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![Page 1: Tianhong Pan, MD, PhD; Wenjie Xie, MD; Pawan Rawal,MD Joseph Jankovic, MD; Weidong Le, MD, PhD Department of Neurology, Baylor College of Medicine, Houston,](https://reader038.fdocuments.in/reader038/viewer/2022102700/5519d558550346047c8b4d6c/html5/thumbnails/1.jpg)
Tianhong Pan, MD, PhD; Wenjie Xie, MD; Pawan Rawal,MD Joseph Jankovic, MD; Weidong Le, MD, PhD
Department of Neurology, Baylor College of Medicine, Houston, TX
Rapamycin Protects Against Rotenone-induced Apoptosis Through Autophagy Induction
AAN, Seattle, 04/30/2009
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Diana Helis Henry Medical Research Foundation
Carolyn Weiss Law Seed Funding
National Parkinson Foundation grant to the
Baylor College of Medicine Center of Excellence
This Study Was Supported by
DISCLOSURE
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BACKGROUND
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ATP Free radicals
-synuclein aggregation
Dopaminergic neuron death
Environmental toxin
Genetic factors(DJ-1, PINK1, LRRK2, etc)
Aging
Mutation in ATP13A2Mutation in
parkin, UCHL1
Mutations in -synuclein
UPS = Ubiquitin Proteasome System
Mitochondrial Dysfunction
Accumulation of Aggregated/ Misfolded Proteins
Pathogenesis of Neurodegeneration in Parkinson’s Disease
UPS dysfunction ALP dysfunction
ALP = Autophagy Lysosome Pathway
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Ubiquitin-proteasome system
(UPS)
Protein Degradation Routes
Autophagy-lysosome pathway
(ALP)
Protein Degradation
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Two concentric membranes engulf cell components or aggregated proteins to be
degraded
Autophagosome/Mitophahgosome Autophagolysosome
Autophagic Vacuoles (AVs)
Misfolded/aggregated proteins or cell
componentsAmino and
fatty acids are released into cytoplasma
Lysosome
Autophagosome fuses with lysosome
3-methyladenine (3MA)
Bafilomycin A1 (Baf1)
Enzymes
Cytosolic protein eg. -synuclein
Cytosolic Chaperon (hsc70)
Microautophagy
Macroautophagy (Autophagy)
CMA
Lamp2a
Cytosolic protein-molecular chaperone complex
ALP in mammalian cells
Pan et al. Brain (2008), 131, 1969-1978
ALP = Autophagy Lysosome Pathway
mTOR
Various Signals
Mitophagy
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Autophagy (Macroautophagy)
Aggregated proteins that fail to be degraded by UPS
Entire organelles (eg. mitochondria)
Long-lived, stable proteins
A process of bulk degradation of
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HYPOTHESIS
Autophagy enhancement may prevent accumulation of aggregated/misfolded proteins and of damaged mitochondria, postulated to be two major pathogenic mechanisms of neurodegeneration associated with PD.
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To explore the potential neuroprotective
effects of autophagy enhancement on
neurotoxin-induced injury and its
possible mechanisms
OBJECTIVE
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Rapamycin FDA-approved antibiotic and immuno-
suppressant Enhances autophagy via inhibition of
mammalian target of rapamycin (mTOR), a
negative regulator of autophagy
Rotenone An inhibitor of mitochondrial complex I,
used as a model for neurotoxin-induced
neurodegeneration in PD
REAGENTS
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METHODS The human neuroblastoma SH-SY5Y cells were treated
with rapamycin at various concentrations for different
time durations The cells were exposed to rotenone with/without
rapamycin pretreatment on both small interference
RNA of Atg5 (Atg5 siRNA)-transfected cells, in which
the autophagy was suppressed, and non-transfected
cells. After specific treatment, the cells were either harvested
for protein isolation for Elisa assay or immunoblotting
assay, or were fixed for immunostaining assay and
electron microscopy analysis.
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RESULTS
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Rapamycin Enhanced Autophagy in SH-SY5Y Cells
LC3: Autophagy Marker ; Con = Control; Rapa = Rapamycin
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Rapamycin Protected Against Rotenone-Induced Apoptosis
Con = control; Rapa = Rapamycin; Rot = Rotenone
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Autophagy Inhibition Blocked Rapamycin’s Neuroprotection
Con = control; Rapa = Rapamycin; Rot = Rotenone
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Rapamycin Protected Mitochondrial Function
Con = control; Rapa = Rapamycin; Rot = Rotenone
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Rapamycin Enhanced Degradation of Ubiquitinated Proteins
Rapa = Rapamycin; Rot = Rotenone
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Injured Mitochondria Cleared via Autophagy
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CONCLUSION 1
Rapamycin exerts a neuroprotective role by
interfering with pro-apoptotic insults via
enhanced clearance of misfolded/aggregated
proteins and/or of dysfunctional mitochondria
through autophagy enhancement.
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Anti-apoptosis via Autophagy Enhancement by Rapamycin
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CONCLUSION 2
Autophagy enhancers, such as
rapamycin, may be considered
potential therapeutic agents for
the treatment of PD.
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Therapeutic Targets for PD
Misfolded/Aggregated
Proteins
Injured Mitochondria
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Novel Therapeutic Strategy for PD
Autophagy
Enhancement
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Parkinson’s Disease Center and Movement Disorder Clinic:
Joseph Jankovic, MD
Parkinson’s Disease Research Lab:
Weidong Le, MD, PhD
Pawan Rawal, MD
Yunchen Wu, MD, PhD
Wenjie Xie, MD
Institutional Core Grant #CA16672 High Resolution Electron Microscopy facility, UTMDACC
Kenneth Dunner
Acknowledgement
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Parkinson’s Disease Center and Movement Disorders Clinic