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Amniotic membrane as a scaffold for melanocyte transplantation in patients with stable vitiligo

2012-12-06T01:44:53Z

Title: Amniotic membrane as a scaffold for melanocyte transplantation in patients with stable vitiligo Author(s) : Redondo, P. (Pedro); Gimenez-de-Azcarate, A. (Ana); Marques, L. (Laura); Garcia-Guzman, M. (María); Andreu, E.J. (Enrique José); Prosper, F. (Felipe) Abstract: Vitiligo is an acquired skin disease that significantly impacts the quality of life of patients. Medical treatment of vitiligo includes the use of melanocyte transplant, but the results are variable. We have treated 4 patients with either focal or generalized stable vitiligo using a graft of autologous melanocytes' culture on a denuded amniotic membrane (AM). A culture biopsy was obtained in every patient and grown in melanocytes' media for 10-14 days after which cells were transferred to a denuded AM and transplanted into the achromic lesions. Patients were followed for up to 6 months using clinical assessment of achromic lesions. Treated areas ranged between 4 cm(2) and 210.6 cm(2). Response to treatment was excellent in all patients with 90-95% repigmentation success rate. Our results demonstrate that transplantation of autologous melanocytes cultured on AM is a new, simple, and effective treatment for stable vitiligo.

Increased concentrations of tumor necrosis factor and interleukin-6 contribute to the hemostatic abnormalities in advanced liver disease

2012-07-03T17:51:35Z

Title: Increased concentrations of tumor necrosis factor and interleukin-6 contribute to the hemostatic abnormalities in advanced liver disease Author(s) : Paramo, J.A. (José Antonio); Sangro, B. (Bruno); Prosper, F. (Felipe); Quiroga, J. (Jorge); Rifon, J. (José); Rocha, E. (Eduardo) Abstract: Abnormal cytokine levels have been described in patients with chronic liver disease, but studies correlating cytokine homeostasis with abnormalities in coagulation and fibrinolysis are lacking. In order to establish a link between cytokines and the hemostatic changes the following parameters were determined in 44 patients with cirrhosis (alcoholic = 15, postnecrotic = 22, others = 7): TNF-alpha, IL-6, thrombin-antithrombin (TAT) complexes, prothrombin fragment 1 + 2 (F1 + 2) and t-PA by using enzyme-linked immunosorbent assays, and PAI-1, plasminogen and alpha 2-antiplasmin (alpha 2-AP) by using chromogenic substrates. All patients were at stages B and C of Child's classification when entering the study. Mean cytokine concentrations were significantly higher in cirrhotic patients as compared to age- and sex-matched controls (p < 0.009). There was a significant increase of TAT (p < 0.02) and F1 + 2 (p < 0.001) in the patients groups, suggesting a grade of intravascular coagulation. A hyperfibrinolytic state as demonstrated by an increase of t-PA and decrease of plasminogen and alpha 2-AP was also observed (p < 0.001). We could define a subgroup of patients with cytokine values higher than 20 pg/ml. Interestingly, in this group there was a significant increase of TAT (p < 0.04) and t-PA (p < 0.02) levels and a decrease of plasminogen and alpha 2-AP (p < 0.02) as compared to values observed in patients with cytokines lower than 20 pg/ml. We conclude that high levels of TNF-alpha and IL-6 may contribute to hyperfibrinolysis and intravascular coagulation in patients with liver cirrhosis, as assessed by the increase of TAT and t-PA levels and the reduction of plasminogen and alpha 2-AP.

Frequent and simultaneous epigenetic inactivation of TP53 pathway genes in acute lymphoblastic leukemia

2012-06-04T00:09:48Z

Title: Frequent and simultaneous epigenetic inactivation of TP53 pathway genes in acute lymphoblastic leukemia Author(s) : Vilas–Zornoza, A. (Amaya); Agirre, X. (Xabier); Martin-Palanco, V. (Vanesa); Martin-Subero, J.I. (José Ignacio); San-Jose-Eneriz, E. (Edurne); Garate, L. (Leire); Alvarez, S. (Sara); Miranda, E. (Estibaliz); Rodriguez-Otero, P. (Paula); Rifon, J. (José); Torres, A. (Antonio); Calasanz, M.J. (María José); Cigudosa, J.C. (Juan Cruz); Roman-Gomez, J. (José); Prosper, F. (Felipe) Abstract: Aberrant DNA methylation is one of the most frequent alterations in patients with Acute Lymphoblastic Leukemia (ALL). Using methylation bead arrays we analyzed the methylation status of 807 genes implicated in cancer in a group of ALL samples at diagnosis (n = 48). We found that 154 genes were methylated in more than 10% of ALL samples. Interestingly, the expression of 13 genes implicated in the TP53 pathway was downregulated by hypermethylation. Direct or indirect activation of TP53 pathway with 5-aza-29-deoxycitidine, Curcumin or Nutlin-3 induced an increase in apoptosis of ALL cells. The results obtained with the initial group of 48 patients was validated retrospectively in a second cohort of 200 newly diagnosed ALL patients. Methylation of at least 1 of the 13 genes implicated in the TP53 pathway was observed in 78% of the patients, which significantly correlated with a higher relapse (p = 0.001) and mortality (p,0.001) rate being an independent prognostic factor for disease-free survival (DFS) (p = 0.006) and overall survival (OS) (p = 0.005) in the multivariate analysis. All these findings indicate that TP53 pathway is altered by epigenetic mechanisms in the majority of ALL patients and correlates with prognosis. Treatments with compounds that may reverse the epigenetic abnormalities or activate directly the p53 pathway represent a new therapeutic alternative for patients with ALL.

Histological and ultrastructural comparison of cauterization and thrombosis stroke models in immune-deficient mice

2012-06-02T00:08:30Z

Title: Histological and ultrastructural comparison of cauterization and thrombosis stroke models in immune-deficient mice Author(s) : Mora-Lee, S. (Silvia); Sirerol-Pique, M.S. (M. Salome); Gutierrez-Perez, M. (María); Lopez, T. (Tania); Casado-Nieto, M. (Maite); Jauquicoam, C. (Carlos); Abizanda, G. (Gloria); Romaguera-Ros, M. (Mirian); Gomez-Pinedo, U. (Ulises); Prosper, F. (Felipe); Garcia-Verdugo, J.M. (José Manuel) Abstract: Background: Stroke models are essential tools in experimental stroke. Although several models of stroke have been developed in a variety of animals, with the development of transgenic mice there is the need to develop a reliable and reproducible stroke model in mice, which mimics as close as possible human stroke. Methods: BALB/Ca-RAG2-/-gc-/- mice were subjected to cauterization or thrombosis stroke model and sacrificed at different time points (48hr, 1wk, 2wk and 4wk) after stroke. Mice received BrdU to estimate activation of cell proliferation in the SVZ. Brains were processed for immunohistochemical and EM. Results: In both stroke models, after inflammation the same glial scar formation process and damage evolution takes place. After stroke, necrotic tissue is progressively removed, and healthy tissue is preserved from injury through the glial scar formation. Cauterization stroke model produced unspecific damage, was less efficient and the infarct was less homogeneous compared to thrombosis infarct. Finally, thrombosis stroke model produces activation of SVZ proliferation. Conclusions: Our results provide an exhaustive analysis of the histopathological changes (inflammation, necrosis, tissue remodeling, scarring...) that occur after stroke in the ischemic boundary zone, which are of key importance for the final stroke outcome. This analysis would allow evaluating how different therapies would affect wound and regeneration. Moreover, this stroke model in RAG 2-/- gC -/- allows cell transplant from different species, even human, to be analyzed.

Mesenchymal stem cells and cardiovascular disease: a bench to bedside roadmap

2012-07-05T11:10:05Z

Title: Mesenchymal stem cells and cardiovascular disease: a bench to bedside roadmap Author(s) : Mazo, M. (Manuel); Araña, M. (Mirian); Pelacho, B. (Beatriz); Prosper, F. (Felipe) Abstract: In recent years, the incredible boost in stem cell research has kindled the expectations of both patients and physicians. Mesenchymal progenitors, owing to their availability, ease of manipulation, and therapeutic potential, have become one of the most attractive options for the treatment of a wide range of diseases, from cartilage defects to cardiac disorders. Moreover, their immunomodulatory capacity has opened up their allogenic use, consequently broadening the possibilities for their application. In this review, we will focus on their use in the therapy of myocardial infarction, looking at their characteristics, in vitro and in vivo mechanisms of action, as well as clinical trials.

Indications for bone marrow transplantation

2012-07-04T08:25:13Z

Title: Indications for bone marrow transplantation Author(s) : Prosper, F. (Felipe); Rifon, J. (José); Cuesta, B. (Braulia); Hermida, J. (José); Panizo, C. (Carlos); Hernandez, M. (M.); Rocha, E. (Eduardo)

Lack of Bcr-Abl point mutations in chronic myeloid leukemia patients in chronic phase before imatinib treatment is not predictive of response

2011-12-19T13:36:00Z

Title: Lack of Bcr-Abl point mutations in chronic myeloid leukemia patients in chronic phase before imatinib treatment is not predictive of response Author(s) : Aguirre, X. (Xavier); Fontalba, A. (A.); Andreu, E.J. (E.J.); Odero, M.D. (María D.); Larrayoz, M.J. (María J.); Montiel-Duarte, C. (Cristina); Calasanz, M.J. (Maria José); Fernandez-Luna, J. (J.L.); Prosper, F. (Felipe)

Past, present and future of anti-idiotype vaccination

2012-07-02T10:13:17Z

Title: Past, present and future of anti-idiotype vaccination Author(s) : Rodriguez-Calvillo, M. (Mercedes); Inoges, S. (Susana); Lopez-Diaz-de-Cerio, A. (Ascensión); Zabalegui, N. (Natalia); Panizo, C. (Carlos); Hernandez, M. (Milagros); Perez-Calvo, J. (Javier); Prosper, F. (Felipe); Melero, I. (Ignacio); Sanchez-Ibarrola, A. (Alfonso); Rocha, E. (Eduardo); Bendandi, M. (Maurizio) Abstract: Cancer vaccines are conceived as therapeutic tools, in contrast to the prophylactic vaccines used to fight against infectious diseases. Among the most potent therapeutic vaccines, anti-idiotype vaccination is directed against the tumor idiotype, the only well-characterized tumor antigen displayed in neoplastic B-cells. Anti-idiotype vaccines have demonstrated clinical benefit against follicular lymphoma and are currently being evaluated in two different phase III clinical trials. Additional emerging strategies, which include the use of dendritic cells and the production of vaccines via molecular means will surely allow us to draw important conclusions concerning the treatment of cancer patients.

RUNX/AML and C/EBP factors regulate CD11a integrin expression in myeloid cells through overlapping regulatory elements

2012-01-14T18:52:03Z

Title: RUNX/AML and C/EBP factors regulate CD11a integrin expression in myeloid cells through overlapping regulatory elements Author(s) : Puig-Kröger, A. (Amaya); Sanchez-Elsner, T. (Tilman); Ruiz, N. (Natividad); Andreu, E.J. (E.J.); Prosper, F. (Felipe); Jensen, U. (U.B.); Gil, J. (Juana); Erickson, P. (P.); Drabkin, H. (H.); Groner, Y. (Y.); Corbi, A.L. (Angel L.) Abstract: The CD11a/CD18 (leukocyte functionassociated antigen 1 [LFA-1]) integrin mediates critical leukocyte adhesive interactions during immune and inflammatory responses. The CD11a promoter directs CD11a/CD18 integrin expression, and its activity in lymphoid cells depends on a functional RUNX1/AML-1–binding site (AML-110) within the MS7 sequence. We now report that MS7 contains a C/EBPbinding site (C/EBP-100), which overlaps with AML-110 and is bound by C/EBP factors in myeloid cells. C/EBP and RUNX/ AML factors compete for binding to their respective cognate elements and bind to the CD11a promoter MS7 sequence in a cell lineage- and differentiation-dependent manner. In myeloid cells MS7 is primarily recognized by C/EBP factors in proliferating cells whereas RUNX/AMLfactors (especially RUNX3/AML-2) bind to MS7 in differentiated cells. RUNX3/AML-2 binding to the CD11a promoter correlates with increased RUNX3/AML-2 protein levels and enhanced CD11a/CD18 cell surface expression. The relevance of the AML-110 element is underscored by the ability of AML-1/ETO to inhibit CD11a promoter activity, thus explaining the low CD11a/CD18 expression in t(8;21)–containing myeloid leukemia cells. Therefore, the expression of the CD11a/CD18 integrin in myeloid cells is determined through the differential occupancy of the CD11a proximal promoter by transcription factors implicated in the pathogenesis of myeloid leukemia.

Autologous human serum for cell culture avoids the implantation of cardioverter-defibrillators in cellular cardiomyoplasty

2012-01-14T18:51:08Z

Title: Autologous human serum for cell culture avoids the implantation of cardioverter-defibrillators in cellular cardiomyoplasty Author(s) : Chachques, J. (J.); Herreros, J. (Jesús); Trainini, J. (Jorge); Juffe, A. (A.); Rendal, E. (Esther); Prosper, F. (Felipe); Genovese, J. (J.) Abstract: Background: Current clinical experience with cellular cardiomyoplasty (using serum bovine-cultivated myoblasts) has demonstrated significant malignant ventricular arrhythmias and sudden deaths in patients. In some ongoing clinical trials the implantation of cardioverterdefibrillator is mandatory. We have hypothesized that contact of human cells with fetal bovine serum results after 3-week fixation of animal proteins on the cell surface, representing an antigenic substrate for immunological and inflammatory adverse events. Methods and Results: Autologous myoblasts were transplanted into infarcted LV in 20 patients (90% males, mean age 62±8 years). Cells were cultivated in a complete human medium during 3 weeks, using the patients' own serum obtained from a blood sample or from plasmapheresis. Injections were performed during CABG (2.1 grafts/pt). All patients had an uneventful recovery. At a mean follow-up of 14±5 months without mortality, no malignant cardiac arrhythmias are reported. LV ejection fraction improved from 28±3% to 52:k4.7% (p = 0.03), and regional wall motion score index (WMSI) from 3.1 to 1.4 (p = 0.04) in the cell-treated segments. Myocardial viability tests showed areas of regeneration. Patients moved from mean NYHA class 2.5 to class 1.2. Conclusions: A total autologous cell culture procedure was used in cellular cardiomyoplasty reducing the risk of arrhythmia. Human-autologous-serum cell expansion avoids the risk of prion, viral or zoonoses contamination. Since patients treated with noncultivated bone marrow cells are free of arrhythmia, the bovine-culture medium seems to be responsible for this complication. Cellular cardiomyoplasty may be efficient to avoid progression of ventricular remodeling and subsequent heart failure in ischemic heart disease.

Chromosomal abnormalities clustering in multiple myeloma reveals cytogenetic subgroups with nonrandom acquisition of chromosomal changes

2013-01-09T17:18:00Z

Title: Chromosomal abnormalities clustering in multiple myeloma reveals cytogenetic subgroups with nonrandom acquisition of chromosomal changes Author(s) : Saez, B. (Borja); Martin-Subero, J.I. (José Ignacio); Guillen-Grima, F. (F.); Odero, M.D. (María D.); Prosper, F. (Felipe); Cigudosa, J.C. (Juan Cruz); Harder, L. (L.); Calasanz, M.J. (Maria José); Siebert, R. (Reiner)

Cellular Cardiomyoplasty: Clinical Application

2012-01-14T12:02:28Z

Title: Cellular Cardiomyoplasty: Clinical Application Author(s) : Chachques, J. (J.); Acar, C. (Christophe); Herreros, J. (Jesús); Trainini, J. (Jorge); Prosper, F. (Felipe); D’Attellis, N. (N.); Fabiani, J.N. (J.N.); Carpentier, A. (A.) Abstract: Myocardial regeneration can be induced with the implantation of a variety of myogenic and angiogenic cell types. More than 150 patients have been treated with cellular cardiomyoplasty worldwide, 18 patients have been treated by our group. Cellular cardiomyoplasty seems to reduce the size and fibrosis of infarct scars, limit postischemic remodelling, and restore regional myocardial contractility. Techniques for skeletal myoblasts culture and ex vivo expansion using autologous patient serum (obtained from plasmapheresis) have been developed by our group. In this article we propose (1) a total autologous cell culture technique and procedures for cell delivery and (2) a clinical trial with appropriate endpoints structured to determine the efficacy of cellular cardiomyoplasty.

A single prior course of BCNU-cisplatin chemotherapy has a significant deleterious effect on mobilization kinetics of otherwise untreated patients

2012-01-14T12:02:26Z

Title: A single prior course of BCNU-cisplatin chemotherapy has a significant deleterious effect on mobilization kinetics of otherwise untreated patients Author(s) : Perez-Calvo, J. (Javier); Fernandez, O. (O.); Aristu, J. (Javier); Aramendia, J.M. (J.M.); Rifon, J. (José); Prosper, F. (Felipe); Bendandi, M. (Maurizio); Rocha, E. (Eduardo); Martin-Algarra, S. (Salvador) Abstract: Extensive prior treatment with cytotoxic agents is associated with impaired mobilization of hematopoietic cells. To assess the effect of a single course of standarddose chemotherapy (CT), we compared the results of filgrastim-induced mobilization among two sequential groups of grade III–IV malignant glioma patients included in a hematopoietic transplantation program. The first group (21 patients) had never been treated with CT until 2 days after surgery, when they received a course of 100 mg/m2 BCNU (IV) and 100 mg intracarotid cisplatin for cytoreduction (not for mobilization). At 1 month after this CT, they were mobilized with 12 lg/kg filgrastim. The second group (22 patients) was mobilized with the same dose of filgrastim directly after the surgery, without having ever received any prior CT. The blood level of CD34þ cells was significantly lower in the CT-treated patients, both on the fourth day of filgrastim (15 vs 36 cells 106/l; P¼0.01) and on the fifth (25 vs 58 cells 106/l; P¼0.003), as it was the number of CD34þ cells collected per apheresis (1.3 vs 3.5 106/l; Po0.0005). The toxic effect of a single course of BCNUcisplatin CT led to significant impairment of the filgrastim-induced mobilization response. Bone Marrow Transplantation advance online

Promoter hypermethylation of cancer-related genes: a strong independent prognostic factor in acute lymphoblastic leukemia

2012-01-14T12:02:22Z

Title: Promoter hypermethylation of cancer-related genes: a strong independent prognostic factor in acute lymphoblastic leukemia Author(s) : Roman-Gomez, J. (José); Jimenez-Velasco, A. (A.); Castillejo, J.A. (J.A.); Aguirre, X. (Xavier); Barrios, M. (M.); Navarro, G. (Germán); Molina, F.J. (Francisco J.); Calasanz, M.J. (Maria José); Prosper, F. (Felipe); Heiniger, A. (A.); Torres, A. (Antonio) Abstract: Promoter hypermethylation plays an important role in the inactivation of cancerrelated genes. This abnormality occurs early in leukemogenesis and seems to be associated with poor prognosis in acute lymphoblastic leukemia (ALL). To determine the extent of hypermethylation in ALL, we analyzed the methylation status of the CDH1, p73, p16, p15, p57, NES-1, DKK-3, CDH13, p14, TMS-1, APAF-1, DAPK, PARKIN, LATS-1, and PTEN genes in 251 consecutive ALL patients.Atotal of 77.3% of samples had at least 1 gene methylated, whereas 35.9% of cases had 4 or more genes methylated. Clinical features and complete remission rate did not differ among patients without methylated genes, patients with 1 to 3 methylated genes (methylated group A), or patients with more than 3 methylated genes (methylated group B). Estimated disease-free survival (DFS) and overall survival (OS) at 11 years were 75.5% and 66.1%, respectively, for the nonmethylated group; 37.2% and 45.5% for methylated group A; and 9.4% and 7.8% for methylated group B (P < .0001 and P .0004, respectively). Multivariate analysis demonstrated that the methylation profile was an independent prognostic factor in predicting DFS (P < .0001) and OS (P .003). Our results suggest that the methylation profile may be a potential new biomarker of risk prediction in ALL

Promoter hypomethylation of the LINE-1 retrotransposable elements activates sense/antisense transcription and marks the progression of chronic myeloid leukemia

2012-01-14T12:02:08Z

Title: Promoter hypomethylation of the LINE-1 retrotransposable elements activates sense/antisense transcription and marks the progression of chronic myeloid leukemia Author(s) : Roman-Gomez, J. (José); Jimenez-Velasco, A. (A.); Aguirre, X. (Xavier); Cervantes, F. (F.); Sanchez, J. (Joaquín); Garate, L. (L.); Barrios, M. (M.); Castillejo, J.A. (J.A.); Navarro, G. (Germán); Colomer, D. (D.); Prosper, F. (Felipe); Heiniger, A. (A.); Torres, A. (Antonio) Abstract: Aberrant genome-wide hypomethylation is thought to be related to tumorigenesis by promoting genomic instability. Since DNA methylation is considered an important mechanism for the silencingof retroelements, hypomethylation in human tumors may lead to their reactivation. However, the role of DNA hypomethylation in chronic myeloid leukemia (CML) remains to be elucidated. In this study, the methylation status of the LINE-1 (L1) retrotransposon promoter was analysed in CML samples from the chronicphase (CP, n¼140) and the blast crisis (BC, n¼47). L1 hypomethylation was significantly more frequent in BC (74.5%) than in CP (38%) (Po0.0001). Furthermore, L1 hypomethylation led to activation of both ORF1 sense transcription (Po0.0001) and c-MET gene antisense transcription (Po0.0001), and was significantly associated with high levels of BCR–ABL (P¼0.02) and DNMT3b4 (P¼0.001) transcripts. Interestingly, in CP-CML, extensive L1 hypomethylation was associated with poorer prognosis in terms of cytogenetic response to interferon (P¼0.004) or imatinib (P¼0.034) and progression-free survival (P¼0.005). The above results strongly suggest that activation of both sense and antisense transcriptions by aberrant promoter hypomethylation of the L1 elements plays a role in the progression and clinical behavior of the CML.