23 results on '"Amalia Capilla"'
Search Results
2. Gut microbiota trajectory in early life may predict development of celiac disease
- Author
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Marta Olivares, Alan W. Walker, Amalia Capilla, Alfonso Benítez-Páez, Francesc Palau, Julian Parkhill, Gemma Castillejo, and Yolanda Sanz
- Subjects
Celiac disease ,Intestinal microbiology ,HLA genes ,Microbial ecology ,QR100-130 - Abstract
Abstract Background To investigate whether alterations in the developing intestinal microbiota and immune markers precede celiac disease (CD) onset in infants at familial risk of developing the disease. Methods A nested case-control study was carried out as part of a larger prospective cohort study, which included healthy full-term newborns (> 200) with at least one first relative with biopsy-verified CD. The present study includes cases of CD (n = 10) and the best-matched controls (n = 10) who did not develop the disease after 5-year follow-up. Fecal microbiota, assessed by high-throughput 16S rRNA gene amplicon sequencing, and immune parameters were profiled at 4 and 6 months of age and related to CD onset. Results The microbiota of infants who remained healthy showed an increase in bacterial diversity over time, characterized by increases in Firmicutes families, but not those who developed CD. Infants who subsequently developed CD showed a significant reduction in sIgA levels over time, while those who remained healthy showed increases in TNF-α correlated to Bifidobacterium spp. An increased relative abundance of Bifidobacterium longum was associated with control children while increased proportions of Bifidobacterium breve and Enterococcus spp. were associated with CD development. Conclusion The findings suggest that alterations in the early trajectory of gut microbiota in infants at CD risk could influence the immune maturation process and predispose to CD, although larger population studies are warranted to confirm this hypothesis.
- Published
- 2018
- Full Text
- View/download PDF
3. Oblique scanning laser microscopy for simultaneously volumetric structural and molecular imaging using only one raster scan
- Author
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Lei Zhang, Amalia Capilla, Weiye Song, Gustavo Mostoslavsky, and Ji Yi
- Subjects
Medicine ,Science - Abstract
Abstract Multi-modal three dimensional (3D) optical imaging combining both structural sensitivity and molecular specificity is highly desirable in biomedical research. In this paper, we present a method termed oblique scanning laser microscopy (OSLM) to combine optical coherence tomography (OCT), for simultaneously volumetric structural and molecular imaging with cellular resolution in all three dimensions. Conventional 3D laser scanning fluorescence microscopy requires repeated optical sectioning to create z-stacks in depth. Here, the use of an obliquely scanning laser eliminates the z-stacking process, then allows highly efficient 3D OCT and fluorescence imaging by using only one raster scan. The current setup provides ~3.6 × 4.2 × 6.5 μm resolution in fluorescence imaging, ~7 × 7 × 3.5 μm in OCT in three dimensions, and the current speed of imaging is up to 100 frames per second (fps) over a volume about 0.8 × 1 × 0.5 mm3. We demonstrate several mechanisms for molecular imaging, including intrinsically expressed GFP fluorescence, autofluorescence from Flavin proteins, and exogenous antibody-conjugated dyes. We also demonstrate potential applications in imaging human intestinal organoids (HIOs), colon mucosa, and retina.
- Published
- 2017
- Full Text
- View/download PDF
4. Modeling APC mutagenesis and familial adenomatous polyposis using human iPS cells.
- Author
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Cesar A Sommer, Amalia Capilla, Francisco J Molina-Estevez, Andreia Gianotti-Sommer, Nicholas Skvir, Ignacio Caballero, Sanjib Chowdhury, and Gustavo Mostoslavsky
- Subjects
Medicine ,Science - Abstract
Mutations in the gene Adenomatous Polyposis Coli or APC appear in most sporadic cases of colorectal cancer and it is the most frequent mutation causing hereditary Familial Adenomatous Polyposis. The detailed molecular mechanism by which APC mutations predispose to the development of colorectal cancer is not completely understood. This is in part due to the lack of accessibility to appropriate models that recapitulate the early events associated with APC mediated intestinal transformation. We have established a novel platform utilizing human induced Pluripotent Stem cells or iPSC from normal or FAP-specific APC mutant individuals and evaluated the effect of the mutation in the cells before and after differentiation into intestinal organoids. In order to minimize genetic background effects, we also established an isogenic platform using TALEN-mediated gene editing. Comparison of normal and APC mutant iPSC revealed a significant defect in cell identity and polarity due to the presence of APC in heterozygosity as well as chromosomal aberrations including abnormal anaphases and centrosome numbers. Importantly, upon specification into intestinal progeny, APC heterozygosity was responsible for a major change in the transcriptional identity of the cells with dysregulation of key signaling pathways, including metabolic reprogramming, abnormal lipid metabolism and intestinal-specific cadherin expression. In conclusion, we have developed a novel iPSC/intestinal model of APC mutagenesis and provide strong evidence that APC in heterozygosity imparts a clear phenotypic and molecular defect, affecting basic cellular functions and integrity, providing novel insights in the earlier events of APC-mediated tumorigenesis.
- Published
- 2018
- Full Text
- View/download PDF
5. Influence of milk-feeding type and genetic risk of developing coeliac disease on intestinal microbiota of infants: the PROFICEL study.
- Author
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Giada De Palma, Amalia Capilla, Esther Nova, Gemma Castillejo, Vicente Varea, Tamara Pozo, José Antonio Garrote, Isabel Polanco, Ana López, Carmen Ribes-Koninckx, Ascensión Marcos, María Dolores García-Novo, Carmen Calvo, Luis Ortigosa, Luis Peña-Quintana, Francesc Palau, and Yolanda Sanz
- Subjects
Medicine ,Science - Abstract
Interactions between environmental factors and predisposing genes could be involved in the development of coeliac disease (CD). This study has assessed whether milk-feeding type and HLA-genotype influence the intestinal microbiota composition of infants with a family history of CD. The study included 164 healthy newborns, with at least one first-degree relative with CD, classified according to their HLA-DQ genotype by PCR-SSP DQB1 and DQA1 typing. Faecal microbiota was analysed by quantitative PCR at 7 days, and at 1 and 4 months of age. Significant interactions between milk-feeding type and HLA-DQ genotype on bacterial numbers were not detected by applying a linear mixed-model analysis for repeated measures. In the whole population, breast-feeding promoted colonization of C. leptum group, B. longum and B. breve, while formula-feeding promoted that of Bacteroides fragilis group, C. coccoides-E. rectale group, E. coli and B. lactis. Moreover, increased numbers of B. fragilis group and Staphylococcus spp., and reduced numbers of Bifidobacterium spp. and B. longum were detected in infants with increased genetic risk of developing CD. Analyses within subgroups of either breast-fed or formula-fed infants indicated that in both cases increased risk of CD was associated with lower numbers of B. longum and/or Bifidobacterium spp. In addition, in breast-fed infants the increased genetic risk of developing CD was associated with increased C. leptum group numbers, while in formula-fed infants it was associated with increased Staphylococcus and B. fragilis group numbers. Overall, milk-feeding type in conjunction with HLA-DQ genotype play a role in establishing infants' gut microbiota; moreover, breast-feeding reduced the genotype-related differences in microbiota composition, which could partly explain the protective role attributed to breast milk in this disorder.
- Published
- 2012
- Full Text
- View/download PDF
6. Variants in Interferon Lambda are Associated with Very Early Onset Inflammatory Bowel Disease
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Jodie D. Ouahed, Achille Broggi, Abigail Glick, Robin T. Haring, Michael Field, Daniel Chinnapen, Michael J. Grey, Wayne Lencer, Steven J. Steiner, Evida Dennis-Heyward, Noah Shroyer, Zachary Criss, Shih-Ching Lin, Xi-Lei Zeng, Sue E. Crawford, Mary K. Estes, Jay R. Thiagarajah, Amalia Capilla, Whitney Scoon, Gustavo Mostoslavsky, Jason Spence, Christoph Klein, Aleixo M. Muise, Bruce Horwitz, Ivan Zanoni, and Scott B. Snapper
- Abstract
Inflammatory bowel diseases (IBD) are chronic inflammatory disorders of the intestine that affect children and adults. The etiology is multifactorial with the contribution of genetic, immune, microbial, and environmental risk factors with a substantial involvement of host:microbial cross-talk in the lumen. Whether and how antiviral responses contribute to IBD pathogenesis is under intense investigation. Here, we identified two unrelated patients diagnosed with very early onset IBD (VEOIBD) with rare variants in type III Interferons (IFNs), also known as interferon lambdas (IFNλs), a group of IFNs that play key roles in the protection against enteric viruses. The first patient was found to have homozygous variants in both IFNL2 and IFNL3, the genes encoding, IFN-λ2 and IFN-λ3, respectively. The second patient was found to have inherited compound heterozygous variants in IFNL3, with one of the two alleles bearing the same variant as the first patient. Functional analyses revealed that the proteins coded for by these variant IFN-λ genes exhibited defects in the induction of IFN signaling. More detailed assessment of the variants identified in Patient 1 demonstrated defects in their ability to bind to IFN-λ receptor 1 (IFNLR1), as well as their ability to induce heterodimerization of IFNLR1 and IL10RB, which together compromise the functional receptor for IFN-λ (IFNLR). These patient-encoded IFN-λ variants also exhibit defects in the ability to induce robust downstream IFN-stimulated genes (ISGs) in patient-derived intestinal organoids. All in all, we demonstrate that VEOIBD is associated with variants in IFN-λs and defective induction of downstream IFN signaling.
- Published
- 2022
7. Generation of mesenchyme free intestinal organoids from human induced pluripotent stem cells
- Author
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Finn Hawkins, Amalia Capilla, Aleksander D. Szymaniak, Alexander Stuffer, Marall Vedaie, Dar Heinze, Aditya Mithal, Carlos Villacorta-Martin, Gustavo Mostoslavsky, Darrell N. Kotton, Megan Peasley, Kristine M. Abo, John Mahoney, Anjali Jacob, and Andrew Berical
- Subjects
0301 basic medicine ,Cystic Fibrosis ,Science ,Mesenchyme ,Cellular differentiation ,Genetic Vectors ,Induced Pluripotent Stem Cells ,Thyroid Nuclear Factor 1 ,Stem-cell differentiation ,General Physics and Astronomy ,Biology ,Cystic fibrosis ,Article ,General Biochemistry, Genetics and Molecular Biology ,Mesoderm ,03 medical and health sciences ,0302 clinical medicine ,Directed differentiation ,Intestine, Small ,medicine ,Organoid ,Humans ,CDX2 Transcription Factor ,Gastrointestinal models ,Gene Knock-In Techniques ,Progenitor cell ,lcsh:Science ,Induced pluripotent stem cell ,CDX2 ,Multidisciplinary ,Disease model ,Cell Differentiation ,Epithelial Cells ,General Chemistry ,medicine.disease ,3. Good health ,Cell biology ,Intestines ,Organoids ,030104 developmental biology ,medicine.anatomical_structure ,030220 oncology & carcinogenesis ,lcsh:Q - Abstract
Efficient generation of human induced pluripotent stem cell (hiPSC)-derived human intestinal organoids (HIOs) would facilitate the development of in vitro models for a variety of diseases that affect the gastrointestinal tract, such as inflammatory bowel disease or Cystic Fibrosis. Here, we report a directed differentiation protocol for the generation of mesenchyme-free HIOs that can be primed towards more colonic or proximal intestinal lineages in serum-free defined conditions. Using a CDX2eGFP iPSC knock-in reporter line to track the emergence of hindgut progenitors, we follow the kinetics of CDX2 expression throughout directed differentiation, enabling the purification of intestinal progenitors and robust generation of mesenchyme-free organoids expressing characteristic markers of small intestinal or colonic epithelium. We employ HIOs generated in this way to measure CFTR function using cystic fibrosis patient-derived iPSC lines before and after correction of the CFTR mutation, demonstrating their future potential for disease modeling and therapeutic screening applications., Human induced pluripotent stem cell-derived intestinal organoids (HIOs) are powerful tools to study development and diseases of the gastrointestinal tract. Here, the authors develop a directed differentiation protocol to generate mesenchyme-free HIOs that can be patterned towards proximal small intestine or colonic epithelium, and demonstrated their utility in modeling CFTR function.
- Published
- 2020
8. Bioengineering of functional human induced pluripotent stem cell-derived intestinal grafts
- Author
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Sarah E. Gilpin, Allan M. Goldstein, Dana M. Schwartz, Cesar Sommer, Amalia Capilla, Harald C. Ott, Douglas J. Mathisen, Gustavo Mostoslavsky, Gregory R. Wojtkiewicz, Kentaro Kitano, Haiyang Zhou, and Xi Ren
- Subjects
Male ,Short Bowel Syndrome ,0301 basic medicine ,Pathology ,medicine.medical_specialty ,Endothelium ,Science ,Induced Pluripotent Stem Cells ,Transplants ,General Physics and Astronomy ,Bioengineering ,Article ,General Biochemistry, Genetics and Molecular Biology ,Rats, Sprague-Dawley ,03 medical and health sciences ,0302 clinical medicine ,Intestinal mucosa ,medicine ,Animals ,Humans ,Intestinal Mucosa ,Progenitor cell ,Induced pluripotent stem cell ,lcsh:Science ,Cells, Cultured ,Cell Proliferation ,Multidisciplinary ,Tissue Engineering ,Tissue Scaffolds ,business.industry ,Fatty Acids ,Endothelial Cells ,Cell Differentiation ,General Chemistry ,Intestinal epithelium ,Epithelium ,Small intestine ,Rats ,3. Good health ,Intestines ,Transplantation ,Glucose ,030104 developmental biology ,medicine.anatomical_structure ,surgical procedures, operative ,030220 oncology & carcinogenesis ,lcsh:Q ,business - Abstract
Patients with short bowel syndrome lack sufficient functional intestine to sustain themselves with enteral intake alone. Transplantable vascularized bioengineered intestine could restore nutrient absorption. Here we report the engineering of humanized intestinal grafts by repopulating decellularized rat intestinal matrix with human induced pluripotent stem cell-derived intestinal epithelium and human endothelium. After 28 days of in vitro culture, hiPSC-derived progenitor cells differentiate into a monolayer of polarized intestinal epithelium. Human endothelial cells seeded via native vasculature restore perfusability. Ex vivo isolated perfusion testing confirms transfer of glucose and medium-chain fatty acids from lumen to venous effluent. Four weeks after transplantation to RNU rats, grafts show survival and maturation of regenerated epithelium. Systemic venous sampling and positron emission tomography confirm uptake of glucose and fatty acids in vivo. Bioengineering intestine on vascularized native scaffolds could bridge the gap between cell/tissue-scale regeneration and whole organ-scale technology needed to treat intestinal failure patients., There is a need for humanised grafts to treat patients with intestinal failure. Here, the authors generate intestinal grafts by recellularizing native intestinal matrix with human induced pluripotent stem cell-derived epithelium and human endothelium, and show nutrient absorption after transplantation in rats.
- Published
- 2017
9. Gut microbiota trajectory in early life may predict development of celiac disease
- Author
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Alan W. Walker, Gemma Castillejo, Alfonso Benítez-Páez, Francesc Palau, Amalia Capilla, Yolanda Sanz, Marta Olivares, Julian Parkhill, Ministerio de Economía y Competitividad (España), Olivares, Marta [0000-0002-7966-2781], and Apollo - University of Cambridge Repository
- Subjects
0301 basic medicine ,Microbiology (medical) ,Male ,Bifidobacterium longum ,HLA genes ,ved/biology.organism_classification_rank.species ,Population ,Firmicutes ,Gut flora ,Microbiology ,lcsh:Microbial ecology ,03 medical and health sciences ,Feces ,0302 clinical medicine ,Immune system ,fluids and secretions ,RNA, Ribosomal, 16S ,Humans ,Prospective Studies ,Prospective cohort study ,education ,Bifidobacterium ,education.field_of_study ,Bifidobacterium breve ,biology ,ved/biology ,Intestinal microbiology ,Research ,Case-control study ,Infant, Newborn ,High-Throughput Nucleotide Sequencing ,Infant ,biology.organism_classification ,3. Good health ,Gastrointestinal Microbiome ,Gastrointestinal Tract ,Celiac Disease ,030104 developmental biology ,Case-Control Studies ,Child, Preschool ,Immunology ,lcsh:QR100-130 ,030211 gastroenterology & hepatology ,Female ,Enterococcus - Abstract
Background To investigate whether alterations in the developing intestinal microbiota and immune markers precede celiac disease (CD) onset in infants at familial risk of developing the disease. Methods A nested case-control study was carried out as part of a larger prospective cohort study, which included healthy full-term newborns (> 200) with at least one first relative with biopsy-verified CD. The present study includes cases of CD (n = 10) and the best-matched controls (n = 10) who did not develop the disease after 5-year follow-up. Fecal microbiota, assessed by high-throughput 16S rRNA gene amplicon sequencing, and immune parameters were profiled at 4 and 6 months of age and related to CD onset. Results The microbiota of infants who remained healthy showed an increase in bacterial diversity over time, characterized by increases in Firmicutes families, but not those who developed CD. Infants who subsequently developed CD showed a significant reduction in sIgA levels over time, while those who remained healthy showed increases in TNF-α correlated to Bifidobacterium spp. An increased relative abundance of Bifidobacterium longum was associated with control children while increased proportions of Bifidobacterium breve and Enterococcus spp. were associated with CD development. Conclusion The findings suggest that alterations in the early trajectory of gut microbiota in infants at CD risk could influence the immune maturation process and predispose to CD, although larger population studies are warranted to confirm this hypothesis., This work was supported by grants AGL2011-25169, AGL2014-52101-P, and AGL2007-66126-C03-03/ALI (YS and FP) from the Spanish Ministry of Economy and Competitiveness (MINECO). Funding for AWW and JP and 16S rRNA gene Olivares et al. Microbiome (2018) 6:36 Page 9 of 11 sequencing was provided by Wellcome Trust (Grant 098051); AWW and The Rowett Institute, University of Aberdeen, receive core funding support from the Scottish Government Rural and Environmental Science and Analysis Service (RESAS). The scholarship to MO from CSIC (JAEpre) and the contract to ABP from the European Union’s Seventh Framework Program under the grant agreement no 613979 (MyNewGut) are also fully acknowledged.
- Published
- 2018
10. Increased prevalence of pathogenic bacteria in the gut microbiota of infants at risk of developing celiac disease: The PROFICEL study
- Author
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Luis Ortigosa, Francesc Palau, Amalia Capilla, Carmen Ribes-Koninckx, Vicente Varea, Gemma Castillejo, Ester Donat, Alfonso Benítez-Páez, Esther Nova, Yolanda Sanz, C. Calvo, José Antonio Garrote, Giada De Palma, Ascensión Marcos, Marta Olivares, Isabel Polanco, Ministerio de Economía y Competitividad (España), and European Research Council
- Subjects
0301 basic medicine ,Disease ,Gut flora ,medicine.disease_cause ,Feces ,0302 clinical medicine ,Genotype ,Celiac disease ,gut microbiota ,Enteropathy ,2. Zero hunger ,biology ,Gastroenterology ,celiac disease ,3. Good health ,Infectious Diseases ,HLA genotype ,PCR ,030211 gastroenterology & hepatology ,Microbiology (medical) ,Risk ,Brief Report - Invited ,Gut microbiota ,Microbiology ,03 medical and health sciences ,HLA-DQ Antigens ,medicine ,Enterotoxigenic Escherichia coli ,Humans ,Genetic Predisposition to Disease ,Clostridium ,Bacteria ,Infant, Newborn ,Gluten intolerance ,Pathogenic bacteria ,Feeding Behavior ,Clostridium perfringens ,medicine.disease ,biology.organism_classification ,Gastrointestinal Microbiome ,030104 developmental biology ,Spain ,Immunology ,pathogenic bacteria - Abstract
8 páginas, 1 figura, 2 tablas, Celiac disease (CD) is an immune-mediated enteropathy involving genetic and environmental factors, whose interaction influences disease risk. The intestinal microbiota, including viruses and bacteria, could play a role in the pathological process leading to gluten intolerance. In this study, we investigated the prevalence of pathogens in the intestinal microbiota of infants at familial risk of developing CD. We included 127 full-term newborns with at least one first-degree relative with CD. Infants were classified according to milk-feeding practice (breastfeeding or formula feeding) and HLA-DQ genotype (low, intermediate or high genetic risk). The prevalence of pathogenic bacteria and viruses was assessed in the faeces of the infants at 7 days, 1 month and 4 months of age. The prevalence of Clostridium perfringens was higher in formula-fed infants than in breast-fed over the study period, and that of C. difficile at 4 months. Among breastfed infants, a higher prevalence of enterotoxigenic E. coli (ETEC) was found in infants with the highest genetic risk compared either to those with a low or intermediate risk. Among formula-fed infants, a higher prevalence of ETEC was also found in infants with a high genetic risk compared to those of intermediate risk. Our results show that specific factors, such as formula feeding and the HLA-DQ2 genotype, previously linked to a higher risk of developing CD, influence the presence of pathogenic bacteria differently in the intestinal microbiota in early life. Further studies are warranted to establish whether these associations are related to CD onset later in life., This work was supported by grants AGL2011-25169, AGL2014-52101-P and AGL2007-66126-C03-03/ALI (YS and FP) from the Spanish Ministry of Economy and Competitiveness (MINECO). The scholarship to MO from CSIC (JAEpre) and the contract to ABP from the European Union's Seventh Framework Program under the grant agreement no 613979 (MyNewGut) are also fully acknowledged.
- Published
- 2018
11. Modeling APC mutagenesis and familial adenomatous polyposis using human iPS cells
- Author
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Gustavo Mostoslavsky, Ignacio S. Caballero, F.J. Molina-Estevez, Andreia Gianotti-Sommer, Nicholas Skvir, Cesar Sommer, Amalia Capilla, and Sanjib Chowdhury
- Subjects
0301 basic medicine ,Cellular differentiation ,Gene Expression ,lcsh:Medicine ,medicine.disease_cause ,Loss of heterozygosity ,Animal Cells ,Medicine and Health Sciences ,Organ Cultures ,Induced pluripotent stem cell ,lcsh:Science ,Cells, Cultured ,Connective Tissue Cells ,Staining ,Mutation ,Multidisciplinary ,biology ,Stem Cells ,Gene Expression Regulation, Developmental ,Cell Staining ,Cell Differentiation ,3. Good health ,Intestines ,Organoids ,Cell Transformation, Neoplastic ,Adenomatous Polyposis Coli ,Connective Tissue ,Biological Cultures ,Cellular Types ,Anatomy ,Signal Transduction ,Research Article ,Adenomatous polyposis coli ,Immune Cells ,Adenomatous Polyposis Coli Protein ,Induced Pluripotent Stem Cells ,Immunology ,Antigen-Presenting Cells ,Mutagenesis (molecular biology technique) ,Research and Analysis Methods ,Familial adenomatous polyposis ,03 medical and health sciences ,Genetics ,medicine ,Humans ,lcsh:R ,Biology and Life Sciences ,Cell Biology ,Fibroblasts ,medicine.disease ,Gastrointestinal Tract ,HEK293 Cells ,Biological Tissue ,030104 developmental biology ,Mutagenesis ,Specimen Preparation and Treatment ,Cancer research ,biology.protein ,lcsh:Q ,Carcinogenesis ,Digestive System ,Developmental Biology - Abstract
Mutations in the gene Adenomatous Polyposis Coli or APC appear in most sporadic cases of colorectal cancer and it is the most frequent mutation causing hereditary Familial Adenomatous Polyposis. The detailed molecular mechanism by which APC mutations predispose to the development of colorectal cancer is not completely understood. This is in part due to the lack of accessibility to appropriate models that recapitulate the early events associated with APC mediated intestinal transformation. We have established a novel platform utilizing human induced Pluripotent Stem cells or iPSC from normal or FAP-specific APC mutant individuals and evaluated the effect of the mutation in the cells before and after differentiation into intestinal organoids. In order to minimize genetic background effects, we also established an isogenic platform using TALEN-mediated gene editing. Comparison of normal and APC mutant iPSC revealed a significant defect in cell identity and polarity due to the presence of APC in heterozygosity as well as chromosomal aberrations including abnormal anaphases and centrosome numbers. Importantly, upon specification into intestinal progeny, APC heterozygosity was responsible for a major change in the transcriptional identity of the cells with dysregulation of key signaling pathways, including metabolic reprogramming, abnormal lipid metabolism and intestinal-specific cadherin expression. In conclusion, we have developed a novel iPSC/intestinal model of APC mutagenesis and provide strong evidence that APC in heterozygosity imparts a clear phenotypic and molecular defect, affecting basic cellular functions and integrity, providing novel insights in the earlier events of APC-mediated tumorigenesis.
- Published
- 2018
12. Oblique scanning laser microscopy for simultaneously volumetric structural and molecular imaging using only one raster scan
- Author
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Ji Yi, Gustavo Mostoslavsky, Weiye Song, Lei Zhang, and Amalia Capilla
- Subjects
0301 basic medicine ,Fluorescence-lifetime imaging microscopy ,Materials science ,Optical sectioning ,Laser scanning ,Science ,Green Fluorescent Proteins ,01 natural sciences ,Fluorescence ,Article ,010309 optics ,Mice ,03 medical and health sciences ,Imaging, Three-Dimensional ,Optical coherence tomography ,0103 physical sciences ,Microscopy ,medicine ,Animals ,Humans ,Computer vision ,Intestinal Mucosa ,Microscopy, Confocal ,Multidisciplinary ,Fourier Analysis ,medicine.diagnostic_test ,Phantoms, Imaging ,business.industry ,Retinal Vessels ,Microspheres ,Molecular Imaging ,Rats ,Intestines ,Organoids ,Autofluorescence ,030104 developmental biology ,Medicine ,sense organs ,Artificial intelligence ,Molecular imaging ,Raster scan ,business ,Biomedical engineering - Abstract
Multi-modal three dimensional (3D) optical imaging combining both structural sensitivity and molecular specificity is highly desirable in biomedical research. In this paper, we present a method termed oblique scanning laser microscopy (OSLM) to combine optical coherence tomography (OCT), for simultaneously volumetric structural and molecular imaging with cellular resolution in all three dimensions. Conventional 3D laser scanning fluorescence microscopy requires repeated optical sectioning to create z-stacks in depth. Here, the use of an obliquely scanning laser eliminates the z-stacking process, then allows highly efficient 3D OCT and fluorescence imaging by using only one raster scan. The current setup provides ~3.6 × 4.2 × 6.5 μm resolution in fluorescence imaging, ~7 × 7 × 3.5 μm in OCT in three dimensions, and the current speed of imaging is up to 100 frames per second (fps) over a volume about 0.8 × 1 × 0.5 mm3. We demonstrate several mechanisms for molecular imaging, including intrinsically expressed GFP fluorescence, autofluorescence from Flavin proteins, and exogenous antibody-conjugated dyes. We also demonstrate potential applications in imaging human intestinal organoids (HIOs), colon mucosa, and retina.
- Published
- 2017
13. Toll pathway is required for wound-induced expression of barrier repair genes in the
- Author
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Amalia, Capilla, Dmitry, Karachentsev, Rachel A, Patterson, Anita, Hermann, Michelle T, Juarez, and William, McGinnis
- Subjects
Wound Healing ,Drosophila melanogaster ,integumentary system ,Gene Expression Regulation ,PNAS Plus ,Toll-Like Receptors ,NF-kappa B ,Animals ,Drosophila Proteins ,Models, Biological ,Signal Transduction - Abstract
After breaks in animal epidermal barriers, repair genes are activated in the cells adjacent to wound sites that help regenerate the barrier. The fruit fly Drosophila melanogaster is a favorable genetic system to find molecular detection systems that sense wounds and activate repair genes. In this paper, we find that the Toll signaling pathway, including the extracellular ligand Spätzle, the Toll receptor, and the Dif transcription factor, form a detection system to sense broken epidermis and then activate regeneration genes. The Toll pathway thus is involved not only in the activation of genes involved in fighting microbial invasion after epidermal breaks, but also in the activation of genes that regenerate epidermal barrier function.
- Published
- 2017
14. 272 - Novel Intestinal Differentiation Approach from Human Induced Pluripotent Stem Cells for Modeling Hereditary Colorectal Cancer
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Amalia Capilla, Aditya Mithal, Gustavo Mostoslavsky, and Dar Heinze
- Subjects
Hepatology ,Colorectal cancer ,Gastroenterology ,medicine ,Cancer research ,Human Induced Pluripotent Stem Cells ,Biology ,medicine.disease - Published
- 2018
15. Genetic analyses of celiac disease in a Spanish population confirm association with CELIAC3 but not with CELIAC4
- Author
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Amalia Capilla, Ester Donat, Dolores Planelles, Carmen Ribes-Koninckx, Carmen Espinós, and Francesc Palau
- Subjects
Male ,Genotype ,Immunology ,Locus (genetics) ,Human leukocyte antigen ,Myosins ,Biology ,Biochemistry ,Antigens, CD ,Genetics ,Genetic predisposition ,Humans ,Immunology and Allergy ,CTLA-4 Antigen ,Genetic Predisposition to Disease ,Allele ,Alleles ,Genetic association ,Polymorphism, Genetic ,Tumor Necrosis Factor-alpha ,Haplotype ,General Medicine ,Transmission disequilibrium test ,Antigens, Differentiation ,Celiac Disease ,Haplotypes ,Spain ,Case-Control Studies ,Female - Abstract
Genetic predisposition to celiac disease (CD) is determined primarily by the human leukocyte antigen (HLA) genes (CELIAC1 region; 6p21), although many loci are involved in disease susceptibility. First, we have analysed a large series of CD patients from the Spanish Mediterranean region who had previously been characterised for the HLA complex. We have investigated how relevant regions contribute to CD susceptibility: CELIAC3 (CD28/CTLA4/ICOS region on 2q33) and CELIAC4 (19p13) as well as the tumour necrosis factor alpha (TNF-alpha) and the linfotoxin loci by case-control and association analyses. We highlight the association with the +49*A allele of cytotoxic T-lymphocyte-associated antigen 4 locus (P = 0.01), and the -308*A of TNF-alpha locus (P = 0.0008) in DQ2 individuals, although an independent role for TNF-alpha as risk factor has not been proven. Moreover, we do not confirm the association with the CELIAC4 region polymorphisms described in other populations.
- Published
- 2007
16. The HLA-DQ2 genotype selects for early intestinal microbiota composition in infants at high risk of developing coeliac disease
- Author
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Carmen Ribes-Koninckx, Isabel Polanco, Luis Ortigosa, Esther Nova, Vicente Varea, Yolanda Sanz, Alexander Neef, Marta Olivares, G De Palma, Gemma Castillejo, Francesc Palau, L Izquierdo, Ascensión Marcos, and Amalia Capilla
- Subjects
Genetic Markers ,Male ,Genotype ,Firmicutes ,Biology ,Gut flora ,Real-Time Polymerase Chain Reaction ,Coeliac disease ,Microbiology ,Intestinal Bacteria ,Feces ,Bifidobacteria ,Risk Factors ,HLA-DQ Antigens ,medicine ,Celiac disease ,Humans ,Prospective Studies ,Bifidobacterium ,Bifidobacteria, Celiac Disease, Intestinal Bacteria ,Clostridium ,Microbiota ,Haplotype ,Gastroenterology ,HLA-DQ2 ,Infant ,medicine.disease ,biology.organism_classification ,Bifidobacteriaceae ,Intestines ,Celiac Disease ,Haplotypes ,Immunology ,Female - Abstract
Objective: Intestinal dysbiosis has been associated with coeliac disease (CD), but whether the alterations are cause or consequence of the disease is unknown. This study investigated whether the human leukocyte antigen (HLA)-DQ2 genotype is an independent factor influencing the early gut microbiota composition of healthy infants at family risk of CD. Design: As part of a larger prospective study, a subset (n=22) of exclusively breastfed and vaginally delivered infants with either high genetic risk (HLA-DQ2 carriers) or low genetic risk (non-HLA-DQ2/8 carriers) of developing CD were selected from a cohort of healthy infants with at least one first-degree relative with CD. Infant faecal microbiota was analysed by 16S rRNA gene pyrosequencing and real time quantitative PCR. Results: Infants with a high genetic risk had significantly higher proportions of Firmicutes and Proteobacteria and lower proportions of Actinobacteria compared with low-risk infants. At genus level, high-risk infants had significantly less Bifidobacterium and unclassified Bifidobacteriaceae proportions and more Corynebacterium, Gemella, Clostridium sensu stricto, unclassified Clostridiaceae, unclassified Enterobacteriaceae and Raoultella proportions. Quantitative real time PCR also revealed lower numbers of Bifidobacterium species in infants with high genetic risk than in those with low genetic risk. In high-risk infants negative correlations were identified between Bifidobacterium species and several genera of Proteobacteria (Escherichia/Shigella) and Firmicutes (Clostridium). Conclusions: The genotype of infants at family risk of developing CD, carrying the HLA-DQ2 haplotypes, influences the early gut microbiota composition. This finding suggests that a specific disease-biased host genotype may also select for the first gut colonisers and could contribute to determining disease risk.
- Published
- 2014
17. Influence of early environmental factors on peripheral lymphocyte subsets and gut microbiota in infants at risk for celiac disease
- Author
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T. Pozo Rubio, Yolanda Sanz, Amalia Capilla, M. D. Acuña, G De Palma, Ascensión Marcos, Isabel Polanco, Esther Nova, and Jorge R. Mujico
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Nutrition and Dietetics ,biology ,business.industry ,Immunology ,Medicine (miscellaneous) ,Medicine ,Disease ,Gut flora ,business ,biology.organism_classification ,Peripheral ,Lymphocyte subsets - Published
- 2013
18. Planar cell polarity controls directional Notch signaling in the Drosophila leg
- Author
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Sarah J. Bray, Maria Benavente, Maki Daniels, Máximo Ibo Galindo, Ruth I. Johnson, and Amalia Capilla
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Frizzled ,Notch ,Endocytic cycle ,Notch signaling pathway ,Dishevelled Proteins ,Biology ,Dishevelled ,Cell polarity ,Animals ,Drosophila Proteins ,Molecular Biology ,Research Articles ,Adaptor Proteins, Signal Transducing ,rab5 GTP-Binding Proteins ,chemistry.chemical_classification ,Receptors, Notch ,Planar Cell Polarity ,Cell Polarity ,Phosphoproteins ,Frizzled Receptors ,Cell biology ,chemistry ,Notch proteins ,Mutation ,Drosophila ,Signal transduction ,Drosophila Protein ,Developmental Biology ,Signal Transduction - Abstract
10 páginas, 8 figuras., The generation of functional structures during development requires tight spatial regulation of signaling pathways. Thus, in Drosophila legs, in which Notch pathway activity is required to specify joints, only cells distal to ligand-producing cells are capable of responding. Here, we show that the asymmetric distribution of planar cell polarity (PCP) proteins correlates with this spatial restriction of Notch activation. Frizzled and Dishevelled are enriched at distal sides of each cell and hence localize at the interface with ligand-expressing cells in the non-responding cells. Elimination of PCP gene function in cells proximal to ligand-expressing cells is sufficient to alleviate the repression, resulting in ectopic Notch activity and ectopic joint formation. Mutations that compromise a direct interaction between Dishevelled and Notch reduce the efficacy of repression. Likewise, increased Rab5 levels or dominant-negative Deltex can suppress the ectopic joints. Together, these results suggest that PCP coordinates the spatial activity of the Notch pathway by regulating endocytic trafficking of the receptor, This work was funded by a programme grant from the UK Medical Research Council [G0300034 to S.J.B.]; a Spanish Ministry of Science and Innovation project grant [BFU2009-07949 to M.I.G.]; and a ‘Ramón y Cajal’ fellowship [to M.I.G.]. M.B. is supported by a Spanish Research Council (CSIC) JAE-Tec contract.
- Published
- 2012
19. Influence of Milk-Feeding Type and Genetic Risk of Developing Coeliac Disease on Intestinal Microbiota of Infants: The PROFICEL Study
- Author
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Luis Ortigosa, Luis Peña-Quintana, Francesc Palau, Isabel Polanco, Carmen Ribes-Koninckx, Giada De Palma, Yolanda Sanz, C. Calvo, Amalia Capilla, José Antonio Garrote, Esther Nova, Gemma Castillejo, T. Pozo, Ana Jeremías López, Ascensión Marcos, Maria Dolores García-Novo, and Vicente Varea
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Genotype ,Science ,Population ,Immunology ,Gastroenterology and Hepatology ,Biology ,Gut flora ,Breast milk ,Microbiology ,Pediatrics ,fluids and secretions ,HLA-DQ Antigens ,Humans ,Family ,Genetic Predisposition to Disease ,education ,Bifidobacterium ,education.field_of_study ,Multidisciplinary ,Milk, Human ,Immunity ,Infant, Newborn ,Obstetrics and Gynecology ,Infant ,food and beverages ,Feeding Behavior ,biology.organism_classification ,Infant Formula ,Intestines ,Celiac Disease ,Breast Feeding ,Metagenome ,Medicine ,Clinical Immunology ,Bacteroides fragilis ,Bacteroides ,Breast feeding ,Research Article - Abstract
10 pages, 10 tables. PMID:22319588[PubMed] PMCID:PMC3272021, Interactions between environmental factors and predisposing genes could be involved in the development of coeliac disease (CD). This study has assessed whether milk-feeding type and HLA-genotype influence the intestinal microbiota composition of infants with a family history of CD. The study included 164 healthy newborns, with at least one first-degree relative with CD, classified according to their HLA-DQ genotype by PCR-SSP DQB1 and DQA1 typing. Faecal microbiota was analysed by quantitative PCR at 7 days, and at 1 and 4 months of age. Significant interactions between milk-feeding type and HLA-DQ genotype on bacterial numbers were not detected by applying a linear mixed-model analysis for repeated measures. In the whole population, breast-feeding promoted colonization of C. leptum group, B. longum and B. breve, while formula-feeding promoted that of Bacteroides fragilis group, C. coccoides-E. rectale group, E. coli and B. lactis. Moreover, increased numbers of B. fragilis group and Staphylococcus spp., and reduced numbers of Bifidobacterium spp. and B. longum were detected in infants with increased genetic risk of developing CD. Analyses within subgroups of either breast-fed or formula-fed infants indicated that in both cases increased risk of CD was associated with lower numbers of B. longum and/or Bifidobacterium spp. In addition, in breast-fed infants the increased genetic risk of developing CD was associated with increased C. leptum group numbers, while in formula-fed infants it was associated with increased Staphylococcus and B. fragilis group numbers. Overall, milk-feeding type in conjunction with HLA-DQ genotype play a role in establishing infants' gut microbiota; moreover, breast-feeding reduced the genotype-related differences in microbiota composition, which could partly explain the protective role attributed to breast milk in this disorder., This study was supported by public grants AGL2007-66126-C03-01-03/ALI and Consolider Fun-C-Food CSD2007-00063 from the Spanish Ministry of Science and Innovation. GDP was recipient of I3P scholarship from Consejo Superior de Investigaciones Científicas (CSIC), Spain
- Published
- 2012
20. Influence of breastfeeding versus formula feeding on lymphocyte subsets in infants at risk of coeliac disease: the PROFICEL study
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Amalia Capilla, Isabel Polanco, Vicente Varea, Tamara Pozo-Rubio, Jorge R. Mujico, Francesc Palau, Gemma Castillejo, Esther Nova, Yolanda Sanz, Maria Dolores García-Novo, Luis Peña-Quintana, Ascensión Marcos, Luis Ortigosa, Carmen Ribes-Koninckx, and Giada De Palma
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CD4-Positive T-Lymphocytes ,Lymphocyte subsets ,Breastfeeding ,Medicine (miscellaneous) ,Coeliac disease ,Formula feeding ,Risk Factors ,HLA-DQ Antigens ,Genotype ,medicine ,Celiac disease ,Humans ,Analysis of Variance ,Nutrition and Dietetics ,business.industry ,Infant ,medicine.disease ,Infant Formula ,Lymphocyte Subsets ,Killer Cells, Natural ,Celiac Disease ,HLA genotype ,Breast Feeding ,Infant formula ,Immunology ,Analysis of variance ,business ,Breast feeding ,Infants - Abstract
10 páginas., Purpose: In addition to genetic risk, environmental factors might influence celiac disease (CD) development. We sought to assess the effect of the interaction between milk-feeding practices and the HLA-DQ genotype, on peripheral lymphocyte subsets and their activation markers in infants at familial risk for CD. Methods: 170 newborns were classified in 3 different genetic risk groups (high risk, HR; intermediate risk, IR; and low risk, LR) after DQB1 and DQA1 typing. Lymphocyte subsets were studied at the age of 4 months by flow cytometry analysis. Results: 79 infants were receiving exclusive breast-feeding (BF), and 91 partial breast-feeding or formula-feeding (FF). Regarding genetic risk, 40 infants were classified in HR group, 75 in IR group and 55 in LR group. Two-way ANOVA did not show significant interactions between the type of milkfeeding and genetic risk group on the lymphocyte subsets analysed. One-way ANOVA for milk-feeding practice alone showed that the percentage of CD4+CD25+ cells was significantly higher in BF group than in FF group (BF, 10.92±2.71; FF, 9.94±2.96; p=0.026), and absolute counts of CD4+CD38+ cells were significantly higher in FF group than in BF group (FF, 2881.23±973.48; BF, 2557.95±977.06; p=0.038). One-way ANOVA for genetic risk alone showed that absolute counts of NK cells were significantly higher in IR group than HR and LR groups (IR, 539.24±340.63; HR, 405.01±239.53; LR, 419.86±262.85; p=0.028). Conclusion: Lymphocyte subset profiles in the early stages of life could be modulated by milk-feeding practices and genetic risk separately. Breast-feeding might have a positive immunomodulatory effect on lymphocyte subsets in infants at risk for CD., supported by grants AGL2007-66126-C03-01/ALI, AGL2007-66126-C03-02/ALI and AGL2007-66126-C03-03/ALI, from the Spanish Ministry of Science and Innovation and grants 200570F0091 and 200570F0093 from CSIC.
- Published
- 2012
21. Influence of Environmental and Genetic Factors Linked to Celiac Disease Risk on Infant Gut Colonization by Bacteroides Species
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José Antonio Garrote, Vicente Varea, T. Pozo, Amalia Capilla, Ester Sánchez, Luis Ortigosa, Carmen Ribes-Koninckx, Giada De Palma, Francesc Palau, Yolanda Sanz, C. Calvo, Esther Nova, Maria Dolores García-Novo, Ana Claudia López, Gemma Castillejo, Isabel Polanco, and Ascensión Marcos
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Physiology ,Disease ,Biology ,Environment ,Applied Microbiology and Biotechnology ,Microbial Ecology ,Eating ,Feces ,Risk Factors ,HLA-DQ Antigens ,RNA, Ribosomal, 16S ,Genotype ,medicine ,Animals ,Bacteroides ,Humans ,Enteropathy ,Colonization ,Ecology ,Infant, Newborn ,Infant ,Feeding Behavior ,medicine.disease ,biology.organism_classification ,Infant Formula ,Intestines ,Celiac Disease ,Breast Feeding ,Milk ,Infant formula ,Immunology ,Infant Food ,Breast feeding ,Food Science ,Biotechnology - Abstract
Celiac disease (CD) is an immune-mediated enteropathy involving genetic and environmental factors whose interaction might influence disease risk. The aim of this study was to determine the effects of milk-feeding practices and the HLA-DQ genotype on intestinal colonization of Bacteroides species in infants at risk of CD development. This study included 75 full-term newborns with at least one first-degree relative suffering from CD. Infants were classified according to milk-feeding practice (breast-feeding or formula feeding) and HLA-DQ genotype (high or low genetic risk). Stools were analyzed at 7 days, 1month, and 4 months by PCR and denaturing gradient gel electrophoresis (DGGE). The Bacteroides species diversity index was higher in formula-fed infants than in breast-fed infants. Breast-fed infants showed a higher prevalence of Bacteroides uniformis at 1 and 4 months of age, while formula-fed infants had a higher prevalence of B. intestinalis at all sampling times, of B. caccae at 7 days and 4 months, and of B. plebeius at 4 months. Infants with high genetic risk showed a higher prevalence of B. vulgatus, while those with low genetic risk showed a higher prevalence of B. ovatus, B. plebeius, and B. uniformis. Among breast-fed infants, the prevalence of B. uniformis was higher in those with low genetic risk than in those with high genetic risk. Among formula-fed infants, the prevalence of B. ovatus and B. plebeius was increased in those with low genetic risk, while the prevalence of B. vulgatus was higher in those with high genetic risk. The results indicate that both the type of milk feeding and the HLA-DQ genotype influence the colonization process of Bacteroides species, and possibly the disease risk. © 2011, American Society for Microbiology.
- Published
- 2011
22. Gut colonisation process of newborns and breast-fed babies at risk of developing coeliac disease
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Inmaculada Nadal, Gemma Castillejo, Amalia Capilla, Esther Nova, Vicente Varea, Ascensión Marcos, Isabel Polanco, José Antonio Garrote, Francesc Palau, C. Ribes-Coninckx, G De Palma, Ana Jeremías López, Yolanda Sanz, C. Calvo, T. Pozo, M. L. Cilleruelo, and Maria Dolores García-Novo
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Colonisation ,Pediatrics ,medicine.medical_specialty ,Nutrition and Dietetics ,business.industry ,Medicine (miscellaneous) ,Medicine ,business ,medicine.disease ,Coeliac disease - Published
- 2010
23. P0443 GENOTYPIC STUDY OF HLA-AND NON HLA ( CTLA-4 ) REGIONS IN PATIENTS WITH CELIAC DISEASE AND THEIR FIRST-DEGREE RELATIVES
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M. Planelles, Amalia Capilla, F. Palau, J. A. Montoro, Carmen Ribes-Koninckx, C. Espinos, and Ester Donat
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CTLA-4 ,business.industry ,Pediatrics, Perinatology and Child Health ,Immunology ,Genotype ,Gastroenterology ,Medicine ,In patient ,Human leukocyte antigen ,Disease ,First-degree relatives ,business - Published
- 2004
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