(Circulation. 2000;101:1578.)
© 2000 American Heart Association, Inc.
Basic Science Reports |
Enhanced Cardiac Contractility After Gene Transfer of V2 Vasopressin Receptors In Vivo by Ultrasound-Guided Injection or Transcoronary Delivery
From the 1. Medizinische Klinik (H.-J.W., K.-L.L., A.M., K.K., M.S., A.S., M.U.), and Institut für Experimentelle Onkologie/Therapieforschung (C.S., S.B., T.B.), Klinikum rechts der Isar, Technische Universität, München, Germany.
Correspondence to PD Dr Ungerer, 1. Medizinische Klinik and Deutsches Herzzentrum, Klinikum rechts der Isar, Ismaninger Straße 22, D-81675 München, Germany. E-mail ungerer{at}med1.med.tu-muenchen.de
 |
Abstract |
|---|
 Top Abstract IntroductionMethodsResultsDiscussionReferences |
|---|
Background—Systemic levels of arginine vasopressin (AVP) are increased in congestive heart failure, resulting in vasoconstriction and reduced cardiac contractility via V1 vasopressin receptors. V2 vasopressin receptors (V2Rs), which promote activation of adenylyl cyclase, are physiologically expressed only in the kidney and are absent in the myocardium. Heterologous expression of V2Rs in the myocardium could result in a positive inotropic effect by using the endogenous high concentrations of AVP in heart failure.
Methods and Results—We tested gene transfer with a recombinant adenovirus for the human V2R (Ad-V2R) to stimulate contractility of rat or rabbit myocardium in vivo. Ultrasound-guided direct injection or transcoronary delivery of adenovirus in vivo resulted in recombinant receptor expression in the myocardial target area, leading to a substantial increase in [3H]AVP binding. In 50% of the cardiomyocytes isolated from the directly injected area, single-cell shortening measurements detected a significant increase in contraction amplitude after exposure to AVP or the V2R-specific desmopressin (DDAVP). Echocardiography of the target myocardial area documented a marked increase in local fractional shortening after systemic administration of DDAVP in V2R-expressing animals but not in control virus–treated hearts. Simultaneous measurement of global contractility (dP/dtmax) confirmed a positive inotropic effect of DDAVP on left ventricular function in the Ad-V2R–injected animals.
Conclusions—Adenoviral gene transfer of the V2R into the myocardium increases cardiac contractility in vivo. Heterologous expression of cAMP-forming receptors in the myocardium could lead to novel strategies in the therapy of congestive heart failure by bypassing the desensitized ß-adrenergic receptor–signaling cascade.
Key Words: adenovirus • gene transfer • vasopressin • myocardium
|
Introduction |
|---|
|
TopAbstractIntroductionMethodsResultsDiscussionReferences |
|---|
Congestive heart failure is characterized by increased sympathoadrenergic drive, stimulation of the renin-angiotensin-aldosterone system, and augmented release of several peptide hormones, primarily arginine vasopressin (AVP) and atrial natriuretic peptide. The increase in AVP and catecholamines is inversely correlated with the prognosis of patients with congestive heart failure.1 2 In response to adrenergic stimulation, however, the failing myocardium cannot build up a sufficient contractile force because ß-adrenergic receptors are functionally inactivated.3 4
The current study tested the feasibility of adenoviral gene transfer in vivo to improve cardiac contractility by overexpressing a positive inotropic receptor, the recombinant V2 vasopressin receptor (rV2R). Physiologically, only vasopressin receptors (V1ARs) are expressed in the myocardium, where they exert a weak, negative inotropic effect. In contrast, V2Rs are expressed exclusively in the kidney, where they couple to cAMP formation. Therefore, we wished to induce a genetic receptor subtype shift in the myocardium by overexpressing the rV2Rs. In our previous study in cardiomyocytes, we had shown that rV2Rs are not downregulated by agonists.5 Therefore, the heterologous receptor can be assumed to retain most of its function even at prolonged agonist exposure. rV2Rs could also be expected to exert a positive inotropic effect in the failing heart, since in this condition, the endogenous concentrations of AVP are markedly increased.
Adenoviral gene transfer into the myocardium in vivo has been accomplished for several marker genes after direct injection into the ventricular wall or after intravasal application into the coronary arteries.6 7 However, all of these approaches were hampered by a relatively low efficiency of gene transfer. No studies have described the expression of transgenes with inotropic action, so that no measurements of transgene function in vivo were carried out. One publication hypothesized increased cardiac vascularization after gene transfer of fibroblast growth factor-5, without providing data on the cell type of transgene expression or the efficacy of gene transfer.8 Recently, Hajjar and colleagues9 demonstrated dampened contractility after adenoviral gene transfer of phospholamban by open-chest direct injection into the ventricle with simultaneous pulmonary and aortic clamping.
For our study, however, we wished to test an easier and hence, more practicable approach for transgene delivery, ie, intracoronary administration after percutaneous catheterization. This concept seemed feasible, after a recent study in a rabbit heart Langendorff preparation showed a much higher efficiency for transcoronary gene transfer ex vivo after pharmacological modification of the endothelial barrier.10 We used a modification of this protocol to achieve transcoronary gene delivery in vivo. The aim of the study was to explore novel approaches for in vivo gene transfer of recombinant adenoviral vectors encoding the human V2R to improve cardiac inotropy in adult myocardium in vivo.
|
Methods |
|---|
|
TopAbstractIntroductionMethodsResultsDiscussionReferences |
|---|
Construction and Purification of Recombinant Adenovirus
Generation of recombinant (E1-deficient) adenoviruses encoding the human V2R (Ad-V2R) or the nuclear-targeted mutant ß-galactosidase (Ad–ß-Gal) has already been described.5 In addition, a bicistronic adenovirus encoding both V2R and green-fluorescent protein (Ad–V2R-GFP) was generated after kind provision of plasmids by Dr He (Howard Hughes Medical Institute, Baltimore, Md).11 All transgenes were driven by cytomegalovirus promoters. The viruses were amplified in large stocks and purified through CsCl gradients. Adenoviral titers were determined by plaque titration on HEK 293 cells.5
Ultrasound-Guided Direct Adenovirus Injection Into Rat
Myocardium
Twelve- to 16-week-old male Wistar rats (weight 420±23 g; from
Charles River, Munich, Germany) were used. The project was
approved by the institutional ethics review board. The rats were
anesthetized with midazolam (2 mg/kg SC) and medetomidin
(150 µg/kg SC) and placed in a supine position. For
echocardiography, a 7.5-MHz probe was fixed on a
tripod. A 31-gauge needle was placed, under ultrasound control, into
the left ventricular inferior wall. The virus
suspension (150 µL, 109 pfu) was slowly
injected in a fractionated manner.
Transcoronary Delivery of Recombinant Adenovirus to
Rabbit Myocardium
New Zealand White rabbits (weight 3.6±0.3 kg; from
Charles River) received medetomidin (100 µg/kg IM), propofol (5
mg · kg-1 ·
h-1 IV), and a bolus of fentanyl (10 µg/kg
IV). They were intubated, ventilated, and monitored for ECG,
echocardiography, and pressure throughout the
experiment. Through a 5F sheath in the carotid artery, a JR-5F human
diagnostic catheter (Cordis) was placed opposite the left
coronary ostium. Then, a 2.4F Microferret (Cook) catheter was
selectively introduced into the proximal circumflex artery. Three
milliliters of Krebs-Ringer solution containing 10 µmol/L
serotonin and 1.25 mmol/L calcium was infused for 2
minutes, followed by infusion of 1.5 mL of 1010 pfu of
purified adenovirus over 2 minutes. No signs of ischemia or
ischemic damage were registered either during the intervention
(continuous ECG recording) or at necropsy.
Myocardial Contractility Measurement by
Echocardiography and
Intraventricular Tip Catheterization
Left ventricular contractility was
examined before and 72 hours after adenoviral gene transfer. The rats
or rabbits were anesthetized as described before.
Echocardiographic M-mode recording was carried
out as described in previous studies.12 In addition, ECG
and blood pressure were monitored continuously. After preparation of
the right carotid artery, a Millar 2.5F tip catheter connected to a
differentiating device (Hugo Sachs) was placed in the left ventricle.
After definition of basal contractility and left
ventricular pressure, 200 µL of NaCl (0.9%) was injected
as a negative control. After a sufficient equilibration period,
desmopressin (DDAVP) was injected in doses of
10-11 mol and 10-10 mol
(rats) or 4x10-12 to
4x10-8 mol (rabbits) to achieve estimated
intracoronary concentrations ranging from
10-9 mol/L to 10-8 mol/L
(rats) or 10-10 to 10-6
mol/L (rabbits), respectively. Measurements were carried out 1 minute
after each injection.
Preparation of Adult Ventricular Cardiomyocytes
As previously described,5 single, calcium-tolerant
ventricular cardiomyocytes were isolated for in
vitro infection or were prepared from the target area after injection
of adenovirus into rat hearts in vivo. In vitro infection (at a
multiplicity of infection of 80 pfu/cell) was carried out after 6 hours
in culture. The contraction amplitude of cardiomyocytes was
determined by using an electro-optical monitoring system as described
before.5
Radioligand Binding
The rat or rabbit hearts that had been examined
physiologically in vivo were excised, cut into
pieces, resuspended in 5 mmol/L Tris-HCl (pH 7.4) and 2
mmol/L EDTA, and homogenized. The homogenate
was centrifuged at 1000g for 15 minutes, and the
supernatant was centrifuged twice at 100 000g for
30 minutes. The resulting membrane pellet was resuspended in
Tris-HCl-buffer, pH 7.4. The radioligand binding experiment
was carried out as described before.5
ß-Gal Expression
Cardiomyocytes infected with Ad–ß-Gal were fixed and stained
as previously described.5 Frozen hearts were cut into
slices at a thickness of 7 µm in a freeze microtome, stained
similarly, and counterstained by hematoxylin-eosin.
Data Analysis
Data represent the mean±SD or, where appropriate,
mean±SEM. Data were compared by ANOVA for repeated measurements with
respect to equal doses in all groups, followed by a post hoc Scheffe’s
test (all results except those of Figure 3
) or by Student’s
t test with a 2-tailed distribution (Figure 3).
|
|
Results |
|---|
|
TopAbstractIntroductionMethodsResultsDiscussionReferences |
|---|
Single Cardiomyocytes Infected In Vitro
Figure 1
shows the effect of the
physiological agonist AVP and of the V2R-specific
agonist DDAVP on externally paced, isolated
ventricular cardiomyocytes infected in vitro.
Neither AVP nor DDAVP changed the contraction amplitude in control
cells, whereas both agents stimulated Ad-V2R–infected cells to the
same extent as ß-adrenergic agonists. For both agents, we observed
subnanomolar EC50 values, well below the
concentrations needed for stimulation of the natively present
V1ARs. Similarly, DDAVP increased contraction amplitude in
green-fluorescent Ad–V2R-GFP–infected cells, whereas no
effect was observed in nonfluorescent cells (positive and
negative cells in Figure 2B).
|
|
Ultrasound-Guided Injection of Adenovirus Into the
Myocardium
Ad-V2R, Ad–ß-Gal, or Ad–V2R-GFP was used for somatic gene
transfer into the myocardium. We established a method to
insert an echogenic needle subxyphoidally, guided by
echocardiography. Needle placement and the correct
injection site could be monitored in real time. Misplaced injections
could be distinguished by the absence of an echo contrast enhancement
in the target area. In a pilot study that compared echo imaging with
histological analysis, we found that
echocardiography showed the correct placement of
gene injection with a specificity of 100% (n=6), whereas misplaced
injections were also correctly predicted (n=6). Figure 2
shows
the macroscopic (Figure 2C) and microscopic (Figures 2D and 2E) images of hearts injected with Ad–ß-Gal (Figures 2C and 2E) or controls (Figure 2D). In a series of
reproducible experiments, approximately one fifth of the total
ventricular myocardium could be reached by a
single, fractionated injection, as estimated from macroscopic staining
of serial slices of the myocardium (Figure 2C).
Specific staining showed a small, central necrotic focus along the
needle track of the injection area and a markedly larger area of viable
transgene-expressing myocardium surrounding this track.
There was no evidence of ß-Gal expression in the livers, spleens,
aortas, or lungs of the animals that received intracardiac injections
of recombinant adenovirus. Fluorescence imaging of sections of
a heart injected with bicistronic Ad–V2R-GFP revealed transgene
expression of GFP and thus, rV2R, throughout the myocardium
(Figure 2F).
Percutaneous Transcoronary Administration
of Adenovirus
Ad-V2R and Ad–ß-Gal were also applied to rabbit
myocardium by transcoronary infusion after prior
intracoronary administration of 10 µmol/L
serotonin at a low calcium concentration (1.25
mmol/L). Sections through the posterior wall of rabbit hearts 3 days
after administration of Ad-V2R or Ad–ß-Gal are shown in Figures 2I and 2H. By using our method, approximately one fifth
of the left ventricular myocardium could be
reached, as estimated from macroscopic staining of serial slices of the
myocardium (Figure 2G). Transgene expression was
reproducibly restricted to the lateroposterior wall, corresponding to
the perfusion bed of the circumflex artery in rabbits.
V2R Expression in the Target Area of Intracardiac Gene
Transfer
Similar to Ad–ß-Gal, Ad-V2R and Ad–V2R-GFP were injected
directly into the inferior wall of rat hearts or applied
transcoronarily into the lateroposterior wall of rabbits in vivo. In
all cases, radioligand binding with
[3H]AVP documented expression of rV2Rs after
infection with Ad-V2R. The average Bmax value in
the target myocardium reached 3 times the value of control
areas in the same animals or in Ad–ß-Gal–infected animals (Figure 3). In the control areas,
[3H]AVP binding results from native expression
of V1AR.
Time Course of Transgene Expression
Adenoviral gene transfer led to maximum transgene expression 6
days after infection (estimated efficiency of 50%), with a subsequent,
gradual reduction of expression for the following weeks (estimated at
25% after 2 weeks, 3% to 4% after 3 weeks, and <1% after 4 weeks).
This was paralleled by an increasing infiltration of inflammatory
cells (negligible 2 days after infection, pronounced after 2 and 3
weeks, and decreasing after 4 weeks).
Isolated Cardiomyocytes From the Target Myocardium
After In Vivo Gene Transfer
Robust expression of the 2 adenoviral transgenes was demonstrated
after myocardial infection of rat hearts in vivo and subsequent
isolation of ventricular cardiomyocytes from
the target myocardial area. After in vivo infection with
109 pfu Ad–ß-Gal and subsequent staining with
X-gal, 
50% of the cardiomyocytes reproducibly showed
positive staining. To investigate the effect of rV2R expression on the
function of single cardiomyocytes after in vivo transfer,
we measured the contraction amplitude of rat cardiomyocytes
isolated from the target and control areas and from control
virus–injected hearts. All investigated cardiomyocytes
showed a clear increase in contraction amplitude after administration
of 10-7 mol/L isoproterenol.
Ad–V2R-GFP–infected cardiomyocytes that displayed green
fluorescence (n=10) showed a marked increase in contraction
amplitude in response to DDAVP, whereas nonfluorescent cells
did not (n=8; not shown). From these experiments, we concluded that the
capacity of cardiomyocytes to be stimulated by DDAVP
clearly depended on expression of rV2R in an individual cell. In 50%
of the cardiomyocytes from the Ad-V2R–injected area, we
detected a significant increase in contraction amplitude after
application of DDAVP, starting at a concentration of
10-11 mol/L (Figure 4). The maximum increase in contraction
amplitude was comparable to the results obtained with in
vitro–infected cardiomyocytes and reached a similar level
as did stimulation of native ß-adrenergic receptors with
isoproterenol. However, the remaining 50% of the
cardiomyocytes isolated from the target area did not
respond to the addition of DDAVP ("negative cells" in Figure 4), demonstrating that they had no or insufficient transgene
expression to produce a functional effect on
contraction.
|
Effect of Ad-V2R Infection on Cardiac Function In Vivo
To examine the effect of V2R expression in the intact
myocardium in vivo, we measured local
echocardiographic fractional shortening before and
after the systemic application of DDAVP. Validation of the
echocardiographic measurements has been extensively
described.12 Neither in rats nor in rabbits did cardiac
frequency and systemic blood pressure differ significantly at any
concentration of DDAVP, as documented by simultaneous ECG
recording (also see the
Table). Figure 5A shows the mean values of local
fractional shortening in the injection area for control- and
Ad-V2R–injected rats. A clear increase in fractional shortening
occurred in the inferior wall of Ad-V2R–injected animals,
whereas no such effect was seen in control virus–injected animals.
Left ventricular dP/dtmax as a
measure of global ventricular contractility
was determined by placing a tip catheter in the left ventricle. This
approach aimed at detecting changes in global left
ventricular contractility to complement the
echocardiographic information on local myocardial
contractility. After injection of DDAVP, an increase in
dP/dtmax was observed in the V2R-expressing rats,
whereas no change was seen in the control group (Figure 5B).
Also after transcoronary administration of Ad-V2R in rabbits,
we observed significant increases in fractional shortening, relative
systolic thickening of the posterior wall (calculated according
to Reference 1313 ), and left ventricular
contractility, which were all absent in rabbits that
had received control virus. The results are shown in the
Table.
|
|
|
Discussion |
|---|
|
TopAbstractIntroductionMethodsResultsDiscussionReferences |
|---|
In the present study, we have demonstrated effective functional expression of rV2Rs in the myocardium after adenoviral gene transfer in vivo by either direct injection or transcoronary administration. Thus, we show that recombinant, heterologous receptors are capable of enhancing cardiac contractility in vivo. These results complement a previous study, in which we had shown that infection of cultured cardiomyocytes with Ad-V2R in vitro led to a robust transgene expression.5
Several previous studies have described the successful transfer of recombinant adenoviruses into the myocardium either by direct injection or by intracoronary infusion.6 7 However, all of these approaches were hampered by a relatively low efficiency of gene transfer. None described the expression of transgenes with inotropic action, so that no measurements of transgene function in vivo were carried out. For the first time, we have shown in vivo somatic gene transfer with sufficient efficacy to achieve a positive inotropic effect of the transgene after either direct intracardiac injection or transcoronary delivery.
For this purpose, we improved both methods. Direct virus injection with high specificity was made possible by using a fractionated injection under control of a highly resolving ultrasound probe. This technique enabled us to control both the injection site and the success of the injection and to exclude misplaced injections by the absence of echo contrast enhancement in the target myocardial area. Transcoronary gene delivery was improved by using the protocol proposed by Donahue and collaborators10 in a recent seminal ex vivo study that showed a 96% transduction efficacy by infusing serotonin in a calcium-depleted solution before virus administration to modify the endothelial barrier.
The success of infection was tested by histological examination and X-gal staining for ß-Gal expression and by radioligand binding for V2R expression. Membranes from noninfected myocardium showed endogenous ligand binding for [3H]AVP due to the cardiac expression of native V1ARs. In contrast, [3H]AVP binding was increased 3-fold in cardiac membranes prepared from the target area of Ad-V2R administration, very similar to the findings after Ad-V2R gene transfer in vitro.5 Our previous study had shown that this increase in specific radioligand binding was due to the additional expression of rV2Rs. Widespread myocardial expression of rV2Rs in situ was documented by using a bicistronic Ad–V2R-GFP. After adenoviral infection with Ad-V2R, we were thus able to induce a genetic receptor subtype shift by overexpressing rV2R versus native V1R in the target myocardial area by a factor of 2 to 3:1.
In addition, we have demonstrated a functional in vivo effect of the recombinant receptor proteins after injection of selective V2 agonists. The V2-selective compound DDAVP had to be used for these in vivo experiments because the physiological agonist, AVP, induces strong, coronary vasoconstriction and hence, negative inotropy, when given intracoronarily.14 In addition, DDAVP did not show any chronotropic effects. Moreover, DDAVP is virtually devoid of peripheral vascular effects, and its effects on renal fluid retention only occur 10 to 20 minutes after systemic administration.
Echocardiographically, we determined a clear increase in regional fractional shortening of the target myocardial area after the administration of the receptor agonist. This increase must have been caused specifically by rV2Rs, because it was absent in control animals or in control virus–injected animals. On measuring dP/dtmax in the left ventricle simultaneously, we also detected a significant increase in global contractility, which was also absent in control animals or in control virus–injected animals.
By studying individual cardiomyocytes isolated from the target injection area of rat hearts, we were able to demonstrate that the observed increase in contractility was mediated by the cardiomyocytes and to exclude artificial effects mediated by other cell types, such as fibrocytes. After exposure to AVP, DDAVP, or both, Ad-V2R–infected ventricular cardiomyocytes showed a dose-dependent increase in their contraction amplitude. In the in vivo–infected myocardial target area, this was true for 50% of the isolated cardiomyocytes, thus proving high efficacy of gene transfer. Because of the better transfection conditions that can be reached in vitro than in vivo, more V2 receptors/myocytes will be expressed after in vitro infection. This fact should explain the difference in maximum average fractional shortening between the in vivo and in vitro groups, since a higher receptor/myocyte density will lead to a more pronounced effect on contractility.
Obviously, rV2Rs were coupled to the endogenous Gs/adenylyl cyclase system with sufficient stringency to affect cardiac function. The effect started at a concentration of 0.5 nmol/L, which is correlated to the physiological AVP concentrations in vivo in the circulation of patients with congestive heart failure.2 In addition, wall stress–triggered local myocardial synthesis of vasopressin has been described recently.15 Overexpressing V2Rs in the myocardium of patients with heart failure might therefore turn the negative inotropic effect of high levels of AVP into a beneficial, positive inotropic effect. This article represents the first report about the functional modulation of the cardiac contractile apparatus in vivo by overexpression of a heterologous, positive inotropic receptor. It might therefore be possible to use this strategy to modulate different functions of target organs in a wide variety of pathological states.
In heart failure, several alterations of the ß-adrenergic pathway occur, including downregulation of ß1-adrenergic receptors3 and uncoupling of the remaining ß-adrenergic receptors from adenylyl cyclase3 by an increased expression of ß-adrenergic receptor kinase (ßARK-1).4 Investigations of transgenic mice have shown that ß-adrenergic signaling can be "resensitized" in the long term by overexpressing the ßARK inhibitor "ßARKmini."16 17 In a transgenic mouse model of heart failure due to disruption of actin filaments (MLP -/- mice), overexpression of ßARKmini prevented the development of heart failure, whereas overexpression of ß2-adrenergic receptors even precipitated it.18 Because of these diverging results for proteins that all increase intracellular cAMP levels, specific transgenic interventions in the G protein–related messenger system might produce beneficial effects in the long run in vivo, despite the well-known failure of some cAMP-raising pharmacological agents to produce better long-term outcomes.19 Our report complements these findings, inasmuch as it shows that somatic gene transfer of a heterologous protein that stimulates endogenous adenylyl cyclase can also increase cardiac contractility.
Limitations of the Study
Intermittent as opposed to continuous cAMP stimulation via
heterologous rV2Rs might therefore be superior to some previous
approaches, although this assumption has to be proved in an ongoing
study on the long-term effects of V2R overexpression. We have shown in
the present study that ectopic expression of the V2R can be a
sufficient, alternative method to bypass the altered ß-adrenergic
signaling pathway in the heart. Moreover, our previous study has shown
that rV2Rs expressed in cardiomyocytes are not subject to
downregulation,5 so that they can maintain their function
even after prolonged agonist exposure. Whether this is also true for
the long-term overexpression of V2Rs in vivo remains to be
determined.
Note Added in Proof
During the review process of the present article, successful
somatic gene transfer of ß2-adrenergic
receptors to rabbit hearts has been reported by Maurice et al
(J Clin Invest. 1999;104:21–29).
|
Acknowledgments |
|---|
This work was supported by the Deutsche Forschungsgemeinschaft (grants Un 103/1-2 and La 1218/1-1) and by Forschungsmittel der TU München (H57-98).
|
Footnotes |
|---|
The first 2 authors contributed equally to this work.
Received August 30, 1999; revision received November 4, 1999; accepted November 8, 1999.
|
References |
|---|
|
TopAbstractIntroductionMethodsResultsDiscussionReferences |
|---|
- Cohn JN, Levine MB, Olivari T, Garberg V, Lura D,
Francis GS, Simon AB, Rector T. Plasma noradrenaline as a
guide to prognosis in patients with chronic congestive heart failure.
N Engl J Med. 1984;311:819–823.[Abstract]
- Landry DW, Levin HR, Gallant EM, Ashton RC, Seo S,
D’Alessandro D, Oz MC, Oliver JA. Vasopressin deficiency contributes
to the vasodilation of septic shock. Circulation. 1997;95:1122–1125.
[Abstract/Free Full Text]
- Bristow MR, Ginsberg R, Umans V, Fowler W,
Minobe W, Rasmussen R, Zera P, Menlove R, Shah P, Jamieson S.
ß1- and ß2-Adrenergic
receptor subpopulation in nonfailing and failing human
ventricular myocardium: coupling of both
receptor subtypes to muscle contraction and selective
ß1-receptor downregulation in heart failure.
Circ Res. 1986;59:297–309.
[Abstract/Free Full Text]
- Ungerer M, Böhm M, Elce JS, Erdmann E, Lohse MJ.
Altered expression of ß-adrenergic receptor kinase and
ß1-adrenergic receptors in failing human heart.
Circulation. 1993;87:454–463.
[Abstract/Free Full Text]
- Laugwitz KL, Ungerer M, Schöneberg T, Weig HJ,
Kronsbein K, Hoffmann K, Moretti A, Schultz G, Schömig A.
Adenoviral gene transfer of the human V2 vasopressin receptor improves
contractile force of rat cardiomyocytes.
Circulation. 1999;99:925–933.
[Abstract/Free Full Text]
- Barr E, Caroll J, Kalynych AM, Tripathy SK, Kozarsky
K, Wilson JM, Leiden JM. Efficient catheter-mediated gene transfer into
the heart using replication-defective adenovirus. Hum Gene
Ther. 1994;1:51–58.
- Guzman RJ, Lemarchand P, Crystal RG, Epstein SE,
Finkel T. Efficient gene transfer into myocardium by direct
injection of adenovirus vectors. Circ Res. 1993;73:1202–1207.
[Abstract/Free Full Text]
- Giordano F, Ping P, McKirnan D, Nozaki S, DeMaria AN,
Dillman WH, Mathieu-Costello O, Hammond K. Intracoronary gene
transfer of fibroblast growth factor-5 increases blood flow and
contractile function in an ischemic region of the heart.
Nat Med. 1996;2:534–539.[Medline]
[Order article via Infotrieve]
- Hajjar RJ, Schmidt U, Matsui T, Guerrero JL, Lee KH,
Gwathmey JK, Dec GW, Semigran MJ, Rosenzweig A. Modulation of
ventricular function through gene transfer in vivo.
Proc Natl Acad Sci U S A. 1998;95:5251–5256.
[Abstract/Free Full Text]
- Donahue JK, Kikkawa K, Thomas AD, Marban E,
Lawrence JH. Acceleration of widespread adenoviral gene transfer to
intact rabbit hearts by coronary perfusion with low calcium and
serotonin. Gene Ther. 1998;5:630–634.[Medline]
[Order article via Infotrieve]
- He TC, Zhou S, da Costa LT, Yu J, Kinzler KW,
Vogelstein B. A simplified system for generating recombinant
adenoviruses. Proc Natl Acad Sci U S A. 1998;95:2509–2514.
[Abstract/Free Full Text]
- Gardin JM, Siri FM, Kitsis RN, Edwards JG, Leinwand LA.
Echocardiographic assessment of left
ventricular mass and systolic function in mice.
Circ Res. 1995;76:907–914.
[Abstract/Free Full Text]
- St John Sutton MG, Reichek N, Kastor JA, Giuliani ER.
Computerized M-mode echocardiographic analysis
of left ventricular dysfunction in cardiac amyloid.
Circulation. 1982;66:790–799.
[Free Full Text]
- Graf BM, Fischer B, Martin E, Bosnjak ZJ, Stowe DJ.
Differential effects of arginine vasopressin on isolated guinea pig
heart function during perfusion at constant flow and constant pressure.
J Cardiovasc Pharmacol. 1996;29:1–7.
- Koch WJ, Rockman HA, Samama P, Hamilton R, Bond RA,
Milano CA, Lefkowitz RJ. Cardiac function in mice overexpressing
the ß-adrenergic receptor kinase or a ßARK
inhibitor. Science. 1995;268:1350–1353.
[Abstract/Free Full Text]
- Koch WJ, Milano CA, Lefkowitz RJ. Transgenic
manipulation of myocardial G protein–coupled receptors and receptor
kinases. Circ Res. 1996;78:511–516.
[Free Full Text]
- Rockman HA, Chien KR, Choi DJ, Iaccarino G,
Hunter JJ, Ross J, Lefkowitz RJ, Koch WJ. Expression of a
ß-adrenergic receptor kinase 1 inhibitor prevents the
development of myocardial failure in gene-targeted mice. Proc
Natl Acad Sci U S A. 1998;95:7000–7005.
[Abstract/Free Full Text]
- Packer M, Carver JR, Rodehoffer RJ. Effect of oral
milrinone on mortality in severe chronic heart failure. N
Engl J Med. 1991;325:1468–1475.[Abstract]
- Hupf H, Grimm D, Riegger GAJ, Schunkert H. Evidence for
a vasopressin system in the rat heart. Circ Res. 1999;84:365–370.
[Abstract/Free Full Text]
This article has been cited by other articles:
 |
|
 |
|
  J. Gutkowska, M. Miszkurka, B. Danalache, N. Gassanov, D. Wang, and M. Jankowski Functional arginine vasopressin system in early heart maturation Am J Physiol Heart Circ Physiol, October 1, 2007; 293(4): H2262 - H2270. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 M. Miyagawa, M. Beyer, B. Wagner, M. Anton, C. Spitzweg, B. Gansbacher, M. Schwaiger, and F. M. Bengel Cardiac reporter gene imaging using the human sodium/iodide symporter gene Cardiovasc Res, January 1, 2005; 65(1): 195 - 202. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 M. Miyagawa, M. Anton, R. Haubner, M. V. Simoes, C. Stadele, W. Erhardt, S. Reder, T. Lehner, B. Wagner, S. Noll, et al. PET of Cardiac Transgene Expression: Comparison of 2 Approaches Based on Herpesviral Thymidine Kinase Reporter Gene J. Nucl. Med., November 1, 2004; 45(11): 1917 - 1923. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
D. M. Roth, N. C. Lai, M. H. Gao, J. D. Drumm, J. Jimenez, J. R. Feramisco, and H. K. Hammond Indirect intracoronary delivery of adenovirus encoding adenylyl cyclase increases left ventricular contractile function in mice Am J Physiol Heart Circ Physiol, July 1, 2004; 287(1): H172 - H177. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 G. Vassalli, S. Fleury, J. Li, J.-J. Goy, L. Kappenberger, and L. K. von Segesser Gene transfer of cytoprotective and immunomodulatory molecules for prevention of cardiac allograft rejection Eur. J. Cardiothorac. Surg., November 1, 2003; 24(5): 794 - 806. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 I. D. Cox, C. A. Thompson, and S. N. Oesterle Biointerventional cardiovascular therapy Eur. Heart J., November 2, 2002; 23(22): 1753 - 1756. [Full Text] [PDF]
|
|
|
|
 |
|
 R. Beeri, J. L. Guerrero, G. Supple, S. Sullivan, R. A. Levine, and R. J. Hajjar New Efficient Catheter-Based System for Myocardial Gene Delivery Circulation, October 1, 2002; 106(14): 1756 - 1759. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
P. M.L Janssen, W. Schillinger, J.K. Donahue, O. Zeitz, S. Emami, S. E Lehnart, J. Weil, T. Eschenhagen, G. Hasenfuss, and J. Prestle Intracellular {beta}-blockade: overexpression of G{alpha}i2 depresses the {beta}-adrenergic response in intact myocardium Cardiovasc Res, August 1, 2002; 55(2): 300 - 308. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 C. A Thompson and S. N Oesterle Biointerventional cardiology: the future interface of interventional cardiovascular medicine and bioengineering Vascular Medicine, May 1, 2002; 7(2): 135 - 140. [Abstract] [PDF]
|
|
|
|
 |
|
 P Menasche and M Desnos Cardiac reparation: fixing the heart with cells, new vessels and genes Eur. Heart J. Suppl., April 1, 2002; 4(suppl_D): D73 - D81. [Abstract] [PDF]
|
|
|
|
|
|
N. C. Lai, D. M. Roth, M. H. Gao, S. Fine, B. P. Head, J. Zhu, M. D. McKirnan, C. Kwong, N. Dalton, K. Urasawa, et al. Intracoronary Delivery of Adenovirus Encoding Adenylyl Cyclase VI Increases Left Ventricular Function and cAMP-Generating Capacity Circulation, November 7, 2000; 102(19): 2396 - 2401. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
E. Marban Gene Therapy for Common Acquired Diseases of the Heart : The Sirens’ Song Circulation, April 4, 2000; 101(13): 1498 - 1499. [Full Text] [PDF]
|
|
|
|
 |
|
 K.-L. Laugwitz, H.-J. Weig, A. Moretti, E. Hoffmann, P. Ueblacker, I. Pragst, K. Rosport, A. Schomig, and M. Ungerer Gene Transfer of Heterologous G Protein-Coupled Receptors to Cardiomyocytes : Differential Effects on Contractility Circ. Res., April 13, 2001; 88(7): 688 - 695. [Abstract] [Full Text] [PDF]
|
|
| |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||