Savage

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Julie Savage presentation made at the Alzheimer Research Forum webinar held November 7, 2012 (www.alzforum.org)

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Savage

  1. 1. Grant Proposal –Epigenetics: driver of amyloid pathology?Julie DelaCruzSandroDaMesquitaXenos MasonJulie SavageJochenDe Vry
  2. 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. 3. Research questions• Are epigenetic changes responsible for altered Aβ deposition?
  4. 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. 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. 6. Aim 1 - Enriched Environment (EE)• 3 weeks EE (Fischer et al., Nature 2007)  Increased Histone-tail acetylation• Similar approach to CMS
  7. 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. 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. 9. TimetableStart End Enriched Environment HDAC2 up/down regulation (Aim 2) Human study (Aim 3) (Aim 1) Chronic mild stress (Aim 1)
  10. 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. 11. Output• 4 papers!!!• Paper: EE• Paper: CMS• Paper: HDAC2 up/down regulation• Paper: human study

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