ROLE OF HEME OXYGENASE-1 (HO) & CARBON MONOXIDE (CO)
IN TRANSPLANTATION
Hun-Taeg Chung, M.D., Ph.D.Meta-inflammation Research Laboratory,
School of Biological Sciences,University of Ulsan, Ulsan 680-749, Korea
CONTENTS
A Introduction
B Gaseous Transmitters
C Physiological Function of HO system
D Therapeutic Effects of HO-1 System in Transplantation
E ConclusionD-1. HO-1 and transplantationD-2. CO and transplantationD-3. Br/Bv and transplantationD-4. HO-1 and IRI
CO
Fe2+
BiliverdinHeme Bilirubin
2NADPH3O2
2NADP+
3H2O
NADPHH+
NADP+
Heme Oxygenase
Biliverdin Reductase
Introduction
Introduction and COCO : Its Chemical Properties
:CO:- +
Carbon monoxide (CO) is an odorless,
tasteless and colorless gas composed
of one carbon atom and one oxygen
atom with a triple bonding.
Due to its formal charges, CO is more
reactive than oxygen molecule, but
not more than nitric oxide.
Degradation of Heme by HO-1and HO-2
Fe
CO
BILIRUBIN
HEME
HEMOGLOBIN
Bone marrowDestruction of maturing
erythroid cells
SpleenDestruction of senescent
erythrocytes
All tissuesTurnover of heme
& hemoprotein
HO-1or
HO-2
Introduction and HO-1 & HO-2
Heme
CO
Fe2+Bilirubin
Iron homeostasisAnti-apoptosisCytoprotection
AntioxidantAnti-apoptosisCytoprotection
Anti-apoptosisCytoprotection
Anti-inflammationAnti-proliferation
HO-1
AntioxidantsAnti-inflammatory cytokines
Heat shock proteins
NOSome drugs
HO-1 as a Therapeutic Funnel
CO/HO-1
Up-regulation
Down-regulation
ROS
Vascular permeability
Adhesion molecules
Cell proliferation
Apoptosis
cGMP
Cytoprotection
Angiogenesis
Tissue repair
Resolution
Trans-
plantation
?
HO-1/CO : Its Biochemical Roles
Introduction and HO-1/CO
Landscape View of Gas Biology
Gaseous Transmitters and General Properties
1. The gases are highly membrane-permeable and thus serve as a substance
conveying the signal from one site to another
Important Gaseous Transmitters in Mammalian Cells:
Nitric Oxide (NO), Carbon Monoxide (CO)
& Hydrogen Sulfide (H2S)
3. The gases could not only exert comparable biological actions but also compete
with and are antagonists with each other
2. The gases could exert their biological actions via interaction with proteins in
multiple ways
A unique class of biomaterials indispensable of maintaining the homeostasis of biological system
Criteria for Aerokines (Gaseous Transmitters)
(1) Small molecules of gas, like NO, CO and H2S.
(2) Freely permeable to membrane.
Their effect will not rely on cognate membrane receptors.
(3) Endogenously and enzymatically generated and their generation
is regulated.
(4) Well-defined specific functions at physiologically relevant concentrations.
(5) Mediated by second messengers, but should have specific cellular and
molecular targets.
Gaseous Transmitters and NO, CO and H2S
Synthesis & Significances
Enzyme SubstrateGaseous Mediator
Cystathionine -Synthase (CBS)
Cystathionine -Lyase (CSE)dHydrogen Sulfide (H2S)c L-Cysteine (L-Cys)
Nitric Oxide (NO)a
Carbon Monoxide (CO)b
NO Synthase (NOS)
[e.g. iNOSd, eNOS, nNOS]
Heme Oxygenase (HO)
[e.g. HO-1d, HO-2, HO-3]
L-Arginine (L-Arg)
Free Heme
Different gases that are synthesized by enzymes in the body could not only exertcomparable biological actions but often compete with and are antagonists witheach other
a Radical species with high reactivity, short half-life, colorless & odorlessb Gas with high binding affinity with hemoglobin, colorless & odorlessc Hydrophilic and flammable gas, colorless & characteristic smell of rotten eggsd Inducible
iNOS knockout
Immune deficiency
Endothelial dysfunction
HO-1 knockout
Chronic inflammation
Endothelial dysfunction
CSE knockout
Hyperhomocysteinemia
Endothelial dysfunction
Gaseous Transmitters and Enzymes and Substrates
Activation of Secondary Messenger
H2S
H3O+H2O +H2S
1. Activation of cAMP
2. Reaction with H2O
CO
Fe
Fe
CO
CO
O2
1. Activation of sGC
2. Inhibition of CYP 450NO
Fe
Fe
NO
NO
ONOO-O2-+NO
RS-ONRSH
NO
3. Modulation of thiol enzymes
1. Activation of sGC
2. Formation of peroxynitrite
CYP 450
Substrate
Product
CO
+ HS-
Adenyl cyclase
HS-
cAMP
AMP
PKA
CBS
CSE
L-Cys
NOS
L-Arg
HO
Biliverdin
HemeFe2+
Reducing cytotoxic homocycteineReducing pathological antigen Reducing inflammatory free heme
Gaseous Transmitters and Signaling Pathways
Hypothetical Interplay among CO, NO and H2S
NO/NOS
H2S/CBSCO/HO-1&CSE
InhibitionStimulation
Gaseous Transmitters and Interactions among Aerokines
Possible Mechanisms underlying HO-1 Inductionby NO and NO Derivatives in Eukaryotic Cells
HO-1 Induction by NO and Its Derivatives
antioxidant
antinitrosative
NO + O2- ONOO-
Transcriptional Activator ?
HO-1
decreased NOS
Protein levels
decreased NOS
activity
cell growth
vessel tone
OS =Oxidative stress
NS =Nitrosative stress
iNOS
induction
NO
donors
Bilirubin CO
cGMP
Thiol depletion
OS
NSOS
NS
Gaseous Transmitters and Interactions among Aerokines
CO Induces Nrf2 activation and HO-1 Expression
Nrf2/CF
Nrf2/NF
HO-1
Actin
-
-
-
-
5
-
-
-
10
-
-
-
20
-
-
-
-
+
-
-
5
+
-
-
10
+
-
-
-
-
5
-
RuCO (M)
Hb
RuCl2 (M)
CO gas
-
-
10
-
-
-
20
-
-
-
-
+
20
+
-
-
RuCO (M)
Hb
RuCl2 (M)
CO gas
5
-
-
-
1
2
0
10
-
-
-
20
-
-
-
-
+
-
-
10
+
-
-
20
+
-
-
-
-
5
-
-
-
10
-
-
-
-
+
-
-
-
-
5
+
-
-
Fold
in
du
ctio
n Nrf2/NF
Nrf2/CFHO-1
-
-
20
-
*
*
* *
*
**
*
*
*
*
*
CO
Nrf2
HO-1
In HUVECs, CO donor and CO gas induce Nrf2 nuclear translocation and HO-1
expression in a dose-dependent manner. However, in the presence of the CO
scavenger, Nrf2 activation and HO-1 expression are not observed.
Gaseous Transmitters and Interactions among Aerokines
Proved Interplay among CO, NO and H2S
H2S
CO
NO
CO
Hypothesized triangular
interactions among CO, H2S and
NO in vascular tissues. The solid
line ( ) indicates the stimulatory
input and the dashed line ( ), the
inhibitory input.
H.T. Chung et al. Current pharmaceutical design, 2008, 14(5):422-428
Gaseous Transmitters and Interactions among Aerokines
NO, CO & H2S ; Their Interplay in Biological System
CO
HO-1
NO
iNOS
H2S
CSE
Gaseous Transmitters and Interactions among Aerokines
Initial Signals of HO-1/CO
Bilban M, et. al. J. Mol. Med. 2008, 86:267-279.
Gaseous Transmitters and Physiological Roles of HO-1/CO
Anti-Apoptosis, -Inflammation, -Proliferation & -Thrombosis
Ryter S.W. et. al. Physiol. Rev. 2006, 86:583-650.
Gaseous Transmitters and Physiological Roles of HO-1/CO
Anti-Inflammation/Up (IL-10) & Down (TNF-α, IL-1β & MIP-1β) Regulation
Otterbein L.E. et. al. Nat. Med. 2003, 6;422-428.
Gaseous Transmitters and Physiological Roles of HO-1/CO
Anti-Proliferation/Up (EC) & Down (SMC) in Balloon Injury
HO-1/COControl
3 Days
2 Weeks
2 Weeks
Gaseous Transmitters and Physiological Roles of HO-1/CO
Pro-Survival Effects on Tissues/Cells
Wang X et. al. J. Biol. Chem. 2007, 282(3):1718-1726.
Gaseous Transmitters and Physiological Roles of HO-1/CO
CO Protects ECs from ER-Stress-Induced Apoptosis by Blocking CHOP via p38
p-JNK
p-p38
Actin
p-ERK1/2
- 1 5 10
RuCO (M)
p-p38
Actin
0 10 20 40 (min)
RuCO 10M
20
40
60
80
120
0
100
* *
TG
RuCO
SB203580
+
-
-
+
+
-
+
+
+
-
-
-
Via
bil
ity (
%)
-
-
+
*
CHOP
Actin
HO-1
+
+
+
TG
RuCO
SB203580
-
-
-
+
-
-
+
+
-
Nrf2/Keap 1eIF2
ATF4 Nrf2
PERK
HO-1CHOP siRNA
TG/HCysCO
Keap 1
Cell SurvivalCell Death
CHOP
p38 Nrf2/Keap 1eIF2
ATF4 Nrf2
PERK
HO-1CHOP siRNA
TG/HCysCO
Keap 1
Cell SurvivalCell Death
CHOP
p38
The p38 inhibitor SB reverses CO-induced inhibition of CHOP expression and apoptosis induced by ER stress.
Gaseous Transmitters and Physiological Roles of HO-1/CO
Integrative Stress Response vs. Integrative Survival Response
PERK PKR GCN2 HRI
ER Stress dsRNA AA Starvation Carbon Monoxide
eIF2 kinase
Anti-apoptotic
Molecules
CHOP
eIF2-P
Heme Deficiency
PERK
eIF2 kinase
ATF4
CHOP
eIF2-P
ATF4
Apoptosis
Anti-apoptotic
Molecules
Apoptosis
Integrative Survival ResponseIntegrative Stress Response
Differential Points
Between
I. Str. R. and I. Sur. R.
Gaseous Transmitters and Physiological Roles of HO-1/CO
HO-1 Expression in Regulatory T Cells
CD4+
CD4+CD25-
CD4+CD25+98.5%
0.5%
7.4%
CD4-FITC
CD
25
-PE
0 3 6 12 24 0 3 6 12 24
CD4+CD25- CD4+CD25+
CD
4+
CD
4+C
D2
5-
CD
4+C
D2
5+
Time (h)
HO-1
-actin
HO-1
-actin
Stimulated
Resting
Resting Stimulated
HO-1
CD4+
CD4+CD25-
CD4+CD25+
0
100
200
300
400
Pro
life
rati
on (
cpm
x 1
03)
Pae HO, et al. Differential expressions of heme oxygenase-1 gene in CD25- and CD25+ subsets
of human CD4+ T cells. Biochem Biophys Res Commun. 2003;306(3):701-705.
HO-1/CO in Immune System and Effects of HO-1/CO on Processing
- +
Actin
-
HO-1
+ CoPP
* *
0
20
40
60
80
100
--
+-
++
CD3/CD28CoPP
[H3]
TdR
Upta
ke
(cpm
/wel
l x
10
-3)
0
10
20
30
40
50
--
+-
* *
++
[H3]
TdR
Upta
ke
(cpm
/wel
l x
10
-3)
CD3/CD28CoPP
CD4+ T cellsPBMCs
(B)(A)
Jurk
at/
pcD
NA
Actin
HO-1
Jurk
at
Jurk
at/
HO
-1
Jurk
at/
pcD
NA
40
30
20
10
0
50
Jurk
at
Jurk
at/
HO
-1
[H3]
TdR
Up
take
(cpm
/wel
l x
10
-3)
*
(C)
Actin
HO-1
CoPP
1. Pae HO, et al. Carbon monoxide produced by heme oxygenase-1 suppresses T cell proliferation via inhibition
of IL-2 production. J Immunol. 2004;172(8):4744-4751.
2. Pae HO, et al. Roles of heme oxygenase-1 in the antiproliferative and antiapoptotic effects of nitric oxide on
Jurkat T cells. Mol Pharmacol. 2004;66(1):122-128.
Anti-proliferative Effect of HO-1 Expression on T Cells
HO-1/CO in Immune System and Effects of HO-1/CO on Processing
Anti-proliferative Effect of CO on TCR-stimulated T Cells
1. Pae HO, et al. Carbon monoxide produced by heme oxygenase-1 suppresses T cell proliferation via inhibition
of IL-2 production. J Immunol. 2004;172(8):4744-4751.
2. Pae HO, et al. Roles of heme oxygenase-1 in the antiproliferative and antiapoptotic effects of nitric oxide on
Jurkat T cells. Mol Pharmacol. 2004;66(1):122-128.
*
0
20
40
60
80
100
[H3]
TdR
Upta
ke
(cpm
/wel
l x
10
-3)
CO gas
Fe2+
Bilirubin
-
-
-
+
-
-
-
+
-
-
-
+
(A) (B)
% I
nhib
itio
n
RuCO (M)
RuCO + HbRuCO
0
20
40
60
80
100
0 20 40 60 80
(C)
0
20
40
60
80
100
CD3/CD28
CoPP
Hb
+
-
-
+
+
-
+
+
+
* *
[H3]
TdR
Upta
ke
(cpm
/wel
l x
10
-3)
HO-1/CO in Immune System and Effects of HO-1/CO on Processing
CO Blocks Early Signals of TCR-stimulated T Cells
0
12
3
4
5
6
7
0
1
2
34
56
7
0
12
3
0
CD3/CD28
RuCO After CD3/CD28
RuCOBefore CD3/CD28
Medium
Even
t
CFSE
(A) (B)Medium CD3/CD28
RuCO Before CD3/CD28
Cel
l N
um
ber
Relative DNA Content
G0/G1
G2/M
RuCO After CD3/CD28
1234567 0
HO
-1
CFSE
Division number
(C)
Pae HO, et al. Carbon monoxide produced by heme oxygenase-1 suppresses T cell proliferation
via inhibition of IL-2 production. J Immunol. 2004;172(8):4744-4751.
HO-1/CO in Immune System and Effects of HO-1/CO on Processing
Transfer of Foxp3 Gene into Tres Mimics Treg Functions
Vec
tor/
Tre
s
Fre
sh T
reg
Fre
sh T
res
FO
Xp
3/T
res
Fre
sh T
reg
+ F
resh
Tre
s
FO
Xp
3/T
res
+F
resh
Tre
s
(Tra
nsw
ell)
FO
Xp
3/T
res
+ F
resh
Tre
s
FOXp3/Tres
Fresh Treg
Fresh Tres
(B)
(C)
(A)
SuppressionAnergy
Yagi H, et al. Crucial role of FOXP3 in the development and function of human CD25+CD4+ regulatory T cells.
Int Immunol. 2004;16(11):1643-1656.
HO-1/CO in Immune System and Effects of HO-1/CO on Regulation
Transfer of Foxp3 Gene Induces HO-1 Expression
0.00Rel
ati
ve
Cel
l P
roli
fera
tio
n
0.50
1.00
Jurk
at
1.25
Jurk
at/
pcD
NA
Jurk
at/
Fo
xp
3
*
0.75
0.25
C
B
Rel
ati
ve
Fo
ld A
ctiv
ity
1
0
Jurk
at
Jurk
at/
Fo
xp
3
Jurk
at/
pcD
NA
*
2
3
4
-actin
Jurk
at
Jurk
at/F
oxp
3
Jurk
at/p
cDN
A
HO-1
D
IL-2
(p
g/m
L)
50
0
Jurk
at
Jurk
at/
pcD
NA
Jurk
at/
Fo
xp
3
*
100
150
200
Foxp3
Jurk
at
Jurk
at/F
oxp
3
Jurk
at/p
cDN
A
-actin
A
IFN
- (p
g/m
L)
25
0
Jurk
at
Jurk
at/
pcD
NA
Jurk
at/
Fo
xp
3
50
100
75
*
Choi BM, et al. Critical role of heme oxygenase-1 in Foxp3-mediated immune suppression.
Biochem Biophys Res Commun. 2005;327(4):1066-71.
HO-1/CO in Immune System and Effects of HO-1/CO on Regulation
HO-1 Induced by Foxp3 Inhibits T Cell Proliferation
Rel
ati
ve
Cel
l P
roli
fera
tion
0.0
1.0
Jurk
at
1.5
Jurk
at/
pcD
NA
Jurk
at/
Fo
xp
3
NoneIL-2
*
0.5
C
Rel
ati
ve
Cel
l P
roli
fera
tion
0.0
1.0
Jurk
at
1.5Ju
rkat
/ p
cDN
A
Jurk
at/
Fo
xp
3
NoneZnPPSiRNA
**
0.5
A
*
0
IL-2
(p
g/m
L)
100
150
Jurk
at
200
Jurk
at/
pcD
NA
Jurk
at/
Fo
xp
3
NoneZnPPSiRNA
50
B
**
*
Anergy
Choi BM, et al. Critical role of heme oxygenase-1 in Foxp3-mediated immune suppression.
Biochem Biophys Res Commun. 2005;327(4):1066-71.
HO-1/CO in Immune System and Effects of HO-1/CO on Regulation
HO-1 Induced by Foxp3 Suppresses T Cell Proliferation
Rel
ati
ve
Cel
l P
roli
fera
tion
0.0
1.0
Jurk
at
alo
ne
1.5
+Ju
rkat
/ F
oxp
3
NoneZnPPSiRNA
*0.5
**
A
Rel
ati
ve
Cel
l P
roli
fera
tion
0.0
1.0
1.5
0.5
*
NoneIL-2
B
Jurk
at
alo
ne
+Ju
rkat
/ F
oxp
3
Rel
ati
ve
Cel
l P
roli
fera
tion
0.0
1.0
1.5
0.5
C
Jurk
at
alo
ne
+Ju
rkat
/ F
oxp
3
+Ju
rkat
/ F
oxp
3
(tra
nsw
ell
)
Suppression
Choi BM, et al. Critical role of heme oxygenase-1 in Foxp3-mediated immune suppression.
Biochem Biophys Res Commun. 2005;327(4):1066-71.
HO-1/CO in Immune System and Effects of HO-1/CO on Regulation
Tres Treg
APC
MHC
TCRB7
CD28
MHC
TCR B7
CTLA-4
Tryptophan
KynureninesIDO
HO-1
Heme
Fe2+
COBiliverdin
IL-2
IFN-
IL-10
TGF-
IL-2
HA
An Integral Role for HO-1/CO in Maintaining Peripheral Tolerance by CD4+CD25+ Regulatory T Cells
Brusko et. al. Journal of Immunology 2005, 174:5181-5186.
HO-1/CO in Immune System and Effects of HO-1/CO on Regulation
Involvement of HO-1/CO in Generation/Function of Treg Cells
HO-1/CO in Immune System and Effects of HO-1/CO on Regulation
T-Cell
APC
Monocyte
Th1
CO
CO
Cytokine
AICD
COCO
HO-1
Enhancing
Blocking
Treg
Tres
CO
CO
HO-1
NO H2S
IFN-
Tres
IL-10
Treg
As Inducer
As Inhibitor
Interactions of Gases, Cytokines & Cells in the Immune Response
HO-1/CO in Immune System and Effects of HO-1/CO on Regulation
Protective Effects of HO-1/CO on RA & Autoimmune Hepatitis
Co
llag
en
+ C
oP
P
Co
ntr
ol
Co
llag
en
SnPP + Con ACORM + Con A
Control Con A
HO-1/CO in Immune System and Effects of HO-1/CO on Disorders
HO-1
Nrf2
MAPKs
Inducers
Heme
DC
T Cell
MC
Eos
Sensitization
Th2 Cell Proliferation
Mediator Release
Inflammation & Oxidative Stress
Allergen
CO/Nrf2
Fe2+/Ferritin
BV,BR/NF-B Antioxidant/Anti-inflammatory
An
ti-a
lle
rgic
B Cell IgE Production
DC: Dendritic Cell
MC: Mast Cell
Eos: Eosinophil
Re-e
xp
os
ure
Multi-targeted Effects of HO-1 on Allergic Inflammation
HO-1/CO in Immune System and Effects of HO-1/CO on Disorders
Inhibitory Effects of HO-1/CO on Contact Hypersensitivity
Control
SnPP + DNFB
CoPP + DNFB
DNFB
0
5
10
15
20
25
E
ar-s
wel
lin
g r
esp
on
se (
1 x
10
-2m
m)
+-+
DNFBSnPP
CoPP/CO
---
-+-
-++
++-
+--
30
GAPDH
TNF-
IL-1
IL-4
IFN-
IL-5
IL-2
DNFBCoPP/CO
SnPP
---
+--
++-
+-+
Actin
HO-1
Veh
icle
Co
PP
Veh
icle
Co
PP
- DNFB + DNFB
HO-1/CO in Immune System and Effects of HO-1/CO on Disorders
Inducersor
DonorsSensors & Signals Biological Effects Tissue Protection
GasCORM
LPSTNF-α
HypoxiaNO & CO
COFromHeme
Oxygenase
Vasoregulation
Anti-inflammation
Anti-proliferation
Anti-thrombosis
Anti-apoptosis
IR-
injury
Endotoxemia
Asthma/COPD
Bleomycin-
and ventilator-
induced lung injury
Oxidative lung injury
Lung transplantation
Hemoprotein
sGC cGMP
MtROS MAPK (p38)
KCa
eNOS
HO-1/CO AS REHABILITATING MOLECULES
CO-induced Pathways of Potential Therapy in Lung Diseases
Potential Preventive Roles in Sepsis
In response to LPS, HO-1-deficient
mice, as compared with wild-type
mice, exhibit greater impairment in
renal hemodynamics.
Also, HO-1-deficient mice exhibit an
exaggerated induction of pro-
inflammatory cytokines in the kidney
in response to LPS.
HO-1 deficiency exaggerates
activation of NF-B, whereas HO-1
over-expression reduces NF-B
activation.
Tracz et. al. Am J Pathol. 2007;170(6):1820–1830.
LPSHO-1
CO/BR
Uncontrolled
Inflammation
NF-B?
HO-1
CORM
CO gas ?
Tissue
Damage
??
Endogenous
Exogenous
HO-1/CO in Immune System and Effects of HO-1/CO on Disorders
Rat lung transplantation increased HO-1 mRNA expression and protein level
A: Total RNA from the transplanted lung (day 4) was extracted and subjected to Northernblot hybridization with a 32P-labeled HO-1 cDNA Probe
B: Protein from the transplanted lung (day 4) was extracted and subjected to Western blothybridization with HO-1 antibody Am J Pathol 2003, 163:231–242
THERAPEUTIC EFFECTS OF HO-1 SYSTEM ON TRANSPLANTATION
Heme Oxygenase-1 and Transplantation
HO-1 induction in, or administering CO or bilirubin to, the donor, prolonged BALB/c isletssurvival in C57BL/6 recipients following transplantation FASEB J. 21, 3450–3457 (2007)
THERAPEUTIC EFFECTS OF HO-1 SYSTEM ON TRANSPLANTATION
Heme Oxygenase-1 and Transplantation
Combined treatments with CoPP, CO, and bilirubin led to long-term survival of isletallograft FASEB J. 21, 3450–3457 (2007)
Heme Oxygenase-1 and Transplantation
THERAPEUTIC EFFECTS OF HO-1 SYSTEM ON TRANSPLANTATION
Expression of Foxp3, TGF-,and IL-10 in spleens and isletgrafts from recipients carryinglong-term surviving grafts
FASEB J. 21, 3450–3457 (2007)
THERAPEUTIC EFFECTS OF HO-1 SYSTEM ON TRANSPLANTATION
Heme Oxygenase-1 and Transplantation
Detection of Foxp3 expression in islet grafts at 7 days following transplantationby staining with the anti-Foxp3 antibody FASEB J. 21, 3450–3457 (2007)
THERAPEUTIC EFFECTS OF HO-1 SYSTEM ON TRANSPLANTATION
Heme Oxygenase-1 and Transplantation
Diabetes 54:1400–1406, 2005
CO administration only to the donor (●, n = 6), the islet (■, n = 6), or the recipient (▲, n= 6) led to long-term survival of allogeneic islets. A high percentage of grafts survivedlong-term when both donor and recipient were exposed to CO (✖, n = 7) compared withthe controls (○, n = 9).
Carbon Monoxide and Transplantation
THERAPEUTIC EFFECTS OF HO-1 SYSTEM ON TRANSPLANTATION
Gross anatomy of lungs from rats 6 days after transplantation (arrow in A, B),and lungs from rats 6 days after transplantation which received 500 ppm COover this time period (arrow in C, D) Am J Pathol 2003, 163:231–242
Carbon Monoxide and Transplantation
THERAPEUTIC EFFECTS OF HO-1 SYSTEM ON TRANSPLANTATION
CO administration only to the donor suppresses macrophage infiltration to theislet grafts in the recipients Diabetes 54:1400–1406, 2005
THERAPEUTIC EFFECTS OF HO-1 SYSTEM ON TRANSPLANTATION
Carbon Monoxide and Transplantation
Myeloperoxidase activity was decreased in the CO-treated lung fromtransplantation Am J Pathol 2003, 163:231–242
THERAPEUTIC EFFECTS OF HO-1 SYSTEM ON TRANSPLANTATION
Carbon Monoxide and Transplantation
Effect of CO on transplantation-induced TUNEL staining. Markedly elevated TUNELstaining (A, arrow and blue staining in B) occurred in lungs from rats 6 days aftertransplantation compared to lungs (C and D) from transplantation which received 500ppm CO over an equivalent time period. Scale bar, 100 m. Am J Pathol 2003, 163:231–242
THERAPEUTIC EFFECTS OF HO-1 SYSTEM ON TRANSPLANTATION
Carbon Monoxide and Transplantation
CO inhibited transplantation-induced IL-6 mRNA expression and serum IL-6
A: Total RNA from the transplanted lung was extracted and subjected to Northern blothybridization with a 32P-labeled IL-6 cDNA probe
B: Serum was collected 4 days after transplantation and analyzed for IL-6 levels by ELISAAm J Pathol 2003, 163:231–242
THERAPEUTIC EFFECTS OF HO-1 SYSTEM ON TRANSPLANTATION
Carbon Monoxide and Transplantation
CO decreased protein levels of multiple chemokines/cytokines following lung transplantation Am J Pathol 2003, 163:231–242
THERAPEUTIC EFFECTS OF HO-1 SYSTEM ON TRANSPLANTATION
Carbon Monoxide and Transplantation
Biliverdin IXa
Bilirubin IXa
Heme
Two steps of heme degradation
THERAPEUTIC EFFECTS OF HO-1 / Br & Bv and Transplantation
Biliverdin Bilirubin
Biliverdin
Mechanisms of bilirubin/biliverdin action on acute rejection
THERAPEUTIC EFFECTS OF HO-1 / Br & Bv and Transplantation
Exogenous biliverdin administration induces donor specific tolerance to cardiac allografts
THERAPEUTIC EFFECTS OF HO-1 / Br & Bv and Transplantation
Bilirubin induces long-term survival to allogeneic islet allografts
THERAPEUTIC EFFECTS OF HO-1 / Br & Bv and Transplantation
Potential mechanism of the beneficial effect of bilirubin on allogeneic transplanted -cell islets
Bilirubin
Bilirubin
Bilirubin
THERAPEUTIC EFFECTS OF HO-1 / Br & Bv and Transplantation
Bilirubin/biliverdin inhibits proliferation of VSMCs
Bilirubin/Biliverdin
Apoptosis
THERAPEUTIC EFFECTS OF HO-1 / Br & Bv and Transplantation
Bilirubin suppresses neointimal hyperplasia associated with balloon injury
THERAPEUTIC EFFECTS OF HO-1 / Br & Bv and Transplantation
Effects of bilirubin/biliverdin in various animal models of IRI
Liver Kidney
Heart Small bowel
THERAPEUTIC EFFECTS OF HO-1 / Br & Bv and Transplantation
Potential mechanisms of bilirubin/biliverdin improving organ function during/after IRI
Bilirubin/Biliverdin
Apoptosis
Inflammation
THERAPEUTIC EFFECTS OF HO-1 / Br & Bv and Transplantation
Br/Bv Inhibits Acute & Chronic Rejections and IRI
Transplantation
Bilirubin/Biliverdin
THERAPEUTIC EFFECTS OF HO-1 / Br & Bv and Transplantation
Schematic representation of HO-1→CO axis, which acts on the proliferative response tothe benefit of the cell type involved so as to best re-establish homeostasis.
[Cell Cycle 7:10, 1379-1384; 15 May 2008
Heme Oxygenase-1 and Ischemia Reperfusion Injury
THERAPEUTIC EFFECTS OF HO-1 / HO-1 and IRI
Balance between immune effectors and protective gene expression: a working paradigmto determine the fate of the graft. X, Y, Z refer to additional protective proteins. EC,endothelial cells; SMC, smooth muscle cells Transplantation 2006;82: S36–S40
HO-1 inhibits Transplant Arteriosclerosis
THERAPEUTIC EFFECTS OF HO-1 / HO-1 and IRI
A mechanistic scheme how HO-1 may affect IRI in the transplantA: The mechanism of IRI can be attributed to local neutrophil accumulation, lymphocyte/macrophage
activation, and release of pro-inflammatory mediators (ROS, cytokines), which lead to cell injury,and culminate in the graft failure
B: HO enzyme activity degrades heme to biliverdin, CO, and Fe 2+ Annals of Transplantation 2004, 9:84–87
Heme Oxygenase-1 and Ischemia Reperfusion Injury
THERAPEUTIC EFFECTS OF HO-1 / HO-1 and IRI
Transplantation. Vol 74, 905–912 (2002)
HO-1 system: mechanisms of cytoprotection
CONCLUSION
Transplantation. Vol 74, 905–912 (2002)
HO-1 dynamics: Local and Systemic Interactions
CONCLUSION
Protective Roles of HO-1, CO & Br/Bv in Transplantation
Decreased Immune Response
Increased Immune Response
Chemokines
Free Heme
Cytokines
Apoptosis
T cell infiltration
HO-1
Inactivation
HO-1
Activation
Graft Survival
Graft Rejection
Nonself
Antigen
I/R injuryCO
BR
HO-1 metabolizes the
pro-oxidant free heme.
CO/BR prevents I/R
injury and apoptosis.
Expression of
chemokines and
cytokines is reduced by
HO-1 expression and/or
CO/BR treatment.
CO/BR inhibits T cell
activation and
infiltrations into graft
tissue.
CONCLUSION
CO-NO-REsearch Group
Dr. PaeDr. Lee
Dr. Kim
Zheng
Thank you