Novel, biologically relevant, in vitro model of the blood-brain tumour barrier (BBTB)

新型、生物学相关的血脑肿瘤屏障（BBTB）体外模型

• 批准号: 2887667
• 金额: --
• 依托单位: University of Liverpool
• 依托单位国家: 英国
• 项目类别: Studentship
• 财政年份: 2023
• 资助国家: 英国
• 起止时间: 2023 至 无数据
• 项目状态: 未结题
• 来源: https://gtr.ukri.org/projects?ref=studentship-2887667
• 关键词: Novel; biologically; relevant; vitro; model

Glioblastoma (GBM) is the most common form of brain cancer. Despite this, treatment has not improved significantly in the past 20 years. Current therapy involves surgery, radiotherapy and chemotherapy, but outcomes remain very poor, with average life expectancy for newly diagnosed patients ranging from 12 to 14 months. The lack of progress in treatment options is multifaceted, but targeted therapy options have yet to overcome the challenge of crossing the blood-brain barrier (BBB) / blood-brain tumour barrier (BBTB). Current models of the BBTB fail to provide an accurate representation of the disease. In vitro 2D cell cultures do not reflect the complexity of the BBTB, and have limited relevance to human physiology. In vivo models may not mimic the clinical disease; novel drugs that work on GBM xenograft models have not been successful in the clinic. The objective of this project is to establish and biologically validate a novel in vitro model of the BBTB by inclusion of GBM cells with the current BBB model. This will be achieved using a hydrogel transwell, consisting of human derived GBM cells, co cultured with brain like endothelial cells derived from induced pluripotent stem cells (iPSCs). The working hypothesis of this project is that this method will generate a biologically relevant model of GBM at the BBB which can then be validated, and used as a screen for novel, targeted therapies. The NC3Rs have funded this project as this model could be used to replace some rodent xenograft studies. Our collaborators at the Open University have recently established a novel BBB model based on deriving brain-like endothelial cells from iPSCs and co-culturing with astrocytes on a hydrogel. This model has been fully validated and forms a tight monolayer with expression of key BBB proteins. The monolayer of BECs on top of the hydrogel transwell culture provides an ideal model where the barrier properties and function can be investigated directly. To achieve the inclusion of GBM cells within the BBB model, our collaborator at University of Edinburgh has provided neural stem cells (NSCs) and patient derived GBM stem-like cells (GSCs). These GBM cells, alongside astrocytes, will be cultured in the hydrogel component of the model. This technique enables the establishment of an in vitro BBTB and a way to study the effects of the GBM cells. The validity of this model will be determined using several criteria: barrier tightness; astrocyte end feet; gene expression; drug crossing and tight junction protein expression will be assessed. Finally, using transcriptomics, the novel in vitro BBTB model will be compared to GBM patient datasets of the BBTB to define the biological relevance of the in vitro model to the human disease. The impact of this model will be to reduce animal usage significantly and in the longer-term have the potential model to replace GBM xenograft models. We estimate an approximate use of at least 4,500 rodents a year for GBM xenograft experiments in the UK. For the worldwide usage of GBM xenograft rodents, a search shows nearly 800 primary publications published in the last year that used GBM xenograft models. From this we estimate at least 60,000 rodents are used per year worldwide for GBM xenograft studies. The use of an in vitro model as a primary screen for novel drug therapies would lead to an estimated 20% reduction in animal work, and therefore approximately 12,000 rodents a year that would not be used in GBM research. In conclusion, this project aims to generate a validated BBTB model that can be rolled out to GBM labs, thus leading to the reduction in the use of GBM xenograft models and improved translation to the human disease.

胶质母细胞瘤（GBM）是脑癌最常见的形式。尽管如此，在过去的20年中，治疗尚未得到显着改善。当前的治疗涉及手术，放疗和化学疗法，但结果仍然很差，新诊断的患者的平均预期寿命为12到14个月。缺乏治疗选择的进展是多方面的，但是有针对性的治疗选择尚未克服跨越血脑屏障（BBB） /血脑肿瘤屏障（BBTB）的挑战。 BBTB的当前模型无法提供该疾病的准确表示。体外2D细胞培养物不能反映BBTB的复杂性，并且与人类生理学相关。体内模型可能不会模仿临床疾病。在GBM异种移植模型上使用的新型药物在诊所尚未成功。该项目的目的是通过将GBM细胞与当前的BBB模型一起建立和生物学验证BBTB的新型体外模型。这将是使用水凝胶Transwell（由人类衍生的GBM细胞组成的）来实现的，该细胞与脑部培养的脑细胞培养，例如源自诱导的多能干细胞（IPSC）的内皮细胞。该项目的工作假设是，该方法将在BBB上生成与GBM的生物学相关模型，然后可以验证，并用作新型靶向疗法的屏幕。 NC3R资助了该项目，因为该模型可用于替代一些啮齿动物异种移植研究。我们在公开大学的合作者最近建立了一种新型的BBB模型，该模型基于从IPSC中得出类似脑的内皮细胞，并在水凝胶上与星形胶质细胞共培养。该模型已得到充分验证，并形成了用关键BBB蛋白表达的紧密单层。 BEC在水凝胶Transwell培养基顶部的单层提供了一个理想的模型，可以直接研究屏障的性能和功能。为了将GBM细胞纳入BBB模型，我们在爱丁堡大学的合作者提供了神经干细胞（NSC）和患者衍生的GBM干细胞（GSC）。这些GBM细胞与星形胶质细胞一起将在模型的水凝胶成分中培养。该技术可以建立体外BBTB和研究GBM细胞效应的方法。该模型的有效性将使用多个标准确定：屏障紧密度；星形胶质细胞末端；基因表达；将评估药物交叉和紧密连接蛋白表达。最后，使用转录组学，将将新型的体外BBTB模型与BBTB的GBM患者数据集进行比较，以定义体外模型与人类疾病的生物学相关性。该模型的影响是显着减少动物使用量，并在长期具有替代GBM异种移植模型的潜在模型。我们估计，在英国，每年至少使用4,500啮齿动物的GBM异种移植实验。对于GBM异种移植啮齿动物的全球用法，搜索显示了使用GBM异种移植模型的近800个主要出版物。由此，我们估计全球每年至少使用60,000个啮齿动物进行GBM异种移植研究。使用体外模型作为新型药物疗法的主要筛查将导致动物工作降低20％，因此每年约有12,000名啮齿动物在GBM研究中不使用。总之，该项目旨在生成经过验证的BBTB模型，该模型可以推广到GBM Labs，从而导致使用GBM异种移植模型的使用减少并改善转化为人类疾病。