# Functional Dissection of Alzheimer's Disease Networks in Drosophila: from Association to   Causal Modulators of Age-Dependent Neurodegeration

> **NIH NIH R01** · BAYLOR COLLEGE OF MEDICINE · 2021 · $136,666

## Abstract

The goal of this supplemental project is to extend the bioinformatic analyses and experimental validation of
AMP-AD target networks, which are the focus of the parent grant. First (AIM1), we will generate single-cell
longitudinal transcriptomic profiles of Drosophila AD models expressing human tau and secreted amyloid-β.
These data will allow to distinguish gene expression profiles from distinct neuronal populations as well as
different glia subtypes. These studies will also strongly complement ongoing AMP-AD single-cell gene
expression profiles from human postmortem tissue. We will include rigorous experimental controls for tau /
amyloid-β expression as well as longitudinal sampling to dissect out the specific contributions of age and AD
pathologic species on cell-type specific gene expression changes in the brain. Initially we will include 2 time
points (early and late) during disease progression. During the first year of the proposed supplement (Y4 of the
parent R01 grant) we will generate Drosophila of the appropriate genotypes, extract mRNA, perform
scRNAseq, and begin analyses of the results. During the second year of the supplement (Y5 of the parent R01
grant) we will complete analysis of the data, integrating with our findings from whole brain RNAseq and
performing cross-species comparisons with AMP-AD scRNAseq. We will also perform independent
experiments to confirm the most promising results from scRNAseq, such as immunofluorescence confocal
microscopy, taking advantage of available antibodies, and reagents. Selected candidate causal drivers will also
be manipulated using cell-type specific drivers, including glia and/or glutamatergic, GABAergic, cholinergic and
dopaminergic neurons within Drosophila CNS to examine requirements for brain maintenance and/or function.
 Second (AIM2), we will extend experimental validation of computationally predicted AD causal genes
from Drosophila to mammalian cells. All causal drivers identified in the tau Drosophila screen, will be tested in
cultured mouse and human neural progenitor cells using shRNAs. Specifically, we will assess the impact of
causal drivers on tau protein levels given the key role of tau accumulation AD pathogenesis. Our data shows
that a subset of the identified tau modifier genes (e.g., Hippo pathway components, Nuak1) lower tau protein
levels in Drosophila. Thus, we hypothesize that a subset of causal drivers identified in the ongoing AMP-AD
screen modulate tau accumulation and this subset may be especially interesting from a therapeutic standpoint.
During the first year of the proposed supplement (Y4 of the parent R01 grant) we will test the Hippo pathway
genes in Neuro2A and human neural precursor cells plus all 44 tau modifier genes for their ability to modulate
tau protein levels in the Drosophila brain using immunoblotting and Homogeneous Time Resolved
Fluorescence approaches. During the second year of the supplement (Y5 of the parent R01 grant) we will test
in Neuro2A and h...

## Key facts

- **NIH application ID:** 10228292
- **Project number:** 3R01AG057339-04S1
- **Recipient organization:** BAYLOR COLLEGE OF MEDICINE
- **Principal Investigator:** Juan Botas
- **Activity code:** R01 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2021
- **Award amount:** $136,666
- **Award type:** 3
- **Project period:** 2017-09-15 → 2022-05-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10228292

## Citation

> US National Institutes of Health, RePORTER application 10228292, Functional Dissection of Alzheimer's Disease Networks in Drosophila: from Association to   Causal Modulators of Age-Dependent Neurodegeration (3R01AG057339-04S1). Retrieved via AI Analytics 2026-05-26 from https://api.ai-analytics.org/grant/nih/10228292. Licensed CC0.

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