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（由申请人提供）：神经干细胞（NSC）在某些情况下在大脑内迁移的天然能力，长期以来使其成为治疗中枢神经系统疾病的候选者。将人类体细胞重编程为组织的方法的开发。特定的干细胞为使用 NSC 疗法治疗许多脑部疾病带来了巨大的希望，在人类疾病的小鼠模型中进行的 NSC 移植实验已证明至少可以纠正几种类型疾病的某些病理学，为基于 NSC 的疗法提供了原理证明。然而，有效翻译的一个重大障碍是由于缺陷的性质，神经遗传疾病在中枢神经系统中具有全局分布的病变，因此治疗需要供体细胞的播散性分布。然而，需要更好地了解 NSC 的移植后特性。我们已将人类患者成纤维细胞重新编程为多能干细胞 (iPSC)，从中衍生出 NSC，并进行基因校正。我们建议进行异种移植研究，以评估改造 hu-iPS-NSC 对移植后分布和存活的影响。人类溶酶体贮积病 (LSD) 的小鼠模型 有超过 50 种 LSD，它们是影响中枢神经系统的所有遗传性儿童遗传病的罪魁祸首。原则上，对于大多数 LSD，基于溶酶体酶从基因校正细胞中输出并被突变细胞吸收以恢复缺失的酶活性的观察，初步研究表明 hu-iPS-NSC 植入了 NOD-SCID 中。我们提出的实验将研究移植后供体细胞分散和存活的基本特征，以解决 NSC 植入不足的问题。数据证明了所提出的实验的可行性，我们将使用定量分析来测量植入量、分化为成熟神经细胞类型以及病理变化。这一系列研究的长期策略是在小鼠中开发 NSC 移植。在未来的转化研究中，它们可以在脑部疾病的大型动物模型中进行测试，并最终进入比小鼠大脑大约 3000 倍的人脑。显然，要在这个问题上取得进展，需要在理解方面取得重大进展。 NSC的生物学植入和制定增强植入的策略。