This grant proposal aims to investigate whether epigenetic changes drive amyloid pathology in Alzheimer's disease (AD). There are three aims: 1) examine how chronic stress and enriched environments induce epigenetic changes and affect amyloid deposition and memory in mouse models, 2) manipulate HDAC2 levels to mimic enriched environment effects, and 3) study related epigenetic markers in human AD cases and controls. The proposal requests $333,000 over three years to study mouse models using techniques like chromatin immunoprecipitation and human brain samples. The expected output is four papers on the topics of enriched environment, chronic stress, HDAC2 manipulation, and human studies.

1. Grant Proposal –
Epigenetics: driver of amyloid
pathology?
Julie DelaCruz
SandroDaMesquita
Xenos Mason
Julie Savage
JochenDe Vry

2. Background
• Age-related increases in DNA methylation in
humans
• AD patients show reduced DNA methylation
• DNMTs/HDACs may be involved in cognitive
decline, changes in learning & memory
• Parallels methylation patterns in AD mouse
models

3. Research questions
• Are epigenetic changes responsible for altered
Aβ deposition?

4. Epigenetic changes
• Environmental influences: negative (chronic
mild stress) AND positive (enriched
environment) …
• … induce epigenetic changes
• Do these epigenetic changes accelerate or
delay Amyloid deposition and memory
performance?
• Wild-type and AD model

5. Aim 1 - Chronic Mild Stress (CMS)
• 3 weeks CMS (Cuadrado-Tejedoret al., J Alzheimer Dis.
2012) in young (3m) and old (12m), wild-type and
APP/PS1 mice
• Examine epigenetic markers in different brain regions
(hipp, EC, PFC, cerebellum)  select brain region with
biggest differences in epigenetic state
• Selected brain region:ChiP focus on genes involved
in epigenetic regulation, validate with qPCR
• Examine identified gene(s) (up- or down-regulation) by
overexpression or RNAi - functional assay  check
alterations in brain pathology and cognitive
performance

6. Aim 1 - Enriched Environment (EE)
• 3 weeks EE (Fischer et al., Nature 2007) 
Increased Histone-tail acetylation
• Similar approach to CMS

7. Aim 2 – Epigenetic manipulation
• Focal up- or down-regulation of HDAC2 in
selected brain region (Aim 1)  mimic EE
• Electroporation HDAC2-siRNA-plasmid,HDAC2-
plasmid, or scrambled plasmid
• After 3-4 weeks: memory performance, IHC
(check pathology)
• Young (3m) and old (12m) – wild-type and
APP/PS1

8. Aim 3 – Comparative human
epigenetics
• Familial cases AD, sporadic AD, age-matched
ctrls brain bank and/or collaboration
• IHC: HDACs, DNMTs, HATs
• Specific brain region: FISH or RT-PCR for
candidate genes Aim 1

9. Timetable
Start End
Enriched Environment HDAC2 up/down regulation
(Aim 2) Human study (Aim 3)
(Aim 1)
Chronic mild stress
(Aim 1)

10. Budget
• Aim 1: $65,000
– Animals: EE and stress  2 x 2 x 2 x 12= 96 animals  $15,000
– CHiP, reagents,…: $50,000
• Aim 2: $38,000
– Animals: $15,000
– Consumables: $10,000
– Stimulator: $13,000
• Aim 3: $50,000
– Consumables: $50,000
• 2 PhD students = 6 years * $30,000 = $180,000
• Total: $333,000

11. Output
• 4 papers!!!
• Paper: EE
• Paper: CMS
• Paper: HDAC2 up/down regulation
• Paper: human study