Addiction Therapy-2014€¦ · Addiction Therapy-2014 Chicago, USA August 4 -6, 2014 Bryan...
Transcript of Addiction Therapy-2014€¦ · Addiction Therapy-2014 Chicago, USA August 4 -6, 2014 Bryan...
![Page 1: Addiction Therapy-2014€¦ · Addiction Therapy-2014 Chicago, USA August 4 -6, 2014 Bryan Yamamoto. Bryan Yamamoto, Ph.D. Department of Neurosciences University of Toledo College](https://reader034.fdocuments.net/reader034/viewer/2022052105/603fe38cd680cb1d34135707/html5/thumbnails/1.jpg)
Addiction Therapy-2014Chicago, USA
August 4 - 6, 2014
Bryan Yamamoto
![Page 2: Addiction Therapy-2014€¦ · Addiction Therapy-2014 Chicago, USA August 4 -6, 2014 Bryan Yamamoto. Bryan Yamamoto, Ph.D. Department of Neurosciences University of Toledo College](https://reader034.fdocuments.net/reader034/viewer/2022052105/603fe38cd680cb1d34135707/html5/thumbnails/2.jpg)
Bryan Yamamoto, Ph.D.
Department of Neurosciences
University of Toledo College of Medicine and Life Sciences
August 4, 2014
![Page 3: Addiction Therapy-2014€¦ · Addiction Therapy-2014 Chicago, USA August 4 -6, 2014 Bryan Yamamoto. Bryan Yamamoto, Ph.D. Department of Neurosciences University of Toledo College](https://reader034.fdocuments.net/reader034/viewer/2022052105/603fe38cd680cb1d34135707/html5/thumbnails/3.jpg)
Volkow et al, Am. J. Psychiatry, 2001
Methamphetamine Neurotoxicity in Humans
Callaghan et al., Drug and Alcohol Dependence, 2011
Meth abuse increases risk of Parkinson’s Disease by 76% compared to age-
matched controls and cocaine-abusers
![Page 4: Addiction Therapy-2014€¦ · Addiction Therapy-2014 Chicago, USA August 4 -6, 2014 Bryan Yamamoto. Bryan Yamamoto, Ph.D. Department of Neurosciences University of Toledo College](https://reader034.fdocuments.net/reader034/viewer/2022052105/603fe38cd680cb1d34135707/html5/thumbnails/4.jpg)
Overarching Hypothesis
Excitotoxicity and oxidation of proteins and lipids are mechanisms underlying the neurotoxicity of Methamphetaminethe neurotoxicity of Methamphetamine
![Page 5: Addiction Therapy-2014€¦ · Addiction Therapy-2014 Chicago, USA August 4 -6, 2014 Bryan Yamamoto. Bryan Yamamoto, Ph.D. Department of Neurosciences University of Toledo College](https://reader034.fdocuments.net/reader034/viewer/2022052105/603fe38cd680cb1d34135707/html5/thumbnails/5.jpg)
Glutamate Release
Calcium
NOS
NO
H2O2
Calpain
Identified Events Leading to a Loss of Dopamine and 5HT
Glu Receptor Activation
Dopamine Release
X
XX
XX
MAO
NO
ONOO-
(peroxynitrite)
O2-
Protein oxidation,
Mitochondrial Dysfunction
Lipid Peroxidation
Calpain
Proteolysis
Fe++
XOH*
![Page 6: Addiction Therapy-2014€¦ · Addiction Therapy-2014 Chicago, USA August 4 -6, 2014 Bryan Yamamoto. Bryan Yamamoto, Ph.D. Department of Neurosciences University of Toledo College](https://reader034.fdocuments.net/reader034/viewer/2022052105/603fe38cd680cb1d34135707/html5/thumbnails/6.jpg)
But, there are caveats to the
proposition that the
neurotoxicity/excitotoxicity of
METH is a result of its direct action METH is a result of its direct action
on the brain……..
![Page 7: Addiction Therapy-2014€¦ · Addiction Therapy-2014 Chicago, USA August 4 -6, 2014 Bryan Yamamoto. Bryan Yamamoto, Ph.D. Department of Neurosciences University of Toledo College](https://reader034.fdocuments.net/reader034/viewer/2022052105/603fe38cd680cb1d34135707/html5/thumbnails/7.jpg)
Local administration of METH does not increase glutamate
0
50
100
150
200
Do
pa
min
e (p
g/2
0 u
l)Systemic MA
Local MA
Systemic and Local Administration of METH:
-1 0 1 2 3 4 5 6 7 8
Time (hours)
Burrows
Glu
tam
ate
(p
g/2
0 µµ µµ
l)
01000
2000300040005000
600070008000
900010000
-1 0 1 2 3 4 5 6 7 8
PerfusionGlu
tam
ate
(p
g/2
0 µµ µµ
l
![Page 8: Addiction Therapy-2014€¦ · Addiction Therapy-2014 Chicago, USA August 4 -6, 2014 Bryan Yamamoto. Bryan Yamamoto, Ph.D. Department of Neurosciences University of Toledo College](https://reader034.fdocuments.net/reader034/viewer/2022052105/603fe38cd680cb1d34135707/html5/thumbnails/8.jpg)
Do
pa
min
e (
pg
/ µµ µµg
pro
tein
)
Glutamate Synergizes with Local METH
To Deplete Dopamine Content
*
#
Do
pa
min
e (
pg
/
Without Glutamate With Glutamate
No MethMeth Meth/Heat No Meth Meth Meth/Heat
#
Burrows
![Page 9: Addiction Therapy-2014€¦ · Addiction Therapy-2014 Chicago, USA August 4 -6, 2014 Bryan Yamamoto. Bryan Yamamoto, Ph.D. Department of Neurosciences University of Toledo College](https://reader034.fdocuments.net/reader034/viewer/2022052105/603fe38cd680cb1d34135707/html5/thumbnails/9.jpg)
�If it is not the direct effect of METH alone on
the brain (striatum), what other systems could
be responsible for the neurotoxicity?
�Another target of METH is the liver
Thinking Outside the Brain……
�Another target of METH is the liver
�Liver damage can produce encephalopathy
(i.e. hepatic encephalopathy)
� Tremor, movement disorders
� Delirium, stupor, confusion, and coma
![Page 10: Addiction Therapy-2014€¦ · Addiction Therapy-2014 Chicago, USA August 4 -6, 2014 Bryan Yamamoto. Bryan Yamamoto, Ph.D. Department of Neurosciences University of Toledo College](https://reader034.fdocuments.net/reader034/viewer/2022052105/603fe38cd680cb1d34135707/html5/thumbnails/10.jpg)
METH and Evidence of Hepatotoxicity
Halpin
![Page 11: Addiction Therapy-2014€¦ · Addiction Therapy-2014 Chicago, USA August 4 -6, 2014 Bryan Yamamoto. Bryan Yamamoto, Ph.D. Department of Neurosciences University of Toledo College](https://reader034.fdocuments.net/reader034/viewer/2022052105/603fe38cd680cb1d34135707/html5/thumbnails/11.jpg)
80
100
120
ALT
*
400
500
600
AST
*
Alanine Aminotransferase Aspartate Aminotransferase
Methamphetamine Increases Liver Enzymes in Plasma
0
20
40
60
ALT
(IU/L)
Saline METH
0
100
200
300
AST
(IU/L)
Saline METH
Halpin
![Page 12: Addiction Therapy-2014€¦ · Addiction Therapy-2014 Chicago, USA August 4 -6, 2014 Bryan Yamamoto. Bryan Yamamoto, Ph.D. Department of Neurosciences University of Toledo College](https://reader034.fdocuments.net/reader034/viewer/2022052105/603fe38cd680cb1d34135707/html5/thumbnails/12.jpg)
Liver Damage and Ammonia
� The liver normally removes excess ammonia from the
blood via the urea cycle.
� This is important because ammonia is a neurotoxic
byproduct of protein metabolism and other byproduct of protein metabolism and other
metabolic reactions.
� If the liver damaged, there is an accumulation of
ammonia that is thought to cause hepatic
encephalopathy
![Page 13: Addiction Therapy-2014€¦ · Addiction Therapy-2014 Chicago, USA August 4 -6, 2014 Bryan Yamamoto. Bryan Yamamoto, Ph.D. Department of Neurosciences University of Toledo College](https://reader034.fdocuments.net/reader034/viewer/2022052105/603fe38cd680cb1d34135707/html5/thumbnails/13.jpg)
Ammonia Toxicity
� Excitotoxicity (Fan and Szerb, 1993, Hermenegildo et
al., 1996)
� Oxidative stress (Kosenko et al., 1997 and Kosenko et
al., 1995)
� Metabolic compromise (Hawkins et
al., 1973, McCandless and Schenker, 1981)al., 1973, McCandless and Schenker, 1981)
∴∴∴∴Similar mechanisms that mediate the toxicity to
ammonia and the neurotoxicity to METH
� Unknown whether peripheral organ (e.g. liver)
damage is produced by METH and if it contributes to
METH neurotoxicity
![Page 14: Addiction Therapy-2014€¦ · Addiction Therapy-2014 Chicago, USA August 4 -6, 2014 Bryan Yamamoto. Bryan Yamamoto, Ph.D. Department of Neurosciences University of Toledo College](https://reader034.fdocuments.net/reader034/viewer/2022052105/603fe38cd680cb1d34135707/html5/thumbnails/14.jpg)
Overarching Hypothesis
Liver Damage
Methamphetamine
Hyperammonemia
Neurotoxicity
LactuloseX
![Page 15: Addiction Therapy-2014€¦ · Addiction Therapy-2014 Chicago, USA August 4 -6, 2014 Bryan Yamamoto. Bryan Yamamoto, Ph.D. Department of Neurosciences University of Toledo College](https://reader034.fdocuments.net/reader034/viewer/2022052105/603fe38cd680cb1d34135707/html5/thumbnails/15.jpg)
120
140
160
180
200
Pla
sma
Am
mo
nia
(µµ µµ
M)
*
#
Methamphetamine Increases Ammonia in Plasma
0
20
40
60
80
100
Saline Lactulose METH METH +
Lactulose
Pla
sma
Am
mo
nia
(
Halpin
![Page 16: Addiction Therapy-2014€¦ · Addiction Therapy-2014 Chicago, USA August 4 -6, 2014 Bryan Yamamoto. Bryan Yamamoto, Ph.D. Department of Neurosciences University of Toledo College](https://reader034.fdocuments.net/reader034/viewer/2022052105/603fe38cd680cb1d34135707/html5/thumbnails/16.jpg)
Bra
in A
mm
on
ia
(Pe
rce
nt
Ba
seli
ne
)
Veh+METH Veh+Sal
Lac+Sal Lac+METH
400
300
500
Methamphetamine Increases Ammonia in Striatum:
Blocked by Lactulose
0 1 2 3 4 5 6 7 8
Bra
in A
mm
on
ia
(Pe
rce
nt
Ba
seli
ne
)
Time (Hr)
300
200
100
Halpin
![Page 17: Addiction Therapy-2014€¦ · Addiction Therapy-2014 Chicago, USA August 4 -6, 2014 Bryan Yamamoto. Bryan Yamamoto, Ph.D. Department of Neurosciences University of Toledo College](https://reader034.fdocuments.net/reader034/viewer/2022052105/603fe38cd680cb1d34135707/html5/thumbnails/17.jpg)
ME
TH
g p
rote
in X
10
-2)
2
3
Lactulose Does Not Affect METH Concentrations in the Brain
(pp
m/ µµ µµ
g p
rote
in X
10
1
METH METH + Lactulose
0
Halpin
![Page 18: Addiction Therapy-2014€¦ · Addiction Therapy-2014 Chicago, USA August 4 -6, 2014 Bryan Yamamoto. Bryan Yamamoto, Ph.D. Department of Neurosciences University of Toledo College](https://reader034.fdocuments.net/reader034/viewer/2022052105/603fe38cd680cb1d34135707/html5/thumbnails/18.jpg)
80
100
120
g p
rote
in)
5
6
7
8
g p
rote
in)
*#
#
Lactulose Attenuates METH-induced
Long-term Depletions of Dopamine and 5HT Content
Dopamine Serotonin
0
20
40
60
1
(pg
/ µµ µµg
pro
tein
)
0
1
2
3
4
5
1Vehicle VehicleVehicleVehicleLactulose Lactulose LactuloseLactulose
Saline Saline SalineSaline METHMETHMETH METH
(pg
/ µµ µµg
pro
tein
)
* *
Halpin
![Page 19: Addiction Therapy-2014€¦ · Addiction Therapy-2014 Chicago, USA August 4 -6, 2014 Bryan Yamamoto. Bryan Yamamoto, Ph.D. Department of Neurosciences University of Toledo College](https://reader034.fdocuments.net/reader034/viewer/2022052105/603fe38cd680cb1d34135707/html5/thumbnails/19.jpg)
1.2
1.0
0.8
0.6
Dopamine Transporter Immunoreactivity
*
% C
on
tro
lLactulose Blocks METH-induced Decreases
in Dopamine Transporter Immmunoreactivity
0.6
0.4
0.2
Vehicle VehicleLactulose Lactulose
Saline Saline METHMETH
% C
on
tro
l
Halpin
![Page 20: Addiction Therapy-2014€¦ · Addiction Therapy-2014 Chicago, USA August 4 -6, 2014 Bryan Yamamoto. Bryan Yamamoto, Ph.D. Department of Neurosciences University of Toledo College](https://reader034.fdocuments.net/reader034/viewer/2022052105/603fe38cd680cb1d34135707/html5/thumbnails/20.jpg)
Glu
tam
ate
(%
Ba
seli
ne
)
400
500
600
300
Vehicle-METH
METH-Induced Increases
in Striatal Extracellular Glutamate:
Blocked by Lactulose
1 2 3 4 5 6 7 8
Glu
tam
ate
(%
Ba
seli
ne
)
200
100
300
B
Time (hrs)
Vehicle-Saline
Lac-METH
Lac-Saline
Halpin, Northrop
![Page 21: Addiction Therapy-2014€¦ · Addiction Therapy-2014 Chicago, USA August 4 -6, 2014 Bryan Yamamoto. Bryan Yamamoto, Ph.D. Department of Neurosciences University of Toledo College](https://reader034.fdocuments.net/reader034/viewer/2022052105/603fe38cd680cb1d34135707/html5/thumbnails/21.jpg)
Lactulose Blocks METH-Induced Increases
in Striatal Spectrin Proteolysis
100
160
140
120
180 *
Pe
rce
nt
Co
ntr
ol
Vehicle Lactulose Vehicle LactuloseSaline Saline METH METH
100
40
20
60
80
Pe
rce
nt
Co
ntr
ol
Halpin, Northrop
![Page 22: Addiction Therapy-2014€¦ · Addiction Therapy-2014 Chicago, USA August 4 -6, 2014 Bryan Yamamoto. Bryan Yamamoto, Ph.D. Department of Neurosciences University of Toledo College](https://reader034.fdocuments.net/reader034/viewer/2022052105/603fe38cd680cb1d34135707/html5/thumbnails/22.jpg)
METH-Induced Astroglyosis (GFAP) is Blocked by Lactulose
100
120
140
160
180
200
Pe
rce
nt
Co
ntr
ol
*
0
20
40
60
80
Vehicle Lactulose Vehicle LactuloseSaline Saline METH METH
Pe
rce
nt
Co
ntr
ol
Halpin, Northrop
![Page 23: Addiction Therapy-2014€¦ · Addiction Therapy-2014 Chicago, USA August 4 -6, 2014 Bryan Yamamoto. Bryan Yamamoto, Ph.D. Department of Neurosciences University of Toledo College](https://reader034.fdocuments.net/reader034/viewer/2022052105/603fe38cd680cb1d34135707/html5/thumbnails/23.jpg)
aCSF TBOA
NH3 NH3+TBOA
Glu
tam
ate
(%
Baseli
ne)
Local Perfusion of Ammonia on Glutamate:Efflux Through Reversal of the Glutamate Transporter
*
300
250
200
150
0 0.5 1 1.5 2 2.5 3 3.5 4 4.5
Glu
tam
ate
(%
Baseli
ne)
Time (hrs)
TBOA or aCSF
NH3 or aCSF
*
100
150
50
Halpin, Northrop
![Page 24: Addiction Therapy-2014€¦ · Addiction Therapy-2014 Chicago, USA August 4 -6, 2014 Bryan Yamamoto. Bryan Yamamoto, Ph.D. Department of Neurosciences University of Toledo College](https://reader034.fdocuments.net/reader034/viewer/2022052105/603fe38cd680cb1d34135707/html5/thumbnails/24.jpg)
60
80
100
120
3
4
5
6
* *
(pg
/ µµ µµg
pro
tein
)
(pg
/ µµ µµg
pro
tein
)
Combined Local Perfusions of Ammonia and METH
Dopamine and 5HT Content in the Striatum
Dopamine 5HT
0
20
40
0
1
2
Vehicle VehicleNH3
Saline Saline METHMETH
Vehicle Vehicle
Saline Saline METHMETH
(pg
/
(pg
/
NH3 NH3 NH3
Halpin, Northrop
METH
GYKI
NH3
METH
GYKI
NH3
![Page 25: Addiction Therapy-2014€¦ · Addiction Therapy-2014 Chicago, USA August 4 -6, 2014 Bryan Yamamoto. Bryan Yamamoto, Ph.D. Department of Neurosciences University of Toledo College](https://reader034.fdocuments.net/reader034/viewer/2022052105/603fe38cd680cb1d34135707/html5/thumbnails/25.jpg)
Are there other targets of ammonia?
![Page 26: Addiction Therapy-2014€¦ · Addiction Therapy-2014 Chicago, USA August 4 -6, 2014 Bryan Yamamoto. Bryan Yamamoto, Ph.D. Department of Neurosciences University of Toledo College](https://reader034.fdocuments.net/reader034/viewer/2022052105/603fe38cd680cb1d34135707/html5/thumbnails/26.jpg)
Amphetamines
The Blood-Brain Barrier: A Protective Barrier of the Brain
![Page 27: Addiction Therapy-2014€¦ · Addiction Therapy-2014 Chicago, USA August 4 -6, 2014 Bryan Yamamoto. Bryan Yamamoto, Ph.D. Department of Neurosciences University of Toledo College](https://reader034.fdocuments.net/reader034/viewer/2022052105/603fe38cd680cb1d34135707/html5/thumbnails/27.jpg)
BBB Function in Cortex After Meth
Vehicle+Saline Lactulose+Saline Vehicle+Meth Lactulose+Meth
Vehicle
Saline
Lactulose
Saline
Lactulose
Meth
Vehicle
Meth
*
#
Quantification of FITC-Dextran Extravasation
Northrop
![Page 28: Addiction Therapy-2014€¦ · Addiction Therapy-2014 Chicago, USA August 4 -6, 2014 Bryan Yamamoto. Bryan Yamamoto, Ph.D. Department of Neurosciences University of Toledo College](https://reader034.fdocuments.net/reader034/viewer/2022052105/603fe38cd680cb1d34135707/html5/thumbnails/28.jpg)
Blood-Brain Barrier
Tight Junctions
Occludin
Claudin 3, 5, 12
JAM
![Page 29: Addiction Therapy-2014€¦ · Addiction Therapy-2014 Chicago, USA August 4 -6, 2014 Bryan Yamamoto. Bryan Yamamoto, Ph.D. Department of Neurosciences University of Toledo College](https://reader034.fdocuments.net/reader034/viewer/2022052105/603fe38cd680cb1d34135707/html5/thumbnails/29.jpg)
*
#
Occludin Claudin-5
*
#
Meth-Induced Disruption of BBB Structure in Cortex
* *
Isolated Brain Capillaries
*
Vehicle
Saline
Lactulose
Saline
Vehicle
Meth
Lactulose
Meth
*
Vehicle
Saline
Lactulose
Saline
Vehicle
Meth
Lactulose
Meth
* *
Northrop
![Page 30: Addiction Therapy-2014€¦ · Addiction Therapy-2014 Chicago, USA August 4 -6, 2014 Bryan Yamamoto. Bryan Yamamoto, Ph.D. Department of Neurosciences University of Toledo College](https://reader034.fdocuments.net/reader034/viewer/2022052105/603fe38cd680cb1d34135707/html5/thumbnails/30.jpg)
Representative Blot for
Nitrotyrosine
Ammonia as a Pro-Oxidant?
Representative Blot for
Tight Junction Proteins
Northrop
Isolated Brain Capillaries from Cortex
![Page 31: Addiction Therapy-2014€¦ · Addiction Therapy-2014 Chicago, USA August 4 -6, 2014 Bryan Yamamoto. Bryan Yamamoto, Ph.D. Department of Neurosciences University of Toledo College](https://reader034.fdocuments.net/reader034/viewer/2022052105/603fe38cd680cb1d34135707/html5/thumbnails/31.jpg)
Cyclooxygenase-2
Ammonia as a Pro-Inflammatory Agent?
*
&
2 Im
mu
no
rea
ctivity
Ve
hic
le +
Sa
line
)
Northrop
Vehicle
Saline
Vehicle
Meth
Lactulose
Meth
Lactulose
Saline
&
CO
X-2
Im
mu
no
rea
ctivity
(%V
eh
icle
+ S
alin
e)
![Page 32: Addiction Therapy-2014€¦ · Addiction Therapy-2014 Chicago, USA August 4 -6, 2014 Bryan Yamamoto. Bryan Yamamoto, Ph.D. Department of Neurosciences University of Toledo College](https://reader034.fdocuments.net/reader034/viewer/2022052105/603fe38cd680cb1d34135707/html5/thumbnails/32.jpg)
Summary and Conclusions
� Increases in extracellular glutamate and evidence of excitotoxicity
and oxidative degradation of proteins
� Causes hepatotoxicity and increases ammonia concentrations in
peripheral plasma and brain to mediate excitotoxicity
Effects of METH:
� Damages the BBB via ammonia, oxidative stress, and
inflammation (COX2-dependent prostaglandin synthesis?)
� Opening of the BBB can render the brain vulnerable to toxins by
that would otherwise by restricted to the periphery.
� Peripheral organ effects should be considered as possible causes
of the neurotoxicity associated with psychostimulant drug use
![Page 33: Addiction Therapy-2014€¦ · Addiction Therapy-2014 Chicago, USA August 4 -6, 2014 Bryan Yamamoto. Bryan Yamamoto, Ph.D. Department of Neurosciences University of Toledo College](https://reader034.fdocuments.net/reader034/viewer/2022052105/603fe38cd680cb1d34135707/html5/thumbnails/33.jpg)
� Jeffrey Brown, Ph.D.
� Kristan Burrows Cline, Ph.D.
� David Eyerman, Ph.D.
� Amy Ferng, M.S.
� Laura Halpin, Ph.D.
� Amanda Blaker
� Veronica Chiu, Ph.D.
� Stuart Collins
� Nicole Harless
� Katelyn Marchal
Collaborators
Methamphetamine ProjectPast and Current
Other Current Lab Members
� Laura Halpin, Ph.D.
� John Irlam, D.O.
� J. F. Nash, Ph.D.
� Arunan Nadarajah, Ph.D.
� Nicole Northrop, Ph.D.
� Robert Staszewski, M.S.
� Despina Tata, Ph.D
� Katelyn Marchal
� Carmen Mitchell
� Reka Natarajan, Ph.D.
� Allen Schroering, M.S.
� Erin Semple
� Branden Stansley
NIH grants DA007606, DA016866, DA035499
![Page 34: Addiction Therapy-2014€¦ · Addiction Therapy-2014 Chicago, USA August 4 -6, 2014 Bryan Yamamoto. Bryan Yamamoto, Ph.D. Department of Neurosciences University of Toledo College](https://reader034.fdocuments.net/reader034/viewer/2022052105/603fe38cd680cb1d34135707/html5/thumbnails/34.jpg)
Meet the eminent gathering once again at
Addiction Therapy-2015Florida, USA
August 3 - 5, 2015
Addiction Therapy – 2015 Website:
addictiontherapy.conferenceseries.com
August 3 - 5, 2015