previous work on a cultured murine mesenchymal GBM cell line showed a time-of-day dependent maximum in DNA double-strand breaks, activation of the apoptotic pathway, and cell death corresponding with the peak of Bmal1 and trough of Per2 expression, two core clock genes. Our preliminary data demonstrate that intracranial GBM tumors have circadian rhythms in gene expression in vivo that can be monitored chronically throughout disease progression. Most notably, our pilot data show that tumor rhythms in gene expression align with those of the host, leading to reversed patterns in tumor gene expression in a reversed light/dark cycle and free-running patterns that align with the host in constant darkness. This proposal will evaluate the hypothesis that GBM tumors have circadian rhythms in vivo that entrain to the host and that those daily rhythms can be leveraged using chronotherapy to improve survival. To address this hypothesis, this proposal will further assess if GBM tumors align with host daily rhythms in vivo as a function of changing light schedule, as described, and circadian genotype by implanting an arrhythmic tumor in a rhythmic mouse and a rhythmic tumor in an arrhythmic mouse and measuring changes in tumor growth rate, as well as synchrony between the host and tumor, according to circadian genotype (Aim 1). Furthermore, this proposal will determine differences in tumor cell death and host survival depending on time of day of TMZ dosing by measuring the effect of TMZ on daily rhythms in gene expression in both the host and tumor, as well as the effect of TMZ on tumor size reduction and severity of hematological toxicity (Aim 2). This proposal will improve outcome for patients with GBM for the first time in 20 years by maximizing tumor destruction while minimizing side effects by personalizing treatment based on patient sex and ideal time of day to treat. It will also serve as an important next step in achieving high-throughput screening for the optimal time to treat diseases beyond Glioblastoma.