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：
10595651
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/10595651
关键词：
Acceleration Address Adult Alleles Animal Model Apoptosis Area Atrophic Award 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 Combined Modality Therapy 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 Length Link Liquid substance Lyssavirus Mesenchymal Methods Mission Mouse Strains Mus National Institute of Neurological Disorders and Stroke Nervous System 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 anxiety reduction cell motility 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 knock-down mouse model mutant nanoengineering nerve stem cell nervous system disorder neurogenesis neuropathology neuroprotection 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.

抽象的亨廷顿氏病（HD）是一种神经退行性疾病，无法治愈，并且对疾病的未满足至关重要 - 修改治疗。我们正在开发用于HD的新型疗法：植入人间充质茎/基质细胞（MSC）设计用于分泌脑衍生的神经营养因子（MSC/BDNF），这是退化的生长因子大脑的纹状体区域，以及基因编辑货物，以特异性降低突变体RNA的生产水平和蛋白质。 BDNF在人类和具有HD的小鼠中很低，并且在转基因啮齿动物模型的大脑中对BDNF的调节高清改善了疾病表型。由于在纹状体神经病理学中的促生物效应，BDNF是一个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型变体开发了靶向基因沉默工具 CAS9（XCAS9 3.7）的构成，可允许宽阔的原始探针相邻基序（PAM）靶向。我们已经证明了这个平台当与强大的转录阻遏物Krab融合时，可以有效地减少亨廷顿。我们已经证明了 MSC是一款出色的送货车。我们正在通过Cisterna Magna作为脊髓/CSF的替代物测试注射我们团队开发的可能会收到这种细胞和基因治疗产品的未来患者的流体递送。细胞纳米工程XCAS9治疗（CellNext）结合了MSC给药对纹状体的有益作用与 BDNF产生的好处是突变亨廷汀的组合降低水平。与BDNF通过直接交付不同向量注射或蛋白质给大脑的蛋白质施用，MSC迁移到损害区域，并具有许多有益的效果。尽管优化的MSC不会持续时间超过几个月，但我们假设 BDNF的神经训练作用将超过MSC的存活。这是由我们实验室的动物数据和其他的。在我们的双盲疗效研究中，人类MSC/BDNF的纹状体递送显着降低了焦虑和与媒介物相比治疗的高清小鼠。我们已经证明，与媒介物相比，使用MSC/BDNF治疗减少了纹状体萎缩治疗的高清小鼠（PMID：26765769）。这种恢复可能是由于刺激内源性神经发生的原因 BDNF并通过MSC对各种互补治疗因素的分泌增强。在计划的研究中我们将进行以下研究，以支持向FDA提交的调查新药：在AIM 1中，我们将评估患者IPSC衍生的NSC和神经元中XCAS9的选择性和耐用性。在AIM 2中，我们将评估剂量和途径在我们的新型HD小鼠模型中施用CellNext。我们的研究将定义可再现技术和方法良好的实验室实践水平，用于评估细胞和基因治疗候选物疾病。我们将定位将CellNext候选人最初移至HD临床试验中，并协助其他人将来将产品用于其他疾病。我们将更好地定义CellNext的作用机理，并将定义效力测定，使用减少突变体狩猎蛋白并保存纹状体萎缩作为关键读数。