Bruce T. Lamb - US grants

DESCRIPTION This is an R29 proposal to use YAC based approaches to presenilin wild-type and mutant transgenics linked to FAD and to breed them to YAC FAD mutant APP transgenics already in hand and examine effects on alterations in cell biology of APP and PS-1, excitotoxicity, LTP and neuropathology.

Alzheimer's disease (AD), the most common cause of dementia in the elderly, is now the fourth major cause of death in the developed world after heart disease, cancer and stroke. AD is strongly influenced by genetics, with current estimates suggesting that greater than 40% of all AD cases are familial (FAD). Genetic investigations have demonstrated that AD is a heterogeneous disorder with several known etiologies including: dosage imbalance for chromosome 21 as occurs in Down syndrome (DS); mutations in the amyloid precursor protein (APP) gene on chromosome 21, the presenilin-1 (PS-1) gene on chromosome 14 and the presenilin-2 (PS-2) gene on chromosome 1 in autosomal dominant early-onset FAD; and inheritance of distinct alpha-2 microglobulin and apolipoprotein E (ApoE) gene alleles on chromosome 12 and 19, respectively, as significant genetic risk factors for late-onset FAD. Additional genetic risk factors for AD are certain to exist. A pathologic hallmark in the brains of individuals with AD is deposits of the beta- amyloid (Abeta) peptide in the parenchyma of amygdala, hippocampus and neocortex. One of the major difficulties in studying the mechanism(s) of Abeta deposition and its impact on AD-related neuropathology, behavior and physiology is the paucity of accurate animal models. To establish an accurate genetic model for AD, we have focused on introducing entire genomic copies of either wild-type (wt) or mutant human APP and PS-1 genes carried on yeast artificial chromosomes (YACs) into the mouse germline and have recently demonstrated that a threshold into the molecular and mechanisms of the disease. The specific aims of the current proposal are to examine the effect of specific human AD genetic risk factors, namely dosage imbalance for genes on chromosome 21 (in the Ts65Dn mouse model of DS) and the various ApoE alleles (in ApoE knock-in mice), across the lifespan of APP YAC transgenic mice on: 1) Localization and timing of Abeta deposition and other structural neuropathology. 2) Learning and memory deficits characteristic of individuals with AD.

DESCRIPTION (provided by applicant): Alzheimer's disease (AD), the most common dementing disorder of late life, is a major cause of disability and death in the elderly. AD is characterized and diagnosed by distinctive neuropathological alterations including extracellular deposits of the ¿-amyloid (A¿) peptide. Epidemiological investigations have demonstrated that AD is a complex, age-related disorder with numerous proposed genetic and environmental etiologies. Human genetic studies have shown that altered dosage and mutations in the amyloid precursor protein gene as well as mutations in presenilin 1 and presenilin 2 genes all cause early-onset AD, while the APOE gene is major risk factor in late-onset AD. Considerable evidence suggests that these genetic factors alter A¿ metabolism and/or A¿ deposition. However, the AD genes identified thus far account for less than 30% of genetic risk for AD. Due to the inherent variability and genetic heterogeneity in late-onset neurodegenerative disorders, the identification of the remainder of the genetic and environmental factors that modulate AD risk have proven extremely difficult. Over the past decade and a half, several groups, including our own, have focused on the development of accurate and defined mouse models of AD, in which both the genetic background and environmental exposure can be precisely and reproducibly modified in an effort to gain insight into factors that modify APP processing, the production and deposition of A¿ peptides, the onset of dementia and the neuropathological abnormalities that occur in AD. In the previous funding period of this grant application, we generated congenic strains, in which a human AD transgene was transferred into different inbred mouse strains. We subsequently characterized the effects of different inbred genetic backgrounds and specific genetic alterations on A¿ metabolism and deposition and other AD phenotypes and identified unique mouse genetic loci that regulate metabolism and deposition. In addition, in preliminary studies we determined that the effects of an environmental factor, namely high-fat/high-cholesterol diets, on brain A¿ levels are dependent upon the genetic background of the mice. The current studies seek to expand and extend these studies with a focus on genetic and environmental factors that regulate A¿ metabolism and deposition in two inbred mouse strains with the most divergent A¿ phenotypes, C57BL/6J and A/J, using the unique genetic resources available for these strains. The specific aims of the current proposal are to: 1) Identify mouse chromosomes with gene(s) that regulate A¿ metabolism and deposition. 2) Identify and characterize genetic loci that regulate A¿ deposition. 3) Characterize genetic loci responsible for diet-induced alterations in A¿ metabolism and deposition. PUBLIC HEALTH RELEVANCE: Human genetic studies have demonstrated that accumulation and aggregation of small peptide fragments, termed beta-amyloid, is central to the disease mechanisms underlying Alzheimer's disease, a major cause of death and disability in the elderly. However, it remains unclear how the multitude of postulated genetic and environmental risk factors for human AD directly influence the production and aggregation of beta-amyloid. The current studies seek to identify and characterize additional genetic and environmental (namely high-fat/high-cholesterol diets) factors influence beta-amyloid generation and deposition within the brains of transgenic mouse models of AD.

DESCRIPTION (provided by applicant): Alzheimer's disease (AD), the most common cause of dementia in the elderly, is now the seventh major cause of death in the United States. AD is characterized and diagnosed by distinctive neuropathological alterations including extracellular deposits of the ß-amyloid (Aß) peptide, intracellular aggregates of the microtubule associated protein tau (MAPT) in neurons and marked neuroinflammation. Similarly in non-AD tauopathies, there is both abundant MAPT pathology and neuroinflammation. However, the exact mechanistic relationship between neuroinflammation and the various brain pathologies remains unclear. Recent studies have implicated neuronal-microglial signaling through the fractalkine receptor (CX3CR1) in neuroprotection and neurodegeneration. To examine the role of CX3CL1-CX3CR1 signaling in Alzheimer's disease and non- AD tauopathies, we conducted preliminary studies to examine the effects of CX3CR1 deficiency on both Aß and MAPT pathologies. Notably, CX3CR1 deficiency resulted in a reduction in Aß pathologies in two different mouse models of AD that was associated with altered microglial activation, while conversely, CX3CR1 deficiency in the hTau mouse model of MAPT pathology resulted in enhanced microglial activation, phosphorylation and aggregation of MAPT and behavioral impairments. Additional studies in both the Aß and MAPT models suggests that IL1 signaling may contribute to the CX3CR1 dependent alterations in AD brain pathologies. The hypothesis to be examined in the current studies is that soluble CX3CL1 released from neurons signals to CX3CR1 within microglia and plays a unique role in AD phenotypes via blocking phagocytic removal of Aß by microglia and reducing phosphorylation and aggregation of MAPT within neurons via mechanisms that involve IL1. These studies will utilize state-of-the art mouse models of Aß and MAPT pathologies, as well as CXC3CR1 knockout mice and CX3CL1 knockouts and transgenic mice to examine the effects of biochemistry, gene expression, neuropathology and behavior. The Specific Aims of this proposal are to: 1. Determine the Role of CX3CL1-CX3CR1 Signaling in a Mouse Model of Aß Deposition. 2. Determine the Role of CX3CL1-CX3CR1 Signaling in a Mouse Model of MAPT Pathology. 3. Determine the Role of IL1 Signaling in CX3CR1 Dependent Alterations in AD Pathologies.

DESCRIPTION (provided by applicant): Traumatic brain injury (TBI) is a very common injury in civilian as well as military populations in the United States and a large number of those that survive live with permanent TBI related disabilities. Notably, individuals exposed to TBI are at a greatly increased risk for developing a number of neurodegenerative diseases termed tauopathies, including Alzheimer's disease (AD) and chronic traumatic encephalopathy (CTE). More specifically, TBI promotes the intracellular aggregation of hyperphosphorylated, microtubule- associated protein tau (MAPT) into neurofibrillary tangles (NFTs) in both CTE and AD. One of the earliest and hallmark features of TBI is induction of neuroinflammation, including infiltration of peripheral monocytes into the site of injury and activation of resident brain microglia. Several lines of evidence from the literature and from our laboratories suggest that altered monocyte infiltration and microglial activation may be directly involved in the pathogenesis of MAPT pathologies. However, assessing the exact role of these cells in TBI-induced MAPT pathologies has proven exceedingly difficult due to the lack of reliable methods to distinguish monocytes and activated microglia within the brain in accurate genetic models of MAPT pathologies. The primary hypothesis to be tested in the current studies is that TBI induces infiltration of peripheral monocytes as well as acute and local activation of brain microglia within the injured brain and that these two cell types play roles distinct from each othe in inducing MAPT phosphorylation and aggregation leading to chronic pathological conditions that pre-dispose individuals exposed to TBI to develop tauopathies later in life. The objectives of this exploratory research grant are to assess the role of infiltrating monocytes and activated microglia in the development of MAPT pathologies following TBI utilizing unique genetic models that fluorescently tag monocytes (CCR2-RFP) and microglia (CX3CR1-GFP), genomic-based mouse models of MAPT (hTau) pathologies and genetic models in which infiltration of monocytes is blocked (CCR2 deficiency). The results of this study will be critical for future preclinical studies designed to therapeutically target either monocytes or microglia to block the effects of TBI on downstream MAPT pathologies. The specific aims of the proposal are: 1. Determine the effect of both mild and moderate TBI on infiltration of peripheral monocytes, activation of microglia and MAPT pathology, cognitive function and neurodegeneration in control and hTau mice. 2. Determine the role of infiltrating monocytes in TBI Induced MAPT pathologies in hTau mice.

PROJECT SUMMARY Alzheimer's disease (AD), the most common cause of dementia in the elderly, is now the third major cause of death in the United States. AD is characterized and diagnosed by distinctive neuropathological alterations including extracellular deposits of the ?-amyloid (A?) peptide intraneuronal aggregates of the microtubule associated protein tau (MAPT) and marked neuroinflammation. However, the exact mechanistic relationship between neuroinflammation and the various brain pathologies and clinical outcomes in AD remains unclear as well as the respective roles of brain resident microglia and infiltrating peripheral immune cells in these processes. Recent genetic and system biology studies have implicated multiple innate immune signaling pathways in late-onset AD. Most importantly, rare heterozygous coding mutations in TREM2, a gene exclusively expressed by myeloid cells were identified that substantially increase risk for AD and other neurodegenerative diseases. Our preliminary findings demonstrate that TREM2 is upregulated in mouse models of AD with A? pathology and human AD. Furthermore, co-localization studies demonstrated that TREM2 is selectively upregulated within myeloid cells in close proximity to A? deposits, but not in myeloid cells further away from A? deposits. Strikingly, careful flow cytometry and immunohistochemical analyses of brains from mouse models of AD revealed that TREM2 is upregulated within cells that bear markers reflective of their potential origin from circulating inflammatory monocytes. Consistent with these findings, preliminary analysis revealed an increase in TREM2+ cells in human AD blood samples. Furthermore, TREM2 deficiency in an AD mouse model leads to a virtual absence of the myeloid cells surrounding A? deposits and an overall reduction in AD-like pathologies. Finally, nuclear receptor agonists selectively target these plaque-associated TREM2+ cells and promote phagocytosis. The hypothesis to be tested in the current collaborative and interdisciplinary studies is that brain resident microglia and blood-derived TREM2+ inflammatory monocytes play distinctive roles in regulating AD pathologies that could provide novel biomarkers/diagnostics and also be targeted therapeutically. These studies will utilize state-of-the art mouse models of AD, constitutive Trem2 knockout mice, transgenic mice enabling deletion of TREM2 in various myeloid cell populations, detailed flow cytometry and genome-wide gene expression analyses as well as neuropathology and behavior to examine the three Specific Aims of the proposal: 1. Examine the Identity, Localization and Gene Expression Profiles of TREM2+ Cells in Mouse Models and Human AD. 2. Determine the Central and Peripheral Role of TREM2 in Regulating AD Pathologies. 3. Therapeutically Targeting TREM2+ Cells

PROJECT SUMMARY OVERALL COMPONENT Alzheimer's disease (AD) is a major cause of dementia, disability and death in the elderly. Despite recent advances in our understanding of basic biological mechanisms underlying AD, we do not yet know how to prevent AD or have an approved disease modifying intervention. Both are essential to slow or stop the growth in dementia prevalence. The National Alzheimer's Project Act (NAPA) seeks to prevent and effectively treat AD by 2025 through innovative research on etiology, early detection, and therapeutics. In support of NAPA's goals, one of the targeted areas of research identified at the NIA sponsored 2015 Alzheimer's Disease Research Summit was the development of the next generation of animal models of AD that will prove more predictive in preclinical studies and thus accelerate the drug testing pipeline. While our current animal models of AD have provided multiple novel insights into AD disease mechanisms, thus far they have not been successfully utilized to predict the effectiveness of therapies that have moved into AD clinical trials. The Indiana University (IU)/Jackson Laboratory (JAX) Alzheimer's Disease Precision Models Center (IU/JAX ADPMC) will leverage IU's strengths in neurodegenerative research including 25 years as an NIA-supported Alzheimer's Disease Center (ADC) and considerable expertise in preclinical drug testing with JAX's eight decades of expertise in mammalian genetics and disease modeling to develop, validate and disseminate new, precise animal models of Alzheimer's disease (AD). In addition, the IU/JAX ADMPC contains Sage Bionetworks to provide expertise in data organization and dissemination. The IU/JAX ADPMC brings together an international, multi-disciplinary team?including geneticists and genetics technology experts, quantitative and computational biologists, clinical experts in AD and neuroimaging, pharmacologists and world leaders in the development of precision animal models of disease?that possesses the collective ability to foresee disease modeling needs as they emerge on the international stage. This will allow the IU/JAX ADPMC to serve the AD scientific community effectively and efficiently. The IU/JAX ADPMC will generate new AD modeling processes and pipelines, data resources, research results and models that will be swiftly shared through JAX's and Sage's proven dissemination pipelines and through the NIA- supported AD Centers, academic medical centers, research institutions and the pharmaceutical industry worldwide. Ultimately, this will accelerate the application of advances in animal models for the greatest possible medical benefit. The Specific Aims of the IU/JAX ADPMC are: 1. Maximize Human Datasets to Identify Putative Variants, Genes and Biomarkers for AD. 2. Generate and Characterize the Next Generation of Mouse Models of AD. 3. Validate the Next Generation of Mouse Models of AD and Develop a Preclinical Testing Pipeline. !

PROJECT SUMMARY ADMINISTRATIVE (ADMIN) CORE The Administration (ADMIN) Core of the Indiana University (IU)/Jackson Laboratory (JAX) Alzheimer's Disease Precision Models Center (IU/JAX ADPMC), led by ADMIN Head Dr. Bruce Lamb at IU and with assistance from multiple Principle Investigators (mPIs) Dr. Paul Territo at IU and Drs. Greg Carter and Gareth Howell at JAX, along with the support of two PhD-level Program co-Managers and full-time administrators at both IU and JAX, will oversee all administrative and scientific functions of the Center. The ADMIN Core will execute these duties with oversight from an External Advisory Board (EAB), composed of leading experts in the field, and a Center Steering Committee (CSC), composed of the PIs, research leaders at both IU and JAX, and program officials from the NIH. The EAB will convene biannually and the CSC monthly to review progress and provide input and feedback (oral and written) to ensure that the IU/JAX ADPMC (1) achieves its mission, aims, milestones and metrics; (2) effectively deals with any unanticipated problems or concerns; and (3) adapts to likely experimental and technical advances in the field. There will be weekly ADMIN meetings and monthly Core and Project meetings. These will be focused on issues unique to specific Cores/Project while ensuring maximal synergy between the various components of the IU/JAX ADPMC. Close interaction will be fostered among the Bioinformatics and Data Management Core, the Preclinical Testing Core and the Disease Model Development and Phenotyping Project, thus facilitating the speed of discovery while reducing overall cost. One of the primary goals of the ADMIN Core will be to ensure that discoveries made in one area of the ADPMC will benefit the whole, leading to efficiencies in time and cost. Finally, the ADMIN Core will also promote interactions and meetings of the IU/JAX ADPMC with other ongoing research efforts in the Alzheimer's research community, including the NIA-supported ADCs/ADRCs, the Alzheimer's Disease Neuroimaging Initiative (ADNI), Alzheimer's Disease Accelerating Medicine's Partnership (AD-AMP), and the Alzheimer's Disease Sequencing Project (ADSP). The Specific Aims of the IU/JAX ADPMC ADMIN Core are: 1. Provide effective and efficient management, oversight and evaluation of the IU/JAX ADPMC and interactions with the AD research community. 2. Ensure timely and successful completion of all project and core aims and milestones. 3. Facilitate interactions between and among the Bioinformatics and Data Management Core, the Preclinical Testing Core and the Disease Model Development and Phenotyping Project. !

PROJECT SUMMARY OVERALL COMPONENT Alzheimer's disease (AD) is a major cause of dementia, disability and death in the elderly. Despite recent advances in our understanding of basic biological mechanisms underlying AD, we do not yet know how to prevent AD or have an approved disease modifying intervention. Both are essential to slow or stop the growth in dementia prevalence. The National Alzheimer's Project Act (NAPA) seeks to prevent and effectively treat AD by 2025 through innovative research on etiology, early detection, and therapeutics. In support of NAPA's goals, one of the targeted areas of research identified at the NIA sponsored 2015 Alzheimer's Disease Research Summit was the development of the next generation of animal models of AD that will prove more predictive in preclinical studies and thus accelerate the drug testing pipeline. While our current animal models of AD have provided multiple novel insights into AD disease mechanisms, thus far they have not been successfully utilized to predict the effectiveness of therapies that have moved into AD clinical trials. The Indiana University (IU)/Jackson Laboratory (JAX) Alzheimer's Disease Precision Models Center (IU/JAX ADPMC) will leverage IU's strengths in neurodegenerative research including 25 years as an NIA-supported Alzheimer's Disease Center (ADC) and considerable expertise in preclinical drug testing with JAX's eight decades of expertise in mammalian genetics and disease modeling to develop, validate and disseminate new, precise animal models of Alzheimer's disease (AD). In addition, the IU/JAX ADMPC contains Sage Bionetworks to provide expertise in data organization and dissemination. The IU/JAX ADPMC brings together an international, multi-disciplinary team?including geneticists and genetics technology experts, quantitative and computational biologists, clinical experts in AD and neuroimaging, pharmacologists and world leaders in the development of precision animal models of disease?that possesses the collective ability to foresee disease modeling needs as they emerge on the international stage. This will allow the IU/JAX ADPMC to serve the AD scientific community effectively and efficiently. The IU/JAX ADPMC will generate new AD modeling processes and pipelines, data resources, research results and models that will be swiftly shared through JAX's and Sage's proven dissemination pipelines and through the NIA- supported AD Centers, academic medical centers, research institutions and the pharmaceutical industry worldwide. Ultimately, this will accelerate the application of advances in animal models for the greatest possible medical benefit. The Specific Aims of the IU/JAX ADPMC are: 1. Maximize Human Datasets to Identify Putative Variants, Genes and Biomarkers for AD. 2. Generate and Characterize the Next Generation of Mouse Models of AD. 3. Validate the Next Generation of Mouse Models of AD and Develop a Preclinical Testing Pipeline. !

PROJECT SUMMARY OVERALL COMPONENT Alzheimer's disease (AD) is a major cause of dementia, disability and death in the elderly. Despite recent advances in our understanding of basic biological mechanisms underlying AD, we do not yet know how to prevent AD or have an approved disease modifying intervention. Both are essential to slow or stop the growth in dementia prevalence. The National Alzheimer's Project Act (NAPA) seeks to prevent and effectively treat AD by 2025 through innovative research on etiology, early detection, and therapeutics. In support of NAPA's goals, one of the targeted areas of research identified at the NIA sponsored 2015 Alzheimer's Disease Research Summit was the development of the next generation of animal models of AD that will prove more predictive in preclinical studies and thus accelerate the drug testing pipeline. While our current animal models of AD have provided multiple novel insights into AD disease mechanisms, thus far they have not been successfully utilized to predict the effectiveness of therapies that have moved into AD clinical trials. The Indiana University (IU)/Jackson Laboratory (JAX) Alzheimer's Disease Precision Models Center (IU/JAX ADPMC) will leverage IU's strengths in neurodegenerative research including 25 years as an NIA-supported Alzheimer's Disease Center (ADC) and considerable expertise in preclinical drug testing with JAX's eight decades of expertise in mammalian genetics and disease modeling to develop, validate and disseminate new, precise animal models of Alzheimer's disease (AD). In addition, the IU/JAX ADMPC contains Sage Bionetworks to provide expertise in data organization and dissemination. The IU/JAX ADPMC brings together an international, multi-disciplinary team?including geneticists and genetics technology experts, quantitative and computational biologists, clinical experts in AD and neuroimaging, pharmacologists and world leaders in the development of precision animal models of disease?that possesses the collective ability to foresee disease modeling needs as they emerge on the international stage. This will allow the IU/JAX ADPMC to serve the AD scientific community effectively and efficiently. The IU/JAX ADPMC will generate new AD modeling processes and pipelines, data resources, research results and models that will be swiftly shared through JAX's and Sage's proven dissemination pipelines and through the NIA- supported AD Centers, academic medical centers, research institutions and the pharmaceutical industry worldwide. Ultimately, this will accelerate the application of advances in animal models for the greatest possible medical benefit. The Specific Aims of the IU/JAX ADPMC are: 1. Maximize Human Datasets to Identify Putative Variants, Genes and Biomarkers for AD. 2. Generate and Characterize the Next Generation of Mouse Models of AD. 3. Validate the Next Generation of Mouse Models of AD and Develop a Preclinical Testing Pipeline. !

Abstract The APOE 4 allele is the most important genetic risk factor for late onset Alzheimer's disease (AD). APOE 4 may contribute to AD risk by altering inflammation, lipid homeostasis and/or amyloid clearance. Recent genetics studies have implicated multiple pathways in innate immunity in late-onset AD, including the triggering receptor expressed on the myeloid cells 2 (TREM2) gene. In contrast, exercise and physical activity (PA) produce anti-inflammatory changes in the periphery and neurogenic, angiogenic, and anti-inflammatory brain changes in animals. The mechanistic relationships between APOE, innate immunity, and AD, and the potential moderating influence of PA, remain to be established. Our published data on cognitively intact, healthy elders followed for 18 months indicate that sedentary 4 carriers demonstrate significantly lower fMRI activation, poorer episodic memory performance, smaller hippocampal volumes, and abnormal white matter diffusion compared to 4 carriers who engage in regular PA. Most importantly, these group differences were not observed between low and high PA non-carriers, suggesting that the neuroprotective effects of PA are especially potent for persons at genetic risk for AD. Additional preliminary studies demonstrate that TREM2+ innate immune cells play a direct role in regulating AD pathologies in both animal models of AD and potentially in human AD patients. For the proposed interdisciplinary project, our overall hypothesis posits that PA counteracts the negative inflammatory effects of the 4 allele, affecting TREM2 and other innate immune pathways implicated in AD, thereby reducing the risk of cognitive decline and AD in 4 carriers. This project has two specific aims. Aim 1 will recruit 150 cognitively intact, healthy elders (ages 65-80): 75 APOE 4 carriers ( 3/ 4) and 75 4 non-carriers ( 3/ 3). Participants will undergo state-of-the-art measurements of PA and fitness; structural and functional 3T MRI; amyloid PET imaging; CSF/blood biomarkers related to AD, inflammation, and exercise; and comprehensive memory/cognition testing on two occasions separated by 24- months. Aim 2 is analogous to the human project and will determine the impact of voluntary wheel running PA across age (3 months, 6 months, and 9 months) in novel transgenic mouse models of AD humanized for 3 and 4 and crossed with APPPS1 (APPPS1; APOE3/3 and APPPS1; APOE4/4) and control strains. Key indices include: TREM2+ cells, expression of proinflammatory markers, brain A?42, and spatial memory performance. Together, these complementary human and animal studies will provide key insights into potential mechanisms linking APOE genotype, exercise, inflammation, AD brain pathology, and cognition that could ultimately be targeted therapeutically.

PROJECT SUMMARY OVERALL COMPONENT Alzheimer's disease (AD) is a major cause of dementia, disability and death in the elderly. Despite recent advances in our understanding of basic biological mechanisms underlying AD, we do not yet know how to prevent AD or have an approved disease modifying intervention. Both are essential to slow or stop the growth in dementia prevalence. The National Alzheimer's Project Act (NAPA) seeks to prevent and effectively treat AD by 2025 through innovative research on etiology, early detection, and therapeutics. In support of NAPA's goals, one of the targeted areas of research identified at the NIA sponsored 2015 Alzheimer's Disease Research Summit was the development of the next generation of animal models of AD that will prove more predictive in preclinical studies and thus accelerate the drug testing pipeline. While our current animal models of AD have provided multiple novel insights into AD disease mechanisms, thus far they have not been successfully utilized to predict the effectiveness of therapies that have moved into AD clinical trials. The Indiana University (IU)/Jackson Laboratory (JAX) Alzheimer's Disease Precision Models Center (IU/JAX ADPMC) will leverage IU's strengths in neurodegenerative research including 25 years as an NIA-supported Alzheimer's Disease Center (ADC) and considerable expertise in preclinical drug testing with JAX's eight decades of expertise in mammalian genetics and disease modeling to develop, validate and disseminate new, precise animal models of Alzheimer's disease (AD). In addition, the IU/JAX ADMPC contains Sage Bionetworks to provide expertise in data organization and dissemination. The IU/JAX ADPMC brings together an international, multi-disciplinary team?including geneticists and genetics technology experts, quantitative and computational biologists, clinical experts in AD and neuroimaging, pharmacologists and world leaders in the development of precision animal models of disease?that possesses the collective ability to foresee disease modeling needs as they emerge on the international stage. This will allow the IU/JAX ADPMC to serve the AD scientific community effectively and efficiently. The IU/JAX ADPMC will generate new AD modeling processes and pipelines, data resources, research results and models that will be swiftly shared through JAX's and Sage's proven dissemination pipelines and through the NIA- supported AD Centers, academic medical centers, research institutions and the pharmaceutical industry worldwide. Ultimately, this will accelerate the application of advances in animal models for the greatest possible medical benefit. The Specific Aims of the IU/JAX ADPMC are: 1. Maximize Human Datasets to Identify Putative Variants, Genes and Biomarkers for AD. 2. Generate and Characterize the Next Generation of Mouse Models of AD. 3. Validate the Next Generation of Mouse Models of AD and Develop a Preclinical Testing Pipeline. !

PROJECT SUMMARY OVERALL COMPONENT Alzheimer's disease (AD) is a major cause of dementia, disability and death in the elderly. Despite recent advances in our understanding of basic biological mechanisms underlying AD, we do not yet know how to prevent AD or have an approved disease modifying intervention. Both are essential to slow or stop the growth in dementia prevalence. The National Alzheimer's Project Act (NAPA) seeks to prevent and effectively treat AD by 2025 through innovative research on etiology, early detection, and therapeutics. In support of NAPA's goals, one of the targeted areas of research identified at the NIA sponsored 2015 Alzheimer's Disease Research Summit was the development of the next generation of animal models of AD that will prove more predictive in preclinical studies and thus accelerate the drug testing pipeline. While our current animal models of AD have provided multiple novel insights into AD disease mechanisms, thus far they have not been successfully utilized to predict the effectiveness of therapies that have moved into AD clinical trials. The Indiana University (IU)/Jackson Laboratory (JAX) Alzheimer's Disease Precision Models Center (IU/JAX ADPMC) will leverage IU's strengths in neurodegenerative research including 25 years as an NIA-supported Alzheimer's Disease Center (ADC) and considerable expertise in preclinical drug testing with JAX's eight decades of expertise in mammalian genetics and disease modeling to develop, validate and disseminate new, precise animal models of Alzheimer's disease (AD). In addition, the IU/JAX ADMPC contains Sage Bionetworks to provide expertise in data organization and dissemination. The IU/JAX ADPMC brings together an international, multi-disciplinary team?including geneticists and genetics technology experts, quantitative and computational biologists, clinical experts in AD and neuroimaging, pharmacologists and world leaders in the development of precision animal models of disease?that possesses the collective ability to foresee disease modeling needs as they emerge on the international stage. This will allow the IU/JAX ADPMC to serve the AD scientific community effectively and efficiently. The IU/JAX ADPMC will generate new AD modeling processes and pipelines, data resources, research results and models that will be swiftly shared through JAX's and Sage's proven dissemination pipelines and through the NIA- supported AD Centers, academic medical centers, research institutions and the pharmaceutical industry worldwide. Ultimately, this will accelerate the application of advances in animal models for the greatest possible medical benefit. The Specific Aims of the IU/JAX ADPMC are: 1. Maximize Human Datasets to Identify Putative Variants, Genes and Biomarkers for AD. 2. Generate and Characterize the Next Generation of Mouse Models of AD. 3. Validate the Next Generation of Mouse Models of AD and Develop a Preclinical Testing Pipeline. !

Abstract The APOE ?4 allele is the most important genetic risk factor for late onset Alzheimer's disease (AD). APOE ?4 may contribute to AD risk by altering inflammation, lipid homeostasis and/or amyloid clearance. Recent genetics studies have implicated multiple pathways in innate immunity in late-onset AD, including the triggering receptor expressed on the myeloid cells 2 (TREM2) gene. In contrast, exercise and physical activity (PA) produce anti-inflammatory changes in the periphery and neurogenic, angiogenic, and anti-inflammatory brain changes in animals. The mechanistic relationships between APOE, innate immunity, and AD, and the potential moderating influence of PA, remain to be established. Our published data on cognitively intact, healthy elders followed for 18 months indicate that sedentary ?4 carriers demonstrate significantly lower fMRI activation, poorer episodic memory performance, smaller hippocampal volumes, and abnormal white matter diffusion compared to ?4 carriers who engage in regular PA. Most importantly, these group differences were not observed between low and high PA non-carriers, suggesting that the neuroprotective effects of PA are especially potent for persons at genetic risk for AD. Additional preliminary studies demonstrate that TREM2+ innate immune cells play a direct role in regulating AD pathologies in both animal models of AD and potentially in human AD patients. For the proposed interdisciplinary project, our overall hypothesis posits that PA counteracts the negative inflammatory effects of the ?4 allele, affecting TREM2 and other innate immune pathways implicated in AD, thereby reducing the risk of cognitive decline and AD in ?4 carriers. This project has two specific aims. Aim 1 will recruit 150 cognitively intact, healthy elders (ages 65-80): 75 APOE ?4 carriers (?3/?4) and 75 ?4 non-carriers (?3/?3). Participants will undergo state-of-the-art measurements of PA and fitness; structural and functional 3T MRI; amyloid PET imaging; CSF/blood biomarkers related to AD, inflammation, and exercise; and comprehensive memory/cognition testing on two occasions separated by 24- months. Aim 2 is analogous to the human project and will determine the impact of voluntary wheel running PA across age (3 months, 6 months, and 9 months) in novel transgenic mouse models of AD humanized for ?3 and ?4 and crossed with APPPS1 (APPPS1; APOE3/3 and APPPS1; APOE4/4) and control strains. Key indices include: TREM2+ cells, expression of proinflammatory markers, brain A?42, and spatial memory performance. Together, these complementary human and animal studies will provide key insights into potential mechanisms linking APOE genotype, exercise, inflammation, AD brain pathology, and cognition that could ultimately be targeted therapeutically.

PROJECT SUMMARY OVERALL COMPONENT Alzheimer's disease (AD) is a major cause of dementia, disability and death in the elderly. Despite recent advances in our understanding of basic biological mechanisms underlying AD, we do not yet know how to prevent AD or have an approved disease modifying intervention. Both are essential to slow or stop the growth in dementia prevalence. The National Alzheimer's Project Act (NAPA) seeks to prevent and effectively treat AD by 2025 through innovative research on etiology, early detection, and therapeutics. In support of NAPA's goals, one of the targeted areas of research identified at the NIA sponsored 2015 Alzheimer's Disease Research Summit was the development of the next generation of animal models of AD that will prove more predictive in preclinical studies and thus accelerate the drug testing pipeline. While our current animal models of AD have provided multiple novel insights into AD disease mechanisms, thus far they have not been successfully utilized to predict the effectiveness of therapies that have moved into AD clinical trials. The Indiana University (IU)/Jackson Laboratory (JAX) Alzheimer's Disease Precision Models Center (IU/JAX ADPMC) will leverage IU's strengths in neurodegenerative research including 25 years as an NIA-supported Alzheimer's Disease Center (ADC) and considerable expertise in preclinical drug testing with JAX's eight decades of expertise in mammalian genetics and disease modeling to develop, validate and disseminate new, precise animal models of Alzheimer's disease (AD). In addition, the IU/JAX ADMPC contains Sage Bionetworks to provide expertise in data organization and dissemination. The IU/JAX ADPMC brings together an international, multi-disciplinary team?including geneticists and genetics technology experts, quantitative and computational biologists, clinical experts in AD and neuroimaging, pharmacologists and world leaders in the development of precision animal models of disease?that possesses the collective ability to foresee disease modeling needs as they emerge on the international stage. This will allow the IU/JAX ADPMC to serve the AD scientific community effectively and efficiently. The IU/JAX ADPMC will generate new AD modeling processes and pipelines, data resources, research results and models that will be swiftly shared through JAX's and Sage's proven dissemination pipelines and through the NIA- supported AD Centers, academic medical centers, research institutions and the pharmaceutical industry worldwide. Ultimately, this will accelerate the application of advances in animal models for the greatest possible medical benefit. The Specific Aims of the IU/JAX ADPMC are: 1. Maximize Human Datasets to Identify Putative Variants, Genes and Biomarkers for AD. 2. Generate and Characterize the Next Generation of Mouse Models of AD. 3. Validate the Next Generation of Mouse Models of AD and Develop a Preclinical Testing Pipeline. !

PROJECT SUMMARY OVERALL COMPONENT Alzheimer's disease (AD) is a major cause of dementia, disability and death in the elderly. Despite recent advances in our understanding of basic biological mechanisms underlying AD, we do not yet know how to prevent AD or have an approved disease modifying intervention. Both are essential to slow or stop the growth in dementia prevalence. The National Alzheimer's Project Act (NAPA) seeks to prevent and effectively treat AD by 2025 through innovative research on etiology, early detection, and therapeutics. In support of NAPA's goals, one of the targeted areas of research identified at the NIA sponsored 2015 Alzheimer's Disease Research Summit was the development of the next generation of animal models of AD that will prove more predictive in preclinical studies and thus accelerate the drug testing pipeline. While our current animal models of AD have provided multiple novel insights into AD disease mechanisms, thus far they have not been successfully utilized to predict the effectiveness of therapies that have moved into AD clinical trials. The Indiana University (IU)/Jackson Laboratory (JAX) Alzheimer's Disease Precision Models Center (IU/JAX ADPMC) will leverage IU's strengths in neurodegenerative research including 25 years as an NIA-supported Alzheimer's Disease Center (ADC) and considerable expertise in preclinical drug testing with JAX's eight decades of expertise in mammalian genetics and disease modeling to develop, validate and disseminate new, precise animal models of Alzheimer's disease (AD). In addition, the IU/JAX ADMPC contains Sage Bionetworks to provide expertise in data organization and dissemination. The IU/JAX ADPMC brings together an international, multi-disciplinary team?including geneticists and genetics technology experts, quantitative and computational biologists, clinical experts in AD and neuroimaging, pharmacologists and world leaders in the development of precision animal models of disease?that possesses the collective ability to foresee disease modeling needs as they emerge on the international stage. This will allow the IU/JAX ADPMC to serve the AD scientific community effectively and efficiently. The IU/JAX ADPMC will generate new AD modeling processes and pipelines, data resources, research results and models that will be swiftly shared through JAX's and Sage's proven dissemination pipelines and through the NIA- supported AD Centers, academic medical centers, research institutions and the pharmaceutical industry worldwide. Ultimately, this will accelerate the application of advances in animal models for the greatest possible medical benefit. The Specific Aims of the IU/JAX ADPMC are: 1. Maximize Human Datasets to Identify Putative Variants, Genes and Biomarkers for AD. 2. Generate and Characterize the Next Generation of Mouse Models of AD. 3. Validate the Next Generation of Mouse Models of AD and Develop a Preclinical Testing Pipeline. !

PROJECT SUMMARY FOR FUNDED PARENT AWARD (OVERALL) Alzheimer's disease (AD) is a major cause of dementia, disability and death in the elderly. Despite recent advances in our understanding of basic biological mechanisms underlying AD, we do not yet know how to prevent AD or have an approved disease modifying intervention. Both are essential to slow or stop the growth in dementia prevalence. The National Alzheimer's Project Act (NAPA) seeks to prevent and effectively treat AD by 2025 through innovative research on etiology, early detection, and therapeutics. In support of NAPA's goals, one of the targeted areas of research identified at the NIA sponsored 2015 Alzheimer's Disease Research Summit was the development of the next generation of animal models of AD that will prove more predictive in preclinical studies and thus accelerate the drug testing pipeline. While our current animal models of AD have provided multiple novel insights into AD disease mechanisms, thus far they have not been successfully utilized to predict the effectiveness of therapies that have moved into AD clinical trials. The Indiana University (IU)/Jackson Laboratory (JAX) Alzheimer's Disease Precision Models Center (IU/JAX ADPMC) will leverage IU's strengths in neurodegenerative research including 25 years as an NIA-supported Alzheimer's Disease Center (ADC) and considerable expertise in preclinical drug testing with JAX's eight decades of expertise in mammalian genetics and disease modeling to develop, validate and disseminate new, precise animal models of Alzheimer's disease (AD). In addition, the IU/JAX ADMPC contains Sage Bionetworks to provide expertise in data organization and dissemination. The IU/JAX ADPMC brings together an international, multi-disciplinary team?including geneticists and genetics technology experts, quantitative and computational biologists, clinical experts in AD and neuroimaging, pharmacologists and world leaders in the development of precision animal models of disease?that possesses the collective ability to foresee disease modeling needs as they emerge on the international stage. This will allow the IU/JAX ADPMC to serve the AD scientific community effectively and efficiently. The IU/JAX ADPMC will generate new AD modeling processes and pipelines, data resources, research results and models that will be swiftly shared through JAX's and Sage's proven dissemination pipelines and through the NIA-supported AD Centers, academic medical centers, research institutions and the pharmaceutical industry worldwide. Ultimately, this will accelerate the application of advances in animal models for the greatest possible medical benefit. The Specific Aims of the IU/JAX ADPMC are: 1. Maximize Human Datasets to Identify Putative Variants, Genes and Biomarkers for AD. 2. Generate and Characterize the Next Generation of Mouse Models of AD. 3. Validate the Next Generation of Mouse Models of AD and Develop a Preclinical Testing Pipeline.

PROJECT SUMMARY OVERALL COMPONENT Alzheimer's disease (AD) is a major cause of dementia, disability and death in the elderly. Despite recent advances in our understanding of basic biological mechanisms underlying AD, we do not yet know how to prevent AD or have an approved disease modifying intervention. Both are essential to slow or stop the growth in dementia prevalence. The National Alzheimer's Project Act (NAPA) seeks to prevent and effectively treat AD by 2025 through innovative research on etiology, early detection, and therapeutics. In support of NAPA's goals, one of the targeted areas of research identified at the NIA sponsored 2015 Alzheimer's Disease Research Summit was the development of the next generation of animal models of AD that will prove more predictive in preclinical studies and thus accelerate the drug testing pipeline. While our current animal models of AD have provided multiple novel insights into AD disease mechanisms, thus far they have not been successfully utilized to predict the effectiveness of therapies that have moved into AD clinical trials. The Indiana University (IU)/Jackson Laboratory (JAX) Alzheimer's Disease Precision Models Center (IU/JAX ADPMC) will leverage IU's strengths in neurodegenerative research including 25 years as an NIA-supported Alzheimer's Disease Center (ADC) and considerable expertise in preclinical drug testing with JAX's eight decades of expertise in mammalian genetics and disease modeling to develop, validate and disseminate new, precise animal models of Alzheimer's disease (AD). In addition, the IU/JAX ADMPC contains Sage Bionetworks to provide expertise in data organization and dissemination. The IU/JAX ADPMC brings together an international, multi-disciplinary team?including geneticists and genetics technology experts, quantitative and computational biologists, clinical experts in AD and neuroimaging, pharmacologists and world leaders in the development of precision animal models of disease?that possesses the collective ability to foresee disease modeling needs as they emerge on the international stage. This will allow the IU/JAX ADPMC to serve the AD scientific community effectively and efficiently. The IU/JAX ADPMC will generate new AD modeling processes and pipelines, data resources, research results and models that will be swiftly shared through JAX's and Sage's proven dissemination pipelines and through the NIA- supported AD Centers, academic medical centers, research institutions and the pharmaceutical industry worldwide. Ultimately, this will accelerate the application of advances in animal models for the greatest possible medical benefit. The Specific Aims of the IU/JAX ADPMC are: 1. Maximize Human Datasets to Identify Putative Variants, Genes and Biomarkers for AD. 2. Generate and Characterize the Next Generation of Mouse Models of AD. 3. Validate the Next Generation of Mouse Models of AD and Develop a Preclinical Testing Pipeline. !

PROJECT SUMMARY FOR FUNDED PARENT AWARD (OVERALL) Alzheimer's disease (AD) is a major cause of dementia, disability and death in the elderly. Despite recent advances in our understanding of basic biological mechanisms underlying AD, we do not yet know how to prevent AD or have an approved disease modifying intervention. Both are essential to slow or stop the growth in dementia prevalence. The National Alzheimer's Project Act (NAPA) seeks to prevent and effectively treat AD by 2025 through innovative research on etiology, early detection, and therapeutics. In support of NAPA's goals, one of the targeted areas of research identified at the NIA sponsored 2015 Alzheimer's Disease Research Summit was the development of the next generation of animal models of AD that will prove more predictive in preclinical studies and thus accelerate the drug testing pipeline. While our current animal models of AD have provided multiple novel insights into AD disease mechanisms, thus far they have not been successfully utilized to predict the effectiveness of therapies that have moved into AD clinical trials. The Indiana University (IU)/Jackson Laboratory (JAX) Alzheimer's Disease Precision Models Center (IU/JAX ADPMC) will leverage IU's strengths in neurodegenerative research including 25 years as an NIA-supported Alzheimer's Disease Center (ADC) and considerable expertise in preclinical drug testing with JAX's eight decades of expertise in mammalian genetics and disease modeling to develop, validate and disseminate new, precise animal models of Alzheimer's disease (AD). In addition, the IU/JAX ADMPC contains Sage Bionetworks to provide expertise in data organization and dissemination. The IU/JAX ADPMC brings together an international, multi-disciplinary team?including geneticists and genetics technology experts, quantitative and computational biologists, clinical experts in AD and neuroimaging, pharmacologists and world leaders in the development of precision animal models of disease?that possesses the collective ability to foresee disease modeling needs as they emerge on the international stage. This will allow the IU/JAX ADPMC to serve the AD scientific community effectively and efficiently. The IU/JAX ADPMC will generate new AD modeling processes and pipelines, data resources, research results and models that will be swiftly shared through JAX's and Sage's proven dissemination pipelines and through the NIA-supported AD Centers, academic medical centers, research institutions and the pharmaceutical industry worldwide. Ultimately, this will accelerate the application of advances in animal models for the greatest possible medical benefit. The Specific Aims of the IU/JAX ADPMC are: 1. Maximize Human Datasets to Identify Putative Variants, Genes and Biomarkers for AD. 2. Generate and Characterize the Next Generation of Mouse Models of AD. 3. Validate the Next Generation of Mouse Models of AD and Develop a Preclinical Testing Pipeline.

PROJECT SUMMARY FOR FUNDED PARENT AWARD (OVERALL) Alzheimer's disease (AD) is a major cause of dementia, disability and death in the elderly. Despite recent advances in our understanding of basic biological mechanisms underlying AD, we do not yet know how to prevent AD or have an approved disease modifying intervention. Both are essential to slow or stop the growth in dementia prevalence. The National Alzheimer's Project Act (NAPA) seeks to prevent and effectively treat AD by 2025 through innovative research on etiology, early detection, and therapeutics. In support of NAPA's goals, one of the targeted areas of research identified at the NIA sponsored 2015 Alzheimer's Disease Research Summit was the development of the next generation of animal models of AD that will prove more predictive in preclinical studies and thus accelerate the drug testing pipeline. While our current animal models of AD have provided multiple novel insights into AD disease mechanisms, thus far they have not been successfully utilized to predict the effectiveness of therapies that have moved into AD clinical trials. The Indiana University (IU)/Jackson Laboratory (JAX) Alzheimer's Disease Precision Models Center (IU/JAX ADPMC) will leverage IU's strengths in neurodegenerative research including 25 years as an NIA-supported Alzheimer's Disease Center (ADC) and considerable expertise in preclinical drug testing with JAX's eight decades of expertise in mammalian genetics and disease modeling to develop, validate and disseminate new, precise animal models of Alzheimer's disease (AD). In addition, the IU/JAX ADMPC contains Sage Bionetworks to provide expertise in data organization and dissemination. The IU/JAX ADPMC brings together an international, multi-disciplinary team?including geneticists and genetics technology experts, quantitative and computational biologists, clinical experts in AD and neuroimaging, pharmacologists and world leaders in the development of precision animal models of disease?that possesses the collective ability to foresee disease modeling needs as they emerge on the international stage. This will allow the IU/JAX ADPMC to serve the AD scientific community effectively and efficiently. The IU/JAX ADPMC will generate new AD modeling processes and pipelines, data resources, research results and models that will be swiftly shared through JAX's and Sage's proven dissemination pipelines and through the NIA-supported AD Centers, academic medical centers, research institutions and the pharmaceutical industry worldwide. Ultimately, this will accelerate the application of advances in animal models for the greatest possible medical benefit. The Specific Aims of the IU/JAX ADPMC are: 1. Maximize Human Datasets to Identify Putative Variants, Genes and Biomarkers for AD. 2. Generate and Characterize the Next Generation of Mouse Models of AD. 3. Validate the Next Generation of Mouse Models of AD and Develop a Preclinical Testing Pipeline.

PROJECT SUMMARY FOR FUNDED PARENT AWARD (OVERALL) Alzheimer's disease (AD) is a major cause of dementia, disability and death in the elderly. Despite recent advances in our understanding of basic biological mechanisms underlying AD, we do not yet know how to prevent AD or have an approved disease modifying intervention. Both are essential to slow or stop the growth in dementia prevalence. The National Alzheimer's Project Act (NAPA) seeks to prevent and effectively treat AD by 2025 through innovative research on etiology, early detection, and therapeutics. In support of NAPA's goals, one of the targeted areas of research identified at the NIA sponsored 2015 Alzheimer's Disease Research Summit was the development of the next generation of animal models of AD that will prove more predictive in preclinical studies and thus accelerate the drug testing pipeline. While our current animal models of AD have provided multiple novel insights into AD disease mechanisms, thus far they have not been successfully utilized to predict the effectiveness of therapies that have moved into AD clinical trials. The Indiana University (IU)/Jackson Laboratory (JAX) Alzheimer's Disease Precision Models Center (IU/JAX ADPMC) will leverage IU's strengths in neurodegenerative research including 25 years as an NIA-supported Alzheimer's Disease Center (ADC) and considerable expertise in preclinical drug testing with JAX's eight decades of expertise in mammalian genetics and disease modeling to develop, validate and disseminate new, precise animal models of Alzheimer's disease (AD). In addition, the IU/JAX ADMPC contains Sage Bionetworks to provide expertise in data organization and dissemination. The IU/JAX ADPMC brings together an international, multi-disciplinary team?including geneticists and genetics technology experts, quantitative and computational biologists, clinical experts in AD and neuroimaging, pharmacologists and world leaders in the development of precision animal models of disease?that possesses the collective ability to foresee disease modeling needs as they emerge on the international stage. This will allow the IU/JAX ADPMC to serve the AD scientific community effectively and efficiently. The IU/JAX ADPMC will generate new AD modeling processes and pipelines, data resources, research results and models that will be swiftly shared through JAX's and Sage's proven dissemination pipelines and through the NIA-supported AD Centers, academic medical centers, research institutions and the pharmaceutical industry worldwide. Ultimately, this will accelerate the application of advances in animal models for the greatest possible medical benefit. The Specific Aims of the IU/JAX ADPMC are: 1. Maximize Human Datasets to Identify Putative Variants, Genes and Biomarkers for AD. 2. Generate and Characterize the Next Generation of Mouse Models of AD. 3. Validate the Next Generation of Mouse Models of AD and Develop a Preclinical Testing Pipeline.

PROJECT SUMMARY FOR FUNDED PARENT AWARD (OVERALL) Alzheimer's disease (AD) is a major cause of dementia, disability and death in the elderly. Despite recent advances in our understanding of basic biological mechanisms underlying AD, we do not yet know how to prevent AD or have an approved disease modifying intervention. Both are essential to slow or stop the growth in dementia prevalence. The National Alzheimer's Project Act (NAPA) seeks to prevent and effectively treat AD by 2025 through innovative research on etiology, early detection, and therapeutics. In support of NAPA's goals, one of the targeted areas of research identified at the NIA sponsored 2015 Alzheimer's Disease Research Summit was the development of the next generation of animal models of AD that will prove more predictive in preclinical studies and thus accelerate the drug testing pipeline. While our current animal models of AD have provided multiple novel insights into AD disease mechanisms, thus far they have not been successfully utilized to predict the effectiveness of therapies that have moved into AD clinical trials. The Indiana University (IU)/Jackson Laboratory (JAX) Alzheimer's Disease Precision Models Center (IU/JAX ADPMC) will leverage IU's strengths in neurodegenerative research including 25 years as an NIA-supported Alzheimer's Disease Center (ADC) and considerable expertise in preclinical drug testing with JAX's eight decades of expertise in mammalian genetics and disease modeling to develop, validate and disseminate new, precise animal models of Alzheimer's disease (AD). In addition, the IU/JAX ADMPC contains Sage Bionetworks to provide expertise in data organization and dissemination. The IU/JAX ADPMC brings together an international, multi-disciplinary team?including geneticists and genetics technology experts, quantitative and computational biologists, clinical experts in AD and neuroimaging, pharmacologists and world leaders in the development of precision animal models of disease?that possesses the collective ability to foresee disease modeling needs as they emerge on the international stage. This will allow the IU/JAX ADPMC to serve the AD scientific community effectively and efficiently. The IU/JAX ADPMC will generate new AD modeling processes and pipelines, data resources, research results and models that will be swiftly shared through JAX's and Sage's proven dissemination pipelines and through the NIA-supported AD Centers, academic medical centers, research institutions and the pharmaceutical industry worldwide. Ultimately, this will accelerate the application of advances in animal models for the greatest possible medical benefit. The Specific Aims of the IU/JAX ADPMC are: 1. Maximize Human Datasets to Identify Putative Variants, Genes and Biomarkers for AD. 2. Generate and Characterize the Next Generation of Mouse Models of AD. 3. Validate the Next Generation of Mouse Models of AD and Develop a Preclinical Testing Pipeline.

PROJECT SUMMARY FOR FUNDED PARENT AWARD (OVERALL) Alzheimer's disease (AD) is a major cause of dementia, disability and death in the elderly. Despite recent advances in our understanding of basic biological mechanisms underlying AD, we do not yet know how to prevent AD or have an approved disease modifying intervention. Both are essential to slow or stop the growth in dementia prevalence. The National Alzheimer's Project Act (NAPA) seeks to prevent and effectively treat AD by 2025 through innovative research on etiology, early detection, and therapeutics. In support of NAPA's goals, one of the targeted areas of research identified at the NIA sponsored 2015 Alzheimer's Disease Research Summit was the development of the next generation of animal models of AD that will prove more predictive in preclinical studies and thus accelerate the drug testing pipeline. While our current animal models of AD have provided multiple novel insights into AD disease mechanisms, thus far they have not been successfully utilized to predict the effectiveness of therapies that have moved into AD clinical trials. The Indiana University (IU)/Jackson Laboratory (JAX) Alzheimer's Disease Precision Models Center (IU/JAX ADPMC) will leverage IU's strengths in neurodegenerative research including 25 years as an NIA-supported Alzheimer's Disease Center (ADC) and considerable expertise in preclinical drug testing with JAX's eight decades of expertise in mammalian genetics and disease modeling to develop, validate and disseminate new, precise animal models of Alzheimer's disease (AD). In addition, the IU/JAX ADMPC contains Sage Bionetworks to provide expertise in data organization and dissemination. The IU/JAX ADPMC brings together an international, multi-disciplinary team?including geneticists and genetics technology experts, quantitative and computational biologists, clinical experts in AD and neuroimaging, pharmacologists and world leaders in the development of precision animal models of disease?that possesses the collective ability to foresee disease modeling needs as they emerge on the international stage. This will allow the IU/JAX ADPMC to serve the AD scientific community effectively and efficiently. The IU/JAX ADPMC will generate new AD modeling processes and pipelines, data resources, research results and models that will be swiftly shared through JAX's and Sage's proven dissemination pipelines and through the NIA-supported AD Centers, academic medical centers, research institutions and the pharmaceutical industry worldwide. Ultimately, this will accelerate the application of advances in animal models for the greatest possible medical benefit. The Specific Aims of the IU/JAX ADPMC are: 1. Maximize Human Datasets to Identify Putative Variants, Genes and Biomarkers for AD. 2. Generate and Characterize the Next Generation of Mouse Models of AD. 3. Validate the Next Generation of Mouse Models of AD and Develop a Preclinical Testing Pipeline.

The strategic goal of the Indiana University School of Medicine Alzheimer?s Disease Drug Discovery (ADDD) Center is to integrate sophisticated capability for early drug discovery and contribute to a broader study of emerging Alzheimer?s disease (AD) target hypotheses (beyond Ab) with the goal of generating new classes of potential therapeutics. Specifically, the ADDD CENTER will establish itself as a strategic and operational partner for the NIA AMP-AD and MODEL-AD initiatives. By design, this will provide drug discovery capability to bridge the foundational work in target discovery (AMP-AD) with newly discovered lead molecules characterized in AD animal models based on human pathology, genetics and translational biomarkers (MODEL-AD). To accomplish the mission of the ADDD CENTER, we have assembled a world-class team of scientists and capabilities from Indiana University, Purdue University, and the Indiana Clinical and Translational Sciences Institute (CTSI). This team has considerable expertise in Alzheimer?s disease biology, CNS pharmaceutical drug discovery and development, and scientific excellence in the core discovery technologies that will drive the ADDD CENTER?s contributions and deliverables. A key advantage and differentiated strength of the proposed ADDD CENTER is the primary scientific coordination and administration through the Indiana University School of Medicine. Specifically, this concentrates a strong and long-standing commitment to neurodegenerative research through co-presence with the NIA-supported Indiana Alzheimer?s Disease Center under the direction of Dr. Andy Saykin, the MODEL-AD consortium under the direction of Dr. Bruce Lamb, and the Longitudinal Early Onset Alzheimer?s Disease (LEAD) study under the direction of Dr. Liana Apostolova. The Specific Aims of the ADDD CENTER are: 1. Create a dynamic portfolio of well-characterized AD drug discovery targets representing novel intervention hypotheses that capitalize on significant investments in basic research and emerging disease understanding. 2. Perform advanced pre-clinical target validation and enablement studies to prioritize the best opportunities for therapeutic discovery. 3. Create high-quality and well-characterized lead molecules for prioritized targets that meet rigorous milestone criteria and are valued opportunities for further translational investment. 4. Create a robust and flexible data sharing platform to enable global researchers with data and target enablement packages to expand on the work of the ADDD CENTER.

The overall goal of the program is to provide multidisciplinary training in Alzheimer?s disease and AD-related dementias (AD/ADRD) to predoctoral and postdoctoral trainees, producing a critically-needed translational workforce for developing effective treatments. The Indiana University School of Medicine (IUSM) is nationally recognized for AD/ADRD research. IUSM-associated centers include the Indiana Alzheimer?s Disease Research Center, the National Cell Repository for Alzheimer?s Disease, the Indiana Center for Neuroimaging, the Regenstrief Institute and the Center for Aging Research. Importantly, the NIA recently funded the MODEL-AD Center, the Alzheimer?s Disease Drug Discovery Center, and the IUSM-directed Longitudinal Early-Onset Alzheimer?s Disease Study. IUSM has invested significantly in state-of-the-art facilities, instrumentation, and the hiring of 10 new AD/ADRD faculty over the past 4 years. This T32 application has enlisted 25 participating faculty with a broad range of expertise including data science, non-invasive imaging, the development and study of animal models, genomics, epidemiology, drug development and discovery, and clinical studies and trials. These faculty are exceptional mentors and are funded at $42M total ($1.69M/faculty annually). This application requests funding for 4 pre- and 4 post-doctoral trainees. We will recruit trainees with diverse backgrounds and train them in AD/ADRD research with contemporary approaches and methodologies using state-of-the-art instrumentation. The predoctoral trainees are graduate students in the Medical Neuroscience Graduate Program, recruited from the IUSM Indiana Biomedical Gateway graduate program and the MD/PhD program. Targeted recruiting of postdoctoral trainees with diverse backgrounds is achieved informally, through our website, and at national and regional meetings. Enrollment in the MedNeuro Graduate Program has doubled since 2017 and the number of postdoctoral fellows with participating faculty has increased from 4 to 27. The predoctoral training program consists of foundational course work in neuroscience, neurodegenerative diseases, and data science along with elective studies. Postdoctoral trainees are offered didactic training in data science. The NIA-sponsored centers and resources at IUSM allow trainees to work on new disease models, advanced drug-discovery technologies, and in clinical settings. This positions trainees to conduct innovative basic, translational, and clinical AD/ADRD research.

We are responding to NOT-AG-18-049 ?Collaborative Studies on AD/ADRD? by establishing a collaborative effort to significantly expand the modeling capacity of the MODEL-AD Center. MODEL-AD was established by the NIA to create, rigorously characterize and ensure the rapid distribution of the next generation of animal models of Late Onset AD. Critical barriers to progress in making these next generation models are technique limitations that have put restrictions on the size of the human genomic context incorporated into each of the new modified alleles in the MODEL-AD mouse lines. Our group at the University of Minnesota (UMN) has developed Gene Replacement (GR) technologies that allow us to routinely replace mouse genes with their full human orthologs up to several hundred kb in size. We used this technology to generate a matched set of Microtubule Associated Protein Tau Gene- Replacement (MAPT-GR) lines of mice in which we replaced the full mouse Mapt genomic coding and regulatory region (156,547bp) with full human MAPT genomic sequences (190,081bp). We have confirmed that mice homozygous for this MAPT-GR allele express human tau at endogenous levels, and that all expected splice variants are found in the appropriate tissues and in ratios expected for the fully functional human MAPT gene. This model set now includes two wt control lines (H1 or H2 MAPT haplotype) and a growing number of experimental lines that precisely match the H1 wt control line except for the pathogenic variant that we specifically introduce into that haplotype. The first of these lines are currently being further characterized by MODEL-AD and are now available to the AD research community without restriction (JAX). Our specific aims for this collaboration are to: 1. Generate Gene-Replacement (GR) sets of mouse lines in which genes involved in the etiology of AD have been replaced by their full human homologs. We are proposing to develop 10 model sets for this collaboration (>20 total lines). 2. Characterize matched sets of GR lines using the established MODEL-AD methods and distribute without restriction. The most translationally relevant alleles will be incorporated into the current MODEL-AD ?base model?. These GR lines will allow us and other AD researchers to evaluate the molecular impact of pathogenic mutations and risk variants within the context of the full human gene sequence in which they occur in patients. These mouse lines will contain all potential human therapeutic targets for each gene, ranging from the full genomic DNA sequences to all RNA transcription and protein products that they encode. Because the genomic sequences of these matched sets will differ only at sequences specifically changed in each line, any significant molecular differences between these lines can confidently be attributed to the risk variant in the experimental lines, and any therapeutic agents found to effectively correct these dysfunctions could be expected to have direct therapeutic value to patients.

PROJECT SUMMARY Alzheimer?s disease (AD) and other neurodegenerative diseases are typified by a robust microglial-mediated immune response. Genetic studies have demonstrated that many of the genes that confer altered risk for AD are those involved in the innate immune response and are expressed primarily in microglia, including phospholipase C gamma 2 (PLCG2). PLCG2 is a critical signaling element for a variety of immune receptors and is a key regulatory hub gene for immune signaling. The primary objective of this proposal is to determine the role of PLCG2 in AD pathogenesis. GWAS studies have demonstrated that the PLCG2 P522R variant is associated with reduced AD risk. Our laboratory has identified a novel SNP (rs617749044) associated with elevated AD risk encoding the PLCG2 M28L variant. The overall objectives in this application are to elucidate the effect of these PLCG2 variants on AD pathogenesis using rodent models of AD and dissect the molecular mechanisms by which PLCG2 variants alter microglia function. The hypotheses are that the M28L variant is a loss of function allele, and conversely the P522R is protective with respect to AD pathogenesis in our murine models. The experiments proposed in this application are entirely novel and allow, for the first time, a unique, comprehensive analysis of an AD risk gene whose genetic variants confer both protection and risk for AD. Preliminary data generated by the applicant suggests that in a rodent model of AD, the M28L variant had accelerated and exacerbated disease related pathology and conversely the P522R variant appeared to attenuate disease severity and progression. The hypotheses will be tested by pursuing three specific aims: 1) Determine AD-related phenotypes altered by loss and gain of function PLCG2 variants in an amyloidogenic model of AD; 2) Identify molecular signatures and pathways in microglia that are associated with protective or risk PLCG2 variants in an amyloidogenic model of AD; and 3) Evaluate the mechanisms through which PLCG2 variants affect intracellular signaling in microglia. These studies are essential prerequisites for the development of PLCG2-directed therapeutics.

Project Summary Overall: The strategic goal of the Indiana University School of Medicine (IUSM) Alzheimer?s Disease Drug Discovery (ADDD) Center is to integrate sophisticated capability for early drug discovery and contribute to a broader study of emerging Alzheimer?s Disease target hypotheses with the goal of generating new classes of potential therapeutics. The ADDD CENTER will establish itself as a strategic and operational partner for the NIA AMP-AD and MODEL-AD initiatives. By design, this will provide drug discovery capability to bridge the work in target discovery (AMP-AD) with newly discovered lead molecules characterized in AD animal models based on human pathology, genetics and translational biomarkers (MODEL-AD). To accomplish the mission of the ADDD CENTER, we have assembled a team of scientists from Indiana University, Purdue University, and the Indiana Clinical and Translational Sciences Institute (CTSI). This team has considerable expertise in Alzheimer?s disease biology, CNS pharmaceutical drug discovery and development, and scientific excellence in the core discovery technologies that will drive the ADDD CENTER?s contributions and deliverables. A key advantage and differentiated strength of the proposed ADDD CENTER is the primary scientific coordination and administration through IUSM. Specific Aims are: 1. Create a portfolio of characterized AD drug discovery targets representing novel intervention hypotheses that capitalizes on significant investments in basic research and emerging disease understanding. 2. Perform advanced pre-clinical target validation and enablement studies to prioritize the best opportunities for therapeutic discovery. 3. Create quality and well characterized Lead molecules for targets that meet milestone criteria and are opportunities for further translational investment. 4. Create a flexible data sharing platform to enable global researchers with data and Target Enablement Packages to expand on the work of the ADDD CENTER.