MSCS ENGINEERED TO PRODUCE BDNF AND GENE EDITING CARGO FOR THE TREATMENT OF HUNTINGTON'S DISEASE

MSCS 旨在生产用于治疗亨廷顿病的 BDNF 和基因编辑货物

基本信息

批准号：
10443416
负责人：
Kyle Fink
金额：
$ 38.89万
依托单位：
UNIVERSITY OF CALIFORNIA AT DAVIS
依托单位国家：
美国
项目类别：
财政年份：
2017
资助国家：
美国
起止时间：
2017-07-01 至 2027-03-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10443416
关键词：
Address Adult Alleles Animal Model Anxiety Apoptosis Area Atrophic Award BDNF gene Binding Biological Assay Brain Brain region Brain-Derived Neurotrophic Factor CRISPR/Cas technology Cell Therapy Cell secretion Cells Chromatin Clinical Trials Clustered Regularly Interspaced Short Palindromic Repeats Corpus striatum structure DNA Binding Domain Data Disease Dose Double-Blind Method Engineering Epigenetic Process Evaluation Funding Future Gene Silencing Gene Targeting Generations Genes Genetic Transcription Glycoproteins Grant Growth Factor Guide RNA Half-Life Heterochromatin Human Huntington Disease Huntington gene Immune Impairment Induced pluripotent stem cell derived neurons Injections Intraventricular Investigational Drugs Knowledge Laboratories Lead Length Link Liquid substance Lyssavirus Mesenchymal Methods Mission Mouse Strains Mus National Institute of Neurological Disorders and Stroke Neurodegenerative Disorders Neurons NuRD complex Pathway interactions Patients Phase III Clinical Trials Positioning Attribute Precipitation Precision therapeutics Production Proteins Protocols documentation Publishing RNA Recovery Repression Reproducibility Research Rodent Model Route Safety Signal Pathway Signal Transduction Spinal Cord Stromal Cells Techniques Technology Testing Therapeutic Transcription Repressor Transgenic Organisms Tropism Up-Regulation Validation Variant Vesicle Vesicular stomatitis Indiana virus Work Zinc Fingers animal data base cell type cellular engineering cisterna magna delivery vehicle design and construction disease phenotype effectiveness evaluation efficacy study engineered stem cells excitotoxicity gene repression gene therapy good laboratory practice implantation induced pluripotent stem cell knock-down mouse model mutant nanoengineering nerve stem cell nervous system disorder neurogenesis neuropathology neurorestoration next generation sequencing novel novel therapeutics oxidative damage phase I trial preservation rabies virus glycoprotein G recruit restoration stem stem cell delivery therapy development tool transcription factor vector

项目摘要

Abstract Huntington's disease (HD) is a neurodegenerative disorder with no cure, and there is a critical unmet need for disease- modifying treatments. We are developing a novel therapy for HD: implantation of human Mesenchymal Stem/Stromal Cells (MSCs) engineered to secrete Brain-Derived Neurotrophic Factor (MSC/BDNF), a growth factor needed in the degenerating striatal regions of the brain, along with gene editing cargo to specifically reduce levels of production of the mutant RNA and protein. BDNF is low in humans and mice with HD, and up- regulation of BDNF in the brains of transgenic rodent models of HD has ameliorated the disease phenotype. Due to pro-survival effects in striatal neuropathology, BDNF is a strong candidate for neuroprotective therapies. Our labs have also developed targeted gene silencing tools using a novel variant of Cas9 (xCas9 3.7) that allows for broad protospacer adjacent motif (PAM) targeting. We have demonstrated this platform is effective in reducing huntingtin when fused to a powerful transcriptional repressor, KRAB. We have demonstrated that MSCs are an excellent delivery vehicle. We are testing injection via the cisterna magna as a surrogate for spinal cord/CSF fluid delivery in future patients who might receive this cell and gene therapy product developed by our team. Cellular Nanoengineered xCas9 Therapy (CellNeXT) combines the beneficial effects of MSC administration to the striata with the benefits of BDNF production while in combination reduction levels of mutant huntingtin. Unlike BDNF delivery via direct vector injection or protein administration into the brain, MSCs migrate into the areas of damage and have numerous beneficial effects. Although optimized MSCs will not persist longer than several months, we hypothesize that the neurorestorative effects of BDNF will outlast the survival of MSCs. This is supported by animal data from our laboratory and others. In our double-blinded efficacy studies, intrastriatal delivery of human MSC/BDNF significantly reduced anxiety and significantly increased neurogenesis in immune suppressed HD mice, with increased survival, in comparison to vehicle treated HD mice. We have demonstrated that treatment with MSC/BDNF decreased striatal atrophy as compared to vehicle treated HD mice (PMID:26765769). This recovery may be due to the stimulation of endogenous neurogenesis promoted by BDNF and enhanced by the secretion of various complementary therapeutic factors by the MSCs. In the planned studies, we will perform the following studies in support of an investigational new drug filing to the FDA: in Aim 1 we will evaluate the selectivity and durability of xCas9 in patient iPSC-derived NSC and neurons. In Aim 2 we will assess dose and route of administration of CellNeXT in our novel HD mouse model. Our studies will define reproducible techniques and methods, at the level of Good Laboratory Practice, for evaluation of cell and gene therapy candidates to be used in neurodegenerative disorders. We will be positioned to move the CellNeXT candidate into clinical trials for HD initially, and to assist others in using the product for additional disorders in the future. We will better define mechanism of action of CellNeXT and will define potency assays, using reduction of mutant huntingtin and preservation of striatal atrophy as key readouts.

抽象的亨廷顿病 (HD) 是一种无法治愈的神经退行性疾病，并且对疾病的迫切需求未得到满足 - 修改治疗方法。我们正在开发一种新的 HD 疗法：植入人类间充质干细胞/基质细胞 (MSC) 被设计用于分泌脑源性神经营养因子 (MSC/BDNF)，这是一种退化细胞所需的生长因子大脑纹状体区域，以及基因编辑货物，专门降低突变 RNA 的产生水平蛋白质。 BDNF 在人类和患有 HD 的小鼠中较低，而在转基因啮齿动物模型的大脑中 BDNF 上调 HD 改善了疾病表型。由于纹状体神经病理学中的促生存作用，BDNF 是一种强大的神经保护疗法的候选者。我们的实验室还使用一种新的变体开发了靶向基因沉默工具 Cas9 (xCas9 3.7) 允许广泛的原型间隔子相邻基序 (PAM) 靶向。我们已经展示了这个平台当与强大的转录抑制因子 KRAB 融合时，可有效减少亨廷顿蛋白。我们已经证明了 MSC 是一种优秀的输送工具。我们正在测试通过小脑延髓池注射作为脊髓/脑脊液的替代物未来可能接受我们团队开发的细胞和基因治疗产品的患者进行液体输送。蜂窝网络纳米工程 xCas9 疗法 (CellNeXT) 将 MSC 给药对纹状体的有益效果与 BDNF 产生的好处，同时减少突变亨廷顿蛋白的水平。与直接传递 BDNF 不同将载体注射或蛋白质注射到大脑中，间充质干细胞迁移到损伤区域并产生大量的有益的影响。尽管优化的 MSC 不会持续超过几个月，但我们假设 BDNF 的神经恢复作用将比 MSC 的存活时间更持久。我们实验室的动物数据支持了这一点其他的。在我们的双盲疗效研究中，纹状体内输送人 MSC/BDNF 显着减少焦虑和与媒介物相比，免疫抑制的 HD 小鼠的神经发生显着增加，存活率增加治疗HD小鼠。我们已经证明，与媒介物相比，MSC/BDNF 治疗减少了纹状体萎缩治疗 HD 小鼠 (PMID:26765769)。这种恢复可能是由于刺激内源性神经发生所致 BDNF 并通过 MSC 分泌的各种补充治疗因子得到增强。在计划的研究中，我们将进行以下研究，以支持向 FDA 提交研究性新药申请：在目标 1 中，我们将评估 xCas9 在患者 iPSC 衍生的 NSC 和神经元中的选择性和持久性。在目标 2 中，我们将评估剂量和途径在我们的新型 HD 小鼠模型中使用 CellNeXT。我们的研究将定义可重复的技术和方法，良好实验室实践水平，用于评估用于神经退行性疾病的细胞和基因疗法候选者失调。我们将首先将 CellNeXT 候选药物转入 HD 临床试验，并协助其他人将来使用该产品治疗其他疾病。我们将更好地定义 CellNeXT 的作用机制并将定义效力测定，使用突变亨廷顿蛋白的减少和纹状体萎缩的保留作为关键读数。