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1 Department of Pathology, Duke University Medical Center, Durham 27710; and 2 Microarray Center and 3 Laboratory of Signal Transduction, National Institute of Environmental Health Sciences, National Institutes of Health, Research Triangle Park, North Carolina 27709
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ABSTRACT |
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 TOP
 ABSTRACT
 INTRODUCTION
METHODS
RESULTS
DISCUSSION
REFERENCES
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Dilated
cardiomyopathy, a disease of unknown etiology and
pathogenesis, is associated with heart failure and compensatory hypertrophy. Although cell and animal models suggest a role for altered
gene expression in the transition to heart failure, there is a paucity
of data derived from the study of human heart tissue. In this study, we
used DNA microarray profiling to investigate changes in the expression
of genes involved in apoptosis that occur in human idiopathic
dilated cardiomyopathic hearts that had progressed to heart failure. We
observed altered gene expression consistent with a proapoptotic
shift in the TNF-
signaling pathway. Specifically, we found
decreased expression of TNF-- and NF-
B-induced antiapoptotic
genes such as growth arrest and DNA damage-inducible (GADD)45
, Flice inhibitory protein
(FLIP), and TNF-induced protein 3 (A20).
Consistent with a role for apoptosis in heart failure, we also
observed a significant decrease in phosphorylation of BAD at Ser-112.
This study identifies several pathways that are altered in human heart
failure and provides new targets for therapy.
TNF-; GADD45; BAD; gene profiling
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INTRODUCTION |
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TOP
ABSTRACT
INTRODUCTION
METHODS
RESULTS
DISCUSSION
REFERENCES
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REDUCED CARDIAC FUNCTION and/or hemodynamic overload lead to cardiac hypertrophy, which is initially a compensatory response but eventually transitions to heart failure. The mechanism(s) responsible for the development of cardiomyopathy, hypertrophy, and the transition to heart failure is poorly understood (12). Although it has been suggested that an increase in apoptosis contributes to the transition to heart failure (21), there are questions about the role of apoptosis in heart failure (14, 23). Apoptosis is regulated by the balance between the activation of survival pathways versus proapoptotic pathways. Examining the apoptotic state in heart failure is complicated because there are a plethora of pro- and antiapoptotic genes. In addition, it is likely that a shift to a more proapoptotic state would result in compensatory changes to oppose apoptosis. Therefore, to assess the role of apoptosis, it is important to examine alterations in gene expression in a large panel of apoptotic genes. To address the role of apoptosis in heart failure, we used DNA microarray profiling to identify patterns of expression changes in a number of apoptotic genes. We reasoned that if increased apoptosis was causally important in the transition to heart failure, then there would be a proapoptotic shift in the expression of key apoptotic genes. Also, because there are multiple apoptotic pathways, an additional aim was to identify which apoptotic pathways are involved. Elucidation of these pathways would allow development of new targets for therapeutic intervention to reduce myocyte loss.
We performed DNA microarray profiling on four idiopathic
cardiomyopathic hearts in failure versus five pooled control hearts. Interestingly, we found decreased expression of several
antiapoptotic genes that are induced by TNF- via activation of
NF-B. TNF- is elevated in heart failure patients
(28), and cardiac-specific overexpression of TNF-
results in a dilated cardiomyopathy (17). Furthermore,
treatment with a soluble TNF receptor lowered TNF- in patients and
improved left ventricular function (4). TNF- can
stimulate apoptosis; however, an increase in TNF- signaling does not lead to myocyte apoptosis when the NF-B pathway is
activated by TNF- (16, 19), presumably because
activation of NF-B leads to increased expression of several
antiapoptotic genes, such as growth arrest and DNA damage-inducible
(GADD)45, Flice inhibitory protein
(FLIP), and TNF-induced protein 3 (A20)
(3). We report that failing human hearts have
decreased expression of the TNF-- and NF-B-induced
antiapoptotic genes GADD45, FLIP, and
A20 and increased expression of TNF receptor superfamily
member (TNFRSM) 10 [TNF-related apoptosis-inducing ligand
(TRAIL)]. These data suggest that decreased expression of
NF-B-induced antiapoptotic genes may be important in the shift
in TNF- signaling toward enhanced apoptosis during heart failure.
We also examined the phosphorylation status of bcl-2 and BAD and found a significant decrease in phospho-BAD (Ser-112) in failing hearts. This decrease in phospho-BAD would also be consistent with a proapoptotic shift in heart failure. To our knowledge, this is the first report of decreased levels of phospho-BAD in failing human myocardium.
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METHODS |
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TOP
ABSTRACT
INTRODUCTION
METHODS
RESULTS
DISCUSSION
REFERENCES
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Tissue collection.
Human left ventricular myocardium was obtained from explanted
cardiomyopathic hearts from patients with end-stage heart failure undergoing heart transplantation, following Institutional Review Board
guidelines. These patients were on a variety of medications that could alter gene expression. Heart tissue was snap frozen in
liquid nitrogen within 15 min of excision and remained frozen in liquid
nitrogen until RNA extraction. The microarray study samples were
obtained from four idiopathic cardiomyopathic hearts (all males, mean
age ± SE = 54 ± 4 yr). Table
1 provides details of the patient's age,
sex, and diagnosis and the assays performed on each sample. One
idiopathic failing heart (heart 004) was from a patient on a
left ventricular assist device (LVAD). For the microarray study, we
also included a pooled sample from seven ischemic
dilated cardiomyopathic hearts (all males, 56 ± 3 yr). Samples were taken from the left ventricle in noninfarcted, nonfibrotic areas. For the real-time PCR validation, we also included an additional two idiopathic cardiomyopathic hearts from patients on LVADs for RT-PCR
analysis (males, age 45 and 60 yr). For the phospho-BAD Western blot
measurement, these two hearts from LVAD patients were also included
along with an additional seven idiopathic cardiomyopathic hearts.
Nonfailing left ventricular myocardium was obtained from donor hearts
(National Disease Resource International; Philadelphia, PA) that could
not be used for transplantation due to technical problems or the age of
the donor. All nonfailing hearts were normal in size and had no grossly
apparent infarcts. Random sections had normal histology. Nonfailing
samples were obtained from five males and three females. Of the eight
donor hearts, an echocardiogram was performed on three hearts and was
normal in two of three hearts. The ejection fraction was 30% in the
heart with the abnormal echocardiogram, and this patient was given
dopamine after an intracranial hemorrhage. RNA was isolated from the
tissue using a Qiagen RNeasy kit. Formaldehyde gel electrophoresis was
run to assess RNA quality; for all samples, the 28S RNA band was twice
the intensity of the 18S RNA band.
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Microarray hybridization experiments. cDNA microarray chips containing 1,920 human cDNAs were prepared using previously described protocols (9). cDNAs (Research Genetics; Huntsville, AL) were derived from the 3' end of the gene and ranged in size from 500 to 2,000 base pairs. Briefly, vector primers were used to amplify insert cDNAs from purified plasmid DNA in a 100-µl PCR. The PCR products were run on 2% agarose gels to ensure quality of the reactions and purified by ethanol precipitation. We routinely perform resequencing of our clones to confirm their identity. Updated clone information is available on the web site http://dir.niehs.nih.gov/mirroarray/clones. The purified cDNAs were resuspended in ArrayIt buffer (Telechem; San Jose, CA) and spotted in house onto poly-L-lysine-coated glass slides using a modified, robotic DNA arrayer (Beecher Instruments; Bethesda, MD). Total RNA (35 µg) was labeled with Cy3- and Cy5-conjugated dUTP (Amersham; Piscataway, NJ) using a reverse transcription reaction and hybridized to the cDNA microarray chip. cDNA chips were scanned using an Axon Scanner (Axon Instruments; Foster City, CA), and images were analyzed using the Array Suite Software (Scanalytics; Fairfax, VA). The relative fluorescence intensity with subtraction of the local background values was measured for each labeled RNA, and a ratio of the values for the intensity of each fluor bound to each probe was calculated. The amount of autofluorescence generated in the Cy3 channel was measured, and a minimum intensity cutoff was set just above this value. The distribution of the ratio of all of the genes was analyzed and normalized based on the median value of a panel of 84 control genes. The distribution of the ratio of all of the genes was then calculated, and intensity ratio values that differed from the median with a confidence interval >95.0% (7) were scored as significant changes.
The probability of detecting a change in gene expression by random chance was calculated by binomial probability (5). With the use of a microarray chip containing 1,920 genes, in a single hybridization, one would expect 240 outliers by random chance; triplicate hybridizations reduces this number to a value <1 occurring in all three hybridizations. Each RNA was labeled and hybridized in three or four independent reactions with each RNA sample labeled with each fluor (Cy3 and Cy5). The coefficient of variance for each hybridization was <0.2. A database tool, MAPS (11), was used to compile the overall list of genes that were consistently and significantly changed across the multiple hybridizations of failing versus nonfailing hearts. To perform hybridizations of all five nonfailing hearts versus all four failing hearts in triplicate would require 60 hybridizations; because this was not feasible, we ran each failing heart versus the pooled nonfailing sample in triplicate or quadruplicate. The pooled nonfailing sample was composed of nonfailing hearts 001, 021, 026, 031, and 032. We evaluated the variability among the nonfailing hearts by hybridizing in duplicate each individual nonfailing heart against RNA from the same cardiomyopathic heart (failing heart 002). Hierarchical cluster analysis confirmed the similarity of the control hearts and was accomplished using the Cluster/Treeview package developed by Eisen et al. (11). We also performed quadruplicate hybridizations on a pooled sample from seven ischemic dilated cardiomyopathic hearts versus the pooled control.Verification.
Quantitative real-time PCR (RT-PCR) was used to verify altered
expression of several of the apoptotic genes, including
GADD45, heat shock protein (HSP)22
(protein kinase H11), TNFRSM 10 (TRAIL), TNF receptor 1A, A20, MAPK-activating death
domain (MADD), p21, and cyclin D. The
primer sequences used in RT-PCR are shown in Table
2. First-strand synthesis was conducted
for 60 min at 48°C in a 10-µl reaction containing 100 ng total RNA,
5.5 mM MgCl2, 2.5 µM random hexamer primers, 2 µM
dNTPs, 4 units RNase inhibitor, and 12.5 units murine leukemia virus
reverse transcriptase. The real-time PCR reaction consisted of
the first-strand synthesis reaction, to which was added 4 mM
MgCl2, 0.8 mM dNTPs, 1× SYBR green PCR buffer (PE
Biosystems), 0.4 µM gene-specific forward and reverse primers, and
2.5 units AmpliTaq Gold DNA polymerase (PE Biosystems). The reaction
was monitored using an Applied Biosystems PRISM 7700 detection system.
mRNA expression was normalized to that of glyceraldehyde-3-phosphate
dehydrogenase (GAPDH).
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RESULTS |
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TOP
ABSTRACT
INTRODUCTION
METHODS
RESULTS
DISCUSSION
REFERENCES
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To determine the pattern of expression of apoptotic genes in failing hearts, we performed cDNA microrarray profiling of human hearts in failure compared with nonfailing hearts. We initially studied idiopathic dilated cardiomyopathic hearts because they lack ischemic/fibrotic areas that could complicate the analysis. However, we also included a pooled RNA group obtained from hearts with ischemic cardiomyopathy.
On the basis of data in the literature, we selected ~75
apoptosis-related genes for evaluation (see Fig.
1). To confirm that there was increased
apoptosis in human failing hearts, we used an antibody that
recognizes activated CPP32 (caspase 3). CPP32 is present in normal
cells as a 32-kDa inactive precursor; cleavage of CPP32 results in an
active p17 caspase (20). As shown in Fig.
2A, the CPP32 antibody
detected the presence of p17 fragments in hearts from patients in heart
failure but little or no detectable p17 fragments in nonfailing hearts.
Figure 2B presents the average densitometry for active
caspase-3 in nonfailing and failing hearts, showing a significant
increase in active caspase-3 in failing hearts. These data are
consistent with previous data showing apoptosis in failing
hearts (20, 21).
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The cluster analysis in Fig. 1 shows that there was no change in
expression of many apoptotic genes, decreased expression of
numerous apoptosis-related genes, and increased expression of a
few genes. Many of the changes in the expression of apoptotic genes
would reduce apoptosis and thus appear to be compensatory to
attenuate the apoptotic response. For example, we found increased expression of the antiapoptotic genes bcl-2 and
v-erb-b2. We also found decreased expression of several
caspases (caspase 3 and 7), several
proapoptotic members of the bcl-2 family [bcl-2 interacting killer
(BIK) and bcl-2 antagonist killer (BAK)], and a
slight decrease in Fas-associated death domain
(FADD). We also found a decrease in p21.
p21 Mediates p53-induced apoptosis and is therefore considered
to be proapoptotic, but a mutant p21 that is resistant to caspase
cleavage is reported to suppress apoptosis. Because a number of
bcl-2 family members showed altered expression, we examined the
phosphorylation status of bcl-2 and BAD because phosphorylation can
regulate the apoptotic activity of these proteins. As shown in Fig.
3A, consistent with the
increase in bcl-2 mRNA, we also observe an increase in the protein
level of the antiapoptotic bcl-2. The densitometry data in Fig.
3B show that compared with nonfailing hearts, failing hearts
had a significant increase in bcl-2. Phosphorylation of bcl-2 (Ser-70),
under most conditions, inactivates bcl-2 and thus increases cell death
(6). We observed no detectable phospho-bcl-2 in failing or
nonfailing hearts (data not shown). To determine whether this increase
in the antiapoptotic bcl-2 was offset by an increase in the
proapoptotic BAD, which binds and sequesters bcl-2, we examined the
levels of total and phospho-BAD. Consistent with an increase in
apoptosis, a striking decrease in phospho-BAD (Ser-112) was
observed in failing hearts (Fig.
4A). As shown in Fig. 4, there
was no consistent difference in total BAD levels between failing and
nonfailing hearts. The averaged densitometry data in Fig. 4B
show a significant decrease in phospho-BAD (Ser-112) but no difference
in the levels of total BAD. When phospho-BAD (Ser-112) was normalized
to total BAD, a significant difference between failing and nonfailing
hearts was also apparent (phospho-BAD/BAD in nonfailing hearts was
1.61 ± 0.40 vs. 0.15 ± 0.03 in failing hearts,
P < 0.05). Antibodies against Ser-155 phospho-BAD
showed only a faint band and no significant difference between failing
and nonfailing hearts (data not shown). We were unable to detect any
bands when we probed with antibodies directed against Ser-136
phospho-BAD (data not shown). Taken together, these data suggest that
the proapoptotic shift in bcl-2 may be offset by a decrease in
phospho-BAD in the failing heart, because dephosphorylated BAD will
bind and inactivate bcl-2.
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We reasoned that if apoptosis is an important contributor
in heart failure, there might be changes in gene expression in multiple signaling pathways that could contribute to a proapoptotic shift. The data in Fig. 1 show an increase in gene expression in
proapoptotic TNFRSM 10 (TRAIL) and a decrease
in the antiapoptotic genes GADD45, FLIP, A20, several mitochondrial HSPs, and
TNF--inducible MADD. These changes in gene expression
strongly suggest a proapoptotic shift in the TNF- pathway during
the transition to heart failure. The TNF- pathway activates both
pro- and antiapoptotic signals, and the balance determines whether
TNF- leads to apoptosis (3, 14, 16, 19).
TNF- activation of NF-B mediates increased expression of
GADD45, FLIP, and A20. Consistent with a lower expression of NF-B-induced genes, we also found increased expression of TNFRSM 10 (TRAIL) and decreased expression of
TNF- receptor 1, which activates NF-B. As illustrated
in Fig. 5, these changes in gene
expression would shift TNF- signaling to a proapoptotic bias.
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To examine whether these changes in gene expression are specific to idiopathic cardiomyopathic hearts or more representative of changes associated with the transition to heart failure, we also examined changes in gene expression in seven pooled ischemic cardiomyopathic hearts versus the pooled control. As shown in Fig. 1, cluster analysis comparing four idiopathic cardiomyopathic hearts versus the nonfailing control and the pooled ischemic cardiomyopathic hearts versus the nonfailing control suggested a similar pattern of gene expression.
We used RT-PCR to confirm altered expression of GADD45,
HSP22, TNFRSM 10, TNF receptor 1A,
A20, p21, MADD, and cyclin
D (see Fig. 6A). We also
confirmed decreased expression of FLIP protein by Western blot analysis
(Fig. 6B).
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One of the idiopathic cardiomyopathic hearts (heart
004) was from a patient on an assist device. Cluster analysis
showed that this heart had a similar overall pattern of gene expression
to other failing hearts. However, as shown in Fig. 1, GADD45,
A20, TNF receptor 1A, and MADD were not
downregulated in the heart on the assist device, and expression of
rel A was higher in the assist device heart than other
failing hearts (see Fig. 1). Expression of other
apoptosis-related genes such as the mitochondrial HSPs, FLIP, and p21 were not different between the assist
device heart and other failing hearts. These data suggest that in the
presence of the LVAD, there is a normalization of NF-B-regulated
gene expression abnormalities observed in failing hearts. To examine whether this is a common response in LVAD hearts, we examined an
additional two LVAD hearts using RT-PCR to measure the mRNA levels of
GADD45, A20, TNF receptor 1A, and MADD. As shown in Fig.
7, mRNA for these genes was higher in
failing LVAD hearts than in non-LVAD hearts.
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We also assessed whether phospho-BAD was increased in LVAD hearts. As
shown in Fig. 8, the three LVAD hearts
showed increased phospho-BAD compared with the non-LVAD failing hearts.
We also included two nonfailing hearts for comparison. Phospho-BAD in the LVAD hearts recovered to a level intermediate between failing and
nonfailing hearts.
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DISCUSSION |
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TOP
ABSTRACT
INTRODUCTION
METHODS
RESULTS
DISCUSSION
REFERENCES
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The data in this study suggest complex changes in gene
expression of pro- and antiapoptotic signaling pathways, including a proapoptotic shift in the TNF- signaling pathway in failing hearts. Several recent studies have shown an increase in the plasma levels of TNF- and the TNF- receptor in patients with
cardiomyopathies (28). TNF- is an important signal in
inflammation, proliferation, differentiation, and apoptosis,
and the balance between these pathways can be modulated by the relative
activation of NF-B (3). The data in the present study
suggest that decreased expression of GADD45,
A20, TNF receptor 1A, and FLIP and the
increased expression of TNFRSM 10 (TRAIL) may be
important in the shift in the TNF- signaling toward enhanced
apoptosis. An increase in TNF- signaling does not lead to
apoptosis when the NF-B pathway is activated by TNF-
(3, 14, 16, 19), because activation of NF-B leads to
increased expression of several antiapoptotic genes, such as
GADD45, FLIP, and A20. However, in
the failing heart, we find decreased expression of these TNF-- and
NF-B-induced antiapoptotic genes. We also find increased
expression of TRAIL and decreased expression of
TNF- receptor 1, which activates NF-B. These data
suggest a shift in TNF- signaling in failing hearts, such that
NF-B is not activated, thereby leading to a proapoptotic bias.
Taken together, these data suggest that decreased expression of these
antiapoptotic genes may be involved in apoptosis in failing
hearts and the transition to heart failure.
As shown in Fig. 5, TNF- binding to its receptor results in
the recruitment of adapter proteins such as TNF receptor I-associated death domain (TRADD) and FADD to the receptor. FADD recruits
pro-caspase 8, which is cleaved to active caspase 8. FLIP (also known
as casper, usurpin, CASH, FLAME-1, I-FLICE, CLARP, and MRIT) inhibits
the activation and/or recruitment of pro-caspase-8 to the complex, thereby antagonizing apoptosis (22). TRADD
recruits receptor-interacting protein (RIP)1 and TNF
receptor-associated factor (TRAF) to the complex and leads to
activation of NF-B and the JNK, ERK, and p38 MAPK pathways.
Activation of NF-B results in transcription of several
antiapoptotic genes such as FLIP, A20, and
GADD45.
Recent studies suggest an important role for GADD45 in
the prosurvival mechanism of NF-B (8). Studies were
done using a "death trap" screening in which NF-B (relA) null
cells were transfected with cDNA libraries derived from TNF--treated
cells. Apoptosis was induced by TNF-, and surviving cells
were selected. GADD45 was identified as a TNF--induced gene that
protected NF-B null cells from apoptosis. De Smaele et al.
(8) further showed that in NF-B null cells, the
TNF--induced activation of JNK was sustained throughout the time
course, whereas in NF-B-containing cells or NF-B null cells with
GADD45, the TNF--induced activation of JNK was transient and
returned to baseline after 40 min. It is suggested that termination of
JNK activation is important in the prosurvival mechanism of NF-B.
Thus decreased expression of GADD45 in failing hearts
would allow TNF- activation of apoptosis.
FLIP is also regulated by NF-B, and it inhibits activation
and/or recruitment of caspase 8 to the death-inducing signaling complex
(22). We found decreased protein levels of FLIP in the failing hearts. MADD is reported to link TNF- signaling with MAPK
activation. Depending on the splice varient, MADD can be pro- or
antiapoptotic (1). These data suggest that the
decrease in FLIP, A20, and GADD45 may shift the
pro-/antiapoptotic balance toward increased apoptosis in
end-stage heart failure.
We also observed a striking decrease in phosphorylation of BAD at
Ser-112 in failing hearts. Unphosphorylated BAD binds to and sequesters
bcl-2 and thereby inhibits its antiapoptotic function. Phosphorylation of BAD results in its dissociation from bcl-2, thereby
promoting cell survival. Baines et al. (2) reported that
overexpression of PKC-
is cardioprotective and leads to increased
phosphorylation of Ser-112 on BAD. BAD can be phosphorylated on
Ser-112, -136, -155, and/or -170 (10, 15, 18, 24, 25, 29).
The relative importance of phosphorylation at different sites is still
unclear. There are data suggesting that phosphorylation at different
sites may be additive (15, 29). Different phosphorylation sites may also be more important in different cell types. Activation of
phosphatidylinositol 3-kinase (PI3K), via the downstream kinases PKB/Akt and/or p70 ribosomal protein S6 kinase, is reported to phosphorylate Ser-136 (13). However, there are
studies reporting that at least in some cell types, activation of the
PI3K-PKB pathway does not lead to phosphorylation of Ser-136 of BAD
(18, 24). PKA appears to be the primary kinase responsible
for phosphorylation of Ser-155 (18, 29). MAPK/p90rsk is
emerging as the primary kinase responsible for phosphorylation of
Ser-112 of BAD (18, 24, 25, 29). Previous studies have
reported an increase in p90rsk activity in failing hearts
(26); these data suggested that p90rsk is unlikely to be
responsible for the decreased phosphorylation of Ser-112 of BAD in
failing hearts. PKA levels are reported also to be similar in failing
and nonfailing hearts. Thus, despite the increased activity of the BAD
kinase p90rsk (26), there is decreased phosphorylation of
Ser-112 or BAD in failing hearts. This raises the possibility that
perhaps there is increased dephosphorylation of BAD. Alternatively,
there could be altered subcellular targeting of the BAD kinase.
Interestingly, hearts that had been on an assist device did not
show decreased expression of the NF-B-regulated genes
GADD45, MADD, A20, or TNF receptor
1A. These data suggest that the assist device might allow
activation of NF-B and production of antiapoptotic factors. We
also observed that hearts from LVAD patients showed a recovery toward
control levels of phospho-BAD.
In summary, this study shows that there is a proapoptotic shift in
TNF- signaling in failing hearts. In addition, LVAD patients have
increased myocardial expression of TNF--regulated NF-B genes,
which may be important in the improvement in these LVAD patients. These
data are consistent with the improvement observed in patients treated
with inhibitors of TNF- (4). Finally, these analyses
indicate that therapies that enhance NF-B activation could be beneficial.
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ACKNOWLEDGEMENTS |
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This study was supported by the Division of Intramural Research, National Institute of Environmental Health Sciences, National Institutes of Health (NIH). C. Steenbergen was supported in part by NIH grant RO1-HL-39752.
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FOOTNOTES |
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* C. Steenbergen and C. A. Afshari contributed equally to this work.
Address for reprint requests and other correspondence: E. Murphy, Laboratory of Signal Transduction, NIEHS, Research Triangle Park, NC 27709 (E-mail: murphy1{at}niehs.nih.gov).
The costs of publication of this article were defrayed in part by the payment of page charges. The article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.
First published September 26, 2002;10.1152/ajpheart.00707.2002
Received 13 August 2002; accepted in final form 20 September 2002.
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REFERENCES |
|---|
|
TOP
ABSTRACT
INTRODUCTION
METHODS
RESULTS
DISCUSSION
REFERENCES
|
|---|
1.
Al-Zoubi, A,
Eflimova E,
Kaithamana S,
Martinez O,
El-Idrissi MEA,
Dogan R,
and
Prabhakar B.
IG20 and its splice isoforms, MADD and DENN-SV, on tumor necrosis factor alpha-induced apoptosis and activation of caspases-8 and -3.
J Biol Chem
276:
47202-47211,
2001
2.
Baines, C,
Zhang J,
Wang G,
Zheng Y,
Xia J,
Cardwell E,
Bolli R,
and
Ping P.
Mitochondrial PKC epsilon and MAPK form signaling modules in the murine heart: enhanced mitochondrial PKC epsilon-MAPK interactions and differential MAPK activatrion in PKC epsilon induced cardioprotection.
Circ Res
90:
390-397,
2002
3.
Baud, V,
and
Karin M.
Signal transduction by tumor necrosis factor and its relatives.
Trends Cell Biol
11:
372-377,
2001[Web of Science][Medline].
4.
Bozkurt, B,
Torre-Amione G,
Warren M,
Whitmore J,
Soran O,
Feldman A,
and
Mann D.
Results of targeted anti-tumor necrosis factor therapy with etanercept (ENBREL) in patients with advanced heart failure.
Circulation
103:
1044-1047,
2001
5.
Bushel, P,
Hamadeh H,
Bennett L,
Sieber S,
Martin K,
Nuwaysir EF,
Johnson K,
Reynolds K,
Paules R,
and
Afshari CA.
A microarray project system for gene expression experiment information and data validation.
Bioinformatics
17:
564-565,
2001
6.
Chao, D,
and
Korsmeyer S.
Bcl-2 family: regulators of cell death.
Annu Rev Immunol
16:
395-419,
1998[Web of Science][Medline].
7.
Chen, Y,
Dougherty ER,
and
Bittner ML.
Ratio-based decisions and the quantitative analysis of cDNA microarray images.
J Biomed Opt
2:
364-374,
1997.
8.
De Smaele, E,
Zazzeroni F,
Papa S,
Nguyen D,
Jin R,
Jones J,
Cong R,
and
Franzoso G.
Induction of gadd45 by NF-B downregulates pro-apoptotic JNK signalling.
Nature
414:
308-313,
2001[Medline].
9.
DeRisi, JL,
Iyer VR,
and
Brown PO.
Exploring the metabolic and genetic control of gene expression on a genomic scale.
Science
278:
680-686,
1997
10.
Dramsi, S,
Scheid M,
Maiti A,
Hojabrpour P,
Chen X,
Schubert K,
Goodlett D,
Awbersold R,
and
Duronio V.
Identification of a novel phosphorylation site, ser 170, as a regulator of BAD pro-apoptotic activity.
J Biol Chem
277:
6399-6405,
2002
11.
Eisen, M,
Spellman P,
Brown P,
and
Botstein D.
Cluster analysis and display of genome-wide expression patterns.
Proc Natl Acad Sci USA
95:
14863-14868,
1998
12.
Felker, G,
Thompson R,
Hare J,
Hruban R,
Clemetson D,
Howard D,
Baughman K,
and
Kasper E.
Underlying causes and long-term survival in patients with initially unexplained cardiomyopathy.
N Engl J Med
242:
2434-2440,
2000.
13.
Harada, H,
Becknell B,
Wilm M,
Mann M,
Huang L,
Taylor S,
Scott J,
and
Korsmeyer S.
Phosphorylation and inactivation of BAD by mitochondrial-anchored protein kinase A.
Mol Cell
3:
413-422,
1999[Web of Science][Medline].
14.
Haunstetter, A,
and
Izumo S.
Future perspective and potential implications of cardiac myocyte apoptosis.
Cardiovasc Res
45:
795-801,
2000
15.
Hayakaw, J,
Ohmichi M,
Kurachi H,
Hisamoto K,
Nishio Y,
Adachi K,
Tasaka K,
Kanzaki T,
and
Murata Y.
Inhibition of BAD phosphorylatin either at serine 112 via extracellular signal related protein kinase cascade or at serine 136 via Akt cascate sensitizes human ovarian cancer cells to cisplatin.
Cancer Res
60:
5988-5994,
2000
16.
Kubota, T, MM,
Frye C,
Alber S,
Bounoutas G,
Kadokami T,
Watkins S,
McTiernan C,
and
Feldman A.
Overexpression of tumor necrosis factor-alpha activates both anti- and pro-apoptotic pathways in the myocardium.
J Mol Cell Cardiol
33:
1331-1344,
2001[Web of Science][Medline].
17.
Kubota, T,
McTiernan C,
Frye C,
Slawson S,
Lemaster B,
Koretsky A,
Demetris A,
and
Feldman A.
Dilated cardiomyopathy in transgenic mice with cardiac-specific overexpression of tumor necrosis factor-.
Circ Res
81:
627-635,
1997
18.
Lizcano, J,
Morrice N,
and
Cohen P.
Regulation of BAD by cAMP-dependent protein kinase is mediated via phosphorylation of a novel site, ser 155.
Biochem J
349:
547-557,
2000[Web of Science][Medline].
19.
Mustapha, S,
Kirshner A,
De Moissac D,
and
Kirshenbaum L.
A direct requirement of nuclear factor-B for suppression of apoptosis in ventricular myocytes.
Am J Physiol Heart Circ Physiol
279:
H939-H945,
2000
20.
Narula, J,
Pandey P,
Arbustini E,
Haider N,
Narula N,
Kolodgie F,
Bello B,
Semigran M,
Bielsa-Masdeu A,
Dec G,
Israels S,
Ballester M,
Virmani R,
Saxena S,
and
Kharbanda S.
Apoptosis in heart failure: release of cytochrome c from mitochondria and activation of caspase-3 in human cardiomyopathy.
Proc Natl Acad Sci USA
96:
8144-8149,
1999
21.
Olivetti, G,
Abbi R,
Quaini F,
Kajsture J,
Cheng W,
Nitahara J,
Quaini E,
DiLoreto C,
Beltrami C,
Reed J,
and
Anversa P.
Apoptosis in the failing human heart.
N Engl J Med
336:
1131-1141,
1997
22.
Scaffidi, C,
Schmitz I,
Krammer P,
and
Peter M.
The role of c-FLIP in modulation of CD95-induced apoptosis.
J Biol Chem
274:
1541-1548,
1999
23.
Schaper, J,
Elsasser A,
and
Kostin S.
The role of cell death in heart failure.
Circ Res
85:
867-869,
1999
24.
Scheid, M,
Schubert K,
and
Duronio V.
Regulation of BAD phosphorylation and association with bcl-xL by the MAPK/Erk kinase.
J Biol Chem
1999:
31108-31113,
1999.
25.
Shimamura, A,
Ballif B,
Richards S,
and
Blennis J.
Rsk1 mediates a MEK-MAP kinase cell survival signal.
Curr Biol
10:
127-135,
2000[Web of Science][Medline].
26.
Takeishi, Y,
Huang Q,
Abe J,
Che W,
Lee JD,
Kawakatsu H,
Hoit B,
Berk B,
and
Walsh R.
Activation of mitogen-activated protein kinases and p90 ribosomal S6 kinase in failing human hearts with dilated cardiomyopathy.
Cardiovasc Res
53:
131-137,
2002
27.
Tong, H,
Chen W,
London R,
Muphy E,
and
Steenbergen C.
Preconditioning enhanced glucose uptake is mediated by p38 MAP kinase not by phosphatidylinositol 3-kinase.
J Biol Chem
275:
11981-11986,
2000
28.
Torre-Amione, G,
Kapadia S,
Lee J,
Durand JB,
Bies R,
Young J,
and
Mann D.
Tumor necrosis factor and tumor necrosis factor receptors in the failing human heart.
Circulation
93:
704-711,
1996
29.
Virdee, K,
Parone P,
and
Tolkovsky A.
Phosphorylation of the pro-apoptotic protein BAD on serine 155, a novel site contributes to cell survival.
Curr Biol
10:
1151-1154,
2000[Web of Science][Medline].
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