Non-Invasive Tracking of Genome-Corrected iPS cells in ALS

使用基因编辑的干细胞，特别是患者来源的iPSC用于细胞替代疗法是一种有效的方法。 这是一种有吸引力的方法，用于疾病相关基因突变的遗传校正。如果这些疗法 在未来的患者中，非常希望有可用的非侵入性细胞跟踪技术， 可以纵向报告移植细胞的分布和存活情况， 它们在体内的命运和优化个性化治疗。我们选择了肌萎缩侧索硬化症（ALS）， 一种毁灭性的疾病，死亡率接近100%，作为目标疾病的一个例子，其中运动功能丧失 神经元是导致肌肉麻痹的关键事件。对于家族性ALS，基因超氧化物突变 已知SOD 1是疾病发展的致病因素。在本建议中，我们的目标是 联合应用两种新的实验成像模式（MPI和PSMA靶向18F-DCFPyL PET） 使用临床新兴技术（1H MRI）回答与疗效相关的几个基本问题 和基因组编辑细胞疗法的安全性。这些是互补的技术，其中MPI和PSMA- 靶向18F-DCFPyL PET可以报告所施用细胞的全身分布，而MRI可以 报告细胞的实时归巢和即时保留。这种三管齐下的成像方法 细胞（MPI和MRI用SPIO标记，PET用18F-DCFPyL标记）将用于跟踪患者- 衍生的天然（39 b-SOD 1 +/AV 4）和基因组校正的（39 b-SOD 1 +/+）iPSC，有和没有分化 在转基因SOD 1G 37 R ALS小鼠模型中，动脉内注射，一种新兴的细胞 输送途径，将用于研究实时图像引导细胞注射的可行性，旨在获得一个 更多的全球性脑细胞分布，而脊髓实质内注射将作为一种 临床上有效的递送技术，将细胞局部递送到受损的运动神经元部位。如果成功， ALS中基因组校正的细胞的该示例性成像应用可以鼓励使用MPI，MRI， 和/或基于PMSA的PET成像，以询问体内其它疾病情况下细胞的命运。

The use of gene-edited stem cells and in particular patient-derived iPSCs for cell replacement therapy is an appealing approach for genetic correction of a disease-associated gene mutation. If such therapies are pursued in future patients, it would be highly desirable to have non-invasive cell tracking techniques available that can report longitudinally on the distribution and survival of transplanted cells, in order to better understand their fate in vivo and to optimize personalized therapies. We have chosen amyotrophic lateral sclerosis (ALS), a devastating disease with near 100% mortality as an example of a target disease, in which loss of motor neurons is a key event leading to muscle paralysis. For familial forms of ALS, mutations in the gene superoxide dismutase 1 (SOD1) are known to be a causative factor for disease development. In this proposal, we aim to apply two novel experimental imaging modalities (MPI and PSMA-targeted 18F-DCFPyL PET) in conjunction with a clinically emerging technique (1H MRI) to answer several basic questions associated with the efficacy and safety of genome-edited cell therapy. These are complementary techniques, where MPI and PSMA- targeted 18F-DCFPyL PET can report on the whole-body distribution of administered cells, whereas MRI can report on real-time homing and immediate retention of cells. This three-pronged approach of imaging the same cell (labeled with SPIO for MPI and MRI, and 18F-DCFPyL for PET) will be applied for tracking of patient- derived native (39b-SOD1+/AV4) and genome-corrected (39b-SOD1+/+) iPSCs, with and without differentiation into motor neurons, in a transgenic SOD1G37R mouse model of ALS. Intra-arterial injection, an emerging cell delivery route, will be used to study the feasibility of real-time image-guided cell injections aimed at obtaining a more global cerebral cell distribution, while intraparenchymal injection in the spinal cord will be applied as a clinically effective delivery technique to deliver cells locally at the site of impaired motor neurons. If successful, this example imaging application of genome-corrected cells in ALS may encourage the use of MPI, MRI, and/or PMSA-based PET imaging to interrogate the fate of cells in other disease scenarios in vivo.