ABSTRACT (PROJECT 1) The underlying goal of Project 1 is to facilitate an integrated neuropathological approach incorporating traditional and quantitative pathology techniques with cell-type and molecular profiling to develop a unique resource to promote next generation AD research. It leverages existing collaborative relationships between UW Neuropathology and the UW ADRC and Kaiser Adult Changes in Thought studies for research autopsies. Project 1 will implement newly developed protocols for modernized autopsy sampling and preservation of brain tissue that are amenable to state-of-the-art cell type and molecular profiling techniques (Aim 1), as planned in Projects 2 and 3. Deceased subjects from both studies with short post-mortem intervals will enter Project 1 tissue pipeline to ensure high quality tissue preservation and availability for -omics Projects, with appropriate quality control measures at each step. Each ACT and ADRC autopsy includes qualitative neuropathological examination with extensive sampling and a battery of immunostains specific to pathologic peptides according to the latest guidelines. Neuropathological data from each case will be reviewed at regular meetings with Project 1 investigators; cases along the spectrum of AD pathology, but lacking co-morbid neuropathologies, will be promoted to eligibility for inclusion in Aim 2 and the Projects 2 and 3 pipelines. Aim 2 expands analysis of selected cases to include all regions of interest for Projects 2 and 3, but also regions of relevance to current AD cognitive subtypes and biomarker studies with a battery of immunohistochemical stains, image analysis, and quantitative assays to characterize neurodegeneration (pathologic peptides), neurotoxicity (neurons, synaptic markers, stains for white matter and oxidative stress), and reactivity (astrocytes, microglia, inflammation). In Aim 3, results from cell-type and molecular profiling studies in Projects 2 and 3 will be prioritized for targets identified in early AD pathogenesis, validated, and extended to the broader autopsy cohort to determine relevance. Quantitative measures for these analyses may include Luminex-based immunoassays and other techniques proteins and metabolites of neurodegeneration, neurotoxicity, and neuroinflammation/gliosis in regions and subjects of interest. We predict, based on the last five years, approximately 30 cases per year that are eligible for inclusion in Aim 2 and Project 2 and 3 pipelines. Each brain that goes through Project 1 pipeline, even if not selected for inclusion in the -omics components of the Center, will be preserved according to these novel protocols and deeply characterized for future studies of AD subtypes, risk variants, related disorders, and exposure profiles.

PROJECT NARRATIVE (PROJECT 1) Detailed analysis of cellular, molecular, and genetic information as well as spatial resolution of affected cell types is a key goal in understanding the cellular processes that give rise to Alzheimer’s disease. The current standard neuropathology protocols for sampling tissue, generally formalin-fixed paraffin-embedded tissue preservation, do not lend themselves to in-depth chemical and molecular analysis, nor does traditional neuropathological characterization of disease states provide sufficient detail to determine the pathophysiology of altered genes and proteins. When successfully completed, this project will have combined traditional and quantitative neuropathologic methods to better characterize AD pathologic stages and identify cases for Projects 2 and 3 and transformed donated brain tissue sampling and preservation in order to promote modern techniques and preserve valuable human tissue in a manner that is amenable to state-of-the-art molecular profiling and cell type analysis.