Novel magnetic core/shell nanoparticle-based stem cell therapy to direct neural s

描述：该应用的长期目标代表了新型磁性核/壳纳米颗粒（MCNPs）的发展，以传递和时空触发干细胞向少突胶质细胞的分化。关于脊髓损伤，神经干/祖细胞（NSPCs）移植已显示出许多有利的治疗效果。然而，移植的NSPCs主要分化为星形胶质细胞，这往往会阻碍移植的有效性。由于这些细胞在轴突周围提供髓鞘，从而使神经冲动在中枢神经系统中快速传播，因此非常需要将移植的NSPCs引导分化为少突胶质细胞。为此，我们的目标是开发新型MCNPs，其具有双重功能，即在热休克启动子下传递编码Olig2的质粒，Olig2可诱导NSPC向少突胶质细胞分化，并通过磁热疗（即使用交变磁场）触发Olig2表达。为应对这些挑战，提出以下具体目标：制备磁性核/壳纳米颗粒和可诱导基因载体，用于转染人诱导多能干细胞衍生的神经干细胞/祖细胞（hiPSC-derived NSPCs）。具体目标2。目的：观察磁热诱导基因表达后，工程NSPCs体外少突胶质细胞分化及再髓鞘再生能力。磁性纳米颗粒先前已应用于MRI，细胞靶向和药物/基因传递。然而，在现有的知识和这些纳米颗粒在干细胞治疗中的临床应用之间存在着一个关键的差距。因此，一种新的基于mcnp的干细胞疗法的发展将证明MNPs在临床相关的脊髓损伤治疗中的多功能特性。特别是，与传统的基因疗法和细胞标记方法相比，基于mcnp的方法将提供许多优势，包括：i)非侵入性磁共振成像（由于磁芯）和拉曼成像（由于金壳）能力，ii)磁场促进基因载体进入干细胞的传递（“磁效应”），iii)磁热疗，这将用于提供被传递基因的激活机制。总的来说，拟议的MCNP方法将带来一种前沿的方法，可以允许用户实现对细胞分化的空间和时间控制，同时潜在地保持干细胞/前体细胞固有的神经保护特性。这样，

DESCRIPTION: The long-term goal of this application represents the development of novel magnetic core/shell nanoparticles (MCNPs) to deliver and spatiotemporally trigger the differentiation of stem cells to oligodendrocytes. With regard to spinal cord injury, neural stem/progenitor cell (NSPCs) transplantation has been shown to afford a number of favorable therapeutic effects. However, grafted NSPCs were found to differentiate primarily into astrocytes, which tend to hinder the effectiveness of transplantation. The guided differentiation of the grafted NSPCs into oligodendrocytes is highly desirable since these cells provide myelin sheaths around axons and thus enable fast propagation of nerve impulses in the CNS. To this end, the objective is to develop novel MCNPs, which have the dual functions of delivering a plasmid encoding Olig2, which has previously been reported to induce NSPC differentiation to oligodendrocytes, under a heat shock promoter and triggering Olig2 expression through magnetic hyperthermia (i.e. using an alternating magnetic field). To address these challenges, the following specific aims are proposed: Specific Aim 1. To prepare magnetic core/shell nanoparticles and inducible gene vectors for delivery into human induced pluirpotent stem cell-derived neural stem/progenitor cells (hiPSC-derived NSPCs). Specific Aim 2. To test the oligodendrocyte differentiation and remyelination ability of the engineered NSPCs in vitro after magnetic hyperthermia-induced gene expression. Magnetic nanoparticles have previously been applied for MRI, cell targeting, and drug/gene delivery. However, there is a critical gap between the existing knowledge and the clinical application of these nanoparticles to stem cell-based therapy. Therefore, the development of a novel MCNP-based stem cell therapy will demonstrate the multifunctional nature of MNPs for a clinically-relevant SCI treatment. In particular, compared to conventional gene therapies and cellular labeling methodologies, a MCNP-based approach would offer many advantages including: i) non-invasive magnetic resonance imaging (due to magnetic core) and Raman imaging (due to the gold shell) capabilities, ii) magnetic field-facilitated delivery ('magnetofection') of gene vectors into the stem cells, and iii) magnetic hyperthermia, which will be used to provide a mechanism for the activation of the delivered gene. Overall, the proposed MCNP approach will bring a methodology to the forefront that can allow the user to achieve spatial and temporal control over cellular differentiation, while potentially maintaining the neuroprotective properties innate to stem/precursor cells. In this way, scientists and clinicians can harness the full potential of stem cells (i.e. intrinsic therapeutic properties and controlled cell fate specification) for an enhanced SCI treatment.