Chemically produced neural progenitors loaded with ferumoxide-protamine sulfate complex for visualization of gliomas

胶质母细胞瘤是一种高度侵袭性的肿瘤，复发率高达98%，多数病例可迅速致死。这个 当前临床和实验治疗未能根除播散性胶质瘤细胞导致肿瘤 复发，中位生存期3-6个月。此外，目前可用的成像技术包括 对可能导致复发的微小肿瘤卫星的检测不够灵敏。近期 研究进展建立了间充质干细胞(MSCs)、神经干细胞(NSCs)和神经细胞 神经前体细胞是治疗神经胶质瘤的有效载体。这些干细胞可以追踪到 肿瘤块中的癌细胞以及迁移到脑实质的癌细胞。正因为如此 作为一种独特的能力，已经探索了使用间充质干细胞和神经干细胞/神经干细胞作为治疗和成像手段。 最近的几项研究表明，神经干细胞比骨髓间充质干细胞具有许多优势。具体地说，NSC有一个独特的 在体内抑制肿瘤生长的能力，并对胶质瘤表现出更大和更特异的趋向性。这个 NSC的缺点是缺乏足够的NSC来源。当前生产的替代方法 通过诱导多能干细胞(IPSC)技术诱导的神经干细胞充满了无法估量的风险。最近，我们 已经能够从人的骨髓和脂肪来源的间充质干细胞中产生神经干细胞/祖细胞 使用纯化学方法(或小分子方法)，这比目前的方法有几个优点 已知的技术。我们最近的研究表明，这些化学诱导的神经前体细胞 (CiNPC)对恶性胶质瘤D54-GM细胞的迁移能力高于hMSCs 预先将这些细胞暴露于D54-MG条件培养液(GCM)中，可使其进一步增加。因此，CiNPC 由我们最近开发的安全、快速、可重现和具有成本效益的重新编程方法产生 被有效地用作治疗脑肿瘤的药物载体和/或磁性载体 磁共振成像(MRI)造影剂用于胶质瘤的可视化。此SBIR第一阶段提案的目标是 为了研究GCM预暴露和磁性标记的CiNPC是否可以用于可视化 MRI检测颅内D54-MG胶质瘤细胞。第二阶段的研究将调查这些细胞用于 不同类型胶质瘤的MRI可视化。商业和临床相容的研究产品 从I/II阶段工作中脱颖而出的技术包括大规模临床级生产磁性材料的技术 负载的CiNPC可用于MRI对不同胶质瘤的可视化。为了实现这些目标， 为第一阶段研究提出了以下具体目标。特定目标1将检验GCM预 暴露的负载有铁氧化物-鱼精蛋白硫酸盐复合体(FE-Pro)的CiNPC将保持其活性和 体外对D54-MG细胞的趋向性。特定目标2将检验以下假设：GCM预暴露和FE-Pro- 负载的CiNPC在体内对D54-MG胶质瘤移植瘤表现出趋向性，这可以通过MRI进行监测。

Glioblastoma is a highly invasive tumor with a recurrence rate of 98%, which is in most cases rapidly fatal. The failure of current clinical and experimental therapies to eradicate disseminated glioma cells results in tumor recurrence and a median survival of 3–6 months. Furthermore, currently available imaging technologies are not sufficiently sensitive for detection of small tumor satellites, which are responsible for recurrence. Recent research advances have established mesenchymal stem cells (MSCs), neural stem cells (NSCs), and neural progenitor cells (NPCs) as promising therapeutic delivery vehicles for gliomas. These stem cells track down cancer cells in the tumor mass as well as migratory cancer cells into the brain parenchyma. Because of this unique ability, the use of MSCs and NSCs/NPCs as therapeutic and imaging modalities has been explored. Several recent studies demonstrated many advantages of NSCs over MSCs. Specifically, NSCs have a unique ability to suppress tumor growth in vivo, and exhibit greater and more specific tropism to gliomas. The disadvantage of NSCs is the lack of an adequate NSC source. Current alternative approaches to produce NSCs by induced pluripotent stem cell (iPSC) technologies are fraught with incalculable risks. Recently, we have been able to generate neural stem/progenitor cells from human bone marrow and adipose derived MSCs using a purely chemical approach (or small molecule approach) that has several advantages over currently known technologies. Our recent studies demonstrated that these chemically induced neural progenitors (CiNPCs) exhibited higher migratory capacity to malignant glioma D54-GM cells compared to hMSCs which was further increased by pre-exposure of these cells to D54-MG conditioned medium (GCM). Thus, CiNPCs produced by our recently developed safe, fast, reproducible, and cost-effective reprogramming approach can be efficiently used as a drug delivery vehicle for the treatment of brain tumors and/or as carriers for magnetic resonance imaging (MRI) contrast agents for visualization of gliomas. The goal of this SBIR phase I proposal is to investigate whether GCM pre-exposed and magnetically labeled CiNPCs could be utilized for visualization of intracranial D54-MG glioma cells by MRI. Phase II studies will investigate the potential use of these cells for MRI visualization of different types of gliomas. Commercial and clinically compatible research products emerging from Phase I/II work includes technology for large-scale clinical grade production of magnetically loaded CiNPCs that can be used for visualization of different gliomas by MRI. To achieve these goals the following Specific Aims are proposed for Phase I studies. Specific Aim 1 will test the hypothesis that GCM pre- exposed CiNPCs loaded with ferumoxide-protamine sulfate complex (FE-Pro) will retain their viability and tropism to D54-MG cells in vitro. Specific Aim 2 will test the hypothesis that GCM pre-exposed and FE-Pro- loaded CiNPCs will exhibit tropism to D54-MG glioma xenografts in vivo, which can be monitored by MRI.