Disseminated gene delivery to the CNS by human iPSC-derived neural stem cells

通过人类 iPSC 衍生的神经干细胞将播散性基因传递至 CNS

• 批准号: 9204865
• 负责人: JOHN H WOLFE
• 金额: $ 36.75万
• 依托单位: CHILDREN'S HOSP OF PHILADELPHIA
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-02-01 至 2020-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9204865
• 关键词: Address; Adult; Affect; Affinity; Animal Model; Biology; Brain; Brain Diseases; Cell Culture Techniques; Cell Therapy; Cell Transplantation; Cell Transplants; Cells; Central Nervous System Diseases; Childhood; Clinical; Data; Defect; Development; Diffuse; Disease; Electrophysiology (science); Engineering; Engraftment; Environment; Enzymes; Fibroblasts; Future; Gene Delivery; Gene Transfer; Goals; Grant; Hereditary Disease; Human; Human Engineering; Human Genetics; In Vitro; Individual; Inherited; Injection of therapeutic agent; Lesion; Life; Lysosomal Storage Diseases; Measures; Mediating; Meta-Analysis; Methods; Mucopolysaccharidosis VII; Mus; NOD/SCID mouse; Nature; Neurons; Notch Signaling Pathway; Outcome; Pathology; Pathway interactions; Patients; Pattern; Pluripotent Stem Cells; Property; Proteins; Research; Safety; Site; Somatic Cell; Stem cells; Structure; Testing; Therapeutic; Time; Tissues; Translating; Translations; Transplantation; Treatment Efficacy; Up-Regulation; Xenograft procedure; base; cell motility; cell type; cellular engineering; clinical development; common treatment; density; dysmyelination; effective therapy; experimental study; gray matter; human disease; improved; in vivo; induced pluripotent stem cell; method development; migration; mouse model; mutant; nerve stem cell; neurogenetics; neuropathology; public health relevance; repaired; stem cell therapy; subventricular zone; therapeutic effectiveness; therapeutic evaluation; transcription factor; transcriptome; translational study; treatment strategy; white matter

 DESCRIPTION (provided by applicant): The natural ability of neural stem cells (NSCs) to migrate within the brain, under certain circumstances, has long made them a candidate for treatment of CNS diseases. The development of methods to reprogram human somatic cells into tissue specific stem cells has provided great hope for using NSC therapies for many brain disorders. NSC transplantation experiments in mouse models of human diseases have demonstrated correction of at least some pathology in several types of diseases, providing proof-of-principle for the NSC- based approach. However, a significant barrier to effective translation is the low level of engraftment that occurs. Neurogenetic diseases have globally distributed lesions in the CNS due to the nature of the defect, thus treatment requires disseminated distribution of the donor cells. Achieving that will, however, require much better understanding of the post-transplantation properties of NSCs. We have reprogrammed human patient fibroblasts into pluripotent stem cells (iPSCs), derived NSCs from them, and genetically corrected the hu-iPS- NSCs. We propose xenograft studies to evaluate the effects of engineering hu-iPS-NSCs on post-transplant distribution and survival. We will test the therapeutic effectiveness of delivering a diffusible protein within the brain, in a well-characterized mouse model of a human lysosomal storage disease (LSD). There are >50 individual LSDs and they are responsible for a large portion of all inherited childhood genetic diseases that affect the CNS. A common treatment strategy can be used, in principle, for most of the LSD's, based on the observation that lysosomal enzymes are exported from genetically corrected cells and taken up by mutant cells to restore the missing enzymatic activity. Preliminary studies indicate the hu-iPS-NSCs engraft in the NOD- SCID mouse brain at low levels. The proposed experiments will investigate basic features of post-transplant dispersion and survival of the donor cells to address the problem of inadequate NSC engraftment. We have obtained compelling preliminary data demonstrating the feasibility of the proposed experiments and we will use quantitative analyses to measure amounts of engraftment, differentiation into mature neural cell types, and changes in pathology. The long-term strategy of this line of research is to develop NSC transplants in the mouse brain to a level where they can be tested in large animal models of brain diseases in future translational studies and eventually into the human brain which is ~3,000 times larger than the mouse brain. It is clear that progress on this problem needs significant advances in understanding the biology of NSC engraftment and development of strategies to enhance engraftment.

 描述（由适用提供）：在某些情况下，神经元干细胞（NSC）在大脑内迁移的自然能力长期以来使它们成为治疗中枢神经系统疾病的候选人。将人类体细胞细胞转化为组织特异性干细胞的方法的发展为用于许多脑部疾病使用NSC疗法提供了巨大的希望。在人类疾病的小鼠模型中进行的NSC移植实验表明，在几种类型的疾病中至少对某些病理进行了纠正，为基于NSC的方法提供了原理证明。但是，有效翻译的重要障碍是发生的低水平的植入水平。由于缺陷的性质，神经遗传疾病在中枢神经系统中具有全球分布的病变，因此治疗需要供体细胞的分布分布。但是，实现这将需要更好地理解NSC的移植后特性。我们已将人类患者的成纤维细胞重新编程为多能干细胞（IPSC），从中得出了NSC，并通过遗传纠正了Hu-ips-NSC。我们提出了异种移植研究，以评估工程HU-IPS-NSC对移植后分布和生存的影响。我们将在人类溶酶体储存疾病（LSD）的特征良好的小鼠模型中测试在大脑内传递可扩散蛋白的治疗有效性。有> 50个单独的LSD，它们负责影响中枢神经系统的所有遗传儿童遗传疾病的大部分。原则上，大多数LSD可以使用一种常见的治疗策略，基于观察结果，即溶酶体酶从遗传校正的细胞中导出并被突变细胞吸收以恢复缺失的酶促活性。初步研究表明，Hu-ips-NSC在低水平中植入了小鼠大脑中的植入。提出的实验将研究移植后分散和供体细胞存活的基本特征，以解决NSC植入不足的问题。我们获得了引人注目的初步数据，证明了提出的实验的可行性，我们将使用定量分析来测量植入量，分化为成熟的神经元细胞类型以及病理学变化。这项研究线的长期策略是将小鼠大脑中的NSC移植物开发到可以在未来翻译研究中大脑疾病的大动物模型中进行测试的水平，最终进入人脑的大脑比小鼠大脑大约3,000倍。显然，在理解NSC植入的生物学和增强植入策略的生物学方面，这一问题的进展需要重大进展。