Engineering stem cell therapies to understand and overcome glioblastoma adaption

项目摘要/摘要 基因工程神经干细胞(NSCs)是治疗高度侵袭性大脑的一种很有前途的疗法。 癌症胶质母细胞瘤(GBM)。经过改造的神经干细胞具有独特的肿瘤归巢能力，使它们能够 抗癌基因产物直接进入局部和侵袭性的基底膜病灶。我们团队和其他人进行的临床前研究 研究表明，抗肿瘤神经干细胞通常可以减少70%-90%的原位GBM异种移植 延长荷瘤小鼠的存活时间。然而，GBM销量最初的大幅下降并不是 维持和治疗耐受性仍然是基于神经干细胞的治疗的主要挑战。发生GBM逃逸 在携带不同治疗有效载荷的神经干细胞治疗后，在固体和 术后肾小球基底膜。我们最近发现，新型肿瘤归巢药物递送载体具有强大的抗肿瘤活性 通过细胞重编程，可以从“诱导神经干细胞”(INSCs)发展出癌症活性。 技术，称为转分化(TD)。抗肿瘤iNSC治疗减少了GBM异种移植230- 在4周内折叠，存活率增加一倍以上。与野生型NSC治疗类似，肿瘤不是 被根除，而GBMS重新发展。调节基底膜再生的事件响应单一- 药物NSC/iNSC的治疗方法尚不清楚。我们的结果显示移植的iNSCs药物载体被清除。 从脑中注入，但重复脑室(Icv)输注可恢复载体水平。我们还有 有证据表明，GBM细胞对iNSC传递的药物产生了抗药性。这使得我们可以假设GBM 对iNSC治疗的耐药性可以通过重复给药来解决载体丢失和多药联用的问题 INSC递送以解决肿瘤耐药性。有了这笔赠款，我们建议测试这一假设，定义 在NSC治疗期间有助于GBM动态适应的事件，并制定战略以转换 最初的肿瘤杀伤转变为持续的GBM抑制。我们将调查携带者清除、寻的和肿瘤 耐药贯穿于GBM的适应和复发过程。然后，我们将通过重复调整iNSC治疗 通过icv输注和运送携带多种药物有效载荷的iNSCs给药，目的是改善治疗 通过克服iNSC丢失和出现对单一药物具有抗药性的GBM焦点来实现持久性 治疗。所有测试都将使用我们的新型小鼠衍生的GBM细胞手术切除模型进行 免疫活性动物和患者来源的CD133+人GBM细胞最大限度地提高临床相关性 了解免疫系统对iNSC治疗耐受性的影响。结果是 这些研究对于创造基于NSC的持久肿瘤疗法至关重要，这些疗法能够产生持久的 在患者试验中抑制GBM。

Project Summary/Abstract Genetically engineered neural stem cells (NSCs) are a promising therapy for the highly aggressive brain cancer Glioblastoma (GBM). Engineered NSCs have unique tumor-homing capacity that allows them to deliver anti-cancer gene products directly into local and invasive GBM foci. Preclinical studies by our group and others have shown tumoricidal NSCs routinely reduce orthotopic GBM xenografts between 70-90% and significantly extend survival of tumor-bearing mice. Yet, these dramatic initial reductions in GBM volumes are not maintained and treatment durability remains a major challenge for NSC-based therapy. GBM escape occurs after treatment with NSCs carrying different therapeutic payloads and in pre-clinical models of both solid and post-surgical GBM. We recently discovered that novel tumor-homing drug delivery vehicles with robust anti- cancer activity can be developed from “induced neural stem cells” (iNSCs) using cellular reprogramming technology, referred to as transdifferentiation (TD). Tumoricidal iNSC therapy reduced GBM xenografts 230- fold in 4 weeks and more than doubled survival. Similar to wild-type NSC therapy, the tumors were not eradicated and the GBMs re-developed. The events mediating the regrowth of GBMs in response to single- agent NSC/iNSC therapy are unknown. Our results show that transplanted iNSCs drug carriers are cleared from the brain, but repeated intracerebroventricular (ICV) infusion restores carrier levels. We also have evidence that GBM cells become resistant to iNSC-delivered drugs. This allows us to hypothesize that GBM resistance to iNSC therapy can be overcome by repeat administration to address carrier loss and multi-agent iNSC delivery to address tumor resistance. With this grant we propose to test this hypothesis, defining the events that contribute to the dynamic adaption of GBM during NSC treatment and develop strategies to convert the initial tumor kill into sustained GBM suppression. We will investigate carrier clearance, homing, and tumor resistance throughout GBM adaption and recurrence. We will then modulate iNSC therapy through repeated dosing via ICV infusion and delivery of iNSCs carrying multi-drug payloads with the goal of improving treatment durability by overcoming iNSC loss and the emergence of GBM foci that are resistant to single-agent treatments. All testing will be done using our novel surgical resection models of murine-derived GBM cells in immune-competent animals and patient-derived CD133+ human GBM cells to maximize the clinical relevancy of our finding and understand the impact of the immune system on iNSC treatment durability. The results of these studies are essential for creating durable NSC-based tumor therapies capable of producing long-lasting GBM suppression in patient trials.