Document text
Principal Investigator: JEFFERY W KELLY
Organization: SCRIPPS RESEARCH INSTITUTE, THE
Fiscal Year: 2024
Award: $898,395
Funding agency: National Institute on Aging
A common molecular phenotype exists for most neurodegenerative diseases, including protein and/or protein-
RNA aggregation, lipid level perturbations, mitochondrial dysfunction, lysosomal dysfunction, and neuro-
inflammation. The literature provides evidence that this pathogenic signature of Alzheimer’s disease and
related dementias can be normalized by genetic and pharmacologic activation of lysosomal flux–one
mechanism to do this is called macroautophagy. To generate mechanistically diverse lysosomal flux activators,
we screened 940,000 small molecules by a cell-based phenotypic screen to identify 108 validated small
molecule hits that hastened lipid droplet clearance. Most known lysosomal flux activators function through
inhibition of mTOR, which suppresses the immune system, putting the already vulnerable elderly population at
higher risk for infectious disease. In this proposal we seek mTOR-independent lysosomal flux activators. In
Aim 1 we employ traditional and novel assays to identify the targets of our hits, as well as their mechanisms of
action. Whether these compounds induce macroautophagy (a cell component recycling pathway) or a
specialized form of autophagy will be revealed by the proposed assays comprising Aim 1. We will also explore
whether lysosomal flux activation occurs by other mechanisms, such as transcriptional reprogramming, or by a
novel mechanism. The data generated in Aim 1 will guide prioritization of lysosomal flux activators that are best
suited for ameliorating Alzheimer’s disease and related dementias. Aim 2 activities will scrutinize lysosomal
flux activator dosing efficacy and dosing regimens in induced pluripotent stem cell-derived neurons, astrocytes
and glial cells from hereditary Alzheimer’s disease patients and in brain organoids derived from these cells, as
well as in brain cells and organoids lacking these mutations. Because autophagy recycles proteins, nucleic
acids, oligosaccharides and lipids into their building blocks for reuse, it is a potential risk that enhancing
lysosomal flux could degrade critical cellular components and therefore lead to on-mechanism toxicity.
Organoids are well-suited for testing lysosomal flux activator multidosing regimens that avoid inducing
cytotoxicity or cellular stress, while also normalizing the pathogenic Alzheimer’s disease-associated
phenotypes present. We will use mass spectrometry-based proteomics and metabolomics / lipidomics to
analyze the organoids after treatment by 20-30 prioritized lysosomal flux activators to learn how to dose so as
to avoid cytotoxicity while normalizing the Alzheimer’s disease-relevant pathobiological phenotypes. Besides
carrying out multiple biological replicates and appropriate statistical analyses, another way to ensure
reproducibility and rigor is that we have distributed our lysosomal flux activators to multiple outside
collaborators to carry out independent cellular assays and and efficacy assessments in models of Alzheimer’s
disease. We hope to deliver a validated, mechanistically diverse, de-risked set of lysosomal flux activators as
candidates to treat Alzheimer’s disease.
Terms: <AD dementia><AD model><AD related dementia><ADRD><Activator Appliances><Activator Orthodontic Appliances><Address><Affinity Chromatography><After Care><After-Treatment><Aftercare><Alkynes><Alzheimer Type Dementia><Alzheimer disease dementia><Alzheimer sclerosis><Alzheimer syndrome><Alzheimer's><Alzheimer's Disease><Alzheimer's and related dementias><Alzheimer's disease and related dementia><Alzheimer's disease and related disorders><Alzheimer's disease model><Alzheimer's disease or a related dementia><Alzheimer's disease or a related disorder><Alzheimer's disease or related dementia><Alzheimer's disease patient><Alzheimer's disease related dementia><Alzheimer's patient><Alzheimers Dementia><American><Assay><Astrocytes><Astrocytus><Astroglia><Autophagocytosis><Bioassay><Biological><Biological Assay><Biology><Biotin><Brain><Brain Nervous System><CNS Degenerative Diseases><CNS Degenerative Disorders><CRISPR approach><CRISPR based approach><CRISPR method><CRISPR methodology><CRISPR technique><CRISPR technology><CRISPR tools><CRISPR-CAS-9><CRISPR-based method><CRISPR-based technique><CRISPR-based technology><CRISPR-based tool><CRISPR/CAS approach><CRISPR/Cas method><CRISPR/Cas technology><CRISPR/Cas9><CRISPR/Cas9 technology><Cas nuclease technology><Cell Body><Cell Components><Cell Line><Cell Structure><Cell model><CellLine><Cells><Cellular Assay><Cellular Stress><Cellular Stress Response><Cellular Structures><Cellular model><Central Nervous System Degenerative Diseases><Cerebral cortex><Chaperone><Chronic><Clustered Regularly Interspaced Short Palindromic Repeats approach><Clustered Regularly Interspaced Short Palindromic Repeats method><Clustered Regularly Interspaced Short Palindromic Repeats methodology><Clustered Regularly Interspaced Short Palindromic Repeats technique><Clustered Regularly Interspaced Short Palindromic Repeats technology><Communicable Diseases><Complex><Data><Degenerative Neurologic Disorders><Diazirine><Diazomethane><Disease><Disorder><Dose><Drugs><Dysfunction><Elderly><Encephalon><Ensure><Enzyme Gene><Enzymes><Esteroproteases><Exhibits><FK506 Binding Protein 12-Rapamycin Associated Protein 1><FKBP12 Rapamycin Complex Associated Protein 1><FRAP1><FRAP1 gene><FRAP2><Function Activator><Functional disorder><Gene Transcription><Generations><Genetic><Genetic Alteration><Genetic Change><Genetic Transcription><Genetic defect><Glia><Glial Cells><Hereditary><Heterozygote><Hortega cell><Hydrolase><Hydrolase Family Gene><Hydrolase Gene><Image><Immune system><Induced pluripotent stem cell derived neurons><Infectious Disease Pathway><Infectious Diseases><Infectious Disorder><Inherited><Knowledge><Kolliker's reticulum><Lead><Learning><Lipids><Literature><Lysosomes><Lytotoxicity><Mass Photometry/Spectrum Analysis><Mass Spectrometry><Mass Spectroscopy><Mass Spectrum><Mass Spectrum Analyses><Mass Spectrum Analysis><Mechanistic Target of Rapamycin><Mediating><Medication><Microglia><Mission><Modeling><Molecular Chaperones><Molecular Fingerprinting><Molecular Profiling><Mutation><NIH><National Institutes of Health><Nervous System Degenerative Diseases><Neural Degenerative Diseases><Neural degenerative Disorders><Neurodegenerative Diseases><Neurodegenerative Disorders><Neuroglia><Neuroglial Cells><Neurologic Degenerative Conditions><Neuron from iPSC><Neuron from induced pluripotent stem cells><Non-Polyadenylated RNA><Non-neuronal cell><Nonneuronal cell><Nucleic Acids><Oligosaccharides><Organoids><Oxidative Stress><Pathogenicity><Pathway interactions><Patients><Pb element><Peptidases><Peptide Hydrolases><Pharmaceutical Agent><Pharmaceutical Preparations><Pharmaceuticals><Pharmacologic Substance><Pharmacological Substance><Phenotype><Physiology><Physiopathology><Population><Primary Senile Degenerative Dementia><Protease Gene><Proteases><Proteinases><Proteins><Proteolytic Enzymes><Proteomics><Protocol><Protocols documentation><Public Health><RAFT1><RNA><RNA Expression><RNA Gene Products><Reaction><Recycling><Regimen><Reproducibility><Research><Ribonucleic Acid><Risk><Signal Pathway><Statistical Data Analyses><Statistical Data Analysis><Statistical Data Interpretation><Strains Cell Lines><Strepavidin><Streptavidin><Stress><TAL effector nuclease><TAL endonuclease><TALE nuclease><TALEN technology><TALENs><Testing><Toxic effect><Toxicities><Transcription><Translating><United States National Institutes of Health><Vitamin H><advanced age><affinity purification><alzheimer model><analog><astrocytic glia><autophagy><biologic><brain cell><cell assay><cell stress><coenzyme R><cultured cell line><cytotoxicity><degenerative diseases of motor and sensory neurons><degenerative neurological diseases><determine efficacy><diazo-methane><disease phenotype><drug/agent><efficacy analysis><efficacy assessment><efficacy determination><efficacy evaluation><efficacy examination><evaluate efficacy><examine efficacy><experiment><experimental research><experimental study><experiments><familial AD><familial Alzheimer><familial Alzheimer disease><genome mutation><geriatric><gitter cell><heavy metal Pb><heavy metal lead><heterozygosity><high risk><human disease><iPS><iPS neurons><iPSC><iPSC derived-neurons><iPSCs><imaging><improved><induced pluripotent cell><induced pluripotent stem cell><induced pluripotent stem cell neurons><inducible pluripotent stem cell><lipidomics><mTOR><mTOR inhibition><mammalian target of rapamycin><mesoglia><metabolism measurement><metabolomics><metabonomics><microglial cell><microgliocyte><mitochondrial dysfunction><molecular phenotype><molecular profile><molecular signature><mutant><nerve cement><neural inflammation><neurodegenerative illness><neuroinflammation><neuroinflammatory><neurons derived from induced pluripotent stem cells><novel><pathophysiology><pathway><patient living with Alzheimer's disease><patient suffering from Alzheimer's disease><patient with Alzheimer's><patient with Alzheimer's disease><perivascular glial cell><pharmaceutical><pharmacologic><post treatment><primary degenerative dementia><programs><screening><screenings><senile dementia of the Alzheimer type><senior citizen><small molecule><statistical analysis><transcription activator-like effector nucleases><transcriptional reprogramming>