Therapeutic Targeting of Malignant Glioma Stem Cells

 描述(申请人提供)：最近对患者胶质母细胞瘤细胞(GBM)进行的大规模基因组和转录组分析表明，肿瘤内涉及胶质母细胞瘤干细胞(GSCs)遗传和表观遗传异常的异质性协调了人类GBM的恶性、复发和化疗辐射耐受。然而，由于GSC靶向药物传递的困难和缺乏有效的遗传靶点，中和GSC的有效策略仍然难以实现。以我们在治疗性干细胞方面的专业知识为基础 (FDA批准了人类神经干细胞用于临床试验/以及间充质干细胞/hMSCs)，在此，我们提出了针对最近发现的上调神经发育转录因子(TF)核心集合-POU3F2、SOX2、SALL2和OLIG2-的新疗法，这些因子共同在人GBM中产生肿瘤增殖性GSC表型。利用已被批准用于临床试验的致瘤NSC/MSCs携带脂聚纳米粒(LpNP)包裹的siRNA药物来敲除GBM中的4个主要转录因子，我们提出了一种新的体内有效的基因治疗策略，如GBM。我们假设新的非病毒基因疗法可以通过抑制控制GSC表型的主TF来设计阻止GSC在胶质瘤中的命运，并提出了以下新策略-策略1：用可定制的表面化学中和促进GSC促进的主TF的产生，以实现细胞类型的选择性和靶向药物输送，并在体外验证它们对具有个性化基因组背景的GSC模型的治疗效果。策略2：开创干细胞介导的体内LpNP递送靶向GSC表型的siRNA的先河，同时利用我们成熟的NSC或MSCs治疗模式。策略3：为了通过体循环加强纳米颗粒向GSC的输送，我们还建议解决NP跨越血液屏障的输送问题。具体地说，我们将研究来自内源性和IPSC来源的内皮细胞的NP循环，作为一种跨越血脑屏障的基于RNA的新传递方法。最后，在策略4：我们将探索新兴的合成基因组编辑工具的新应用，如转录激活物样效应核酸酶(TALEN)技术和CRISPR/Cas9技术，以形成LpNP包裹的基因组编辑工具(GET)，以改变胶质母细胞瘤干细胞样细胞的体内行为，并通过实验性脑瘤模型检测其对动物生存的影响。

 DESCRIPTION (provided by applicant): Recent large scale genomic and transcriptome analyses of patient glioblastoma cells (GBM) indicate that the intratumoral heterogeneity involving the genetic and epigenetic aberrations of glioblastoma stem-like cells (GSCs) orchestrate human GBM malignancy, recurrence, and chemo- radiation resistance. However, effective strategies to neutralize GSCs remain elusive due to difficulty in GSC-targeted drug delivery and lack of effective genetic targets. Building on our expertise in therapeutic stem cells (FDA approved human neural stem cells for clinical trials / as well as mesenchymal stem cells / hMSCs), here we propose novel therapies targeting the recently identified core set of up-regulated neurodevelopmental transcriptional factors (TFs) - POU3F2, SOX2, SALL2, and OLIG2 - which collectively engender the tumor propagating GSC phenotypes in human GBMs. Using tumortropic NSC/MSCs approved for clinical trials to deliver lipo-polymeric nanoparticle (LPNP)-encapsulated siRNA medicine to knockdown the 4 master TFs in GBM, we propose a novel in vivo strategy for effective gene therapies for heterogeneous cancers such as GBM. We hypothesize that novel non-viral gene therapies can be designed to arrest GSC fate in gliomas by suppressing the master TFs that control GSC phenotypes, and propose the following novel strategies - Strategy 1: To neutralize the GSC- promoting production of master TFs with customizable surface chemistry to achieve cell type selectivity and targeted drug delivery and validate their therapeutic efficacy in vitro against GSC models bearing personalized genomic background. Strategy 2: To pioneer stem cell-mediated in vivo LPNP delivery of siRNA targeting GSC phenotypes promoted by the master TFs while leveraging our well established model of NSC or MSCs- based therapies. Strategy 3: To enhance nanoparticle delivery to GSC via systemic circulation, we also propose to tackle the delivery of NP across the blood briar barrier. Specifically, we will examine NP-recycling by endothelial cells from both endogenous and iPSC origins, as a novel RNA-based delivery approach across the BBB. Finally, in Strategy 4: we will explore novel applications of the emerging synthetic genome editing tools, such as the transcription activator like effector nuclease (TALEN) technology and CRISPR/cas9 technology, to formulate LPNP-encapsulated delivery for genome editing tools (GETs) in order to modify the in vivo behaviors of glioblastoma stem-like cells and examine the impact on survival in animals with experimental brain tumor models.