# Uncovering Alzheimer's disease risk mechanisms through neuron-specific analysis of autophagy and endosomal-lysosomal function

> **NIH NIH P01** · NATHAN S. KLINE INSTITUTE FOR PSYCH RES · 2021 · $350,418

## Abstract

ABSTRACT
This PPG has established that dysfunction at multiple stages throughout the lysosomal network (LN),
comprised of autophagy and the endosomal-lysosomal (EL) pathway, occurs in Alzheimer’s disease (AD)
pathogenesis. A hypothesis throughout the PPG is that increasing the levels of β-site cleaved carboxyl-
terminal fragment of the amyloid-β precursor protein (βCTF) through genetic and environmental AD risk
factors, including the ApoE4 allele, cholesterol, and recently identified risk genes found by GWAS, disrupts LN
function in early-onset AD and in the more common late-onset AD form. In this project 4 (P4), we focus on
autophagy alterations driven by late-onset AD risk factors, investigating particularly the role of the βCTF and its
effects on the LN mediated through rab5 activation. The goals will be attained through a comprehensive
transcriptomic approach that evaluates all stages of autophagy in homogeneous vulnerable neuronal
populations using RNA-sequencing (RNA-seq) and bioinformatics applied to custom-designed pipelines of
autophagy and lysosomal genes. These informative and functionally predictive gene analysis pipelines have
been confirmed, and are complemented by immunological and biochemical functional analyses of autophagy,
including the use of a novel neuron-specific autophagy reporter-construct mouse. Our experimental design
overcomes issues associated with the marked cellular heterogeneity of brain tissue, where vulnerable neurons
are intermixed with spared neuronal populations and glia, which we show in our preliminary data to have
different autophagy pathway programs. The strong validation of this approach derives from our longstanding
investigations of autophagy and lysosomal pathways in AD and supports its innovative application to in vivo
studies involving specific vulnerable neuronal populations. In conjunction with P1-P3, we will employ a cohort
of models, including a wild-type APP overexpressing mouse, a novel rab5 transgenic mouse that displays
endosome morphologic and behavioral abnormalities, and humanized ApoE4 mice, among others that display
LN deficits relevant to AD, enabling us to test multiple hypotheses related to the pathogenic effects of βCTF on
the autophagy and EL pathways. In Aim 1 we will evaluate the function of all stages of autophagy in vivo
through a novel combination of cell-specific transcriptomics, tracking of fluorescence reporter constructs, and
parallel immunochemical functional analyses in vulnerable cholinergic, hippocampal, and cortical neurons.
Increasing autophagy turnover and inducing lysosomal biogenesis is an appealing therapeutic approach for
improving the efficiency of the LN in vulnerable cells, and modalities known to induce autophagy include
calorie restriction (CR). In Aim 2 we will test the hypothesis that the benefits of calorie restriction (CR) and an
established CR mimetic, epigallocatechin-3-gallate (EGCG), involve increased autophagy turnover and
lysosomal biogenesis in vi...

## Key facts

- **NIH application ID:** 10126781
- **Project number:** 5P01AG017617-20
- **Recipient organization:** NATHAN S. KLINE INSTITUTE FOR PSYCH RES
- **Principal Investigator:** STEPHEN D GINSBERG
- **Activity code:** P01 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2021
- **Award amount:** $350,418
- **Award type:** 5
- **Project period:** 2000-02-15 → 2025-03-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10126781

## Citation

> US National Institutes of Health, RePORTER application 10126781, Uncovering Alzheimer's disease risk mechanisms through neuron-specific analysis of autophagy and endosomal-lysosomal function (5P01AG017617-20). Retrieved via AI Analytics 2026-05-27 from https://api.ai-analytics.org/grant/nih/10126781. Licensed CC0.

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