Generation of a Large Animal Model of Sialidosis to Enable Future Translation of Novel Therapeutics

生成唾液酸贮积症的大型动物模型，以实现新疗法的未来转化

基本信息

批准号：
10578286
负责人：
Heather L Gray-Edwards
金额：
$ 50.56万
依托单位：
UNIV OF MASSACHUSETTS MED SCH WORCESTER
依托单位国家：
美国
项目类别：
财政年份：
2023
资助国家：
美国
起止时间：
2023-01-01 至 2024-12-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10578286
关键词：
Adolescent Advanced Development Animal Model Animals Ataxia Attenuated Autopsy Biochemical Biological Assay Biological Markers Blindness Breeding CRISPR/Cas technology Cells Cessation of life Chemistry Childhood Clinic Clinical Clinical Trials Clustered Regularly Interspaced Short Palindromic Repeats Collaborations Complete Blood Count Data Data Set Databases Dependovirus Development Developmental Delay Disorders Disease Dose Electroencephalography Electroporation Embryo Ensure Enzymes Evaluation Exhibits Felis catus Fibroblasts Functional Magnetic Resonance Imaging Future Gangliosidosis GM1 Generations Genes Genetic Genetic Diseases Germ Lines Glycopeptides Goals Guide RNA Heterozygote Histopathology Human Image Implant Learning Life Lipomucopolysaccharidoses Lysosomal Storage Diseases Magnetic Resonance Imaging Medical Modeling Mus Mutation Mutation Analysis Myoclonus National Human Genome Research Institute Natural History Neuraminidase Neurodegenerative Disorders Neurologic Neurologic Examination Oligosaccharides Organ Outcome Measure Pathogenesis Pathologic Patients Peripheral Persons Phenotype Physical Examination Physicians Point Mutation Research Ribonucleoproteins Route Sampling Sandhoff Disease Seizures Serum Sheep Sialic Acids Standardization Tay-Sachs Disease Technology Testing Therapeutic Translating Translational Research Translations United States National Institutes of Health Validation Veterinarians Viral Genes access restrictions clinical trial implementation clinically relevant cognitive testing disease phenotype enzyme activity experience experimental study falls fetal first-in-human gene therapy genome editing human data human model infancy loss of function mouse model nervous system disorder novel therapeutics pre-clinical preservation prime editing programs rapid testing research clinical testing sheep model sialylation somatic cell nuclear transfer success therapeutic development therapeutic evaluation treatment strategy trial design

项目摘要

Project Summary Sialidosis is a rare, fatal, neurological disorder caused by a mutation in the NEU1 gene resulting in vision loss, seizures, involuntary myoclonus, and ataxia. Our team has a strong track record in brining gene therapies to the clinic and has plans to develop a gene therapy to treat sialidosis. Our previous experience has shown that testing in large animal models of human genetic diseases better approximates what will happen and patients and use of these models increases the likelihood of efficacy. We have developed a founder sheep with a mutation like type 1 sialidosis patients and will breed him to generate a colony of animals. Since the last submission we have created several severe mutations using CRISPR/spCas9 editing, therefore in this aim we will use Prime genome editing of embryos, to recreate two human mutations (Type 1 and Type 2) with the end goal of a mutation that recapitulates the human condition (Aim 1). We will evaluate each model for its ability to reliably mimic sialidosis then select the best model (Aim 2). This phenotyping includes in-life clinical metrics like MRI, EEG, EMG, neurological and cognitive testing as well as in depth post-mortem assays to determine if it reproduces biochemical and histopathological aspects of disease. External evaluation of in-life clinical testing will be performed by our clinical collaborator Dr. Tifft. Additionally, Dr. Tifft will make human samples available for comparison with the new sheep model. The biochemical aspects of disease will be externally validated by by Dr. d’Azzo (the leader in field of sialidosis) and pathological features characterized by Dr. Koehler a veterinary neuropathologist. After completion of these studies, this fully validated model will be used to learn more about sialidosis as a disorder, and also used in the development of an adeno associated viral gene therapy or other future treatment strategies for sialidosis.

项目概要唾液酸贮积症是一种罕见的致命性神经系统疾病，由 NEU1 基因突变引起，导致视力丧失，我们的团队在基因疗法治疗癫痫、不自主肌阵挛和共济失调方面拥有良好的记录。诊所并计划开发一种治疗唾液酸贮积症的基因疗法。在人类遗传疾病的大型动物模型中进行测试可以更好地模拟将发生的情况和患者并且使用这些模型增加了功效的可能性。突变就像 1 型唾液酸中毒患者一样，并且会繁殖他以产生一群动物。我们使用 CRISPR/spCas9 编辑创建了几个严重的突变，因此为了这个目标，我们将使用胚胎的 Prime 基因组编辑，重建两种人类突变（1 型和 2 型），以达到最终目标概括人类状况的突变（目标 1）我们将评估每个模型的能力。可靠地模拟唾液酸中毒，然后选择最佳模型（目标 2）。 MRI、EEG、EMG、神经学和认知测试以及深入的尸检分析以确定是否再现疾病的生化和组织病理学方面。将由我们的临床合作者 Tifft 博士进行，此外，Tifft 博士还将提供人体样本。与新的绵羊模型进行比较，疾病的生化方面将通过外部验证。由 d’Azzo 博士（唾液酸贮积病领域的领导者）提出，并由兽医 Koehler 博士描述病理特征完成这些研究后，这个经过充分验证的模型将用于了解更多信息。唾液酸贮积症作为一种疾病，也用于开发腺相关病毒基因疗法或其他唾液酸贮积症的未来治疗策略。