GlioPrevent: PiezoMagnetic Nanoparticles to Prevent Glioma Invasion in Human Brain Organoids

多形性胶质母细胞瘤（GBM）是英国最常见的恶性脑肿瘤，占每五个脑肿瘤诊断中的一个以上。GBM患者的预后很差，只有20%的患者存活超过一年，3%的患者存活超过三年。GBM肿瘤扩散迅速，具有侵略性的手指状生长到重要的大脑位置。这使得治疗具有挑战性，GBM肿瘤众所周知难以手术切除。正因为如此，在手术过程中只切除容易接触到的癌症，并且在手术后立即进行放疗和化疗以抑制肿瘤生长。因此，需要开发一种更有效的治疗GBM的方法，可以阻止手指样GBM细胞生长/侵入正常脑区域。研究人员一直在寻找一种简单的方法来修复DNA错误来治疗疾病，因为他们知道DNA变化会导致癌症。该领域最近的一个游戏规则改变者是CRISPR基因编辑系统。CRISPR可用于切割致病基因，以细胞修复系统无法逆转的方式改变它们。这使得细胞对治疗剂更敏感。不幸的是，CRISPR有一些局限性，限制了它作为一种新型治疗方法的使用。1)将CRISPR组分导入细胞的最常见方法是使用修饰的病毒，这可能会导致不必要的遗传变化，2）CRISPR可能会切割预期基因之外的DNA并导致副作用（脱靶编辑）。在这个项目中，我们提出了一种新的递送系统（PiezoMagnetic nanoparticles，PMNPs）来克服这些问题，并使CRISPR能够靶向递送到模拟人脑的系统中。PMNPs是可用于将治疗剂转运到细胞中的材料。将PMNP用治疗剂包被，注射到体内并使用磁体或超声引导到靶细胞。在这项工作中，我们将生产一种涂有CRISPR系统的PMNP，该系统旨在靶向与GBM相关的PLK 1和NeK 2基因。我们相信，阻止这些基因发挥作用将阻止GBM细胞的生长，为未来的治疗提供一个潜在的途径。为了增加PMNP对GBM的特异性，还将向载体施加独特且临床相关的靶向剂。这些将在被称为类器官的人脑小模型中进行测试。将处理的PMNP应用于含有GBM和正常细胞的模型，并测量GBM细胞随时间的生长。研究结果将为PMNPs作为递送系统和类器官作为模型测试系统的进一步发展奠定基础。合作者（布拉德福德大学和杜塞尔多夫大学医院）还将召集一个胶质母细胞瘤专家小组，为研究提供建议，并形成胶质母细胞瘤研究网络的基础。合作者将共同努力，领导会议和活动，以促进和推动进一步的胶质母细胞瘤研究。

Glioblastoma multiforme (GBM) is the most common malignant brain tumour in the UK, accounting for more than one in every five brain tumour diagnoses. GBM patients have a poor prognosis, with just 20% surviving more than a year and 3% surviving more than three years. GBM tumours spread quickly, with aggressive finger-like growth into important brain locations. This makes treatment challenging, with GBM tumours notoriously difficult to remove surgically. Because of this, only easily accessible cancers are removed during surgery, and radiation and chemotherapy are delivered immediately after surgery to suppress tumour growth. Hence developing a more effective therapy for GBM, that can stop the growth/invasion of finger-like GBM cells into normal brain areas is needed. Researchers have been looking for a simple way to fix DNA mistakes to treat diseases ever since they learned that DNA changes cause cancer. A recent game-changer in this field is the CRISPR gene editing system. CRISPR can be used to cut the disease-causing genes, changing them in a way that can't be reversed by cell repair systems. This makes the cell more susceptible to therapeutic agents. Unfortunately, CRISPR has some limitations that restrict its use as a novel therapeutic. 1) The most common way to get CRISPR components into cells is with a modified virus, which can cause unwanted genetic changes, and 2) CRISPR may cut DNA outside of the intended genes and cause side effects (off-target editing). In this project we propose a novel delivery system (PiezoMagnetic nanoparticles, PMNPs) to overcome these problems and enable targeted delivery of CRISPR to systems modelling the human brain.PMNPs are materials that can be used to transport therapeutic agents into cells. The PMNPs are coated with the therapeutic agent, injected into the body and directed to the target cells using a magnet or ultrasound. In this work, we will produce a PMNP coated with a CRISPR system designed to target PLK1 and NeK 2 genes, which are associated with GBM. We believe preventing these genes from functioning will stop the growth of GBM cells, offering a potential avenue for future treatments. To increase the specificity of the PMNP to the GBM, a unique and clinically relevant targeting agent will also be applied to the carriers. These will be tested in small models of the human brain known as organoids. The treated PMNPs will be applied to models containing GBM and normal cells and the growth of the GBM cells will be measured over time. The results will lay the groundwork for the further development of PMNPs as a delivery system and organoids as a model testing system.The collaborators (the University of Bradford and University Hospital Dusseldorf) will also convene a panel of glioblastoma experts to advise on the research and to form the basis of a glioblastoma research network. The collaborators will work together to lead meetings and events to promote and drive forward further glioblastoma research.