Project Summary/Abstract The overall goal of this proposal is to develop a pig model of early-onset Alzheimer’s disease (AD). Alzheimer’s disease is the leading cause of dementia and affects 10% of the U.S. population. Unfortunately, there is currently no treatment option that can reverse the trajectory of the disease. Mouse models of AD have been critical to identify the underlying pathophysiology of this disorder. However, these models often do not recapitulate all symptoms of the human condition, indicating that additional animal models of AD are needed to facilitate our understanding of this disorder and accelerate drug discovery efforts. Pigs are readily available and are considered to be an excellent biomedical model due to anatomical and physiological similarities to humans. In addition, in comparison to rodent models, pigs have a more similar brain structure to humans and can perform more complicated cognitive tasks that better model human behavior, suggesting that they could serve as an ideal large animal model of AD. Here, we propose to develop the first pig models of AD in the U.S and provide the models to biomedical community. Diversifying animal models reflecting phenotypes of human AD patients should be invaluable to design novel remedies that can secure longevity and quality of life in humans. Under this application, we propose to use genome editing technology to develop genetically engineered pigs and cells carrying modified genes that are linked to AD patients. Specifically, polymorphisms in APP and PSEN1 are known to be responsible for early onset AD disease, i.e. Familial Alzheimer's disease. We propose two aims. In Aim 1, we will generate genetically engineered pigs carrying modified APP or PSEN1, and explore the cellular and molecular signs of AD in the pig models. The models will serve as a base that can be used to incorporate more complicated modifications, such as overexpression of other human AD-linked genes in the future. In Aim 2, we will generate pig cells lines carrying disrupted APP/PSEN1 while expressing human AD APP and PSEN1 genes under the control of a brain-specific promoter. The cell lines will be used to generate pig models mimicking phenotypes of the leading mouse model through cloning in the future. Conventional mouse AD models do capture some AD symptoms; however, due to differences in physiology and size of brain, it is rare that findings from the models are translated to the clinic. The development of pig models for early-onset Alzheimer’s disease can serve as a novel resource to the biomedical community and help advance our understanding and potential treatment of this major neurodegenerative disorder. We have expertise to complete the proposed experiment within the grant timeframe and our activities will be valuable to investigators who works on neurodegenerative diseases and memory decline.