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Ann Thorac Surg 2007;84:134-141
© 2007 The Society of Thoracic Surgeons

Original Articles: Cardiovascular

Combined Strategy Using Myoblasts and Hepatocyte Growth Factor in Dilated Cardiomyopathic Hamsters

Haruhiko Kondoh, MDa, Yoshiki Sawa, MDa,*, Norihide Fukushima, MDa, Goro Matsumiya, MDa, Shigeru Miyagawa, MDa, Satoru Kitagawa-Sakakida, MDb, Yukiko Imanishia, Naomasa Kawaguchi, MDc, Nariaki Matsuura, MDc, Hikaru Matsuda, MDa

a Department of Surgery, Division of Cardiovascular Surgery, Osaka University Graduate School of Medicine, Suita, Osaka, Japan
b Department of Molecular Therapeutics, Division of Molecular Therapeutics, Osaka University Graduate School of Medicine, Suita, Osaka, Japan
c Department of Pathology, School of Allied Health Science, Faculty of Medicine, Osaka University Graduate School of Medicine, Suita, Osaka, Japan

Accepted for publication March 19, 2007.

* Address correspondence to Dr Sawa, Division of Cardiovascular Surgery, Department of Surgery, Osaka University Graduate School of Medicine, 2-2 Yamada-oka, Suita, Osaka, 565-0871, Japan (Email: sawa{at}surg1.med.osaka-u.ac.jp).


   Abstract
Top
 Abstract
Introduction
 Material and Methods
 Results
 Comment
 Acknowledgments
 References
 
Background: There are few reports on treating dilated cardiomyopathy (DCM) with myoblast transplantation, and these show limited efficacy. Hepatocyte growth factor has cardioprotective effects on failed myocardium. Here, we combined these two treatments and analyzed cardiac function in DCM hamsters.

Methods: Twenty-seven-week-old BIO TO-2 hamsters, which show moderate cardiac remodeling, were divided into four treatment groups: myoblast transplantation (T group, n = 24), human hepatocyte growth factor gene transfection (H group, n = 29), combined treatment (T+H group, n = 21), and medium alone (C group, n = 26).

Results: Significantly better fractional shortening was observed in the T+H group compared with the others (14.9% ± 1.0%, 11.7% ± 1.5%, 11.3% ± 1.3%, and 8.6% ± 1.1 %, in the T+H, H, T, and C groups, respectively). Immunohistochemical analysis showed alpha- and beta-sarcoglycan expression in the hearts of the H and T+H groups but not in the other groups. There was less myocardial fibrosis in the H and T+H groups than in the other two, and neovascularization in the T+H group was significantly greater than in the other groups (266 ± 24, 209 ± 27, 199 ± 36, and 96 ± 17 vessels/mm2, in the T+H, H, T, and C groups, respectively). Survival was significantly prolonged in the H and T+H groups compared with the other groups.

Conclusions: Hepatocyte growth factor gene transfection and myoblast transplantation preserved the cardiac function of DCM hamsters, probably through different mechanisms, and the combined treatments preserved cardiac performance better than either treatment alone. The combined therapy is a promising strategy for treating DCM.


   Introduction
Top
 Abstract
 Introduction
Material and Methods
 Results
 Comment
 Acknowledgments
 References
 
Dilated cardiomyopathy (DCM) is a common cause of chronic heart failure [1]. Contemporary medical therapy has dramatically improved the prognosis of heart failure [2]. However, mortality is still high, and cardiac transplantation is the best treatment for advanced cases [3, 4]. Unfortunately, cardiac transplantation has limitations, such as donor shortages, rejection, and infection [5]. Therefore, novel strategies are desired.

Dilated cardiomyopathy pathology is characterized by myocardial remodeling, which mainly involves myocardial fibrosis with changes in the cytoskeletal and sarcolemmal proteins in cardiomyocytes, which reduces both their number and function [6]. Consequently, cardiac remodeling causes ventricular dilatation and thinning, leading to progressive congestive heart failure.

Currently, ischemic cardiomyopathy is the primary disease target for skeletal myoblast transplantation. Its functional benefit has been established in animals, and a clinical trial of autologous myoblast transplantation has begun [7–9]. In contrast, although the few preclinical reports on myoblast transplantation in DCM have shown improved left ventricular function, none has documented reduced myocardial fibrosis or cardiac remodeling [10–12]. The mechanism underlying the improved ventricular function is not fully understood, and this approach may require additional treatments.

Noncellular treatments for chronic heart failure, such as hepatocyte growth factor (HGF) gene therapy, are promising. Hepatocyte growth factor is a pleiotropic growth factor that is angiogenic, antifibrotic, and antiapoptotic [13, 14] . It has improved cardiac performance in animal models of ischemic and dilated cardiomyopathy, including through the reduction of myocardial fibrosis and remodeling [15–18] . We recently reported that, compared with either treatment alone, combining cellular cardiomyoplasty and HGF gene transfection significantly improves neovascularization, reduces fibrosis, and increases the expression of cytoskeletal proteins in infarcted rat myocardium [19].

Here, we tested the effect of combining myoblast transplantation and HGF gene transfection on the preservation of DCM heart function, compared with that of each therapy used alone.


   Material and Methods
Top
 Abstract
 Introduction
 Material and Methods
Results
 Comment
 Acknowledgments
 References
 
Experimental Animals
Twenty-seven-week-old male BIO TO-2 hamsters (Bio Breeders, Fitchburg, Massachusetts) were used and cared for in compliance with the "Principles of Laboratory Animal Care," by the National Society for Medical Research, and the "Guide for the Care and Use of Laboratory Animals," by the National Institutes of Health (NIH Publication No. 85-23, revised 1996). The animals were randomly divided into four treatment groups, which received injections of myoblasts (T group, n = 24), hemagglutinating virus of Japan (HVJ) liposomes containing hHGFcDNA (H group, n = 29), both myoblasts and HVJ-liposomes containing hHGFcDNA (T+H group, n = 21), or culture medium alone (C group, n = 26). Ten hamsters in each group were used for histological analyses 2 and 4 weeks after the operation (n = 5 for each point). The remaining hamsters (n = 14, T; n = 19, H; n = 11, T+H; n = 16, C) were used to evaluate cardiac function and prognosis.

Human HGF Gene Transfection
Human HGF cDNA was inserted into the NotI site of the pCU-SR{alpha} expression vector [20]. Preparation of the liposome complex with HVJ was described previously [21].

Isolation of Skeletal Myoblasts
Myoblasts were isolated from the skeletal muscles of the inferior limbs of BIO F1B hamsters as described [12]. The hamsters were euthanized and the muscles were excised and washed with phosphate-buffered saline. After meticulous removal of nonmuscle tissues, the muscles were weighed, minced, and enzymatically digested with collagenase (Gibco BRL, Rockville, Maryland), 5 mg/mL, at 37° for 40 minutes. The cells were collected and resuspended in DMEM (Gibco BRL) with 20% FBS and 1% penicillin-streptomycin (Gibco BRL). Cells were plated in 100-mm collagen-coated culture dishes (IWAKI, Tokyo, Japan) and cultured at 37° in a humidified atmosphere containing 5% CO2. The next day, unattached cells were removed by changing the culture medium, and the cells were grown to about 70% confluence. After 5 days, the cells were harvested, washed with DMEM, and kept on ice until transplantation. Usually, 20 x 106 cells were obtained from 1 hamster, and approximately 50% of the cultured cells were desmin-positive.

Operative Procedures
Hamsters were anesthetized by intraperitoneal injection of pentobarbital (50 mg/kg body weight), intubated, and a ventilator was used to maintain positive pressure (SN-480-7, Shinano, Tokyo, Japan). The heart was exposed through a 2.5-cm left lateral thoracotomy. All animals received injections from a 30G tuberculin syringe into one site in the right ventricle and 3 sites in the left ventricle wall (anterior free wall, apex, and posterior wall). The T group received 0.2 mL cell suspension (5 x 106 cultured cells) at each injection site. The H group received 0.2 mL HVJ-liposome-plasmid complex (320 µg human HGF cDNA). In the T+H group received injections of the same amounts of cell suspension and HVJ-liposome-plasmid complex. The C group received 0.2 mL DMEM per site. This procedure results in a peak HGF concentration in the myocardium at 1 week, and HGF expression persists for at least 3 weeks [18].

Measurement of Hamster Body Weight and Cardiac Function
The hamsters were weighed biweekly, and echocardiographic measurements were made using isoflurane as anesthesia. The isoflurane was reduced to 1% and maintained for 20 minutes to stabilize the hemodynamics before examination. Cardiac function measurements were performed with a SONOS 5500 echocardiograph (Agilent Technologies). A 12-MHz annular array transducer was used, and M-mode echocardiograms were recorded; the heart rate, left ventricular end-systolic dimension, left ventricular end-diastolic dimension, and fractional shortening were determined within about 10 minutes. The echocardiograph reader was blinded to the treatment group.

Histological Analyses
Left ventricular myocardium specimens were obtained 2 and 4 weeks after the operation. Transverse sections of hearts (2-µm thick) were fixed with 10% buffered formalin, embedded in paraffin, and subjected to hematoxylin and eosin and Masson’s trichrome staining. To label vascular endothelial cells, we used antifactor VIII–related antigen coupled with horseradish peroxidase (EPOS Anti-Human von Willebrand factor/HRP; DAKO, Carpinteria, California), following the manufacturer’s protocol. Signals were visualized with diaminobenzidine/hydrogen peroxide.

Frozen sections (4-µm thick) were used for immunohistochemical staining of alpha- and beta-sarcoglycans, performed as described [22, 23] . The primary antibodies were anti-alpha-sarcoglycan and anti-beta-sarcoglycan (clone: Ad1/20A6 and bSarc/5B1, respectively; Novo Castra, United Kingdom). The second antibody was biotinylated rabbit anti-mouse antibody (K0675; DAKO), followed by staining with fluorescein isothiocyanate (FITC)-conjugated streptavidin (F0422; DAKO). Immunofluorescence was viewed by confocal microscopy (ECLIPSE TE 2000-U; Nikon, Japan).

More than five sections were prepared per specimen, and every stained sample was evaluated independently by two pathologists who were blinded to the treatment groups. Percentage of total fibrotic area, determined by Masson’s trichrome staining, was calculated by image analysis of the sections, using a planimetric method with Windows MetaMorph software (Universal Imaging Corporation, Downingtown, Pennsylvania). Factor VIII-positive arteries and capillaries under 100 µm in diameter were counted under light microscopy at x100 magnification, and expressed as the number of vessels/mm2. Immunohistochemical signals were also quantified by the planimetric method, and expressed as a percentage of the value in F1B controls [18]. At least 10 low-power fields per section were analyzed.

Evaluation of Hamster Prognosis After Treatment
We evaluated the life-saving effect of the treatments on the TO-2 hamsters, which were maintained until their death from heart failure. The survival rates of animals in the C, T, H, and T+H groups were calculated by the Kaplan-Meier method using SPSS Ver. 11.0 (SPSS, Chicago, Illinois), and the significance of the differences among groups was tested by log-rank analysis.

Statistical Analysis
All values were expressed as the mean ± SEM and subjected to multiple analysis of variance (ANOVA) using the StatView 5.0 program (Abacus Concepts, Berkeley, California). Echocardiographic data were first analyzed by two-way repeated measurement ANOVA for differences across the whole time course, and one-way ANOVA with the Tukey-Kramer post-hoc test was used to verify the significance for the specific comparison at each time point. Other numerical data, except for survival, were analyzed by one-way ANOVA with the Tukey-Kramer post-hoc test. A p value less than 0.05 was considered significant.


   Results
Top
 Abstract
 Introduction
 Material and Methods
 Results
Comment
 Acknowledgments
 References
 
Operative Mortality, Body Weight, and Heart Rate After Treatment
Of the 100 hamsters operated on, none died within 6 days after surgery. Table 1 shows the hamsters’ body weight measured biweekly. The C group showed a slightly lower body weight after the operation, but there was no difference among the other three groups. There was no difference in the biweekly heart rate among the four groups (Table 1).


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  		Table 1 Body Weight, Heart Rate, and Echocardiographic Measurements of Cardiomyopathic Hamsters
 
Cardiac Function After Treatment
Table 1 shows cardiac function, which was evaluated biweekly by echocardiography. The left ventricular end-diastolic dimension, left ventricular end-systolic dimension, and fractional shortening at age 27 weeks were not significantly different among the four groups. After the operation, in the C group, the left ventricular end-diastolic dimension and end-systolic dimension gradually and progressively enlarged, and the fractional shortening decreased. The other three groups showed a relatively delayed progression in the enlargement of the left ventricular end-systolic dimension. In the T group, the end-systolic dimension continuously enlarged, but at a slower rate than in the C group. In the H group, no end-systolic dimension enlargement was observed in the first 2 weeks, but thereafter it increased at the same rate as in the C group, and the sizes at 35 weeks were similar. Although the end-systolic dimension of the T+H group was slightly (and significantly) smaller than that of the C group, it gradually enlarged as well. The fractional shortening of the H and T groups was significantly better than that of the C group after the operation. The T+H group exhibited the combined effects of the individual treatments, and at 33 or 35 weeks, their fractional shortening was significantly better than that of the other groups.

Histologic Analyses of Hearts After the Operation
Figure 1A shows representative immunohistochemical staining for alpha- and beta-sarcoglycan in the myocardium of treated hamsters 2 weeks after the operation. Alpha- and beta-sarcoglycans were clearly detected in the sarcolemma of F1B (ie, normal) hamsters. As expected from a previous study on the mature TO-2 hamster, in which the promoter region of the delta-sarcoglycan gene is deleted, we confirmed in the C group that no delta-, alpha-, beta-, or gamma-sarcoglycan was expressed [23]. Interestingly, in the H and T+H groups, alpha- and beta-sarcoglycans were readily detected 2 weeks after the operation. However, neither of these proteins was detectable in the T group.


Figure 1
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  		Fig 1. Immunohistochemical staining for alpha- and beta-sarcoglycan. Representative photographs of immunohistochemical staining 2 weeks after the operation (A) and quantitative assessments of the alpha-sarcoglycan (B) and beta-sarcoglycan (C) signals are shown. Sections from age-matched F1B hamsters are shown in (A), and the quantitative results are expressed as the percentage of the signal strength normalized to F1B (n = 5 for each group, *p < 0.05). (C group = received culture medium alone; H group = received hemagglutinating virus of Japan [HVJ] liposomes containing hHGFcDNA; T group = received myoblasts; T+H group = received both myoblasts and HVJ liposomes containing hHGFcDNA.)

 
Figure 1B and C show quantitative analyses of the results in 1A. The area of the above-background green signal in each group was compared with that of the F1B hamsters, and the relative level of expression of each protein is shown. Although no signals reached normal levels, observed in F1B hamsters, they were significantly higher in the H and T+H groups, which showed similar, weak expression of the two cytoskeletal proteins, but at significantly higher levels than in the C and T groups. This confirmed our previous observation that HGF gene transfection reorganizes cytoskeletal proteins in the myocardium of DCM hamsters [18].

Figure 2A shows typical examples of hematoxylin and eosin and Masson-trichrome staining of myocardium from the F1B-strain and from treated TO-2 hamsters, 4 weeks after the operation. Although the hematoxylin and eosin staining was similar among the groups, the Masson’s trichrome staining indicated less fibrosis in the H and T+H hearts. Quantification confirmed that there was significantly less myocardial fibrosis in the H and T+H groups than in the C and T groups (Fig 2B). However, there were no differences between the C and T or between the H and T+H groups. Thus, the reorganization of cytoskeletal proteins and reduction of myocardial fibrosis were similar between the T+H and H groups.


Figure 2
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  		Fig 2. Myocardial morphology of hamsters treated with myoblast transplantation and hepatocyte growth factor (HGF) transfection. (A) Hematoxylin and eosin (HE) and Masson’s trichrome staining of the myocardium from F1B hamsters and treated TO-2 hamsters 4 weeks after the operation. (B) Quantitative results of the fibrotic change in the left ventricle (n = 5 for each group, *p < 0.05). (C group = received culture medium alone; H group = received hemagglutinating virus of Japan [HVJ] liposomes containing hHGFcDNA; T group = received myoblasts; T+H group = received both myoblasts and HVJ liposomes containing hHGFcDNA.)

 
Figure 3A shows typical examples of factor VIII staining in the myocardium from hamsters in the four groups 4 weeks after the operation. A marked increase in the number of factor VIII-positive arterioles and capillaries was observed in the T, H, and T+H groups, compared with the C group (Fig 3B).


Figure 3
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  		Fig 3. Assessment of capillary density in the hearts of treated hamsters. (A) Representative staining for factor VIII in the myocardium 4 weeks after the operation. (B) Quantitative results of the density of arterioles and capillaries (n = 5 for each group, *p < 0.05). (C group = received culture medium alone; H group = received hemagglutinating virus of Japan [HVJ] liposomes containing hHGFcDNA; T group = received myoblasts; T+H group = received both myoblasts and HVJ liposomes containing hHGFcDNA.)

 
Quantification of the staining density of the arterioles and capillaries indicated a modest increase in capillary vessels in the H and T groups and a significantly greater number of vessels in the T+H group (Fig 2B). Thus, the combined strategy significantly increased neovascularization.

Prolonged Life Expectancy After the Operation
In the C group, the number of surviving animals decreased gradually, and all the hamsters died before 48 weeks of age. The mortality rate and number surviving was similar in the T group (p = 0.75). In contrast, most of the hamsters in the H group survived beyond 42 weeks, and showed significantly better survival than the C and T groups (p = 0.005 versus C, p = 0.043 versus T). The T+H group also showed significantly better survival than the C and T groups (p = 0.018 versus C, p = 0.039 versus T). The difference in survival between the H and T+H groups did not reach significance (p = 0.28), but the median survival in the T+H group was longer than that in the other three groups (aged 37, 36, 45, and 51 weeks, in the C, T, H and T+H group, respectively).


   Comment
Top
 Abstract
 Introduction
 Material and Methods
 Results
 Comment
Acknowledgments
 References
 
The present study demonstrated that cardiac performance of adult DCM hamsters was preserved by the combination of HGF gene transfection and myoblast transplantation. The improvement was associated with enhanced neovascularization caused by the HGF plus myoblast transplantation and with reduced myocardial fibrosis and reorganization of the cytoskeletal proteins caused by the HGF transfection. The survival was significantly prolonged in the HGF and combined therapy groups. These findings suggest that the combined treatment may be a useful therapeutic strategy for cardiac restoration in human DCM.

The TO-2 hamster is a model of human hereditary DCM, in which the number of cardiomyocytes decreases progressively after birth, and cardiac remodeling, mainly myocardial fibrosis, occurs with ventricular dilatation and thinning [22, 24]. Congestive heart failure begins at about age 20 weeks, and progresses gradually until the animals start to die of heart failure at around 30 weeks [25]. Therefore, we speculated that an effective therapeutic strategy for DCM would be to reduce the myocardial remodeling and prevent the ventricular dilatation caused by the numerical and functional loss of cardiomyocytes.

Functional benefits from myoblast transplantation in animal models of ischemic cardiomyopathy were recently established [7, 8] . Skeletal myoblasts transplanted into infarcted heart reduce remodeling and probably prevent myocardial cell loss. However, studies on its effects in DCM are rare, and show only improvement in systolic function, in cardiomyopathic hamsters [9–12]. Here, myoblast transplantation enhanced neovascularization and preserved the left ventricle function of BIO TO-2 hamsters, but it neither reduced nor enhanced myocardial fibrosis. Although some other reports have not shown neovascularization from myoblast transplantation, our results confirmed most of previous observations and provide new insights into the benefits of using myoblast transplantation to treat DCM [26, 27] .

We previously reported that HGF has a protective effect on the ischemic myocardium [13, 14]. More recently, Taniyama and coworkers [17] and we [18] showed that in vivo HGF transfection reduces myocardial fibrosis and reorganizes the impaired cytoskeletal proteins in cardiomyopathic hamsters. Therefore, we speculated that HGF transfection and myoblast transplantation work on cardiomyopathic hearts through different mechanisms, and therefore might be even more effective when used in combination.

We observed alpha- and beta-sarcoglycan expression and reduced myocardial fibrosis in the H and T+H groups but not in the C and T groups. Capillary density was increased by either HGF or myoblasts, and neovascularization was enhanced by the combination therapy. Interestingly, the reorganization of cytoskeletal proteins was associated with a transient delay in the deterioration of systolic function of DCM hearts in the H and T+H groups. The end-systolic dimension of the H group, however, deteriorated at the same speed as that of the C group after the delay, probably because the transfected HGF gene is expressed for only a few weeks [18]. Most importantly, the end-systolic dimension of the T+H group showed a slower deterioration, and the fractional shortening of the T+H group remained significantly higher than that of the other three groups, even at age 35 weeks. Therefore, HGF transfection and myoblast transplantation worked in different ways, and the combination preserved systolic function better than either treatment alone, in cardiomyopathic hamsters.

There are at least two reports on the efficacy of combining cellular cardiomyoplasty with angiogenic growth factors in a myocardial infarction model. Suzuki and associates [28] used vascular endothelial growth factor-expressing myoblasts, and Miyagawa and colleagues [19] showed HGF transfection and skeletal myoblast transplantation improved cardiac performance and increased neovascularization. Both authors speculated that the combined strategy worked better than either treatment alone because the growth factors provided a better environment for the transplanted cells by increasing the blood supply or improving the cytoskeletal organization, or both, in ischemic hearts. Although only low numbers of myoblasts survived after transplantation, and we could not show a significant difference in the number of transplanted myoblasts between the T and T+H groups (data not shown), our study suggested similar synergistic effects in DCM as in ischemia (Fig 2) [12, 28].

One important issue in genetic cardiomyopathy is the origin of the cells used for myoblast transplantation. Although autologous skeletal myoblasts do not cause immunological rejection, they have the same genetic predisposition as the heart muscle. Therefore, we used F1B hamsters as the myoblast source. Although F1B strains are frequently used as controls for TO-2 hamsters, and we did not observe signs of immune rejection in our histologic examinations, it is still possible that F1B myoblasts transplanted into our T and T+H groups were at a disadvantage for survival in TO-2 hearts.

As we reported previously, even transient transfection with HGF prolonged the survival of adult cardiomyopathic hamsters, but it is still unclear whether survival is prolonged by autologous myoblast transplantation [18]. Pouly and associates found myoblast transplantation to have little survival benefit, at least soon after transplantation [12]. Our results showed F1B myoblast transplantation significantly improved cardiac performance but failed to prolong adult TO-2 hamster survival. Nevertheless, the T+H and H groups showed significant survival prolongation compared with the C and T groups, and the median survival time of the T+H group was 6 weeks longer than that of the H group. Therefore, a therapeutic strategy that includes myoblast transplantation might enhance the survival of DCM patients.

There are two limitations of this study we wish to address. First, cardiac function was evaluated by measuring the left ventricle dimension by two-dimensional echocardiography, because more sensitive techniques, such as three-dimensional analysis, are very expensive. However, our analysis allowed for repeated monitoring, and the biweekly evaluations strengthen the reliability of our assessment. Second, a definitive measurement of the survival rate of the transplanted cells could not be performed because of a lack of suitable reagents; therefore, we could not determine precisely how the combination of HGF transfection and myoblast transplantation improved the cardiac function.

In summary, HGF gene transfection and myoblast transplantation improved the cardiac function of DCM hamsters, probably through different mechanisms, and the combination therapy preserved the cardiac performance better than either treatment alone. Thus, combination therapy seems promising for the treatment of DCM.


   Acknowledgments
Top
 Abstract
 Introduction
 Material and Methods
 Results
 Comment
 Acknowledgments
References
 
This work was supported by a Grant-in-Aid for Scientific Research in Japan No. 15659324. We thank Akiko Nishimura, Masako Yokoyama, and Aiko Miki for their excellent technical assistance.


   References
Top
 Abstract
 Introduction
 Material and Methods
 Results
 Comment
 Acknowledgments
 References
 

  1. Thiene G, Angelini A, Basso C, Calabrese F, Valente M. The new definition and classification of cardiomyopathies Adv Clin Pathol 2000;4:53-57.[Medline]
  2. Towbin JA, Bowles NE. The failing heart Nature 2002;415:227-233.[Medline]
  3. Canver CC, Chanda J. Heart transplantation Ann Thorac Surg 2001;72:658-660.[Free Full Text]
  4. Hosenpud JD, Bennett LE, Keck BM, Fiol B, Boucek MM, Novick RJ. The Registry of the International Society for Heart and Lung Transplantation: sixteenth official report—1999 J Heart Lung Transplant 1999;18:611-626.[Medline]
  5. Rose EA, Gelijns AC, Moskowitz AJ, et al. Long-term mechanical left ventricular assistance for end-stage heart failure N Engl J Med 2001;345:1435-1443.[Abstract/Free Full Text]
  6. Bowles NE, Bowles KR, Towbin JA. The "final common pathway" hypothesis and inherited cardiovascular diseaseThe role of cytoskeletal proteins in dilated cardiomyopathy. Herz 2000;25:168-175.[Medline]
  7. Murry CE, Wiseman RW, Schwartz SM, Hauschka SD. Skeletal myoblast transplantation for repair of myocardial necrosis J Clin Invest 1996;98:2512-2523.[Medline]
  8. Taylor DA, Atkins BZ, Hungspreugs P, et al. Regenerating functional myocardium: improved performance after skeletal myoblast transplantation Nat Med 1998;4:929-933.[Medline]
  9. Chachques JC, Acar C, Herreros J, et al. Cellular cardiomyoplasty: clinical application Ann Thorac Surg 2004;77:1121-1130.[Abstract/Free Full Text]
  10. Suzuki K, Murtuza B, Suzuki N, Smolenski RT, Yacoub MH. Intracoronary infusion of skeletal myoblasts improves cardiac function in doxorubicin-induced heart failure Circulation 2001;104(Suppl 1):213-217.
  11. Ohno N, Fedak PW, Weisel RD, Mickle DA, Fujii T, Li RK. Transplantation of cryopreserved muscle cells in dilated cardiomyopathy: effects on left ventricular geometry and function J Thorac Cardiovasc Surg 2003;126:1537-1548.[Abstract/Free Full Text]
  12. Pouly J, Hagege AA, Vilquin JT, et al. Does the functional efficacy of skeletal myoblast transplantation extend to nonischemic cardiomyopathy? Circulation 2004;110:1626-1631.[Abstract/Free Full Text]
  13. Taniyama Y, Morishita R, Aoki M, et al. Therapeutic angiogenesis induced by human hepatocyte growth factor gene in rat and rabbit hindlimb ischemia models: preclinical study for treatment of peripheral arterial disease Gene Ther 2001;8:181-189.[Medline]
  14. Liu Y, Rajur K, Tolbert E, Dworkin LD. Endogenous hepatocyte growth factor ameliorates chronic renal injury by activating matrix degradation pathways Kidney Int 2000;582048–3.
  15. Funatsu T, Sawa Y, Ohtake S, et al. Therapeutic angiogenesis in the ischemic canine heart induced by myocardial injection of naked complementary DNA plasmid encoding hepatocyte growth factor J Thorac Cardiovasc Surg 2002;124:1099-1105.[Abstract/Free Full Text]
  16. Ahmet I, Sawa Y, Yamaguchi T, Matsuda H. Gene transfer of hepatocyte growth factor improves angiogenesis and function of chronic ischemic myocardium in canine heart.
  17. Taniyama Y, Morishita R, Aoki M, et al. Angiogenesis and antifibrotic action by hepatocyte growth factor in cardiomyopathy Hypertension 2002;40:47-53.[Abstract/Free Full Text]
  18. Kondoh H, Sawa Y, Fukushima N, et al. Re-organization of cytoskeletal proteins and prolonged life expectancy caused by hepatocyte growth factor in a hamster model of late-phase dilated cardiomyopathy J Thorac Cardiovasc Surg 2005;130:295-302.[Abstract/Free Full Text]
  19. Miyagawa S, Sawa Y, Taketani S, et al. Myocardial regeneration therapy for heart failure: hepatocyte growth factor enhances the effect of cellular cardiomyoplasty Circulation 2002;105:2556-2561.[Abstract/Free Full Text]
  20. Seki T, Hagiya M, Shimonishi M, Nakamura T, Shimizu S. Organization of the human hepatocyte growth factor-encoding gene Gene 1991;102:213-219.[Medline]
  21. Kaneda Y, Iwai K, Uchida T. Expression of DNA cointroduced with nuclear protein in adult rat liver Science 1989;243:375-378.[Abstract/Free Full Text]
  22. Sakamoto A, Ono K, Abe M, et al. Both hypertrophic and dilated cardiomyopathies are caused by mutation of the same gene, delta-sarcoglycan, in hamster: an animal model of disrupted dystrophin-associated glycoprotein complex Proc Natl Acad Sci USA 1997;94:13873-13878.[Abstract/Free Full Text]
  23. Kawada T, Nakazawa M, Nakauchi S, et al. Rescue of hereditary form of dilated cardiomyopathy by rAAV-mediated somatic gene therapy: amelioration of morphological findings, sarcolemmal permeability, cardiac performances, and the prognosis of TO-2 hamsters Proc Natl Acad Sci USA 2002;99:901-906.[Abstract/Free Full Text]
  24. Bajusz E, Baker JR, Nixon CW, Homburger F. Spontaneous, hereditary myocardial degeneration and congestive heart failure in a strain of Syrian hamsters Ann NY Acad Sci 1969;156:105-129.[Medline]
  25. Shimizu T, Okamoto H, Chiba S, et al. VEGF-mediated angiogenesis is impaired by angiotensin type 1 receptor blockade in cardiomyopathic hamster hearts Cardiovasc Res 2003;58:203-212.[Abstract/Free Full Text]
  26. Chachques JC, Duarte F, Cattadori B, et al. Angiogenic growth factors and/or cellular therapy for myocardial regeneration: a comparative study J Thorac Cardiovasc Surg 2004;128:245-253.[Abstract/Free Full Text]
  27. Pagani FD, DerSimonian H, Zawadzka A, et al. Autologous skeletal myoblasts transplanted to ischemia-damaged myocardium in humansHistological analysis of cell survival and differentiation. J Am Coll Cardiol 2003;41:879-888.[Abstract/Free Full Text]
  28. Suzuki K, Murtuza B, Smolenski RT, et al. Cell transplantation for the treatment of acute myocardial infarction using vascular endothelial growth factor–expressing skeletal myoblasts Circulation 2001;104(Suppl 1):207-212.



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N. A. Gude, G. Emmanuel, W. Wu, C. T. Cottage, K. Fischer, P. Quijada, J. A. Muraski, R. Alvarez, M. Rubio, E. Schaefer, et al.
Activation of Notch-Mediated Protective Signaling in the Myocardium
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