Aurora Astudillo - Academia.edu (original) (raw)

Papers by Aurora Astudillo

Research paper thumbnail of Loss of collagenase-2 confers increased skin tumor susceptibility to male mice

Nature Genetics, 2003

We generated mice deficient in collagenase-2 (Mmp8), an MMP mainly produced by neutrophils in inf... more We generated mice deficient in collagenase-2 (Mmp8), an MMP mainly produced by neutrophils in inflammatory reactions and detected in some malignant tumors 1,4 . Loss of Mmp8 did not cause abnormalities during embryonic development or in adult mice. Contrary to previous studies with MMP-deficient mice, however, the absence of Mmp8 strongly increased the incidence of skin tumors in male Mmp8 -/mice. Female Mmp8 -/mice whose ovaries were removed or were treated with tamoxifen were also more susceptible to tumors compared with wild-type mice. Bone marrow transplantation experiments confirmed that Mmp8 supplied by neutrophils was sufficient to restore the natural protection against tumor development mediated by this protease in male mice. Histopathological analysis showed that mutant mice had abnormalities in the inflammatory response induced by carcinogens. Our study identifies a paradoxical protective role for Mmp8 in cancer and provides a genetic model to evaluate the molecular basis of gender differences in cancer susceptibility.

Research paper thumbnail of Reply to ‘Aberrant cytoplasmic expression of C-terminal truncated NPM leukaemic mutant is dictated by tryptophans loss and a new NES motif’ by Falini et al

Leukemia, 2007

Martelli MF et al. Identification and functional characterization of a cytoplasmic nucleophosmin ... more Martelli MF et al. Identification and functional characterization of a cytoplasmic nucleophosmin leukaemic mutant generated by a novel exon-11 NPM1 mutation. Leukemia 2007; 21: 1099-1103. 4 Falini B, Bolli N, Shan J, Martelli MP, Liso A, Pucciarini A et al. Both carboxy-terminus NES motif and mutated tryptophan(s) are crucial for aberrant nuclear export of nucleophosmin leukemic mutants in NPMc+ AML. Blood 2006; 107: 4514-4523.

Research paper thumbnail of Treatment of Chronic Viral Hepatitis in Woodchucks by Prolonged Intrahepatic Expression of Interleukin-12

Journal of Virology, 2011

Chronic hepatitis B is a major cause of liver-related death worldwide. Interleukin-12 (IL-12) ind... more Chronic hepatitis B is a major cause of liver-related death worldwide. Interleukin-12 (IL-12) induction accompanies viral clearance in chronic hepatitis B virus infection. Here, we tested the therapeutic potential of IL-12 gene therapy in woodchucks chronically infected with woodchuck hepatitis virus (WHV), an infection that closely resembles chronic hepatitis B. The woodchucks were treated by intrahepatic injection of a helper-dependent adenoviral vector encoding IL-12 under the control of a liver-specific RU486-responsive promoter. All woodchucks with viral loads below 10 10 viral genomes (vg)/ml showed a marked and sustained reduction of viremia that was accompanied by a reduction in hepatic WHV DNA, a loss of e antigen and surface antigen, and improved liver histology. In contrast, none of the woodchucks with higher viremia levels responded to therapy. The antiviral effect was associated with the induction of T-cell immunity against viral antigens and a reduction of hepatic expression of Foxp3 in the responsive animals. Studies were performed in vitro to elucidate the resistance to therapy in highly viremic woodchucks. These studies showed that lymphocytes from healthy woodchucks or from animals with low viremia levels produced gamma interferon (IFN-␥) upon IL-12 stimulation, while lymphocytes from woodchucks with high viremia failed to upregulate IFN-␥ in response to IL-12. In conclusion, IL-12-based gene therapy is an efficient approach to treat chronic hepadnavirus infection in woodchucks with viral loads below 10 10 vg/ml. Interestingly, this therapy is able to break immunological tolerance to viral antigens in chronic WHV carriers.

Research paper thumbnail of A new type of NPM1 gene mutation in AML leading to a C-terminal truncated protein

Leukemia : official journal of the Leukemia Society of America, Leukemia Research Fund, U.K, 2007

by Thomas et al. 5 and observed by Crossman et al. 6 in patients with chronic myeloid leukemia.

Research paper thumbnail of Stride parameters and hindlimb length in horses fatigued on a treadmill and at an endurance ride

Equine Veterinary Journal, 2006

The relationship between fatigue and stride and/or muscle stiffness requires further study. Objec... more The relationship between fatigue and stride and/or muscle stiffness requires further study. Objective: To measure stride parameters in horses undergoing fatigue associated with running at submaximal speeds both on a treadmill and in an endurance ride. Hypothesis: Stride frequencies and estimates of hindlimb stiffness would he decreased in fatigued horses.

Research paper thumbnail of The relation between hypoxia-inducible factor (HIF)-1α and HIF-2α expression with anemia and outcome in surgically treated head and neck cancer

Research paper thumbnail of Prognostic significance of lymphangiogenesis in pharyngolaryngeal carcinoma patients

BMC Cancer, 2010

Background: Lymphatic vessel spread is considered a major route for head and neck squamous cell c... more Background: Lymphatic vessel spread is considered a major route for head and neck squamous cell carcinoma metastasis. Formation of new lymphatic vessels could facilitate the process, raising the malignant potential of these tumours. Recent identification of lymphatic markers allows the study of the lymphangiogenesis phenomenon. We searched for molecular events involved in the lymphangiogenic process that could have prognostic value in laryngeal/pharyngeal carcinoma patients. Methods: 104 paraffin-embedded pharyngeal/laryngeal tumour samples were studied. Immunohistochemical analysis of podoplanin and double immunofluorescence analysis of Ki-67 and D2-40 were performed. Lymph vessel density (inside the tumour mass, at its periphery or considered as a whole) and the presence of tumour emboli inside lymphatics were recorded. The proliferative state of endothelial lymphatic cells was evaluated.

Research paper thumbnail of Effects of melatonin in an experimental model of ventilator-induced lung injury

AJP: Lung Cellular and Molecular Physiology, 2008

all 10 cities including the rural areas of the province of Kerman. All data were finally analyzed... more all 10 cities including the rural areas of the province of Kerman. All data were finally analyzed by SPSS software (version 11.5). Results On the basis of recorded statistical analysis, the mortality cases of human rabies in the province of Kerman during one decade was 10 persons (eight males and two females). One-half of them (50%) were bitten by dogs and the others (50%) by foxes. Among the reported deaths, 40% were from Kahnooj county (Jiroft region). The reported data indicated that 21,546 persons were bitten by animals during 10 years in the province of Kerman. The mean of age of the people who were bitten by dogs was 24.80 years (SD = ±14.6), while the mean age of the people who were bitten by foxes was 57.25 years (SD = ±1.50). There was a significant difference between the mean age of these two groups of the people (P < 0.05). The most frequent rate of injured people was reported in the age group 10-19 years old and the frequency rate of males (76.00%) was more than females (24.00%). Therefore, there was a statistically significant difference between males and females in this study (P < 0.01). About 60% of all persons that were bitten by animals were from rural areas and 40% of them were from urban areas (P < 0.05). Among the people who were bitten and injured by animals during one decade in the province of Kerman, 85.70% of them were not treated by the rabies prophylaxis treatment regimen. Among all of them who were bitten by animals, 50% were injured through hands and feet, 40%

Research paper thumbnail of Lack of matrix metalloproteinase-9 worsens ventilator-induced lung injury

AJP: Lung Cellular and Molecular Physiology, 2007

Matrix metalloproteinase-9 (MMP-9) is released by neutrophils at the sites of acute inflammation.... more Matrix metalloproteinase-9 (MMP-9) is released by neutrophils at the sites of acute inflammation. This enzyme modulates matrix turnover and inflammatory response and its activity has been found increased after ventilator-induced lung injury. To clarify the role of MMP-9, mice lacking this enzyme and their wildtype counterparts were ventilated for two hours with high or low peak inspiratory pressures (25 and 15 cmH 2 O, respectively). Lung injury was evaluated by gas exchange, respiratory mechanics, wet-to-dry weight ratio and histologic analysis. The activity of MMP-9 and levels of interleukin-1 , interleukin-4 and macrophage inflammatory protein (MIP-2) were measured in lung tissue and bronchoalveolar lavage fluid (BALF). Cell count and myeloperoxidase activity were measured in BALF. There were no differences between wildtype and Mmp-9 -/animals after low-pressure ventilation. After high pressure ventilation, wildtype mice exhibited an increase in MMP-9 in tissue and BALF. Mice lacking MMP-9 developed more severe lung injury than wildtype mice, in terms of impaired oxygenation and lung mechanics, and higher damage in the histological study. These effects correlated with an increase in both cell count and myeloperoxidase activity in the BALF, suggesting an increased neutrophilic influx in response to ventilation. An increase in IL-1 and IL-4 in the BALF only in knockout mice could be responsible for the differences. There were no differences between genotypes in MMP-2, MMP-8 or tissue inhibitors of metalloproteinases.

Research paper thumbnail of Significado pronóstico de la angiogénesis en los carcinomas epidermoides de laringe supraglótica tratados con cirugía

Acta Otorrinolaringológica Española, 2009

It is now generally accepted that angiogenesis is crucial in tumour growth. However, controversy ... more It is now generally accepted that angiogenesis is crucial in tumour growth. However, controversy still exists regarding the prognostic significance of angiogenesis in head and neck carcinomas. The aim of this paper is to determine the prognostic significance of angiogenesis in a homogeneously treated group of supraglottic squamous cell carcinomas. 108 patients surgically treated for squamous cell carcinoma of the supraglottic larynx were studied. Angiogenesis was estimated in the primary tumour and in the nodal metastases by determining microvessel density using the &amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;quot;hot spot&amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;quot; method. Anti-CD34 antibody was used to stain blood vessels. The mean microvessel density in primary tumours was 72+/-34 vessels/mm(2) and 58.5+/-31.5 vessels/mm(2) in nodal metastases. No correlation was found between microvessel density in the primary tumours and the corresponding nodal metastasis (P=0.195). No significant differences in microvessel density were observed in relation to clinico-pathological parameters or survival (P=0.19). Our results suggest that microvessel density is not a useful prognostic marker in surgically treated supraglottic squamous cell carcinomas.

Research paper thumbnail of Clinical significance of Anoctamin-1 gene at 11q13 in the development and progression of head and neck squamous cell carcinomas

Scientific reports, Jan 26, 2015

This study investigates the clinical significance of Anoctamin-1 gene mapping at 11q13 amplicon i... more This study investigates the clinical significance of Anoctamin-1 gene mapping at 11q13 amplicon in both the development and progression of head and neck squamous cell carcinomas (HNSCC). ANO1 protein expression was evaluated by immunohistochemistry in a cohort of 372 surgically treated HNSCC patients and also in 35 laryngeal precancerous lesions. ANO1 gene amplification was determined by real-time PCR in all the laryngeal premalignancies and 60 of the HNSCCs, and molecular data correlated with clinical outcome. ANO1 gene amplification was frequently detected in both premalignant lesions (63%) and HNSCC tumours (58%), whereas concomitant ANO1 expression occurred at a much lower frequency (20 and 22%). Interestingly, laryngeal dysplasias harbouring ANO1 gene amplification showed a higher risk of malignant transformation (HR = 3.62; 95% CI 0.79-16.57; P = 0.097; Cox regression). ANO1 expression and gene amplification showed no significant associations with clinicopathological parameter...

Research paper thumbnail of Heparan sulfate proteoglycans undergo differential expression alterations in right sided colorectal cancer, depending on their metastatic character

BMC Cancer, 2015

Heparan sulfate proteoglycans (HSPGs) are complex molecules involved in the growth, invasion and ... more Heparan sulfate proteoglycans (HSPGs) are complex molecules involved in the growth, invasion and metastatic properties of cancerous cells. This study analyses the alterations in the expression patterns of these molecules in right sided colorectal cancer (CRC), both metastatic and non-metastatic. Twenty right sided CRCs were studied. A transcriptomic approach was used, employing qPCR to analyze both the expression of the enzymes involved in heparan sulfate (HS) chains biosynthesis, as well as the proteoglycan core proteins. Since some of these proteoglycans can also carry chondroitin sulfate (CS) chains, we include the study of the genes involved in the biosynthesis of these glycosaminoglycans. Immunohistochemical techniques were also used to analyze tissue expression of particular genes showing significant expression differences, of potential interest. Changes in proteoglycan core proteins differ depending on their location; those located intracellularly or in the extracellular matrix show very similar alteration patterns, while those located on the cell surface vary greatly depending on the nature of the tumor: glypicans 1, 3, 6 and betaglycan are affected in the non-metastatic tumors, whereas in the metastatic, only glypican-1 and syndecan-1 are modified, the latter showing opposing alterations in levels of RNA and of protein, suggesting post-transcriptional regulation in these tumors. Furthermore, in non-metastatic tumors, polymerization of glycosaminoglycan chains is modified, particularly affecting the synthesis of the tetrasaccharide linker and the initiation and elongation of CS chains, HS chains being less affected. Regarding the enzymes responsible for the modificaton of the HS chains, alterations were only found in non-metastatic tumors, affecting N-sulfation and the isoforms HS6ST1, HS3ST3B and HS3ST5. In contrast, synthesis of the CS chains suggests changes in epimerization and sulfation of the C4 and C2 in both types of tumor. Right sided CRCs show alterations in the expression of HSPGs, including the expression of the cell surface core proteins, many glycosiltransferases and some enzymes that modify the HS chains depending on the metastatic nature of the tumor, resulting more affected in non-metastatic ones. However, matrix proteoglycans and enzymes involved in CS fine structure synthesis are extensively modified independetly of the presence of lymph node metastasis.

Research paper thumbnail of The role of pneumolysin in mediating lung damage in a lethal pneumococcal pneumonia murine model

Respiratory research, Jan 26, 2007

Intranasal inoculation of Streptococcus pneumoniae D39 serotype 2 causes fatal pneumonia in mice.... more Intranasal inoculation of Streptococcus pneumoniae D39 serotype 2 causes fatal pneumonia in mice. The cytotoxic and inflammatory properties of pneumolysin (PLY) have been implicated in the pathogenesis of pneumococcal pneumonia. To examine the role of PLY in this experimental model we performed ELISA assays for PLY quantification. The distribution patterns of PLY and apoptosis were established by immunohistochemical detection of PLY, caspase-9 activity and TUNEL assay on tissue sections from mice lungs at various times, and the results were quantified with image analysis. Inflammatory and apoptotic cells were also quantified on lung tissue sections from antibody treated mice. In bronchoalveolar lavages (BAL), total PLY was found at sublytic concentrations which were located in alveolar macrophages and leukocytes. The bronchoalveolar epithelium was PLY-positive, while the vascular endothelium was not PLY reactive. The pattern and extension of cellular apoptosis was similar. Anti-PLY ...

Research paper thumbnail of Increased inflammation delays wound healing in mice deficient in collagenase-2 (MMP-8)

FASEB Journal, 2007

Matrix metalloproteinases (MMPs) have been implicated in numerous tissue-remodeling processes. Th... more Matrix metalloproteinases (MMPs) have been implicated in numerous tissue-remodeling processes. The finding that mice deficient in collagenase-2 (MMP-8) are more susceptible to develop skin cancer, prompted us to investigate the role of this protease in cutaneous wound healing. We have observed a significant delay in wound closure in MMP8 Ϫ/Ϫ mice and an altered inflammatory response in their wounds, with a delay of neutrophil infiltration during the first days and a persistent inflammation at later time points. These changes were accompanied by alterations in the TGF-␤1 signaling pathway and by an apoptosis defect in MMP8 Ϫ/Ϫ mice. The delay in wound healing observed in MMP8 ؊/؊ mice was rescued by bone marrow transplantation from wildtype mice. Analysis of other MMPs showed that MMP8 Ϫ/Ϫ mice had a significant increase in the expression of MMP-9, suggesting that both proteases might act coordinately in this process. This possibility was further supported by the novel finding that MMP-8 and MMP-9 form specific complexes in vivo. Taken together, these data indicate that MMP-8 participates in wound repair by contributing to the resolution of inflammation and open the possibility to develop new strategies for treating wound healing defects.-Gutiérrez-Fernández, A., Inada,

Research paper thumbnail of Defective prelamin A processing and muscular and adipocyte alterations in Zmpste24 metalloproteinase-deficient mice

Nature Genetics, 2002

The mouse ortholog of human FACE-1, Zmpste24, is a multispanning membrane protein widely distribu... more The mouse ortholog of human FACE-1, Zmpste24, is a multispanning membrane protein widely distributed in mammalian tissues 1,2 and structurally related to Afc1p/ste24p, a yeast metalloproteinase involved in the maturation of fungal pheromones 3 . Disruption of the gene Zmpste24 caused severe growth retardation and premature death in homozygous-null mice. Histopathological analysis of the mutant mice revealed several abnormalities, including dilated cardiomyopathy, muscular dystrophy and lipodystrophy. These alterations are similar to those developed by mice deficient in A-type lamin 4 , a major component of the nuclear lamina 5 , and phenocopy most defects observed in humans with diverse congenital laminopathies 6-8 . In agreement with this finding, Zmpste24-null mice are defective in the proteolytic processing of prelamin A. This deficiency in prelamin A maturation leads to the generation of abnormalities in nuclear architecture that probably underlie the many phenotypes observed in both mice and humans with mutations in the lamin A gene. These results indicate that prelamin A is a specific substrate for Zmpste24 and demonstrate the usefulness of genetic approaches for identifying the in vivo substrates of proteolytic enzymes.

Research paper thumbnail of 672 Dissecting the protective role of vitamin D3 on colon cancer: new targets from the protein degradation machinery

European Journal of Cancer Supplements, 2010

Sox2. However, very little is known about Sox2 regulators. It is well-established that mir-21 has... more Sox2. However, very little is known about Sox2 regulators. It is well-established that mir-21 has an anti-apoptotic effect in various cancer cells. Conclusions: The Sox2 suppressing effect of mir-21 suggests a hitherto unknown novel pathway. These findings could be implicated in anti-glioma therapy. Targeting mir-21 would not only lead to increased apoptosis, as has previously been demonstrated by several investigators, but also to decreased expression of a transcription factor which is required for the maintenance of stemness.

Research paper thumbnail of Neuroendocrine Tumors Show Altered Expression of Chondroitin Sulfate, Glypican 1, Glypican 5, and Syndecan 2 Depending on Their Differentiation Grade

Frontiers in Oncology, 2014

Neuroendocrine tumors (NETs) are found throughout the body and are important as they give rise to... more Neuroendocrine tumors (NETs) are found throughout the body and are important as they give rise to distinct clinical syndromes. Glycosaminoglycans, in proteoglycan (PG) form or as free chains, play vital roles in every step of tumor progression. Analyzing tumor samples with different degrees of histological differentiation we determined the existence of important alterations in chondroitin sulfate (CS) chains. Analysis of the transcription of the genes responsible for the production of CS showed a decline in the expression of some genes in poorly differentiated compared to well-differentiated tumors. Using anti-CS antibodies, normal stroma was always negative whereas tumoral stroma always showed a positive staining, more intense in the highest grade carcinomas, while tumor cells were negative. Moreover, certain specific cell surface PGs experienced a drastic decrease in expression depending on tumor differentiation. Syndecan 2 levels were very low or undetectable in healthy tissues, increasing significantly in well-differentiated tumors, and decreasing in poorly differentiated NETs, and its expression levels showed a positive correlation with patient survival. Glypican 5 appeared overexpressed in high-grade tumors with epithelial differentiation, and not in those that displayed a neuroendocrine phenotype. In contrast, normal neuroendocrine cells were positive for glypican 1, displaying intense staining in cytoplasm and membrane. Low-grade NETs had increased expression of this PG, but this reduced as tumor grade increased, its expression correlating positively with patient survival. Whilst elevated glypican 1 expression has been documented in different tumors, the downregulation in high-grade tumors observed in this work suggests that this proteoglycan could be involved in cancer development in a more complex and context-dependent manner than previously thought. Citation: García-Suárez O, García B, Fernández-Vega I, Astudillo A and Quirós LM (2014) Neuroendocrine tumors show altered expression of chondroitin sulfate, glypican 1, glypican 5, and syndecan 2 depending on their differentiation grade. Front. Oncol. 4:15.

Research paper thumbnail of Resistance to Bleomycin-Induced Lung Fibrosis in MMP-8 Deficient Mice Is Mediated by Interleukin-10

PLoS ONE, 2010

Background: Matrix metalloproteinases (MMPs) may have pro and antifibrotic roles within the lungs... more Background: Matrix metalloproteinases (MMPs) may have pro and antifibrotic roles within the lungs, due to its ability to modulate collagen turnover and immune mediators. MMP-8 is a collagenase that also cleaves a number of cytokines and chemokines.

Research paper thumbnail of Epigenetic repression of ROR2 has a Wnt-mediated, pro-tumourigenic role in colon cancer

Molecular Cancer, 2010

Background: Wnt factors control cell differentiation through semi-independent molecular cascades ... more Background: Wnt factors control cell differentiation through semi-independent molecular cascades known as the βcatenin-dependent (canonical) and -independent (non-canonical) Wnt signalling pathways. Genetic and epigenetic alteration of components of the canonical Wnt signalling pathway is one of the primary mechanisms underlying colon cancer. Despite increasing evidence of the role of the non-canonical pathways in tumourigenesis, however, the underlying molecular mechanisms are poorly understood.

Research paper thumbnail of Diet-Induced Obesity and Reduced Skin Cancer Susceptibility in Matrix Metalloproteinase 19-Deficient Mice

Molecular and Cellular Biology, 2004

to most MMPs, is widely expressed in human tissues under normal quiescent conditions. MMP-19 has ... more to most MMPs, is widely expressed in human tissues under normal quiescent conditions. MMP-19 has been found to be associated with ovulation and angiogenic processes and is deregulated in diverse pathological conditions such as rheumatoid arthritis and cancer. To gain further insights into the in vivo functions of this protease, we have generated mutant mice deficient in Mmp19. These mice are viable and fertile and do not display any obvious abnormalities. However, Mmp19-null mice develop a diet-induced obesity due to adipocyte hypertrophy and exhibit decreased susceptibility to skin tumors induced by chemical carcinogens. Based on these results, we suggest that this enzyme plays an in vivo role in some of the tissue remodeling events associated with adipogenesis, as well as in pathological processes such as tumor progression.

Research paper thumbnail of Loss of collagenase-2 confers increased skin tumor susceptibility to male mice

Nature Genetics, 2003

We generated mice deficient in collagenase-2 (Mmp8), an MMP mainly produced by neutrophils in inf... more We generated mice deficient in collagenase-2 (Mmp8), an MMP mainly produced by neutrophils in inflammatory reactions and detected in some malignant tumors 1,4 . Loss of Mmp8 did not cause abnormalities during embryonic development or in adult mice. Contrary to previous studies with MMP-deficient mice, however, the absence of Mmp8 strongly increased the incidence of skin tumors in male Mmp8 -/mice. Female Mmp8 -/mice whose ovaries were removed or were treated with tamoxifen were also more susceptible to tumors compared with wild-type mice. Bone marrow transplantation experiments confirmed that Mmp8 supplied by neutrophils was sufficient to restore the natural protection against tumor development mediated by this protease in male mice. Histopathological analysis showed that mutant mice had abnormalities in the inflammatory response induced by carcinogens. Our study identifies a paradoxical protective role for Mmp8 in cancer and provides a genetic model to evaluate the molecular basis of gender differences in cancer susceptibility.

Research paper thumbnail of Reply to ‘Aberrant cytoplasmic expression of C-terminal truncated NPM leukaemic mutant is dictated by tryptophans loss and a new NES motif’ by Falini et al

Leukemia, 2007

Martelli MF et al. Identification and functional characterization of a cytoplasmic nucleophosmin ... more Martelli MF et al. Identification and functional characterization of a cytoplasmic nucleophosmin leukaemic mutant generated by a novel exon-11 NPM1 mutation. Leukemia 2007; 21: 1099-1103. 4 Falini B, Bolli N, Shan J, Martelli MP, Liso A, Pucciarini A et al. Both carboxy-terminus NES motif and mutated tryptophan(s) are crucial for aberrant nuclear export of nucleophosmin leukemic mutants in NPMc+ AML. Blood 2006; 107: 4514-4523.

Research paper thumbnail of Treatment of Chronic Viral Hepatitis in Woodchucks by Prolonged Intrahepatic Expression of Interleukin-12

Journal of Virology, 2011

Chronic hepatitis B is a major cause of liver-related death worldwide. Interleukin-12 (IL-12) ind... more Chronic hepatitis B is a major cause of liver-related death worldwide. Interleukin-12 (IL-12) induction accompanies viral clearance in chronic hepatitis B virus infection. Here, we tested the therapeutic potential of IL-12 gene therapy in woodchucks chronically infected with woodchuck hepatitis virus (WHV), an infection that closely resembles chronic hepatitis B. The woodchucks were treated by intrahepatic injection of a helper-dependent adenoviral vector encoding IL-12 under the control of a liver-specific RU486-responsive promoter. All woodchucks with viral loads below 10 10 viral genomes (vg)/ml showed a marked and sustained reduction of viremia that was accompanied by a reduction in hepatic WHV DNA, a loss of e antigen and surface antigen, and improved liver histology. In contrast, none of the woodchucks with higher viremia levels responded to therapy. The antiviral effect was associated with the induction of T-cell immunity against viral antigens and a reduction of hepatic expression of Foxp3 in the responsive animals. Studies were performed in vitro to elucidate the resistance to therapy in highly viremic woodchucks. These studies showed that lymphocytes from healthy woodchucks or from animals with low viremia levels produced gamma interferon (IFN-␥) upon IL-12 stimulation, while lymphocytes from woodchucks with high viremia failed to upregulate IFN-␥ in response to IL-12. In conclusion, IL-12-based gene therapy is an efficient approach to treat chronic hepadnavirus infection in woodchucks with viral loads below 10 10 vg/ml. Interestingly, this therapy is able to break immunological tolerance to viral antigens in chronic WHV carriers.

Research paper thumbnail of A new type of NPM1 gene mutation in AML leading to a C-terminal truncated protein

Leukemia : official journal of the Leukemia Society of America, Leukemia Research Fund, U.K, 2007

by Thomas et al. 5 and observed by Crossman et al. 6 in patients with chronic myeloid leukemia.

Research paper thumbnail of Stride parameters and hindlimb length in horses fatigued on a treadmill and at an endurance ride

Equine Veterinary Journal, 2006

The relationship between fatigue and stride and/or muscle stiffness requires further study. Objec... more The relationship between fatigue and stride and/or muscle stiffness requires further study. Objective: To measure stride parameters in horses undergoing fatigue associated with running at submaximal speeds both on a treadmill and in an endurance ride. Hypothesis: Stride frequencies and estimates of hindlimb stiffness would he decreased in fatigued horses.

Research paper thumbnail of The relation between hypoxia-inducible factor (HIF)-1α and HIF-2α expression with anemia and outcome in surgically treated head and neck cancer

Research paper thumbnail of Prognostic significance of lymphangiogenesis in pharyngolaryngeal carcinoma patients

BMC Cancer, 2010

Background: Lymphatic vessel spread is considered a major route for head and neck squamous cell c... more Background: Lymphatic vessel spread is considered a major route for head and neck squamous cell carcinoma metastasis. Formation of new lymphatic vessels could facilitate the process, raising the malignant potential of these tumours. Recent identification of lymphatic markers allows the study of the lymphangiogenesis phenomenon. We searched for molecular events involved in the lymphangiogenic process that could have prognostic value in laryngeal/pharyngeal carcinoma patients. Methods: 104 paraffin-embedded pharyngeal/laryngeal tumour samples were studied. Immunohistochemical analysis of podoplanin and double immunofluorescence analysis of Ki-67 and D2-40 were performed. Lymph vessel density (inside the tumour mass, at its periphery or considered as a whole) and the presence of tumour emboli inside lymphatics were recorded. The proliferative state of endothelial lymphatic cells was evaluated.

Research paper thumbnail of Effects of melatonin in an experimental model of ventilator-induced lung injury

AJP: Lung Cellular and Molecular Physiology, 2008

all 10 cities including the rural areas of the province of Kerman. All data were finally analyzed... more all 10 cities including the rural areas of the province of Kerman. All data were finally analyzed by SPSS software (version 11.5). Results On the basis of recorded statistical analysis, the mortality cases of human rabies in the province of Kerman during one decade was 10 persons (eight males and two females). One-half of them (50%) were bitten by dogs and the others (50%) by foxes. Among the reported deaths, 40% were from Kahnooj county (Jiroft region). The reported data indicated that 21,546 persons were bitten by animals during 10 years in the province of Kerman. The mean of age of the people who were bitten by dogs was 24.80 years (SD = ±14.6), while the mean age of the people who were bitten by foxes was 57.25 years (SD = ±1.50). There was a significant difference between the mean age of these two groups of the people (P < 0.05). The most frequent rate of injured people was reported in the age group 10-19 years old and the frequency rate of males (76.00%) was more than females (24.00%). Therefore, there was a statistically significant difference between males and females in this study (P < 0.01). About 60% of all persons that were bitten by animals were from rural areas and 40% of them were from urban areas (P < 0.05). Among the people who were bitten and injured by animals during one decade in the province of Kerman, 85.70% of them were not treated by the rabies prophylaxis treatment regimen. Among all of them who were bitten by animals, 50% were injured through hands and feet, 40%

Research paper thumbnail of Lack of matrix metalloproteinase-9 worsens ventilator-induced lung injury

AJP: Lung Cellular and Molecular Physiology, 2007

Matrix metalloproteinase-9 (MMP-9) is released by neutrophils at the sites of acute inflammation.... more Matrix metalloproteinase-9 (MMP-9) is released by neutrophils at the sites of acute inflammation. This enzyme modulates matrix turnover and inflammatory response and its activity has been found increased after ventilator-induced lung injury. To clarify the role of MMP-9, mice lacking this enzyme and their wildtype counterparts were ventilated for two hours with high or low peak inspiratory pressures (25 and 15 cmH 2 O, respectively). Lung injury was evaluated by gas exchange, respiratory mechanics, wet-to-dry weight ratio and histologic analysis. The activity of MMP-9 and levels of interleukin-1 , interleukin-4 and macrophage inflammatory protein (MIP-2) were measured in lung tissue and bronchoalveolar lavage fluid (BALF). Cell count and myeloperoxidase activity were measured in BALF. There were no differences between wildtype and Mmp-9 -/animals after low-pressure ventilation. After high pressure ventilation, wildtype mice exhibited an increase in MMP-9 in tissue and BALF. Mice lacking MMP-9 developed more severe lung injury than wildtype mice, in terms of impaired oxygenation and lung mechanics, and higher damage in the histological study. These effects correlated with an increase in both cell count and myeloperoxidase activity in the BALF, suggesting an increased neutrophilic influx in response to ventilation. An increase in IL-1 and IL-4 in the BALF only in knockout mice could be responsible for the differences. There were no differences between genotypes in MMP-2, MMP-8 or tissue inhibitors of metalloproteinases.

Research paper thumbnail of Significado pronóstico de la angiogénesis en los carcinomas epidermoides de laringe supraglótica tratados con cirugía

Acta Otorrinolaringológica Española, 2009

It is now generally accepted that angiogenesis is crucial in tumour growth. However, controversy ... more It is now generally accepted that angiogenesis is crucial in tumour growth. However, controversy still exists regarding the prognostic significance of angiogenesis in head and neck carcinomas. The aim of this paper is to determine the prognostic significance of angiogenesis in a homogeneously treated group of supraglottic squamous cell carcinomas. 108 patients surgically treated for squamous cell carcinoma of the supraglottic larynx were studied. Angiogenesis was estimated in the primary tumour and in the nodal metastases by determining microvessel density using the &amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;quot;hot spot&amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;quot; method. Anti-CD34 antibody was used to stain blood vessels. The mean microvessel density in primary tumours was 72+/-34 vessels/mm(2) and 58.5+/-31.5 vessels/mm(2) in nodal metastases. No correlation was found between microvessel density in the primary tumours and the corresponding nodal metastasis (P=0.195). No significant differences in microvessel density were observed in relation to clinico-pathological parameters or survival (P=0.19). Our results suggest that microvessel density is not a useful prognostic marker in surgically treated supraglottic squamous cell carcinomas.

Research paper thumbnail of Clinical significance of Anoctamin-1 gene at 11q13 in the development and progression of head and neck squamous cell carcinomas

Scientific reports, Jan 26, 2015

This study investigates the clinical significance of Anoctamin-1 gene mapping at 11q13 amplicon i... more This study investigates the clinical significance of Anoctamin-1 gene mapping at 11q13 amplicon in both the development and progression of head and neck squamous cell carcinomas (HNSCC). ANO1 protein expression was evaluated by immunohistochemistry in a cohort of 372 surgically treated HNSCC patients and also in 35 laryngeal precancerous lesions. ANO1 gene amplification was determined by real-time PCR in all the laryngeal premalignancies and 60 of the HNSCCs, and molecular data correlated with clinical outcome. ANO1 gene amplification was frequently detected in both premalignant lesions (63%) and HNSCC tumours (58%), whereas concomitant ANO1 expression occurred at a much lower frequency (20 and 22%). Interestingly, laryngeal dysplasias harbouring ANO1 gene amplification showed a higher risk of malignant transformation (HR = 3.62; 95% CI 0.79-16.57; P = 0.097; Cox regression). ANO1 expression and gene amplification showed no significant associations with clinicopathological parameter...

Research paper thumbnail of Heparan sulfate proteoglycans undergo differential expression alterations in right sided colorectal cancer, depending on their metastatic character

BMC Cancer, 2015

Heparan sulfate proteoglycans (HSPGs) are complex molecules involved in the growth, invasion and ... more Heparan sulfate proteoglycans (HSPGs) are complex molecules involved in the growth, invasion and metastatic properties of cancerous cells. This study analyses the alterations in the expression patterns of these molecules in right sided colorectal cancer (CRC), both metastatic and non-metastatic. Twenty right sided CRCs were studied. A transcriptomic approach was used, employing qPCR to analyze both the expression of the enzymes involved in heparan sulfate (HS) chains biosynthesis, as well as the proteoglycan core proteins. Since some of these proteoglycans can also carry chondroitin sulfate (CS) chains, we include the study of the genes involved in the biosynthesis of these glycosaminoglycans. Immunohistochemical techniques were also used to analyze tissue expression of particular genes showing significant expression differences, of potential interest. Changes in proteoglycan core proteins differ depending on their location; those located intracellularly or in the extracellular matrix show very similar alteration patterns, while those located on the cell surface vary greatly depending on the nature of the tumor: glypicans 1, 3, 6 and betaglycan are affected in the non-metastatic tumors, whereas in the metastatic, only glypican-1 and syndecan-1 are modified, the latter showing opposing alterations in levels of RNA and of protein, suggesting post-transcriptional regulation in these tumors. Furthermore, in non-metastatic tumors, polymerization of glycosaminoglycan chains is modified, particularly affecting the synthesis of the tetrasaccharide linker and the initiation and elongation of CS chains, HS chains being less affected. Regarding the enzymes responsible for the modificaton of the HS chains, alterations were only found in non-metastatic tumors, affecting N-sulfation and the isoforms HS6ST1, HS3ST3B and HS3ST5. In contrast, synthesis of the CS chains suggests changes in epimerization and sulfation of the C4 and C2 in both types of tumor. Right sided CRCs show alterations in the expression of HSPGs, including the expression of the cell surface core proteins, many glycosiltransferases and some enzymes that modify the HS chains depending on the metastatic nature of the tumor, resulting more affected in non-metastatic ones. However, matrix proteoglycans and enzymes involved in CS fine structure synthesis are extensively modified independetly of the presence of lymph node metastasis.

Research paper thumbnail of The role of pneumolysin in mediating lung damage in a lethal pneumococcal pneumonia murine model

Respiratory research, Jan 26, 2007

Intranasal inoculation of Streptococcus pneumoniae D39 serotype 2 causes fatal pneumonia in mice.... more Intranasal inoculation of Streptococcus pneumoniae D39 serotype 2 causes fatal pneumonia in mice. The cytotoxic and inflammatory properties of pneumolysin (PLY) have been implicated in the pathogenesis of pneumococcal pneumonia. To examine the role of PLY in this experimental model we performed ELISA assays for PLY quantification. The distribution patterns of PLY and apoptosis were established by immunohistochemical detection of PLY, caspase-9 activity and TUNEL assay on tissue sections from mice lungs at various times, and the results were quantified with image analysis. Inflammatory and apoptotic cells were also quantified on lung tissue sections from antibody treated mice. In bronchoalveolar lavages (BAL), total PLY was found at sublytic concentrations which were located in alveolar macrophages and leukocytes. The bronchoalveolar epithelium was PLY-positive, while the vascular endothelium was not PLY reactive. The pattern and extension of cellular apoptosis was similar. Anti-PLY ...

Research paper thumbnail of Increased inflammation delays wound healing in mice deficient in collagenase-2 (MMP-8)

FASEB Journal, 2007

Matrix metalloproteinases (MMPs) have been implicated in numerous tissue-remodeling processes. Th... more Matrix metalloproteinases (MMPs) have been implicated in numerous tissue-remodeling processes. The finding that mice deficient in collagenase-2 (MMP-8) are more susceptible to develop skin cancer, prompted us to investigate the role of this protease in cutaneous wound healing. We have observed a significant delay in wound closure in MMP8 Ϫ/Ϫ mice and an altered inflammatory response in their wounds, with a delay of neutrophil infiltration during the first days and a persistent inflammation at later time points. These changes were accompanied by alterations in the TGF-␤1 signaling pathway and by an apoptosis defect in MMP8 Ϫ/Ϫ mice. The delay in wound healing observed in MMP8 ؊/؊ mice was rescued by bone marrow transplantation from wildtype mice. Analysis of other MMPs showed that MMP8 Ϫ/Ϫ mice had a significant increase in the expression of MMP-9, suggesting that both proteases might act coordinately in this process. This possibility was further supported by the novel finding that MMP-8 and MMP-9 form specific complexes in vivo. Taken together, these data indicate that MMP-8 participates in wound repair by contributing to the resolution of inflammation and open the possibility to develop new strategies for treating wound healing defects.-Gutiérrez-Fernández, A., Inada,

Research paper thumbnail of Defective prelamin A processing and muscular and adipocyte alterations in Zmpste24 metalloproteinase-deficient mice

Nature Genetics, 2002

The mouse ortholog of human FACE-1, Zmpste24, is a multispanning membrane protein widely distribu... more The mouse ortholog of human FACE-1, Zmpste24, is a multispanning membrane protein widely distributed in mammalian tissues 1,2 and structurally related to Afc1p/ste24p, a yeast metalloproteinase involved in the maturation of fungal pheromones 3 . Disruption of the gene Zmpste24 caused severe growth retardation and premature death in homozygous-null mice. Histopathological analysis of the mutant mice revealed several abnormalities, including dilated cardiomyopathy, muscular dystrophy and lipodystrophy. These alterations are similar to those developed by mice deficient in A-type lamin 4 , a major component of the nuclear lamina 5 , and phenocopy most defects observed in humans with diverse congenital laminopathies 6-8 . In agreement with this finding, Zmpste24-null mice are defective in the proteolytic processing of prelamin A. This deficiency in prelamin A maturation leads to the generation of abnormalities in nuclear architecture that probably underlie the many phenotypes observed in both mice and humans with mutations in the lamin A gene. These results indicate that prelamin A is a specific substrate for Zmpste24 and demonstrate the usefulness of genetic approaches for identifying the in vivo substrates of proteolytic enzymes.

Research paper thumbnail of 672 Dissecting the protective role of vitamin D3 on colon cancer: new targets from the protein degradation machinery

European Journal of Cancer Supplements, 2010

Sox2. However, very little is known about Sox2 regulators. It is well-established that mir-21 has... more Sox2. However, very little is known about Sox2 regulators. It is well-established that mir-21 has an anti-apoptotic effect in various cancer cells. Conclusions: The Sox2 suppressing effect of mir-21 suggests a hitherto unknown novel pathway. These findings could be implicated in anti-glioma therapy. Targeting mir-21 would not only lead to increased apoptosis, as has previously been demonstrated by several investigators, but also to decreased expression of a transcription factor which is required for the maintenance of stemness.

Research paper thumbnail of Neuroendocrine Tumors Show Altered Expression of Chondroitin Sulfate, Glypican 1, Glypican 5, and Syndecan 2 Depending on Their Differentiation Grade

Frontiers in Oncology, 2014

Neuroendocrine tumors (NETs) are found throughout the body and are important as they give rise to... more Neuroendocrine tumors (NETs) are found throughout the body and are important as they give rise to distinct clinical syndromes. Glycosaminoglycans, in proteoglycan (PG) form or as free chains, play vital roles in every step of tumor progression. Analyzing tumor samples with different degrees of histological differentiation we determined the existence of important alterations in chondroitin sulfate (CS) chains. Analysis of the transcription of the genes responsible for the production of CS showed a decline in the expression of some genes in poorly differentiated compared to well-differentiated tumors. Using anti-CS antibodies, normal stroma was always negative whereas tumoral stroma always showed a positive staining, more intense in the highest grade carcinomas, while tumor cells were negative. Moreover, certain specific cell surface PGs experienced a drastic decrease in expression depending on tumor differentiation. Syndecan 2 levels were very low or undetectable in healthy tissues, increasing significantly in well-differentiated tumors, and decreasing in poorly differentiated NETs, and its expression levels showed a positive correlation with patient survival. Glypican 5 appeared overexpressed in high-grade tumors with epithelial differentiation, and not in those that displayed a neuroendocrine phenotype. In contrast, normal neuroendocrine cells were positive for glypican 1, displaying intense staining in cytoplasm and membrane. Low-grade NETs had increased expression of this PG, but this reduced as tumor grade increased, its expression correlating positively with patient survival. Whilst elevated glypican 1 expression has been documented in different tumors, the downregulation in high-grade tumors observed in this work suggests that this proteoglycan could be involved in cancer development in a more complex and context-dependent manner than previously thought. Citation: García-Suárez O, García B, Fernández-Vega I, Astudillo A and Quirós LM (2014) Neuroendocrine tumors show altered expression of chondroitin sulfate, glypican 1, glypican 5, and syndecan 2 depending on their differentiation grade. Front. Oncol. 4:15.

Research paper thumbnail of Resistance to Bleomycin-Induced Lung Fibrosis in MMP-8 Deficient Mice Is Mediated by Interleukin-10

PLoS ONE, 2010

Background: Matrix metalloproteinases (MMPs) may have pro and antifibrotic roles within the lungs... more Background: Matrix metalloproteinases (MMPs) may have pro and antifibrotic roles within the lungs, due to its ability to modulate collagen turnover and immune mediators. MMP-8 is a collagenase that also cleaves a number of cytokines and chemokines.

Research paper thumbnail of Epigenetic repression of ROR2 has a Wnt-mediated, pro-tumourigenic role in colon cancer

Molecular Cancer, 2010

Background: Wnt factors control cell differentiation through semi-independent molecular cascades ... more Background: Wnt factors control cell differentiation through semi-independent molecular cascades known as the βcatenin-dependent (canonical) and -independent (non-canonical) Wnt signalling pathways. Genetic and epigenetic alteration of components of the canonical Wnt signalling pathway is one of the primary mechanisms underlying colon cancer. Despite increasing evidence of the role of the non-canonical pathways in tumourigenesis, however, the underlying molecular mechanisms are poorly understood.

Research paper thumbnail of Diet-Induced Obesity and Reduced Skin Cancer Susceptibility in Matrix Metalloproteinase 19-Deficient Mice

Molecular and Cellular Biology, 2004

to most MMPs, is widely expressed in human tissues under normal quiescent conditions. MMP-19 has ... more to most MMPs, is widely expressed in human tissues under normal quiescent conditions. MMP-19 has been found to be associated with ovulation and angiogenic processes and is deregulated in diverse pathological conditions such as rheumatoid arthritis and cancer. To gain further insights into the in vivo functions of this protease, we have generated mutant mice deficient in Mmp19. These mice are viable and fertile and do not display any obvious abnormalities. However, Mmp19-null mice develop a diet-induced obesity due to adipocyte hypertrophy and exhibit decreased susceptibility to skin tumors induced by chemical carcinogens. Based on these results, we suggest that this enzyme plays an in vivo role in some of the tissue remodeling events associated with adipogenesis, as well as in pathological processes such as tumor progression.