TY - JOUR T1 - Fibroblast activation and abnormal extracellular matrix remodelling as common hallmarks in three cancer-prone genodermatoses. JF - Br J Dermatol Y1 - 2019 A1 - Chacón-Solano, E A1 - León, C A1 - Díaz, F A1 - García-García, F A1 - García, M A1 - Escámez, M J A1 - Guerrero-Aspizua, S A1 - Conti, C J A1 - Mencía, Á A1 - Martínez-Santamaría, L A1 - Llames, S A1 - Pévida, M A1 - Carbonell-Caballero, J A1 - Puig-Butillé, J A A1 - Maseda, R A1 - Puig, S A1 - de Lucas, R A1 - Baselga, E A1 - Larcher, F A1 - Dopazo, J A1 - Del Rio, M KW - Adolescent KW - Adult KW - Biopsy KW - Blister KW - Case-Control Studies KW - Cells, Cultured KW - Child KW - Child, Preschool KW - Epidermolysis Bullosa KW - Epidermolysis Bullosa Dystrophica KW - Extracellular Matrix KW - Extracellular Matrix Proteins KW - Female KW - Fibroblasts KW - Fibrosis KW - Gene Expression Regulation KW - Healthy Volunteers KW - Humans KW - Infant KW - Infant, Newborn KW - Male KW - Middle Aged KW - mutation KW - Periodontal Diseases KW - Photosensitivity Disorders KW - Primary Cell Culture KW - RNA-seq KW - Skin KW - Xeroderma Pigmentosum KW - Young Adult AB -

BACKGROUND: Recessive dystrophic epidermolysis bullosa (RDEB), Kindler syndrome (KS) and xeroderma pigmentosum complementation group C (XPC) are three cancer-prone genodermatoses whose causal genetic mutations cannot fully explain, on their own, the array of associated phenotypic manifestations. Recent evidence highlights the role of the stromal microenvironment in the pathology of these disorders.

OBJECTIVES: To investigate, by means of comparative gene expression analysis, the role played by dermal fibroblasts in the pathogenesis of RDEB, KS and XPC.

METHODS: We conducted RNA-Seq analysis, which included a thorough examination of the differentially expressed genes, a functional enrichment analysis and a description of affected signalling circuits. Transcriptomic data were validated at the protein level in cell cultures, serum samples and skin biopsies.

RESULTS: Interdisease comparisons against control fibroblasts revealed a unifying signature of 186 differentially expressed genes and four signalling pathways in the three genodermatoses. Remarkably, some of the uncovered expression changes suggest a synthetic fibroblast phenotype characterized by the aberrant expression of extracellular matrix (ECM) proteins. Western blot and immunofluorescence in situ analyses validated the RNA-Seq data. In addition, enzyme-linked immunosorbent assay revealed increased circulating levels of periostin in patients with RDEB.

CONCLUSIONS: Our results suggest that the different causal genetic defects converge into common changes in gene expression, possibly due to injury-sensitive events. These, in turn, trigger a cascade of reactions involving abnormal ECM deposition and underexpression of antioxidant enzymes. The elucidated expression signature provides new potential biomarkers and common therapeutic targets in RDEB, XPC and KS. What's already known about this topic? Recessive dystrophic epidermolysis bullosa (RDEB), Kindler syndrome (KS) and xeroderma pigmentosum complementation group C (XPC) are three genodermatoses with high predisposition to cancer development. Although their causal genetic mutations mainly affect epithelia, the dermal microenvironment likely contributes to the physiopathology of these disorders. What does this study add? We disclose a large overlapping transcription profile between XPC, KS and RDEB fibroblasts that points towards an activated phenotype with high matrix-synthetic capacity. This common signature seems to be independent of the primary causal deficiency, but reflects an underlying derangement of the extracellular matrix via transforming growth factor-β signalling activation and oxidative state imbalance. What is the translational message? This study broadens the current knowledge about the pathology of these diseases and highlights new targets and biomarkers for effective therapeutic intervention. It is suggested that high levels of circulating periostin could represent a potential biomarker in RDEB.

VL - 181 IS - 3 U1 - https://www.ncbi.nlm.nih.gov/pubmed/30693469?dopt=Abstract ER - TY - JOUR T1 - Global Transcriptome Analysis of Primary Cerebrocortical Cells: Identification of Genes Regulated by Triiodothyronine in Specific Cell Types. JF - Cereb Cortex Y1 - 2017 A1 - Gil-Ibañez, Pilar A1 - Garcia-Garcia, Francisco A1 - Dopazo, Joaquin A1 - Bernal, Juan A1 - Morte, Beatriz KW - Animals KW - Astrocytes KW - Cells, Cultured KW - Cerebral Cortex KW - Fluorescent Antibody Technique KW - Gene Expression Profiling KW - Mice, 129 Strain KW - Mice, Inbred BALB C KW - Mice, Inbred C57BL KW - Neurons KW - Piperazines KW - Transcriptome KW - Triiodothyronine AB -

Thyroid hormones, thyroxine, and triiodothyronine (T3) are crucial for cerebral cortex development acting through regulation of gene expression. To define the transcriptional program under T3 regulation, we have performed RNA-Seq of T3-treated and untreated primary mouse cerebrocortical cells. The expression of 1145 genes or 7.7% of expressed genes was changed upon T3 addition, of which 371 responded to T3 in the presence of cycloheximide indicating direct transcriptional regulation. The results were compared with available transcriptomic datasets of defined cellular types. In this way, we could identify targets of T3 within genes enriched in astrocytes and neurons, in specific layers including the subplate, and in specific neurons such as prepronociceptin, cholecystokinin, or cortistatin neurons. The subplate and the prepronociceptin neurons appear as potentially major targets of T3 action. T3 upregulates mostly genes related to cell membrane events, such as G-protein signaling, neurotransmission, and ion transport and downregulates genes involved in nuclear events associated with the M phase of cell cycle, such as chromosome organization and segregation. Remarkably, the transcriptomic changes induced by T3 sustain the transition from fetal to adult patterns of gene expression. The results allow defining in molecular terms the elusive role of thyroid hormones on neocortical development.

VL - 27 IS - 1 U1 - https://www.ncbi.nlm.nih.gov/pubmed/26534908?dopt=Abstract ER - TY - JOUR T1 - Early transcriptional defense responses in Arabidopsis cell suspension culture under high-light conditions. JF - Plant Physiol Y1 - 2011 A1 - González-Pérez, Sergio A1 - Gutiérrez, Jorge A1 - Garcia-Garcia, Francisco A1 - Osuna, Daniel A1 - Dopazo, Joaquin A1 - Lorenzo, Oscar A1 - Revuelta, José L A1 - Arellano, Juan B KW - Arabidopsis KW - Blotting, Western KW - Cell Culture Techniques KW - Cells, Cultured KW - Chloroplasts KW - Cluster Analysis KW - Gene Expression Profiling KW - Gene Expression Regulation, Plant KW - Hydrogen Peroxide KW - Light KW - mutation KW - Oligonucleotide Array Sequence Analysis KW - Photosystem II Protein Complex KW - Plant Growth Regulators KW - Reproducibility of Results KW - Reverse Transcriptase Polymerase Chain Reaction KW - RNA, Messenger KW - Signal Transduction KW - Stress, Physiological KW - Transcription, Genetic AB -

The early transcriptional defense responses and reactive oxygen species (ROS) production in Arabidopsis (Arabidopsis thaliana) cell suspension culture (ACSC), containing functional chloroplasts, were examined at high light (HL). The transcriptional analysis revealed that most of the ROS markers identified among the 449 transcripts with significant differential expression were transcripts specifically up-regulated by singlet oxygen ((1)O(2)). On the contrary, minimal correlation was established with transcripts specifically up-regulated by superoxide radical or hydrogen peroxide. The transcriptional analysis was supported by fluorescence microscopy experiments. The incubation of ACSC with the (1)O(2) sensor green reagent and 2',7'-dichlorofluorescein diacetate showed that the 30-min-HL-treated cultures emitted fluorescence that corresponded with the production of (1)O(2) but not of hydrogen peroxide. Furthermore, the in vivo photodamage of the D1 protein of photosystem II indicated that the photogeneration of (1)O(2) took place within the photosystem II reaction center. Functional enrichment analyses identified transcripts that are key components of the ROS signaling transduction pathway in plants as well as others encoding transcription factors that regulate both ROS scavenging and water deficit stress. A meta-analysis examining the transcriptional profiles of mutants and hormone treatments in Arabidopsis showed a high correlation between ACSC at HL and the fluorescent mutant family of Arabidopsis, a producer of (1)O(2) in plastids. Intriguingly, a high correlation was also observed with ABA deficient1 and more axillary growth4, two mutants with defects in the biosynthesis pathways of two key (apo)carotenoid-derived plant hormones (i.e. abscisic acid and strigolactones, respectively). ACSC has proven to be a valuable system for studying early transcriptional responses to HL stress.

VL - 156 IS - 3 U1 - https://www.ncbi.nlm.nih.gov/pubmed/21531897?dopt=Abstract ER - TY - JOUR T1 - Hypoxia promotes efficient differentiation of human embryonic stem cells to functional endothelium. JF - Stem Cells Y1 - 2010 A1 - Prado-Lopez, Sonia A1 - Conesa, Ana A1 - Armiñán, Ana A1 - Martínez-Losa, Magdalena A1 - Escobedo-Lucea, Carmen A1 - Gandia, Carolina A1 - Tarazona, Sonia A1 - Melguizo, Dario A1 - Blesa, David A1 - Montaner, David A1 - Sanz-González, Silvia A1 - Sepúlveda, Pilar A1 - Götz, Stefan A1 - O'Connor, José Enrique A1 - Moreno, Ruben A1 - Dopazo, Joaquin A1 - Burks, Deborah J A1 - Stojkovic, Miodrag KW - Angiopoietin-1 KW - Animals KW - biomarkers KW - Cell Culture Techniques KW - Cell Differentiation KW - Cell Hypoxia KW - Cell Transplantation KW - Cells, Cultured KW - Down-Regulation KW - Embryonic Stem Cells KW - Endothelial Cells KW - Gene Expression Profiling KW - Gene Expression Regulation KW - Humans KW - Male KW - Myocardial Infarction KW - Neovascularization, Physiologic KW - Oxygen KW - Pluripotent Stem Cells KW - Rats KW - Rats, Nude KW - Vascular Endothelial Growth Factor A AB -

Early development of mammalian embryos occurs in an environment of relative hypoxia. Nevertheless, human embryonic stem cells (hESC), which are derived from the inner cell mass of blastocyst, are routinely cultured under the same atmospheric conditions (21% O(2)) as somatic cells. We hypothesized that O(2) levels modulate gene expression and differentiation potential of hESC, and thus, we performed gene profiling of hESC maintained under normoxic or hypoxic (1% or 5% O(2)) conditions. Our analysis revealed that hypoxia downregulates expression of pluripotency markers in hESC but increases significantly the expression of genes associated with angio- and vasculogenesis including vascular endothelial growth factor and angiopoitein-like proteins. Consequently, we were able to efficiently differentiate hESC to functional endothelial cells (EC) by varying O(2) levels; after 24 hours at 5% O(2), more than 50% of cells were CD34+. Transplantation of resulting endothelial-like cells improved both systolic function and fractional shortening in a rodent model of myocardial infarction. Moreover, analysis of the infarcted zone revealed that transplanted EC reduced the area of fibrous scar tissue by 50%. Thus, use of hypoxic conditions to specify the endothelial lineage suggests a novel strategy for cellular therapies aimed at repair of damaged vasculature in pathologies such as cerebral ischemia and myocardial infarction.

VL - 28 IS - 3 U1 - https://www.ncbi.nlm.nih.gov/pubmed/20049902?dopt=Abstract ER -