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Domenico D'Amario,
Mauricio C Cabral-Da-Silva, Hanqiao Zheng,
Claudia Fiorini,
Polina Goichberg,
Elisabeth Steadman,
João Ferreira-Martins,
Fumihiro Sanada,
Marco Piccoli,
Donato Cappetta,
David A D'Alessandro,
Robert E Michler,
Toru Hosoda,
Luigi Anastasia,
Marcello Rota,
Annarosa Leri,
Piero Anversa,
Jan Kajstura
[show abstract]
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ABSTRACT: Age and coronary artery disease may negatively affect the function of human cardiac stem cells (hCSCs) and their potential therapeutic efficacy for autologous cell transplantation in the failing heart.
Insulin-like growth factor (IGF)-1, IGF-2, and angiotensin II (Ang II), as well as their receptors, IGF-1R, IGF-2R, and AT1R, were characterized in c-kit(+) hCSCs to establish whether these systems would allow us to separate hCSC classes with different growth reserve in the aging and diseased myocardium.
C-kit(+) hCSCs were collected from myocardial samples obtained from 24 patients, 48 to 86 years of age, undergoing elective cardiac surgery for coronary artery disease. The expression of IGF-1R in hCSCs recognized a young cell phenotype defined by long telomeres, high telomerase activity, enhanced cell proliferation, and attenuated apoptosis. In addition to IGF-1, IGF-1R(+) hCSCs secreted IGF-2 that promoted myocyte differentiation. Conversely, the presence of IGF-2R and AT1R, in the absence of IGF-1R, identified senescent hCSCs with impaired growth reserve and increased susceptibility to apoptosis. The ability of IGF-1R(+) hCSCs to regenerate infarcted myocardium was then compared with that of unselected c-kit(+) hCSCs. IGF-1R(+) hCSCs improved cardiomyogenesis and vasculogenesis. Pretreatment of IGF-1R(+) hCSCs with IGF-2 resulted in the formation of more mature myocytes and superior recovery of ventricular structure.
hCSCs expressing only IGF-1R synthesize both IGF-1 and IGF-2, which are potent modulators of stem cell replication, commitment to the myocyte lineage, and myocyte differentiation, which points to this hCSC subset as the ideal candidate cell for the management of human heart failure.
Circulation Research 06/2011; 108(12):1467-81. · 9.49 Impact Factor
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Jan Kajstura,
Marcello Rota,
Sean R Hall,
Toru Hosoda,
Domenico D'Amario,
Fumihiro Sanada, Hanqiao Zheng,
Barbara Ogórek,
Carlos Rondon-Clavo,
João Ferreira-Martins, [......],
Kathleen J Haley,
Silvana Bardelli,
Hussein Rayatzadeh,
Xiaoli Liu,
Federico Quaini,
Ronglih Liao,
Annarosa Leri,
Mark A Perrella,
Joseph Loscalzo,
Piero Anversa
[show abstract]
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ABSTRACT: Although progenitor cells have been described in distinct anatomical regions of the lung, description of resident stem cells has remained elusive.
Surgical lung-tissue specimens were studied in situ to identify and characterize human lung stem cells. We defined their phenotype and functional properties in vitro and in vivo.
Human lungs contain undifferentiated human lung stem cells nested in niches in the distal airways. These cells are self-renewing, clonogenic, and multipotent in vitro. After injection into damaged mouse lung in vivo, human lung stem cells form human bronchioles, alveoli, and pulmonary vessels integrated structurally and functionally with the damaged organ. The formation of a chimeric lung was confirmed by detection of human transcripts for epithelial and vascular genes. In addition, the self-renewal and long-term proliferation of human lung stem cells was shown in serial-transplantation assays.
Human lungs contain identifiable stem cells. In animal models, these cells participate in tissue homeostasis and regeneration. They have the undemonstrated potential to promote tissue restoration in patients with lung disease. (Funded by the National Institutes of Health.).
New England Journal of Medicine 05/2011; 364(19):1795-806. · 53.30 Impact Factor
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Toru Hosoda, Hanqiao Zheng,
Mauricio Cabral-da-Silva,
Fumihiro Sanada,
Noriko Ide-Iwata,
Barbara Ogórek,
João Ferreira-Martins,
Christian Arranto,
Domenico D'Amario,
Federica del Monte,
Konrad Urbanek,
David A D'Alessandro,
Robert E Michler,
Piero Anversa,
Marcello Rota,
Jan Kajstura,
Annarosa Leri
[show abstract]
[hide abstract]
ABSTRACT: Cardiac stem cells (CSCs) delivered to the infarcted heart generate a large number of small fetal-neonatal cardiomyocytes that fail to acquire the differentiated phenotype. However, the interaction of CSCs with postmitotic myocytes results in the formation of cells with adult characteristics.
On the basis of results of in vitro and in vivo assays, we report that the commitment of human CSCs (hCSCs) to the myocyte lineage and the generation of mature working cardiomyocytes are influenced by microRNA-499 (miR-499), which is barely detectable in hCSCs but is highly expressed in postmitotic human cardiomyocytes. miR-499 traverses gap junction channels and translocates to structurally coupled hCSCs favoring their differentiation into functionally competent cells. Expression of miR-499 in hCSCs represses the miR-499 target genes Sox6 and Rod1, enhancing cardiomyogenesis in vitro and after infarction in vivo. Although cardiac repair was detected in all cell-treated infarcted hearts, the aggregate volume of the regenerated myocyte mass and myocyte cell volume were greater in animals injected with hCSCs overexpressing miR-499. Treatment with hCSCs resulted in an improvement in ventricular function, consisting of a better preservation of developed pressure and positive and negative dP/dt after infarction. An additional positive effect on cardiac performance occurred with miR-499, pointing to enhanced myocyte differentiation/hypertrophy as the mechanism by which miR-499 potentiated the restoration of myocardial mass and function in the infarcted heart.
The recognition that miR-499 promotes the differentiation of hCSCs into mechanically integrated cardiomyocytes has important clinical implications for the treatment of human heart failure.
Circulation 03/2011; 123(12):1287-96. · 14.74 Impact Factor
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Polina Goichberg,
Yingnan Bai,
Domenico D'Amario,
João Ferreira-Martins,
Claudia Fiorini, Hanqiao Zheng,
Sergio Signore,
Federica del Monte,
Sergio Ottolenghi,
David A D'Alessandro,
Robert E Michler,
Toru Hosoda,
Piero Anversa,
Jan Kajstura,
Marcello Rota,
Annarosa Leri
[show abstract]
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ABSTRACT: Understanding the mechanisms that regulate trafficking of human cardiac stem cells (hCSCs) may lead to development of new therapeutic approaches for the failing heart.
We tested whether the motility of hCSCs in immunosuppressed infarcted animals is controlled by the guidance system that involves the interaction of Eph receptors with ephrin ligands.
Within the cardiac niches, cardiomyocytes expressed preferentially the ephrin A1 ligand, whereas hCSCs possessed the EphA2 receptor. Treatment of hCSCs with ephrin A1 resulted in the rapid internalization of the ephrin A1-EphA2 complex, posttranslational modifications of Src kinases, and morphological changes consistent with the acquisition of a motile cell phenotype. Ephrin A1 enhanced the motility of hCSCs in vitro, and their migration in vivo following acute myocardial infarction. At 2 weeks after infarction, the volume of the regenerated myocardium was 2-fold larger in animals injected with ephrin A1-activated hCSCs than in animals receiving control hCSCs; this difference was dictated by a greater number of newly formed cardiomyocytes and coronary vessels. The increased recovery in myocardial mass with ephrin A1-treated hCSCs was characterized by further restoration of cardiac function and by a reduction in arrhythmic events.
Ephrin A1 promotes the motility of EphA2-positive hCSCs, facilitates their migration to the area of damage, and enhances cardiac repair. Thus, in situ stimulation of resident hCSCs with ephrin A1 or their ex vivo activation before myocardial delivery improves cell targeting to sites of injury, possibly providing a novel strategy for the management of the diseased heart.
Circulation Research 03/2011; 108(9):1071-83. · 9.49 Impact Factor
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Domenico D'Amario,
Claudia Fiorini,
Patricia M Campbell,
Polina Goichberg,
Fumihiro Sanada, Hanqiao Zheng,
Toru Hosoda,
Marcello Rota,
John M Connell,
Robert P Gallegos,
Frederick G Welt,
Michael M Givertz,
Richard N Mitchell,
Annarosa Leri,
Jan Kajstura,
Marc A Pfeffer,
Piero Anversa
[show abstract]
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ABSTRACT: Two categories of cardiac stem cells (CSCs) with predominantly myogenic (mCSC) and vasculogenic (vCSC) properties have been characterized in the human heart. However, it is unknown whether functionally competent CSCs of both classes are present in the myocardium of patients affected by end-stage cardiac failure, and whether these cells can be harvested from relatively small myocardial samples.
To establish whether a clinically relevant number of mCSCs and vCSCs can be isolated and expanded from endomyocardial biopsies of patients undergoing cardiac transplantation or left ventricular assist device implantation.
Endomyocardial biopsies were collected with a bioptome from the right side of the septum of explanted hearts or the apical LV core at the time of left ventricular assist device implantation. Two to 5 biopsies from each patient were enzymatically dissociated, and, after expansion, cells were sorted for c-kit (mCSCs) or c-kit and KDR (vCSCs) and characterized. mCSCs and vCSCs constituted 97% and 3% of the c-kit population, respectively. Population doubling time averaged 27 hours in mCSCs and vCSCs; 5×10(6) mCSCs and vCSCs were obtained in 28 and 41 days, respectively. Both CSC classes possessed significant growth reserve as documented by high telomerase activity and relatively long telomeres. mCSCs formed mostly cardiomyocytes, and vCSCs endothelial and smooth muscle cells.
The growth properties of mCSCs and vCSCs isolated from endomyocardial biopsies from patients with advanced heart failure were comparable to those obtained previously from larger myocardial samples of patients undergoing elective cardiac surgery.
Circulation Research 02/2011; 108(7):857-61. · 9.49 Impact Factor
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Konrad Urbanek,
Mauricio Castro Cabral-da-Silva,
Noriko Ide-Iwata,
Silvia Maestroni,
Francesca Delucchi, Hanqiao Zheng,
João Ferreira-Martins,
Barbara Ogórek,
Domenico D'Amario,
Michael Bauer,
Gianpaolo Zerbini,
Marcello Rota,
Toru Hosoda,
Ronglih Liao,
Piero Anversa,
Jan Kajstura,
Annarosa Leri
[show abstract]
[hide abstract]
ABSTRACT: Physiological hypertrophy in the developing heart has been considered the product of an increase in volume of preexisting fetal cardiomyocytes in the absence of myocyte formation.
In this study, we tested whether the mouse heart at birth has a pool of cardiac stem cells (CSCs) that differentiate into myocytes contributing to the postnatal expansion of the parenchymal cell compartment.
We have found that the newborn heart contains a population of c-kit-positive CSCs that are lineage negative, self-renewing, and multipotent. CSCs express the Notch1 receptor and show the nuclear localization of its active fragment, N1ICD. In 60% of cases, N1ICD was coupled with the presence of Nkx2.5, indicating that the commitment of CSCs to the myocyte lineage is regulated by Notch1. Importantly, overexpression of N1ICD in neonatal CSCs significantly expanded the proportion of transit-amplifying myocytes. To establish whether these in vitro findings had a functional counterpart in vivo, the Notch pathway was blocked in newborn mice by administration of a gamma-secretase inhibitor. This intervention resulted in the development of a dilated myopathy and high mortality rates. Ventricular decompensation was characterized by a 62% reduction in amplifying myocytes, which resulted in a 54% decrease in myocyte number. After cessation of Notch blockade and recovery of myocyte regeneration, cardiac anatomy and function were largely restored.
Notch1 signaling is a critical determinant of CSC growth and differentiation; when this cascade of events is altered, cardiomyogenesis is impaired, physiological cardiac hypertrophy is prevented, and a life-threatening myopathy supervenes.
Circulation Research 08/2010; 107(3):429-41. · 9.49 Impact Factor
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Jan Kajstura,
Konrad Urbanek,
Shira Perl,
Toru Hosoda, Hanqiao Zheng,
Barbara Ogórek,
João Ferreira-Martins,
Polina Goichberg,
Carlos Rondon-Clavo,
Fumihiro Sanada,
Domenico D'Amario,
Marcello Rota,
Federica Del Monte,
Donald Orlic,
John Tisdale,
Annarosa Leri,
Piero Anversa
[show abstract]
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ABSTRACT: The ability of the human heart to regenerate large quantities of myocytes remains controversial, and the extent of myocyte renewal claimed by different laboratories varies from none to nearly 20% per year.
To address this issue, we examined the percentage of myocytes, endothelial cells, and fibroblasts labeled by iododeoxyuridine in postmortem samples obtained from cancer patients who received the thymidine analog for therapeutic purposes. Additionally, the potential contribution of DNA repair, polyploidy, and cell fusion to the measurement of myocyte regeneration was determined.
The fraction of myocytes labeled by iododeoxyuridine ranged from 2.5% to 46%, and similar values were found in fibroblasts and endothelial cells. An average 22%, 20%, and 13% new myocytes, fibroblasts, and endothelial cells were generated per year, suggesting that the lifespan of these cells was approximately 4.5, 5, and 8 years, respectively. The newly formed cardiac cells showed a fully differentiated adult phenotype and did not express the senescence-associated protein p16(INK4a). Moreover, measurements by confocal microscopy and flow cytometry documented that the human heart is composed predominantly of myocytes with 2n diploid DNA content and that tetraploid and octaploid nuclei constitute only a small fraction of the parenchymal cell pool. Importantly, DNA repair, ploidy formation, and cell fusion were not implicated in the assessment of myocyte regeneration.
Our findings indicate that the human heart has a significant growth reserve and replaces its myocyte and nonmyocyte compartment several times during the course of life.
Circulation Research 07/2010; 107(2):305-15. · 9.49 Impact Factor
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Antonella De Angelis,
Elena Piegari,
Donato Cappetta,
Laura Marino,
Amelia Filippelli,
Liberato Berrino,
João Ferreira-Martins, Hanqiao Zheng,
Toru Hosoda,
Marcello Rota,
Konrad Urbanek,
Jan Kajstura,
Annarosa Leri,
Francesco Rossi,
Piero Anversa
[show abstract]
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ABSTRACT: Anthracyclines are the most effective drugs available in the treatment of neoplastic diseases; however, they have profound consequences on the structure and function of the heart, which over time cause a cardiomyopathy that leads to congestive heart failure.
Administration of doxorubicin in rats led to a dilated myopathy, heart failure, and death. To test whether the effects of doxorubicin on cardiac anatomy and function were mediated by alterations in cardiac progenitor cells (CPCs), these cells were exposed to the anthracycline, which increased the formation of reactive oxygen species and caused increases in DNA damage, expression of p53, telomere attrition, and apoptosis. Additionally, doxorubicin resulted in cell-cycle arrest at the G2/M transition, which led to a significant decrease in CPC growth. Doxorubicin elicited multiple molecular adaptations; the massive apoptotic death that occurred in CPCs in the presence of anthracycline imposed on the surviving CPC pool the activation of several pathways aimed at preservation of the primitive state, cell division, lineage differentiation, and repair of damaged DNA. To establish whether delivery of syngeneic progenitor cells opposed the progression of doxorubicin cardiotoxicity, enhanced green fluorescent protein-labeled CPCs were injected in the failing myocardium; this treatment promoted regeneration of cardiomyocytes and vascular structures, which improved ventricular performance and rate of animal survival.
Our results raise the possibility that autologous CPCs can be obtained before antineoplastic drugs are given to cancer patients and subsequently administered to individuals who are particularly sensitive to the cardiotoxicity of these agents for prevention or management of heart failure.
Circulation 01/2010; 121(2):276-92. · 14.74 Impact Factor
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David A D'Alessandro,
Jan Kajstura,
Toru Hosoda,
Alessandro Gatti,
Ricardo Bello,
Federico Mosna,
Silvana Bardelli, Hanqiao Zheng,
Domenico D'Amario,
M Elena Padin-Iruegas,
Adriana Bastos Carvalho,
Marcello Rota,
Michael O Zembala,
David Stern,
Ornella Rimoldi,
Konrad Urbanek,
Robert E Michler,
Annarosa Leri,
Piero Anversa
[show abstract]
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ABSTRACT: Chronic rejection, accelerated coronary atherosclerosis, myocardial infarction, and ischemic heart failure determine the unfavorable evolution of the transplanted heart in humans.
Here we tested whether the pathological manifestations of the transplanted heart can be corrected partly by a strategy that implements the use of cardiac progenitor cells from the recipient to repopulate the donor heart with immunocompatible cardiomyocytes and coronary vessels.
A large number of cardiomyocytes and coronary vessels were created in a rather short period of time from the delivery, engraftment, and differentiation of cardiac progenitor cells from the recipient. A proportion of newly formed cardiomyocytes acquired adult characteristics and was integrated structurally and functionally within the transplant. Similarly, the regenerated arteries, arterioles, and capillaries were operative and contributed to the oxygenation of the chimeric myocardium. Attenuation in the extent of acute damage by repopulating cardiomyocytes and vessels decreased significantly the magnitude of myocardial scarring preserving partly the integrity of the donor heart.
Our data suggest that tissue regeneration by differentiation of recipient cardiac progenitor cells restored a significant portion of the rejected donor myocardium. Ultimately, immunosuppressive therapy may be only partially required improving quality of life and lifespan of patients with cardiac transplantation.
Circulation Research 10/2009; 105(11):1128-40. · 9.49 Impact Factor
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Toru Hosoda,
Domenico D'Amario,
Mauricio Castro Cabral-Da-Silva, Hanqiao Zheng,
M Elena Padin-Iruegas,
Barbara Ogorek,
João Ferreira-Martins,
Saori Yasuzawa-Amano,
Katsuya Amano,
Noriko Ide-Iwata,
Wei Cheng,
Marcello Rota,
Konrad Urbanek,
Jan Kajstura,
Piero Anversa,
Annarosa Leri
[show abstract]
[hide abstract]
ABSTRACT: An analysis of the clonality of cardiac progenitor cells (CPCs) and myocyte turnover in vivo requires genetic tagging of the undifferentiated cells so that the clonal marker of individual mother cells is traced in the specialized progeny. CPC niches in the atria and apex of the mouse heart were infected with a lentivirus carrying EGFP, and the destiny of the tagged cells was determined 1-5 months later. A common integration site was identified in isolated CPCs, cardiomyocytes, endothelial cells (ECs), and fibroblasts, documenting CPC self-renewal and multipotentiality and the clonal origin of the differentiated cell populations. Subsequently, the degree of EGFP-lentiviral infection of CPCs was evaluated 2-4 days after injection, and the number of myocytes expressing the reporter gene was measured 6 months later. A BrdU pulse-chasing protocol was also introduced as an additional assay for the analysis of myocyte turnover. Over a period of 6 months, each EGFP-positive CPC divided approximately eight times generating 230 cardiomyocytes; this value was consistent with the number of newly formed cells labeled by BrdU. To determine whether, human CPCs (hCPCs) are self-renewing and multipotent, these cells were transduced with the EGFP-lentivirus and injected after acute myocardial infarction in immunosuppressed rats. hCPCs, myocytes, ECs, and fibroblasts collected from the regenerated myocardium showed common viral integration sites in the human genome. Thus, our results indicate that the adult heart contains a pool of resident stem cells that regulate cardiac homeostasis and repair.
Proceedings of the National Academy of Sciences 10/2009; 106(40):17169-74. · 9.68 Impact Factor
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Alessandro Boni,
Konrad Urbanek,
Angelo Nascimbene,
Toru Hosoda, Hanqiao Zheng,
Francesca Delucchi,
Katsuya Amano,
Arantxa Gonzalez,
Serena Vitale,
Caroline Ojaimi,
Roberto Rizzi,
Roberto Bolli,
Katherine E Yutzey,
Marcello Rota,
Jan Kajstura,
Piero Anversa,
Annarosa Leri
[show abstract]
[hide abstract]
ABSTRACT: The Notch receptor mediates cell fate decision in multiple organs. In the current work we tested the hypothesis that Nkx2.5 is a target gene of Notch1 and raised the possibility that Notch1 regulates myocyte commitment in the adult heart. Cardiac progenitor cells (CPCs) in the niches express Notch1 receptor, and the supporting cells exhibit the Notch ligand Jagged1. The nuclear translocation of Notch1 intracellular domain (N1ICD) up-regulates Nkx2.5 in CPCs and promotes the formation of cycling myocytes in vitro. N1ICD and RBP-Jk form a protein complex, which in turn binds to the Nkx2.5 promoter initiating transcription and myocyte differentiation. In contrast, transcription factors of vascular cells are down-regulated by Jagged1 activation of the Notch1 pathway. Importantly, inhibition of Notch1 in infarcted mice impairs the commitment of resident CPCs to the myocyte lineage opposing cardiomyogenesis. These observations indicate that Notch1 favors the early specification of CPCs to the myocyte phenotype but maintains the newly formed cells in a highly proliferative state. Dividing Nkx2.5-positive myocytes correspond to transit amplifying cells, which condition the replicative capacity of the heart. In conclusion, Notch1 may have critical implications in the control of heart homeostasis and its adaptation to pathologic states.
Proceedings of the National Academy of Sciences 11/2008; 105(40):15529-34. · 9.68 Impact Factor
