About
108
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Introduction
Current institution
Current position
- Principal Investigator/Associate Professor
Additional affiliations
February 2017 - present
Position
- Professor (Associate)
Description
- Biochemistry and Molecular Biology I Department, Faculty of Science Degree in Biology Degree in Biochemistry Master in Traslational Medicine (TransMed) Master in Tissular Engineering
September 2011 - present
April 2009 - August 2011
Andalusian Stem Cell Bank
Position
- Universidad de Granada
Education
October 2000 - April 2005
October 1994 - June 1999
Publications
Publications (108)
Background/Objectives: The treatment of choice for prostate cancer is androgen deprivation (ADT) and novel hormonal agents such as abiraterone, Enzalutamide or Apalutamide. Initially, this therapy is highly effective, but a significant challenge arises as most patients eventually develop resistance, resulting in castration-resistant prostate cancer...
Platelets are pivotal in tumor progression, dissemination, and metastasis in non-small cell lung cancer (NSCLC). Recent investigations have revealed a complex, bidirectional crosstalk between tumor cells and platelets involving the transfer of biomolecules between these cellular entities. To elucidate the potential role of this interaction in promo...
In the tumor context, platelets play a significant role in both primary tumor progression, dissemination and metastasis. Analysis of this interaction in various cancers, such as non-small cell lung cancer (NSCLC), demonstrate that platelets can both transfer and receive biomolecules (e. g. RNA and proteins) to and from the tumor at different stages...
Pancreatic cancer is a highly lethal and metastatic malignancy, mainly because it often remains undetected until advanced stages due to the limitations of current diagnostic methods, rendering currently available therapies ineffective. Therefore, it is imperative to identify useful biomarkers for early diagnosis and new therapeutic targets for panc...
Bernard-Soulier Syndrome (BSS) is a rare congenital disease characterized by macrothrombocytopenia and frequent bleeding. It is caused by pathogenic variants in three genes (GP1BA, GP1BB or GP9) that encode for the GPIbα, GPIbβ and GPIX subunits of the GPIb-V-IX complex, the main platelet surface receptor for von Willebrand Factor, being essential...
Pediatric Acute Myeloid Leukemia (AML) is a rare and heterogeneous disease characterized by a high prevalence of gene fusions as driver mutations. Despite the improvement of survival in the last years, about 50% of patients still experience a relapse. It is not possible to improve prognosis only with further intensification of chemotherapy, as come...
Pediatric acute myeloid leukemia (AML) is a rare and heterogeneous disease that remains the major cause of mortality in children with leukemia. To improve the outcome of pediatric AML we need to gain knowledge on the biological bases of this disease. NUP98-KDM5A (NK5A) fusion protein is present in a particular subgroup of young pediatric patients w...
Hypophosphatasia (HPP) a rare disease caused by mutations in the ALPL gene encoding for the tissue-nonspecific alkaline phosphatase protein (TNSALP), has been identified as a potentially under-diagnosed condition worldwide which may have higher prevalence than currently established. This is largely due to the overlapping of its symptomatology with...
R-loops are three-stranded nucleic acid structures that accumulate on chromatin in neurological diseases and cancers and contribute to genome instability. Using a proximity-dependent labeling system, we identified distinct classes of proteins that regulate R-loops in vivo through different mechanisms. We show that ATRX suppresses R-loops by interac...
Background: Platelets are active players in tumorigenesis, although the exact interactive mechanisms and their direct impact on tumor cells remain largely unknown.
Methods: Bidirectional transference of lipids, proteins and RNA between platelets and tumor cells and its impact on tumor cell behavior and tumor process are analyzed in this work. Pheno...
Integration-deficient lentiviral vectors (IDLVs) have recently generated increasing interest, not only as a tool for transient gene delivery, but also as a technique for detecting off-target cleavage in gene-editing methodologies which rely on customized endonucleases (ENs). Despite their broad potential applications, the efficacy of IDLVs has hist...
Background
Platelets are active players in tumorigenesis, although the exact interactive mechanisms and their direct impact on tumor cells remain largely unknown. Methods
Bidirectional transference of lipids, proteins and RNA between platelets and tumor cells and its impact on tumor cell behavior and tumor process are analyzed in this work. Phenoty...
Androgen deprivation therapy (ADT) and novel hormonal agents (NHAs) (Abiraterone and Enzalutamide) are the goal standard for metastatic prostate cancer (PCa) treatment. Although ADT is initially effective, a subsequent castration resistance status (CRPC) is commonly developed. The expression of androgen receptor (AR) alternative splicing isoforms (...
Familial Platelet Disorder with associated Myeloid Malignancy (FPDMM) is a rare platelet disorder caused by mutations inRUNX1. We generated an iPSC line(GENYOi005-A) from a FPDMM patient with a non-previously reported variant p.Thr196Ala. Non-integrative Sendai viruses expressing the Yamanaka repro-gramming factors were used to reprogram peripheral...
Familial Platelet Disorder with associated Myeloid Malignancy (FPDMM) is a rare platelet disorder caused by mutations in RUNX1. We generated an iPSC line (GENYOi005-A) from a FPDMM patient with a non-previously reported variant p.Thr196Ala. Non-integrative Sendai viruses expressing the Yamanaka reprogramming factors were used to reprogram periphera...
Hypophosphatasia (HPP) is a genetic disease caused by one or several mutations in ALPL gene encoding the tissue-nonspecific alkaline phosphatase affecting the mineralization process. Due to its low prevalence and lack of recognition, this metabolic disorder is generally confused with other more frequent bone disorders. An assessment of serum total...
ADNP syndrome is an intellectual disability associated with Autism spectrum disorder caused by mutations in ADNP. We generated an iPSC line from an ADNP syndrome pediatric patient harboring the mutation p.Trp719* (GENYOi004-A). Peripheral blood mononuclear cells were reprogrammed using a non-transmissible form of Sendai viruses expressing the four...
Mutations in ADNP have been recently associated with intellectual disability and autism spectrum disorder. However, the clinical features of patients with this syndrome are not fully identified, and no treatment currently exists for these patients. Here, we extended the ADNP syndrome phenotype describing skin abnormalities in both a patient with AD...
The t(4;11)(q21;q23) translocation is associated with high-risk infant pro-B-cell acute lymphoblastic leukemia and arises prenatally during embryonic/fetal hematopoiesis. The developmental/pathogenic contribution of the t(4;11)-resulting MLL-AF4 (MA4) and AF4-MLL (A4M) fusions remains unclear; MA4 is always expressed in patients with t(4;11)+ B-cel...
Integration-defective lentiviral vectors (IDLVs) have become an important
alternative tool for gene therapy applications and basic research. Unfortunately, IDLVs
show lower transgene expression as compared to their integrating counterparts. In this
study, we aimed to improve the expression levels of IDLVs by inserting the IS2
element, which harbors...
Runx1 is a master hematopoietic transcription factor essential for hematopoietic stem cell (HSC) emergence. Runx1-deficient mice die during early embryogenesis due to the inability to establish definitive hematopoiesis. Here we have used hPSCs as model to study the role of RUNX1 in human embryonic hematopoiesis. Although the three RUNX1 isoforms a,...
Decellularized vascular scaffolds are promising materials for vessel replacements. However, despite the natural origin of decellularized vessels, issues such as biomechanical incompatibility, immunogenicity risks and the hazards of thrombus formation, still need to be addressed. In this study, we coated decellularized vessels obtained from porcine...
Bernard Soulier Syndrome (BSS) is a rare autosomal platelet disorder characterized by mutations in the von Willebrand factor platelet receptor complex GPIb-V-IX. In this work we have generated an induced pluripotent stem cell (BSS3-PBMC-iPS4F8) from peripheral blood mononuclear cells of a BSS patient with a p.Phe55Ser mutation in the GPIX gene. Cha...
Bernard Soulier Syndrome (BSS) is an inherited rare platelet disorder characterized by mutations in the platelet glycoprotein complex GPIb-IX-V. We generated an induced pluripotent stem cell (iPSC) line from a BSS patient with a mutation p.Asn45Ser in the GPIX locus (BSS2-PBMC-iPS4F24). Peripheral blood mononuclear cells were reprogrammed using non...
We generated an induced pluripotent stem cell (iPSC) line from a Bernard-Soulier Syndrome (BSS) patient carrying the mutation p.Trp71Arg in the GPIX locus (BSS1-PBMC-iPS4F4). Peripheral blood mononuclear cells (PBMCs) were reprogrammed using heat sensitive non integrative Sendai viruses containing the reprogramming factors Oct3/4, SOX2, KLF4 and c-...
Studies with different animal models have shown that the Notch ligand DLL4 has a key role in the development of the embryonic vasculature. Here we describe the generation and characterization of a human embryonic stem cell line and an induced pluripotent stem cell line that constitutively express short hairpin RNAs targeting DLL4 mRNA. These cells...
Here we describe the generation and characterization of the human induced pluripotent stem cell (iPSC) line PBMC1-iPS4F1 from peripheral blood mononuclear cells from a healthy female with Spanish background. We used heat sensitive, non-integrative Sendai viruses containing the reprogramming factors Oct3/4, Sox2, Klf4 and c-Myc, whose expression was...
The Wiskott-Aldrich syndrome (WAS) is an X-linked primary immunodeficiency caused by mutations in the WAS gene and characterized by severe thrombocytopenia. Although the role of WASp in terminally differentiated lymphocytes and myeloid cells is well characterized, its role in early hematopoietic differentiation and in platelets (Plts) biology is po...
We have generated iPSCs from peripheral blood mononuclear cells (PBMCs) of a healthy man using heat sensitive and non-integrative Sendai virus containing Sox2, Oct3/4, c-Myc and Klf4. Human GRX-MCiPS4F-A2 cell line was established and characterized through this study.
Copyright © 2015. Published by Elsevier B.V.
Notch signaling is essential for definitive hematopoiesis, but its role in human embryonic hematopoiesis is largely unknown. We show that in hESCs the expression of the Notch ligand DLL4 is induced during hematopoietic differentiation. We found that DLL4 is only expressed in a subpopulation of bipotent hemato-endothelial progenitors (HEPs) and segr...
Human embryonic stem cells (hESCs) are a unique in vitro model for studying human developmental biology and represent a potential source for cell replacement strategies. Platelets can be generated from cord blood progenitors and hESCs; however, the molecular mechanisms and determinants controlling the in vitro megakaryocytic specification of hESCs...
Key Points
HOXA9 parallels blood development, but is restricted to HEP, and diminishes as they differentiate into blood cells. Functional assays reveal how HOXA9 enhances blood formation by promoting commitment of HEP to CD45+ cells with higher clonogenic potential.
Megakaryocytes (MKs) are bone marrow cells originating from hematopoietic stem cells (HSC) by a differentiation pro- cess known as megakaryopoiesis. Subsequently, these cells mature to produce of platelets, a process denominated throm- bopoiesis. Different labs have generated MKs and platelets in vitro from HSCs purified from bone marrow, umbilical...
La presente invención se encuentra dentro del campo de la biología y la medicina, y
5 más concretamente en el ámbito de las terapias avanzadas. Específicamente, se
refiere al uso de las células madre para la obtención de megacariocitos productores de
plaquetas funcionales, para su empleo frente a enfermedades derivadas de anomalías
hematopoyéticas....
Human embryonic stem cells (hESCs) are a unique in vitro model for studying human developmental biology and represent a potential source for cell replacement strategies. Platelets can be generated from cord blood progenitors and hESCs; however, the molecular mechanisms and determinants controlling the in vitro megakaryocytic specification of hESCs...
The molecular determinants regulating the specification of human embryonic stem cells (hESCs) into hematopoietic cells remain elusive. HOXA9 plays a relevant role in leukemogenesis and hematopoiesis. It is highly expressed in hematopoietic stem/progenitor cells (HSPCs) and is downregulated upon differentiation. Hoxa9-deficient mice display impaired...
The present invention relates to methods and compositions for preventing and/or treating various conditions in a patient, including for example, T-cell lymphoblastic leukemia and lymphoma as well as neurodegenerative diseases, such as for example, Alzheimer's disease. In one preferred embodiment of the invention, such methods include providing a pa...
MLL-AF4 fusion is hallmark in high-risk infant pro-B-acute lymphoblastic leukemia (pro-B-ALL). Our limited understanding of MLL-AF4-mediated transformation reflects the absence of human models reproducing this leukemia. Hematopoietic stem/progenitor cells (HSPCs) constitute likely targets for transformation. We previously reported that MLL-AF4 enha...
Mixed-lineage leukemia (MLL)-AF4 fusion arises prenatally in high-risk infant acute pro-B-lymphoblastic leukemia (pro-B-ALL). In human embryonic stem cells (hESCs), MLL-AF4 skewed hematoendothelial specification but was insufficient for transformation, suggesting that additional oncogenic insults seem required for MLL-AF4-mediated transformation. M...
Genetic manipulation of human embryonic stem cells (hESCs) is instrumental for tracing lineage commitment and to studying human development. Here we used hematopoietic-specific Wiskott-Aldrich syndrome gene (WAS)-promoter driven lentiviral vectors (LVs) to achieve highly specific gene expression in hESCs-derived hematopoietic cells. We first demons...
WAS-promoter driven LVs efficiently express eGFP in hESC-derived myeloid colonies. Transmission (left panels) and fluorescence (right panels) microphotographs from untransduced (NT), pLVTHM-, AWE- and WE-transduced hESCs. The different hESCs were incubated in EB hematopoietic differentiation media for 15 days and then incubated in methylcellulose H...
Lentiviral transduction does not affect hematopoietic differentiation potential of hESCs. Untransduced nESCs (NT) and AWE- and WE-transduced H9 cells were induced towards hematopoiesis by EBs formation. At day 15 of differentiation (top), the EBs were dissociated and analyzed for CD31 and CD45 expression to determine the percentage of CD31+CD45- he...
Phenotypic analysis of the CD45−eGFP+cells in AWE-transduced H9 and AND-1 hESCs at days 10–15 of EB differentiation. pLVTHM- and AWE-transduced H9 and AWE-transduced AND-1 cells were incubated in hematopoietic differentiation media and analyzed for CD45 and eGFP expression (middle plots) after 10 (H9) or 15 (AND-1) days. CD45−eGFP− (left plots) and...
Schematic diagram showing the procedure for hematopoietic differentiation of hESCs. hESC are incubated in non-adherent plates in EBs medium (see M&M for details). Once the EBs are formed, the media is replaced for EB medium supplemented with hematopoietic cytokines and incubated for 22days. Total RNA was obtained at different the days (0, 1, 3, 5,...
Phenotypic analysis of eGFP+ cells in WE-transduced H9 cells at days 10, 15 and 22 of differentiation. WE-transduced H9 cells were incubated in hematopoietic differentiation media and analyzed for CD45, CD31 and CD34 expression at days 10, 15 and 22. eGFP+ (right) and eGFP− (left) populations were first analyzed for expression of CD45 and CD31. CD3...
Phenotypoic analysis of sorted populations. The AWE-transduced H9 cells were induced towards hematopoiesis by EBs formation (see M&M). At day 10 of differentiation, the EBs were dissociated and the different populations sorted. A) Cells were separated based on the expression of eGFP (left plot, arrows) or CD34 (right plot, arrows). eGFP- and eGFP+...
Expression pattern of WAS-promoter driven lentiviral vectors parallel CD45 expression during hESCs hematopoietic development. Graphs showing the percentage of eGFP+ (A) and CD45+ (B) cells in AWE and WE-transduced H9 cells at different days of hematopoietic differentiation. Untransduced cells (NT) were used as negative controls. Data are average +/...
Human induced pluripotent stem cells (hiPSC) have been generated from different tissues, with the age of the donor, tissue source and specific cell type influencing the reprogramming process. Reprogramming hematopoietic progenitors to hiPSC may provide a very useful cellular system for modelling blood diseases. We report the generation and complete...
hiPSC lines display in vitro potential for three germ layer differentiation. Histological analysis of EBs showing spontaneous in vitro differentiation into ectoderm (pan-cytokeratin+), mesoderm (smooth muscle Actin+) and endoderm (neuroephitelium).
(TIF)
Phenotypic and molecular characterization of CB-iPSC after Cre-mediated excision of the reprogramming transgenes. After Cre-mediated excision of the reprogramming transgene, the CB-iPSC express the pluripotent surface markers SSEA-3, SSEA-4, Tra-1-60 and Tra-1-81 as well as nuclear Oct4 by flow cytometry (A) and retain expression of the pluripotenc...
Determining the molecular regulators/pathways responsible for the specification of human embryonic stem cells (hESCs) into hematopoietic precursors has far-reaching implications for potential cell therapies and disease modeling. Mouse models lacking SCL/TAL1 (stem cell leukemia/T-cell acute lymphocytic leukemia 1) do not survive beyond early embryo...
The MLL-AF4 fusion gene is a hallmark genomic aberration in high-risk acute lymphoblastic leukemia in infants. Although it is well established that MLL-AF4 arises prenatally during human development, its effects on hematopoietic development in utero remain unexplored. We have created a human-specific cellular system to study early hemato-endothelia...
Despite the improvements in the human embryonic stem cell (hESC) culture systems, very similar conditions to those used to maintain hESCs on mouse feeders are broadly applied to culture methods based on human feeders. Indeed, basic fibroblast growth factor (bFGF), a master hESC-sustaining factor, is still added in nearly all medium formulations for...
The realization of human embryonic stem cells (hESC) as a model for human developmental hematopoiesis and in potential cell replacement strategies relies on an improved understanding of the extrinsic and intrinsic factors regulating hematopoietic-specific hESC differentiation. Human mesenchymal stem cells (hMSCs) are multipotent cells of mesodermal...
Understanding the transcriptional cues that direct differentiation of human embryonic stem cells (hESCs) and human-induced
pluripotent stem cells to defined and functional cell types is essential for future clinical applications. In this study,
we have compared transcriptional profiles of haematopoietic progenitors derived from hESCs at various dev...
Human ESCs provide access to the earliest stages of human development and may serve as an unlimited source of functional cells for future cell therapies. The optimization of methods directing the differentiation of human embryonic stem cells (hESCs) into tissue-specific precursors becomes crucial. We report an efficient enrichment of mesenchymal st...
Lineage-specific differentiation potential varies among different human pluripotent stem cell (hPSC) lines, becoming therefore highly desirable to prospectively know which hPSC lines exhibit the highest differentiation potential for a certain lineage. We have compared the hematopoietic potential of 14 human embryonic stem cell (hESC)/induced plurip...
11
Glucocorticoids (GC) play a fundamental role in the treatment of all lymphoid tumors because of their capacity to induce apoptosis in lymphoid progenitor cells. The importance of GC therapy in lymphoid malignancies is underscored by the strong association of GC response with prognosis in childhood acute lymphoblastic leukemia (ALL). Thus, resist...
471
T-cell acute lymphoblastic leukemia (T-ALL) is an aggressive malignancy associated with the activation of oncogenic transcription factors. Thus, 5% to 10% of pediatric and up to 30% of adult T-ALL patients show aberrant expression of the TLX1 transcription factor oncogene. Aberrant expression of TLX1 in Lck-TLX1 transgenics induces transformati...
The TLX1 oncogene (encoding the transcription factor T cell leukemia homeobox protein-1) has a major role in the pathogenesis of T cell acute lymphoblastic leukemia (T-ALL). However, the specific mechanisms of T cell transformation downstream of TLX1 remain to be elucidated. Here we show that transgenic expression of human TLX1 in mice induces T-AL...
Supplementary results
Supplementary methods
Supplementary Figure 1 Numerical and structural chromosomal alterations in TLX-induced mouse T-ALL.
Supplementary Figure 2 Mitotic checkpoint analysis in mouse and human T-ALL cells.
Supplementary Figure 3 Cell proliferation and disruption of the mitotic spindle in mouse T-ALLs.
Supplementary Figure 4 TLX...
676
T-cell acute lymphoblastic leukemia (T-ALL) is an aggressive malignancy associated with the activation of transcription factor oncogenes. TLX1/HOX11 was originally isolated from the recurrent t(10;14)(q24;q11) translocation and is aberrantly expressed in 5% to 10% of pediatric and up to 30% of adult T-ALL. The TLX3/HOX11L2 oncogene is closely r...
142
T-cell acute lymphoblastic leukemia (T-ALL) is an aggressive malignancy associated with the activation of transcription factor oncogenes. TLX1/HOX11 was originally isolated from the recurrent t(10;14)(q24;q11) in T-ALL and is aberrantly expressed in 5% to 10% of pediatric and up to 30% of adult T-ALL cases. Tlx1 plays an important role during e...
The molecular mechanisms involved in disease progression and relapse in T-cell acute lymphoblastic leukemia (T-ALL) are poorly understood. We used single nucleotide polymorphism array analysis to analyze paired diagnostic and relapsed T-ALL samples to identify recurrent genetic alterations in T-ALL. This analysis showed that diagnosis and relapsed...
Inhibition of NOTCH1 signaling with gamma-secretase inhibitors (GSIs) has been proposed as a molecularly targeted therapy in T-cell acute lymphoblastic leukemia (T-ALL). However, GSIs seem to have limited antileukemic activity in human T-ALL and are associated with severe gastrointestinal toxicity resulting from inhibition of NOTCH signaling in the...
Gamma-secretase inhibitors (GSIs) block the activation of the oncogenic protein Notch homolog-1 (NOTCH1) in T cell acute lymphoblastic leukemia (T-ALL). However, limited antileukemic cytotoxicity and severe gastrointestinal toxicity have restricted the clinical application of these targeted drugs. Here we show that combination therapy with GSIs plu...
Activating mutations in NOTCH1 are present in over 50% of T-cell acute lymphoblastic leukemias (T-ALL). Consequently, inhibition of NOTCH1 signaling with small molecule γ-secretase inhibitors (GSIs) has been proposed as a targeted therapy for this disease. However, GSIs fail to induce robust apoptosis in T-ALL cells and their clinical application h...
Heterodimerization domain (HD) mutations in NOTCH1 induce ligand-independent activation of the receptor and contribute to the pathogenesis of one-third of human T-cell lymphoblastic leukemias (T-ALLs). Here we report a novel class of activating mutations in NOTCH1 leading to aberrant activation of NOTCH1 signaling in T-cell lymphoblasts. These so-c...
Activating mutations in NOTCH1 are common in T-cell lymphoblastic leukemias (T-ALL), making this receptor a promising target for drugs such as gamma-secretase inhibitors (GSIs), which block NOTCH1 activation. However, enthusiasm for these therapies has been tempered by tumor resistance and the paucity of information on the oncogenic programs regula...
Gamma-secretase inhibitors (GSIs), which block the activation of the NOTCH1 receptor, are currently being tested in the treatment of T-cell lymphoblastic leukemias (T-ALL) with activating mutations in the NOTCH1 gene. However, inhibition of NOTCH1 signaling induces only a delayed cytostatic antileukemic effect with little or no apoptosis and GSI tr...
Gain-of-function mutations in NOTCH1 are common in T-cell lymphoblastic leukemias and lymphomas (T-ALL), making this receptor a promising target for drugs such as gamma-secretase inhibitors, which block a proteolytic cleavage required for NOTCH1 activation. However, the enthusiasm for these therapies has been tempered by tumor resistance and the pa...
Melanoma causes the greatest number of skin cancer-related deaths worldwide. Despite intensive research, prevention and early detection are the only effective measures against melanoma, so new therapeutic strategies are necessary for the management of this devastating disease. Here, we evaluated the efficacy of cannabinoid receptor agonists, a new...
Proline- and acid-rich (PAR) basic region leucine zipper (bZIP) proteins thyrotroph embryonic factor (TEF), D-site-binding
protein (DBP), and hepatic leukemia factor have been involved in neurotransmitter homeostasis and amino acid metabolism. Here
we demonstrate a novel role for these proteins in the transcriptional control of a BH3-only gene. PAR...
The NOTCH1 signaling pathway directly links extracellular signals with transcriptional responses in the cell nucleus and plays a critical role during T cell development and in the pathogenesis over 50% of human T cell lymphoblastic leukemia (T-ALL) cases. However, little is known about the transcriptional programs activated by NOTCH1. Using an inte...
