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

项目总结

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.