Tianhong Pan, MD, PhD; Wenjie Xie, MD; Pawan Rawal,MD Joseph Jankovic, MD; Weidong Le, MD, PhD...

25
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

TAGS:

Transcript of Tianhong Pan, MD, PhD; Wenjie Xie, MD; Pawan Rawal,MD Joseph Jankovic, MD; Weidong Le, MD, PhD...

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,

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

Page 2: 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,

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

Page 3: 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,

BACKGROUND

Page 4: 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,

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

Page 5: 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,

Ubiquitin-proteasome system

(UPS)

Protein Degradation Routes

Autophagy-lysosome pathway

(ALP)

Protein Degradation

Page 6: 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,

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

Page 7: 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,

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

Page 8: 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,

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.

Page 9: 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,

To explore the potential neuroprotective

effects of autophagy enhancement on

neurotoxin-induced injury and its

possible mechanisms

OBJECTIVE

Page 10: 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,

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

Page 11: 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,

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.

Page 12: 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,

RESULTS

Page 13: 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,

Rapamycin Enhanced Autophagy in SH-SY5Y Cells

LC3: Autophagy Marker ; Con = Control; Rapa = Rapamycin

Page 14: 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,

Rapamycin Protected Against Rotenone-Induced Apoptosis

Con = control; Rapa = Rapamycin; Rot = Rotenone

Page 15: 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,

Autophagy Inhibition Blocked Rapamycin’s Neuroprotection

Con = control; Rapa = Rapamycin; Rot = Rotenone

Page 16: 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,

Rapamycin Protected Mitochondrial Function

Con = control; Rapa = Rapamycin; Rot = Rotenone

Page 17: 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,

Rapamycin Enhanced Degradation of Ubiquitinated Proteins

Rapa = Rapamycin; Rot = Rotenone

Page 18: 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,

Injured Mitochondria Cleared via Autophagy

Page 19: 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,

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.

Page 20: 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,

Anti-apoptosis via Autophagy Enhancement by Rapamycin

Page 21: 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,

CONCLUSION 2

Autophagy enhancers, such as

rapamycin, may be considered

potential therapeutic agents for

the treatment of PD.

Page 22: 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,

Therapeutic Targets for PD

Misfolded/Aggregated

Proteins

Injured Mitochondria

Page 23: 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,

Novel Therapeutic Strategy for PD

Autophagy

Enhancement

Page 24: 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,

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

Page 25: 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,

Parkinson’s Disease Center and Movement Disorders Clinic


Chest Pain (GERD) Dimitrios Stefanidis, MD, PhD Steven B. Goldin, MD, PhD.

Chest Pain (GERD) Dimitrios Stefanidis, MD, PhD Steven B. Goldin, MD, PhD.

MD/PhD Program

MD/PhD Program

Szabolcs Péter , MD, PhD Peter Weber, MD, PhD

Szabolcs Péter , MD, PhD Peter Weber, MD, PhD

Prof. Petrisor GEAVLETE, MD, PhD. Razvan Multescu, MD, PhD. Bogdan Geavlete, MD, PhD. Saint John Emergency Clinical Hospital Bucharest, Romania Prof. Petrisor.

Prof. Petrisor GEAVLETE, MD, PhD. Razvan Multescu, MD, PhD. Bogdan Geavlete, MD, PhD. Saint John Emergency Clinical Hospital Bucharest, Romania Prof. Petrisor.

Philippine Kiès, MD, Marianne Bootsma, MD, PhD, Jeroen Bax, MD, PhD, Martin J. Schalij, MD, PhD, Ernst E. van der Wall, MD, PhD Arrhythmogenic right ventricular.

Philippine Kiès, MD, Marianne Bootsma, MD, PhD, Jeroen Bax, MD, PhD, Martin J. Schalij, MD, PhD, Ernst E. van der Wall, MD, PhD Arrhythmogenic right ventricular.

MD, MS, MDS, Phd, DIPLOMAS, M.Phil, PhD

MD, MS, MDS, Phd, DIPLOMAS, M.Phil, PhD

M. Karan MD P. Vulekovic MD PhD Dj. Jajic MD PhD T. Cigic Md PhD V. Papic MD PhD Dj. Djilvesi MD B. Jelaca MD Neurosurgery Clinic Clinical Centre of Vojvodina,

M. Karan MD P. Vulekovic MD PhD Dj. Jajic MD PhD T. Cigic Md PhD V. Papic MD PhD Dj. Djilvesi MD B. Jelaca MD Neurosurgery Clinic Clinical Centre of Vojvodina,

Measuring diseases… Liliana Iliescu, MD, PhD. MORBIDITY Liliana Iliescu, MD, PhD.

Measuring diseases… Liliana Iliescu, MD, PhD. MORBIDITY Liliana Iliescu, MD, PhD.

Translational Research Program (TRP) Chair: Adam P. Dicker, MD, PhD Brain Arnab Chakravarti, MD Breast Wendy Woodward, MD, PhD Steven Chimura, MD, PhD.

Translational Research Program (TRP) Chair: Adam P. Dicker, MD, PhD Brain Arnab Chakravarti, MD Breast Wendy Woodward, MD, PhD Steven Chimura, MD, PhD.

Bleeding time,clotting time, PT, and PTT Ostadrahimi MD,PhD Ostadrahimi MD,PhD.

Bleeding time,clotting time, PT, and PTT Ostadrahimi MD,PhD Ostadrahimi MD,PhD.

OUR MISSION The mission of the Good Neighbor … · Damian Almiron MD-PhD Laurie Delatour MD-PhD Ana Maria Dumitru MD-PhD Chris Rees MD-PhD ... Ann Raynolds PsyD Brenda Haynes RN

OUR MISSION The mission of the Good Neighbor … · Damian Almiron MD-PhD Laurie Delatour MD-PhD Ana Maria Dumitru MD-PhD Chris Rees MD-PhD ... Ann Raynolds PsyD Brenda Haynes RN

Adriaan Coumou MD Dion Paridaens MD PhD

Adriaan Coumou MD Dion Paridaens MD PhD

Olivia Huston Kendall Lee MD, PhD Robert Watson MD, PhD Matt Bernstein PhD John Huston MD Kiaran McGee PhD Intraoperative MRI Imaging Strategies to Evaluate.

Olivia Huston Kendall Lee MD, PhD Robert Watson MD, PhD Matt Bernstein PhD John Huston MD Kiaran McGee PhD Intraoperative MRI Imaging Strategies to Evaluate.

President - SNS · President Maria Francisca Moraes Fontes, MD, PhD Scientific Committee António Coutinho, MD, PhD Nuno Riso, MD Carlos Vasconcelos, MD, PhD Luís Campos, MD

President - SNS · President Maria Francisca Moraes Fontes, MD, PhD Scientific Committee António Coutinho, MD, PhD Nuno Riso, MD Carlos Vasconcelos, MD, PhD Luís Campos, MD

A.D’Hoore MD PhD, A. Wolthuis MD, F. Penninckx MD PhD K. Haustermans MD PhD*, E. Van Cutsem MD PhD** V. Vandecaveye MD PhD*** Department of Abdominal Surgery,

A.D’Hoore MD PhD, A. Wolthuis MD, F. Penninckx MD PhD K. Haustermans MD PhD*, E. Van Cutsem MD PhD** V. Vandecaveye MD PhD*** Department of Abdominal Surgery,

Leonardo Trasande, MD, MPP Teresa Attina, MD, PhD, MPH Jan Blustein, MD, PhD

Leonardo Trasande, MD, MPP Teresa Attina, MD, PhD, MPH Jan Blustein, MD, PhD

Challenging Cases in Diagnosis and Management … MD, PhD, Luis Aguinaga, MD, PhD, Joseph G. Akar, MD, PhD, Vinay Badhwar, MD, Josep Brugada, MD, PhD, John Camm, MD, Peng-Sheng Chen,

Challenging Cases in Diagnosis and Management … MD, PhD, Luis Aguinaga, MD, PhD, Joseph G. Akar, MD, PhD, Vinay Badhwar, MD, Josep Brugada, MD, PhD, John Camm, MD, Peng-Sheng Chen,

Deep anterior lamellar keratoplasty (DALK) in Tohoku University, Japan T Hariya MD, A Kubota MD PhD, M Uematsu MD, S Yokokura MD PhD,K Nishida MD PhD,

Deep anterior lamellar keratoplasty (DALK) in Tohoku University, Japan T Hariya MD, A Kubota MD PhD, M Uematsu MD, S Yokokura MD PhD,K Nishida MD PhD,

oncologyupdates.ro · Călin Buzea, Phys., PhD Laura Mihalache, MD, PhD Simona Ruxandra Volovă˙, MD, PhD Constantin Volovă˙, MD, PhD ORGANIZING COMMITTEE Mihai Andronic, MD Ana

oncologyupdates.ro · Călin Buzea, Phys., PhD Laura Mihalache, MD, PhD Simona Ruxandra Volovă˙, MD, PhD Constantin Volovă˙, MD, PhD ORGANIZING COMMITTEE Mihai Andronic, MD Ana

William B. Cutrer, MD, MEd; Lourdes Estrada, PhD; Bonnie Miller, MD; Lillian Nanney, PhD; Neil Osheroff, PhD; Donald Moore, PhD.

William B. Cutrer, MD, MEd; Lourdes Estrada, PhD; Bonnie Miller, MD; Lillian Nanney, PhD; Neil Osheroff, PhD; Donald Moore, PhD.