Therapeutic Targeting of Malignant Glioma Stem Cells

 描述（由申请人提供）：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，涉及胶质母细胞瘤干细胞样细胞（GSC）遗传和表观遗传畸变的肿瘤内异质性协调了人GBM恶性肿瘤、复发和化疗-放射抗性。然而，由于GSC靶向药物递送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捉摸。基于我们在治疗性干细胞领域的专业知识 (FDA在此，我们提出了靶向最近鉴定的上调的神经发育转录因子（TF）-POU 3F 2、SOX 2、SALL 2和0 LIG 2-的核心组的新疗法，所述核心组共同产生人GBM中的肿瘤传播GSC表型。使用批准用于临床试验的肿瘤性NSC/MSC来递送脂质聚合物纳米颗粒（LPNP）封装的siRNA药物以敲除GBM中的4个主TF，我们提出了一种用于异质性癌症（如GBM）的有效基因疗法的新的体内策略。我们假设可以设计新型非病毒基因疗法，通过抑制控制GSC表型的主TF来阻止胶质瘤中的GSC命运，并提出以下新策略-策略1：为了中和毒气室-促进具有可定制的表面化学的主TF的产生，以实现细胞类型选择性和靶向药物递送，并在体外验证其针对携带个性化的GSC模型的治疗功效。基因组背景策略二：开拓干细胞介导的靶向由主TF促进的GSC表型的siRNA的体内LPNP递送，同时利用我们完善的基于NSC或MSC的疗法的模型。策略三：为了增强纳米颗粒通过体循环递送到GSC，我们还提出解决NP穿过血荆棘屏障的递送。具体来说，我们将研究内皮细胞从内源性和iPSC来源的NP再循环，作为一种新的基于RNA的跨BBB递送方法。最后，在战略4中：我们将探索新兴的合成基因组编辑工具的新应用，例如转录激活因子样效应核酸酶（TALEN）技术和CRISPR/cas9技术，以配制用于基因组编辑工具的LPNP包封递送（Glucose），以改变胶质母细胞瘤干细胞样细胞的体内行为，并检查对实验性脑肿瘤模型动物存活率的影响。

 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.