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Cardiotoxicity of Hyperamylinemia
β-Cell Dysfunction & Apoptosis
Pancreatic β-CellOD
AMYLIN
BLOO AMYLIN
OLIGOMERIZATION HYPERGLYCEMIAAmylin Oligomers
HYPERAMYLINEMIA HYPERINSULINEMIACalcium
d sreg lation
Hypertrophy, Remodeling Heart Failure
dysregulation
Florin Despa Department of PharmacologyUniversity of California, Davis
Patients with Obesity and Type-2 Diabetes Present Cardiac Amylin AccumulationA source of cardiomyocyte Ca2+ dysregulationA source of cardiomyocyte Ca dysregulation
Collaborators Lab MembersCollaboratorsKenneth B. Margulies (UPenn)Heinrich Taegtmeyer (UTHS)
Lab MembersKaleena JacksonKatie GuglielminoA d NDonald Steiner (U of Chicago)
Sanda Despa (UC Davis)Donald M Bers (UC Davis)
Andy NguenTracy TangTerry PangDonald M. Bers (UC Davis)
Anne Knowlton (UC Davis)Peter Havel (UC Davis)
Terry Pang
Funds: AHA, NSF, Vision Grant - UC Davis
sr seas
e3
HYPOTHESIS
Type
-2
iabe
tes
sulin
Ris
k fo
rdi
ac D
is
2
ulinulin tanc
e
T Di
InRC
ard 1
Ctl InsuInsu
Res
isHyperglycemiaDyslipidemiay
cell
HF
eatic
β-c
Hyperinsulinemia
panc
re Hyperamylinemia
BLOODToxic Amylin Oligomers
RATIONALE: selection of human heart samples
LeanNon-Failing Heartsg
Lean, Non-diabetesFailing Hearts
AmylinOligomers
Failing Hearts
Obese/OverweightNon-Failing Hearts
Obese/OverweightFailing Hearts
AmylinOligomers
gdistinct amylin oligomer
size distributions
Type-2 DiabetesFailing Hearts
Failing Hearts
Failing Hearts
Amylin Oligomers Accumulate in HeartFailing vs. Non-failing
L NF OW/OB-HF
Anti-Amylin Antibody TAmylin Trimers
400
500 **
**Con
trol
)
L-NF OW/OB-HF 50
15 tetrameroctamer
200
300
400
B-N
F
B-H
F
F
** **
rimer
s (%
C
10 kDa
15
trimer
tet a e
0
100
L-N
F
L-H
F
OW
/OB
OW
/OB
DM
-HF
Am
ylin
T
16-MER FAILING
Larger Amylin Oligomers
400
500
**
Con
trol
)OCTAMER
TETRAMERNON-FAILING
200
300
F
*
OW
/OB
-N
F
OB
-HF
n Le
vel (
% C
DIMERTRIMER NON-FAILING
0
100L-
NF
L-H
F
DM
-H
O
OW
/O
Am
ylin
Cardiac Amylin Deposition in Type-2 Diabetic Humans
PANCREAS HEARTControlHEART HEART
20 µm20 µm 20 µm20 µm
tibod
y
aini
ng
HEARTHEART
myl
in A
nt
o R
ed S
ta
Ant
i-Am
Con
g
20 µm20 µm
Amylin Oligomers Circulate Through the Blood Obese Diabetes and Kidney FailureBlood Obese, Diabetes and Kidney Failure
Obesity and Type-2 Diabetes With Kidney Failure
160 *
%)
160**
%)tro
l)
150
175
)
T2DM
T2DM
T2DM
T2DM
T2DM
obese
Obesity and Type 2 Diabetes With Kidney Failure
80
120
BMW
IAPP
(%
80
120
MW
IAPP
(%ev
el (%
Con
75
100
125
150
P le
vel (
a.u.
)
HealtyBMI < 25
M M MM
0
40
Ctl OW
/OB
Hig
h M
0
40
Ctl DMH
igh
MIA
PP L
e
L
L
CE12
0E16
0E17
0E18
0E19
0E11
0E13
0E15
0
0
25
50
IAPP
E E E E E E E E
Acute Effect of Amylin on Isolated Cardiac MyocytesMyocytes
Human: KCNTATCATQRLANFLVHSSNNFGAILSSTNVGSNTY
Rat: KCNTATCATQRLANFLVRSSNNLGPVLPPTNVGSNTY
0 6
0.8
cay
(s)
1500 Controlh-amylinr-amylin)
***
0.4
0.6
nient
Dec
500
1000 Systolic***
Ca]
i (nM
)
******
0 0
0.2
Ctl
h-am
ylin
r-am
ylin
a Tr
ansi
0
500
Diastolic
[C
0.0Ca0.0 0.5 1.0 1.5 2.0
Frequency (Hz)
Amylin Oligomers Alter the Structure of Sarcolemma in Isolated Cardiac Myocytes
Thaps+0Ca/0Na200 +carboxyeosin
4
5
*/s)
Sarcolemma in Isolated Cardiac MyocytesPassive Ca2+ Leak
Ctl+ ] i (n
M)
2
3
4
eak
(nM
/
501 min
CtlAmylin
[Ca2
0
1
2
Ctl
Am
ylin
Ca2+
l
2.5 µm 2.5 µm
control
Animal Models
Rat Amylin
pancreas
Human Amylin
pancreas pa c eas
UCD-T2DM rat
pa c eas
HIP rat
Non-fasting Blood Glucose Level
180
200
cose
l)
Non fasting Blood Glucose Level
120
140
160
CD
-T2D
M
IPBlo
od G
lu(m
g/d l
100 UC
HIB
Amylin Accumulates in the Heart of HIP Ratst
UC
DH
IP ra
t D-T2D
M
20 µm20 µm
M rat
HeartPancreas Heart Pancreas
25 kDa64
Lysates DMPD
10 kDa
15 kDa
16kD
32
HIP UCD-T2TM
kDaHIP rat
Myocardial Insulin Signaling in HIP vs. UCD T2DM ratsUCD-T2DM rats
2.0
1.5t (
a.u.
)
0 5
1.0
Akt
/Akt
0.0
0.5
Ctl
UC
DHI
P
0
p-A
100 10Insulin: 0 0 10 10 0 0 10 10 10 10 10 100 0
Ctl HIP UCD
AktpAkt
Akt
Cardiac Amylin Oligomer Accumulation Alters Ca2+ Cycling in HIP Rats
1500 CtlHIP
*** 1500 CtlUCD
HIP rat UCD raty g
1000
HIP
Systolic******
a]i (
nM)
1000Systolic
UCD
a]i (
nM)
0 0 0 5 1 0 1 5 2 00
500
Diastolic***
***[Ca
0
500Diastolic
[Ca
diastolic 0.0 0.5 1.0 1.5 2.0
Frequency (Hz)0.0 0.5 1.0 1.5 2.0
Frequency (Hz)
0 8 * 0 8
dysfunction
0.4
0.6
0.8
ay ti
me
(s) *
0.4
0.6
0.8
ay ti
me
(s)
0.0
0.2Dec
a
Ctl
HIP
0.0
0.2Dec
a
Ctl
UC
D
Contractile Dysfunction in Pre-diabetic HIP RatsHIP Rats
80
120
*
P (m
mH
g)
6000
8000
mH
g/s)
Maximum Rate of P Ri
40
80
IP IPd Sy
stol
ic P
2000
4000
IP IPP/dt
max
(mmPressure Rise
0 RI HI
End
Left-Ventricular End Systolic Pressure
0 RI HId
/s)
8mH
g)
Left-Ventricular End Diastolic Pressure
4000
6000*
min
(mm
Hg/
Maximum Rate of Pressure Fall
4
6
8
tolic
P (m
m0
2000
RIP
HIP
-dP/
dtm Pressure Fall
0
2
RIP
HIP
End
Dia
stcollab. with A. Knowlton (UC Davis)
Cardiac Amylin Oligomer Accumulation Triggers Remodeling of SERCA
CtlPDSERCA *
Ctl DMPD
Triggers Remodeling of SERCA
DMPDSERCA
PLB
**DMCtl PD
HIP rat
NCX*
DMCtl PD
0 40 80 120
*
% vs. Ctl120
Ctl)
40
80
RC
A (%
vs.
C
DMCtl PDUCD rat
0 DMCtl
PDSER