Targeting cardiac fibrosis with engineered T cells.

Fibrosis is observed in nearly every form of myocardial disease1. Upon injury, cardiac fibroblasts in the heart begin to remodel the myocardium by depositing excess extracellular matrix, resulting in increased stiffness and reduced compliance of the tissue. Excessive cardiac fibrosis is an important factor in the progression of various forms of cardiac disease and heart failure2. However, clinical interventions and therapies that target fibrosis remain limited3. Here we demonstrate the efficacy of redirected T cell immunotherapy to specifically target pathological cardiac fibrosis in mice. We find that cardiac fibroblasts that express a xenogeneic antigen can be effectively targeted and ablated by adoptive transfer of antigen-specific CD8+ T cells. Through expression analysis of the gene signatures of cardiac fibroblasts obtained from healthy and diseased human hearts, we identify an endogenous target of cardiac fibroblasts-fibroblast activation protein. Adoptive transfer of T cells that express a chimeric antigen receptor against fibroblast activation protein results in a significant reduction in cardiac fibrosis and restoration of function after injury in mice. These results provide proof-of-principle for the development of immunotherapeutic drugs for the treatment of cardiac disease.

Cardiac fibrosis: Cell biological mechanisms, molecular pathways and therapeutic opportunities.

Cardiac fibrosis is a common pathophysiologic companion of most myocardial diseases, and is associated with systolic and diastolic dysfunction, arrhythmogenesis, and adverse outcome. Because the adult mammalian heart has negligible regenerative capacity, death of a large number of cardiomyocytes results in reparative fibrosis, a process that is critical for preservation of the structural integrity of the infarcted ventricle. On the other hand, pathophysiologic stimuli, such as pressure overload, volume overload, metabolic dysfunction, and aging may cause interstitial and perivascular fibrosis in the absence of infarction. Activated myofibroblasts are the main effector cells in cardiac fibrosis; their expansion following myocardial injury is primarily driven through activation of resident interstitial cell populations. Several other cell types, including cardiomyocytes, endothelial cells, pericytes, macrophages, lymphocytes and mast cells may contribute to the fibrotic process, by producing proteases that participate in matrix metabolism, by secreting fibrogenic mediators and matricellular proteins, or by exerting contact-dependent actions on fibroblast phenotype. The mechanisms of induction of fibrogenic signals are dependent on the type of primary myocardial injury. Activation of neurohumoral pathways stimulates fibroblasts both directly, and through effects on immune cell populations. Cytokines and growth factors, such as Tumor Necrosis Factor-α, Interleukin (IL)-1, IL-10, chemokines, members of the Transforming Growth Factor-ß family, IL-11, and Platelet-Derived Growth Factors are secreted in the cardiac interstitium and play distinct roles in activating specific aspects of the fibrotic response. Secreted fibrogenic mediators and matricellular proteins bind to cell surface receptors in fibroblasts, such as cytokine receptors, integrins, syndecans and CD44, and transduce intracellular signaling cascades that regulate genes involved in synthesis, processing and metabolism of the extracellular matrix. Endogenous pathways involved in negative regulation of fibrosis are critical for cardiac repair and may protect the myocardium from excessive fibrogenic responses. Due to the reparative nature of many forms of cardiac fibrosis, targeting fibrotic remodeling following myocardial injury poses major challenges. Development of effective therapies will require careful dissection of the cell biological mechanisms, study of the functional consequences of fibrotic changes on the myocardium, and identification of heart failure patient subsets with overactive fibrotic responses.

Cardiac manifestations of parasitic diseases.

The heart may be affected directly or indirectly by a variety of protozoa and helminths. This involvement may manifest in different ways, but the syndromes resulting from impairment of the myocardium and pericardium are the most frequent. The myocardium may be invaded by parasites that trigger local inflammatory response with subsequent myocarditis or cardiomyopathy, as occurs in Chagas disease, African trypanosomiasis, toxoplasmosis, trichinellosis and infection with free-living amoebae. In amoebiasis and echinococcosis, the pericardium is the structure most frequently involved with consequent pericardial effusion, acute pericarditis, cardiac tamponade or constrictive pericarditis. Chronic hypereosinophilia due to helminth infections, especially filarial infections, has been associated with the development of tropical endomyocardial fibrosis, a severe form of restrictive cardiomyopathy. Schistosomiasis-associated lung vasculature involvement may cause pulmonary hypertension (PH) and cor pulmonale Tropical pulmonary eosinophilia, which is characterised by progressive interstitial fibrosis and restrictive lung disease, may lead to PH and its consequences may occur in the course of filarial infections. Intracardiac rupture of an Echinococcus cyst can cause membrane or secondary cysts embolisation to the lungs or organs supplied by the systemic circulation. Although unusual causes of cardiac disease outside the endemic areas, heart involvement by parasites should be considered in the differential diagnosis especially of myocardial and/or pericardial diseases of unknown aetiology in both immunocompetent and immunocompromised individuals. In this review, we updated and summarised the current knowledge on the major heart diseases caused by protozoan and metazoan parasites, which either involve the heart directly or otherwise influence the heart adversely.

AAV micro-dystrophin gene therapy alleviates stress-induced cardiac death but not myocardial fibrosis in >21-m-old mdx mice, an end-stage model of Duchenne muscular dystrophy cardiomyopathy.

Duchenne muscular dystrophy (DMD) is a fatal genetic disease caused by the absence of the sarcolemmal protein dystrophin. Dilated cardiomyopathy leading to heart failure is a significant source of morbidity and mortality in DMD. We recently demonstrated amelioration of DMD heart disease in 16 to 20-m-old dystrophin-null mdx mice using adeno-associated virus (AAV) mediated micro-dystrophin gene therapy. DMD patients show severe heart disease near the end of their life expectancy. Similarly, mdx mice exhibit profoundly worsening heart disease when they reach beyond 21 months of age. To more rigorously test micro-dystrophin therapy, we treated mdx mice that were between 21.2 and 22.7-m-old (average, 22.1 ± 0.2 months; N=8). The ∆R4-23/∆C micro-dystrophin gene was packaged in the cardiotropic AAV-9 virus. 5×10(12) viral genome particles/mouse were delivered to mdx mice via the tail vein. AAV transduction, myocardial fibrosis and heart function were examined 1.7 ± 0.2 months after gene therapy. Efficient micro-dystrophin expression was observed in the myocardium of treated mice. Despite the robust dystrophin expression, myocardial fibrosis was not mitigated. Most hemodynamic parameters were not improved either. However, ECG abnormalities were partially corrected. Importantly, treated mice became more resistant to dobutamine-induced cardiac death. In summary, we have revealed for the first time the potential benefits and limitations of AAV micro-dystrophin therapy in end-stage Duchenne dilated cardiomyopathy. Our findings have important implications for the use of AAV gene therapy in dilated cardiomyopathy and heart failure.

Intracardiac masses in young Africans: case reports and a brief review of the literature.

Intracardiac masses in the young occur in some conditions that are prevalent in Africa. Although usually non-malignant, they may present with refractory heart failure and other complications that can be fatal. In the majority of cases, the aetiologic differentiation can be achieved by careful history, physical examination, basic laboratory tests, and transthoracic echocardiography. We report three cases in young Africans and discuss the aetiology, clinical presentation, diagnosis, management, and outcome of selected conditions in resource-limited settings.

Therapeutic approaches to patients with hypereosinophilic syndromes.

Hypereosinophilic syndromes (HES) are a heterogeneous group of disorders that range from asymptomatic eosinophilia > 1,500/mL to aggressive disease complicated by life-threatening end organ involvement, including endomyocardial fibrosis and thromboembolism. To complicate matters further, similar clinical manifestations can occur in the setting of marked eosinophilia due to helminth infection, drug hypersensitivity, and other causes. In the past, therapy was guided only by the exclusion of these secondary causes of eosinophilia and the severity of the clinical manifestations. More recently, the availability of novel targeted therapies and a better understanding of the etiologies of some subtypes of HES have necessitated a more structured approach.

Unusual cardiovascular events in Behçet's disease.

OBJECTIVES: To report on cardiovascular involvement in Behçet's disease (BD). METHODS: A retrospective analysis of clinical, EKG, echodoppler, CT-scan, MRI, conventional angiography, treatment and follow-up data was undertaken in 4 patients suffering BD. RESULTS: Cardiac specific complications included coronary artery involvement (n=3), endomyocardial fibrosis (n=1), left ventricle spontaneous rupture with giant wall pseudo-aneurysm (n=1), and massive left ventricle thrombosis (n=1). Follow-up ranged from 1 month to 17 years. Surgery was complicated with vascular patch leakage, recurrent pseudo-aneurysm or upper-limb venous thrombosis in 2 patients who did not receive pre-operative specific treatment because of delayed BD diagnosis. High-dose steroids (n=4), colchicine (n=4), immunosuppressants (n=3) and anticoagulants (n=4) were eventually prescribed and stabilised cardiac disease in all cases. CONCLUSIONS: At time of life-threatening cardiac complications, BD was often overlooked. Prompt initiation of steroids and immunosuppressive treatment may prevent post-operative complications, recurrences and death.

Cardiac manifestation of the hypereosinophilic syndrome: new insights.

Cardiac manifestation is the major cause of morbidity in patients with hypereosinophilic syndrome (HES). Clinical features range from heart failure to arterial embolism, which are caused by thickening of the endocardium and mural left ventricular thrombosis. Modern magnetic resonance imaging and echocardiography are able to detect fibrosis, eosinophilic infiltrate and thrombi to stage the fibrotic evolution of the disease. Treatment of HES involves standard medication for heart failure, anticoagulant therapy, immunosuppressive therapy and potentially surgical resection. The outcome of HES depends on both the progression of endocardial fibrosis and associated complications and the 5-year mortality is estimated at 30%.

Adenovirus-mediated stromal cell-derived factor-1 alpha gene transfer improves cardiac structure and function after experimental myocardial infarction through angiogenic and antifibrotic actions.

UNLABELLED: Stromal cell-derived factor 1 alpha (SDF-1) is not only a major chemotactic factor, but also an inducer of angiogenesis. The effects of SDF-1 alpha on the left ventricular remodeling in a rat myocardial infarction (MI) model were analyzed. Myocardial infarction was induced by ligation of the left coronary artery in rats. 0.5 x 10(10) pfu/ml AdV-SDF-1 or 0.5 x 10(10) pfu/ml Adv-LacZ were immediately injected into the infarcted myocardium, 120 microl cell-free PBS were injected into the infarcted region or the myocardial wall in control, and sham group, respectively. We found that AdV-SDF-1 group had higher LVSP and +/-dP/dt(max), lower LVEDP compared to control or Adv-LacZ group. The number of c-Kit(+) stem cells, and gene expression of SDF-1, VEGF and bFGF were obviously increased, which was associated with reduced infarct size, thicker left ventricle wall, greater vascular density and cardiocytes density in infarcted hearts of AdV-SDF-1 group. Furthermore, the expression of collagen type I and type III mRNA, and collagen accumulation in the infarcted area was lower, which was associated with decreased TGF-beta1, TIMP-1 and TIMP-2 expression in AdV-SDF-1 group. CONCLUSION: SDF-1 alpha could improve cardiac structure and function after Myocardial infarction through angiogenic and anti-fibrotic actions.

[Endomyocardial fibrosis: a rare case of restrictive cardiomyopathy in a Caucasian female]. / Fibrose endomyocardique: une cause rare de cardiomyopathie restrictive chez une patiente caucasienne.

The authors report the case of endomyocardial fibrosis diagnosed in a young Caucasian female presenting with progressive congestive cardiac failure. The diagnosis was suspected on the echocardiographic, magnetic resonance imaging and cardiac catheterisation findings in association with the clinical presentation. After a short course of symptomatic medical therapy, the patient underwent the only curative treatment of this pathology, surgical endocardectomy and combined valvular surgery. The confirmation of the diagnosis was obtained a posteriori by histopathological examination of the operative findings which showed appearances of endomyocardial fibrosis similar to those observed in tropical regions. The patient was discharged on the eighth postoperative day, much improved clinically, and follow-up at one year was very satisfactory.

[Idiopathic hypereosinophilia with cardiac involvement]. / Idiopathische Hypereosinophilie mit kardialer Beteiligung.

HISTORY AND CLINICAL FINDINGS: A 34-year-old previously healthy woman was admitted to another hospital because of abdominal pain, cough and dyspnea. Peripheral eosinophilia was present. Two months later she was admitted to the cardiology department with signs of mitral regurgitation. Dyspnea, fatigue, skin rashes with pruritus and a systolic murmur were noted. INVESTIGATIONS: Laboratory tests showed 11.4/nl leukocytes (normal range 4.8-10.8/nl) with an eosinophilia of 19% (normal range < 4%) corresponding to 2.2/nl. Cardiac magnetic resonance imaging revealed endomyocardial fibrosis involving the posterior mitral leaflet with resulting valvular regurgitation. Doppler ultrasound showed restrictive heart failure. DIAGNOSIS, THERAPY AND FURTHER COURSE: The diagnosis of idiopathic hypereosinophilia most likely as part of hypereosinophilic syndrome with cardiac involvement was made. The patient was treated with digitalis, diuretics and peptidyl dipeptidase (PDP) inhibitor. The treatment with glucocorticoids and cytotoxic agent to achieve a reduction of eosinophil count was ended by the patient a few weeks later. CONCLUSION: The hypereosinophilic syndrome with endomyocardial fibrosis is rare, and its prognosis is grave. The pathophysiological mechanisms are not entirely clear, nearly 70 years after Löffler first described fibrous endocarditis with eosinophilia. Patients receive symptomatic medical therapies. Additional surgical treatment has been reported,. Antihypereosinophilic therapy is used to control the disease.

[Löffler fibroblastic endocarditis in the thrombotic stages in isolated right ventricular tissue eosinophilia]. / Endokarditis fibroplastica Löffler im thrombotischen Stadium bei isolierter rechtsventrikulärer Gewebe-Eosinophilie.

We report on a male, 31 year old, Turkish patient with an intracardiac mass in the right ventricle, reduction of performance and weight, as well as intermittent fever. No eosinophilia was documented in the peripheral blood; cardiac function was primarily normal. Besides the differential diagnosis of Löffer's endocarditis (endomyocardial fibrosis) an inflammatory disease and a malignant cardiac tumor were suggested. The diagnosis of Löffler's endocarditis could not be confirmed morphologically by echocardiography nor histologically by right ventricular biopsy. Operative removal of the mass lesion was necessary because of fast tumor progression, fulminant pulmonary embolism, and infiltration of the tricuspid valve. Only then, histologically Löffler's eosinophilic endocarditis of thrombotic stage was diagnosed. Antiphlogistic therapy with cortisone was initially performed. With a dose reduction after 6 months, a relapse of the thrombotic mass occurred. Therefore, continuous treatment with cortisone and azathioprine was induced followed by further tumor regression and further clinical stabilization since 8 months of treatment.

[The heart and the eosinophil]. / Le coeur et l'eosinophile.

The beneficial effects of polynuclear eosinophils (PE) are well known. However, under certain circumstances, PE can be harmful. The heart is a prime target for PE toxicity which is due to release of basic proteins by eosinophils including major basic protein, cationic protein, and peroxidase. The most common manifestation of PE toxicity is chronic parietal endocarditis (CPE) which regroups two entities: Loeffler's fibroplastic endocarditis and Davies' endomyocardial fibrosis. Loeffler's fibroplastic endocarditis occurs mainly in temperate climates. Patients present high, persistent eosinophil levels similar to those observed in essential hypereosinophilic syndrome (EHS) or Chusid syndrome. Davies' endomyocardial fibrosis occurs in tropical countries where eosinophilic helminthiasis are endemic. The incidence of eosinophilic myocarditis (EM) is low but probably underestimated. EM can be observed in any case involving PE and has been described in many cases of drug-induced atopy, in Churg and Strauss syndrome, and in EHS. The most common cause of death is short-term occurrence of cardiogenic shock or dilated hypokinetic cardiomyopathy. Some patients have been successfully treated by early, intensive corticosteroid therapy and/or heart transplantation. The nosological classification of EM and CPE remains controversial. The two disorders may form a continuum with CPE as the second phase. Other authors have suggested that EM and CPE result from the action of PE on two distinct targets, i.e. endothelial cells for EM and myocytes for CPE. In the future, it may be possible to identify subjects with a predisposition to PE-induced heart disease by studying of genes coding for interleukins (IL-5, IL-4, IL-3) and GM-CSF in the 5q31-q33 region of chromosome 5.

Endomiocardiofibrose: modelo de miocardiopatia restritiva / Endomyocardial fibrosis: model of restrictive myocardiopathy

Os autores, nesta revisao literária, analisam uma importante causa de miocardiopatia tipo restritiva no nosso meio, a endomiocardiofibrose. Descrevem a etiopatogenia, patologia, diagnóstico clínico e complementar e seu tratamento. Concluem que a etiologia ainda é desconhecida, que seu diagnóstico tem evoluido às custas da ecocardiografia Doppler e que o tratamento permanece pouco eficaz, uma vez que a mortalidade cirúrgica, apesar do avanço nas técnicas, é maior que a esperada e a história natural dos pacientes tratados clinicamente também demonstra alta taxa de óbitos em curto espaço de tempo.

Endomiocardiofibrose. Evoluçäo de pacientes com tratamento clínico e cirúrgico / Endomyocardial fibrosis: outcome according to the kind of treatment

Objetivo: Observar a evoluçäo de um grupo de pacientes, durante determinado período, conforme o tratamento instituído. Casuística e Métodos: Cento e vinte e um pacientes com endomiocardiofibrose (EMF) foram seguidos por um período de 11 anos (média = 32 meses). As idades oscilaram entre cinco e 64 anos (méida = 30 anos) e 41 homens e 80 mulheres. Observou-se, durante esse período, a evoluçäo conforme o tratamento instituido. A lesäo era biventricular em 70 casos, ventricular esquerda em 36 e ventricular direita em 15. Os pacientes foram divididos em dois grupos: 62 tratados clinicamente e 59 submetidos a cirurgia para ressecçäo da fibrose e substituiçäo ou reconstruçäo das valvas atrioventriculares. Todos os casos operados apresentavam classe funcional III ou IV (NYHA). Resultados: No grupo clínico houve 24 óbitos, 21 em pacientes com classe IV, um com classe III e dois com classe II. No grupo cirúrgico houve 18 óbitos, 12 precoces e seis tardios. Conclusäo: 1) Todos os pacientes que sobreviveram a operaçäo mostraram melhora da classe funcional; 2) a mortalidade cirúrgica diminuiu nos últimos anos; 3) o tratamento clínico é indicado em pacientes em classe funcional I ou II, por ser a mortalidade baixa neste grupo; 4) o tratamento cirúrgico é indicado em pacientes em classe funcional III ou IV, por ser a mortalidade com o tratamento clínico elevada neste grupos

Purpose: To follow-up a group of patients during a period of time after either clinical or surgical approach. Patients and Methods: A hundred and twenty-one patients were studied retrospectively with endomyocardial fibrosis (EMF) for a period that varied from one month to 11 years (mean = 32 months). Upon entrance, patients had from 5 to 64 years of age (mean = 30), being 41 male and 80 female. Biventricular involvement were present in 70 cases, whereas 36 showed pure left and 15 pure right ventricular involvement. The patients were devided in two grupos. 62 treated clinically, and 59 surgically, the latest defined as fibrosis resection added to atrioventricular valve rebilding or replacement. All surgical cases had a prior III to IV functional class (NYHA). Results: In the clinical group there were 24 fatalities, 21 who were in class IV, 1 in class III, and 2 in class II. In the surgical group there were 18 fatalalities, 12 early and 6 late. Conclusions: (1) All the patients who survived the operation showed functional class improvement; (2) surgical mortality decreased in the latest years; (3) clinical treatment is indicated to patients in classes I and II, groups with low mortality rates; (4) surgical treatment is indicated to patients in classes III and IV, provided that clinical mortality is high in these groups of patients.

Endomiocardiofibrose / Endomyocardial fibrosis

Baseado na experiência do Instituto do Coraçäo, fundamentada no estudo de 135 casos, discute-se o papel dos exames complementares no diagnóstico da endomiocardiofibrose. Clínica de insuficiência cardíaca congestiva importante é a forma de apresentaçäo mais freqüente. No diagnóstico diferencial deve-se considerar outras síndromes restritivas, especialmente hipertensäo portal. Ecocardiografia e estudo hemodinâmico säo os exames complementares que permitem caracterizar a doença. A conduta clásica é a indicaçäo de ressecçäo da fibrose para todos os pacientes sintomáticos, melhorando-se com esta a sobrevida e a qualidade de vida. A conduta clínica pode ser mantida nos pacientes pouco sintomáticos. O comprometimento ventricular esquerdo é melhor tolerado, ficando os pacientes sintomáticos quando apresentam insuficiência mitral. A lesäo do ventrículo dieito é usualmente menos tolerada e pode determinar alteraçöes sistêmicas que alterariam o prognóstico (como insuficiência hepática) e, portanto, devem ser acompanhadas cuidadosamente. A correçäo cirúrgica deve ser adotada sempre que houver deterioraçäo clínica