Artículos (Cirugía)

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Veratridine-sensitive Na+ channels regulate human sperm fertilization capacity
(Elsevier, 2018-03-01) Cárdenas, L.; Pinto, F. M.; Cejudo Román, Antonio; González Ravina, Cristina; Fernández Sánchez, Manuel; Pérez Hernández, Natalia; Irazusta, Jon; Subirán, Nerea; Cirugía; Gobierno Vasco; Ministerio de Economia y Competitividad; Universidad del País Vasco (UPV/EHU), España; CTS607: Salud Reproductiva de la Mujer
Aims: The sperm plasma membrane contains specific ion channels and transporters that initiate changes in Ca2+, Na+, K+ and H+ ions in the sperm cytoplasm. Ion channels are key regulators of the sperm membrane potential, cytoplasmic Ca2+ and intracellular pH (pHi), which leads to regulate motility, capacitation, acrosome reaction and other physiological processes crucial for successful fertilization. Expression of epithelial sodium channels (ENaC) and voltage-gated sodium channels (Nav) in human spermatozoa has been reported, but the role of Na+ fluxes sodium channels in the regulation of sperm cell function remains poorly understood. In this context, we aimed to analyze the physiological role of Nav channels in human sperm. Main methods: Motility and hyperactivation analysis was conducted by CASA analysis. Flow cytometry and spectrophotometry approaches were carried out to measure Capacitation, Acrosome reaction, immunohistochemistry for Tyr-residues phosporylation, [Ca2+]i levels and membrane potential. Key findings: Functional studies showed that veratridine, a voltage-gated sodium channel activator, increased sperm progressive motility without producing hyperactivation while the Nav antagonist lidocaine did induce hyperactivated motility. Veratridine increased protein tyrosine phosphorylation, an event occurring during capacitation, and its effects were inhibited in the presence of lidocaine and tetrodotoxin. Veratridine had no effect on the acrosome reaction by itself, but was able to block the progesterone-induced acrosome reaction. Moreover, veratridine caused a membrane depolarization and modified the effect of progesterone on [Ca2+]i and sperm membrane potential. Significance: Our results suggest that veratridine-sensitive Nav channels are involved on human sperm fertility acquisition regulating motility, capacitation and the progesterone-induced acrosome reaction in human sperm.
Unveiling balanced prenatal microbial colonization in amniotic fluid through an integrated culture and sequencing approach
(Springer Nature, 2026-01-09) González Rovira, María; Sáinz Bueno, José Antonio; García Díaz, Lutgardo; Martínez-Pancorbo, C.; Sánchez, J.; Gutiérrez Pozo, Gabriel; Magoutas, K.; Mesías-Pérez, A.; Mellado Durán, María Encarnación; Payne, M.; Sousa Martín, Carolina; Moreno Amador, María de Lourdes; Microbiología y Parasitología; Cirugía; Genética; Ministerio de Ciencia, Innovación y Universidades (MICIU). España; Federación de Asociaciones de Celíacos de España (FACE); Universidad de Sevilla
Background The evidence of a low-biomass microbial community in the amniotic fluid (AF) is challenging the traditional concept of a sterile womb. To clarify microbial presence and host responses, a comprehensive, multi-methodological approach is required. Methods We designed an optimized culturing strategy that maximized microorganism recovery by implementing differential centrifugation and concentration of AF samples, followed by plating onto four distinct selective media types and incubation under both stringent aerobic (up to two weeks) and prolonged anaerobic (up to four weeks) conditions, including an initial pre-enrichment step in Brain Heart Infusion (BHI) broth for low-abundance organisms. These results were combined with PacBio 16S rRNA gene sequencing, Illumina shotgun metagenomics, and antimicrobial peptides (AMP) detection. Using this approach, we characterized microbial presence in 154 AF samples across gestational stages. Data normality was assessed with the Shapiro-Wilk test, guiding the selection of both parametric and non-parametric tests, and a p-value of < 0.05 was considered statistically significant. Results We detected culturable microorganisms in 33.1% of samples, with a higher proportion in elective caesarean Sect. (55.0%) compared to amniocentesis (29.5%), suggesting increased microbial load toward term. We applied stringent contamination controls, and repeatedly recovered viable microorganisms Bacillus, Cutibacterium, Micrococcus, and Staphylococcus, with Cutibacterium acnes and Staphylococcus epidermidis common. Both sequencing methods revealed a low-biomass, low-diversity microbial community with high inter-individual variability. Notably, striking microbial discordance in diamniotic twin pregnancies, challenged intrauterine homogeneity. Higher Human Beta Defensin (HBD) -1 levels correlated with absence of culturable bacteria or microbial DNA, while levels of HBD-1, HBD-3, and LL-37 were reduced in Staphylococcus-positive samples, suggesting a dynamic interplay between specific bacteria and host defences. Conclusions Our findings indicate that viable bacteria and/or DNA can transiently access the prenatal environment microbial balance. We propose a novel perspective of a potential regulatory axis between microorganisms and AMP.
Multicompartmental prolapse: a comparative study between clinical examination and ultrasound
(Elsevier ireland LTD; Wiley, 2026-02-21) García Mejido, José Antonio; Salas‐Álvarez, Olaya; Bugatto‐Gonzalez, Fernando; Fernández Palacín, Ana; Fernández Palacín, Fernando; Sáinz Bueno, José Antonio; Cirugía; Medicina Preventiva y Salud Pública; CTS312: Análisis de la Demanda Sanitaria
Objective The accurate diagnosis of multicompartment pelvic organ prolapse (POP) is fundamental to surgical success. This study aims to compare the diagnostic performance of transperineal ultrasound against clinical examination (pelvic organ prolapse quantification, POP-Q) for the detection of compartmental defects in patients with multicompartment POP, using assessment under spinal anesthesia as the reference standard. Method A prospective randomized diagnostic accuracy study was designed, including 129 patients scheduled for multicompartment POP surgery. Patients were randomly assigned to undergo either a preoperative POP-Q two-dimensional transperineal ultrasound assessment. The reference standard for all patients was the intraoperative POP-Q assessment, conducted under spinal anesthesia immediately before surgery. Sensitivity, specificity, and likelihood ratios (LR) were calculated for each method and compartment. Results Both techniques demonstrated high sensitivity for the diagnosis of cystocele (100% vs 98.3%). However, their performance varied across the other compartments. Ultrasound showed superior specificity for uterine prolapse (73.4% vs 45.4%) and rectocele (86.3% vs 66.0%) and was particularly robust in confirming enterocele (LR+ of 10.5). In turn, clinical examination had a higher sensitivity for detecting rectocele (61.5% vs 21.4%) and was highly reliable for ruling out cystocele and uterine prolapse (LR− of 0). Conclusion Clinical examination and ultrasound are complementary in the diagnosis of prolapse. Their combined use is key to accurate surgical planning.
Effect of dietary supplementation with a highly pure and concentrated docosahexaenoic acid (DHA) supplement on human sperm function
(Elsevier, 2018-06-18) González-Ravina, Cristina; Aguirre-Lipperheide, Mercedes; Pinto, Francisco; Martín-Lozano, David; Fernández Sánchez, Manuel; Blasco, Víctor; Santamaría-López, Esther; Candenas, Luz; Cirugía; CTS607: Salud Reproductiva de la Mujer
The aim of this study was to evaluate the possible beneficial effects of diet supplementation with a highly concentrated and purified docosahexaenoic acid (DHA) formula on human sperm function. We performed a prospective, randomized, double blind, placebo-controlled intervention study. One-hundred eighty human semen samples from sixty infertile patients recruited in a private assisted reproduction center were included. All samples were examined according to World Health Organization guidelines. We analyzed macroscopic and microscopic sperm parameters, oxidative stress, apoptosis, lipid peroxidation, mitochondrial membrane potential and DNA fragmentation before and after supplementation with different DHA daily doses (0.5, 1 and 2 g) or placebo for 1 and 3 months. No differences were found in traditional sperm parameters except for progressive sperm motility, with a significant increase after DHA ingestion after the first month with 1 or 2 g doses and after 3 months with 0.5 g of DHA. This effect was more evident in asthenozoospermic patients. No differences were found in any molecular semen parameter except oxidative stress, in which a slight benefit was observed after DHA treatment. In conclusion, this study support previous indications that highlight the importance of DHA supplementation as a means of improving sperm quality in asthenozoospermic men.
Ovarian response to recombinant human follicle-stimulating hormone: a randomized, antimüllerian hormone–stratified, dose–response trial in women undergoing in vitro fertilization/intracytoplasmic sperm injection
(Elsevier, 2014-12) Arce, Joan-Carles; Nyboe Andersen, Anders; Fernández Sánchez, Manuel; Visnova, Hana; Bosch, Ernesto; García Velasco, Juan Antonio; Barri, Pedro; De Sutter, Petra; Klein, Bjarke M.; Fauser, Bart C.J.M; Cirugía; ; CTS607: Salud Reproductiva de la Mujer
Objective: To evaluate the dose–response relationship of a novel recombinant human FSH (rhFSH; FE 999049) with respect to ovarian response in patients undergoing IVF/intracytoplasmic sperm injection treatment; and prospectively study the influence of initial antimüllerian hormone (AMH) concentrations. Design: Randomized, controlled, assessor-blinded, AMH-stratified (low: 5.0–14.9 pmol/L [0.7–<2.1 ng/mL]; high: 15.0–44.9 pmol/L [2.1–6.3 ng/mL]) trial. Setting: Seven infertility centers in four countries. Patient(s): Two hundred sixty-five women aged ≤37 years. Intervention(s): Controlled ovarian stimulation with either 5.2, 6.9, 8.6, 10.3, or 12.1 μg of rhFSH, or 11 μg (150 IU) of follitropin alfa in a GnRH antagonist cycle. Main Outcome Measure(s): Number of oocytes retrieved. Result(s): The number of oocytes retrieved increased in an rhFSH dose–dependent manner, from 5.2 ± 3.3 oocytes with 5.2 μg/d to 12.2 ± 5.9 with 12.1 μg/d. The slopes of the rhFSH dose–response curves differed significantly between the two AMH strata, demonstrating that a 10% increase in dose resulted in 0.5 (95% confidence interval 0.2–0.7) and 1.0 (95% confidence interval 0.7–1.3) more oocytes in the low and high AMH stratum, respectively. Fertilization rate and blastocyst/oocyte ratio decreased significantly with increasing rhFSH doses in both AMH strata. No linear relationship was observed between rhFSH dose and number of blastocysts overall or by AMH strata. Five cases of ovarian hyperstimulation syndrome were reported for the three highest rhFSH doses and in the high AMH stratum. Conclusion(s): Increasing rhFSH doses results in a linear increase in number of oocytes retrieved in an AMH-dependent manner. The availability of blastocysts is less influenced by the rhFSH dose and AMH level.
Follicular and endocrine dose responses according to anti-mullerian hormone levels in IVF patients treated with a novel human recombinant FSH (FE 999049)
(Wiley-Blackwell Publishing, Inc., 2015-09-03) Bosch, Ernesto; Nyboe Andersen, Anders; Barri, Pedro; García-Velasco, Juan Antonio; De Sutter, Petra; Fernández Sánchez, Manuel; Arce, Joan Carles; Cirugía; CTS607: Salud Reproductiva de la Mujer
Objective: To study the association between serum anti-Müllerian hormone (AMH) levels and follicular development and endocrine responses induced by increasing doses (5·2-12·1 μg/day) of a novel recombinant human FSH (rhFSH, FE 999049) in patients undergoing in vitro fertilization (IVF)/intracytoplasmic sperm injection (ICSI) in a GnRH antagonist protocol. Design: Secondary analysis of a randomized controlled trial with stratified randomization according to AMH (lower stratum: 5·0-14·9 pmol/l; higher stratum: 15·0-44·9 pmol/l). Patients: Infertile women of good prognosis (n = 265). Measurements: Follicular development and endocrine parameters during controlled ovarian stimulation (COS) with rhFSH. Results: Serum FSH levels increased with increasing rhFSH doses and steady-state levels for each dose were similar in both AMH strata. In the whole study population, significant (P < 0·001) positive dose responses were observed for the number of follicles ≥ 12 mm, and serum levels of oestradiol, inhibin B, inhibin A and progesterone at end of stimulation. In comparison with the higher AMH stratum, patients in the lower AMH stratum had significantly different slopes of the dose-response curves for these hormones, and no clear dose-related increase was observed for the number of follicles in these patients. Conclusions: Dose-response relationships between rhFSH and follicular development and endocrine parameters are significantly different for IVF/ICSI patients with lower and higher serum AMH levels at start of COS.
The voltage-gated sodium channel Na(v)1.8 is expressed in human sperm
(Public library science (PLoS), 2013-09-13) Cejudo-Roman, Antonio; Pinto, Francisco M.; Subirán, Nerea; Ravina, Cristina G.; Fernández Sánchez, Manuel; Perez Hernandez, Natalia; Candenas, Luz; Cirugía; Unión Europea; Junta de Andalucía; Ministerio de Economía y Competitividad, España; CTS607: Salud Reproductiva de la Mujer
The role of Na(+) fluxes through voltage-gated sodium channels in the regulation of sperm cell function remains poorly understood. Previously, we reported that several genes encoding voltage-gated Na(+) channels were expressed in human testis and mature spermatozoa. In this study, we analyzed the presence and function of the TTX-resistant VGSC α subunit Nav1.8 in human capacitated sperm cells. Using an RT-PCR assay, we found that the mRNA of the gene SCN10A, that encode Na v1.8, was abundantly and specifically expressed in human testis and ejaculated spermatozoa. The Na v1.8 protein was detected in capacitated sperm cells using three different specific antibodies against this channel. Positive immunoreactivity was mainly located in the neck and the principal piece of the flagellum. The presence of Na v1.8 in sperm cells was confirmed by Western blot. Functional studies demonstrated that the increases in progressive motility produced by veratridine, a voltage-gated sodium channel activator, were reduced in sperm cells preincubated with TTX (10 μM), the Na v1.8 antagonist A-803467, or a specific Na v1.8 antibody. Veratridine elicited similar percentage increases in progressive motility in sperm cells maintained in Ca(2+)-containing or Ca(2+)-free solution and did not induce hyperactivation or the acrosome reaction. Veratridine caused a rise in sperm intracellular Na(+), [Na(+)]i, and the sustained phase of the response was inhibited in the presence of A-803467. These results verify that the Na(+) channel Na v1.8 is present in human sperm cells and demonstrate that this channel participates in the regulation of sperm function.
Three novel and the common Arg677Ter RP1 protein truncating mutations causing autosomal dominant retinitis pigmentosa in a Spanish population
(Springer Nature, 2006-04-05) Gamundi, María José; Imma, Hernán; Martínez-Gimeno, María; Maseras, Miquel; García-Sandoval, Blanca; Ayuso, Carmen; Antiñolo Gil, Guillermo; Baiget, Montserrat; Carballo, Miguel; Cirugía; Fondo de Investigaciones Sanitarias (FIS); Fundación ONCE
Background: Retinitis pigmentosa (RP), a clinically and genetically heterogeneous group of retinal degeneration disorders affecting the photoreceptor cells, is one of the leading causes of genetic blindness. Mutations in the photoreceptor-specific gene RP1 account for 3–10% of cases of autosomal dominant RP (adRP). Most of these mutations are clustered in a 500 bp region of exon 4 of RP1. Methods: Denaturing gradient gel electrophoresis (DGGE) analysis and direct genomic sequencing were used to evaluate the 5' coding region of exon 4 of the RP1 gene for mutations in 150 unrelated index adRP patients. Ophthalmic and electrophysiological examination of RP patients and relatives according to pre-existing protocols were carried out. Results: Three novel disease-causing mutations in RP1 were detected: Q686X, K705fsX712 and K722fsX737, predicting truncated proteins. One novel missense mutation, Thr752Met, was detected in one family but the mutation does not co-segregate in the family, thereby excluding this amino acid variation in the protein as a cause of the disease. We found the Arg677Ter mutation, previously reported in other populations, in two independent families, confirming that this mutation is also present in a Spanish population. Conclusion: Most of the mutations reported in the RP1 gene associated with adRP are expected to encode mutant truncated proteins that are approximately one third or half of the size of wild type protein. Patients with mutations in RP1 showed mild RP with variability in phenotype severity. We also observed several cases of non-penetrant mutations.
A novel study of Copy Number Variations in Hirschsprung disease using the Multiple Ligation-dependent Probe Amplification (MLPA) technique
(Springer Nature, 2009-11-19) Núñez-Torres, Rocío; Fernández, Raquel M.; López Alonso, Manuel; Antiñolo Gil, Guillermo; Borrego, Salud; Cirugía; Junta de Andalucía; Fondo de Investigacion Sanitaria, España
Background: Hirschsprung disease (HSCR) is a congenital malformation of the hindgut produced by a disruption in neural crest cell migration during embryonic development. HSCR has a complex genetic etiology and mutations in several genes, mainly the RET proto-oncogene, have been related to the disease. There is a clear predominance of missense/nonsense mutations in these genes whereas copy number variations (CNVs) have been seldom described, probably due to the limitations of conventional techniques usually employed for mutational analysis. Methods: In this study we have aimed to analyze the presence of CNVs in some HSCR genes (RET, EDN3, GDNF and ZFHX1B) using the Multiple Ligation-dependent Probe Amplification (MLPA) approach. Results: Two alterations in the MLPA profiles of RET and EDN3 were detected, but a detailed inspection showed that the decrease in the corresponding dosages were due to point mutations affecting the hybridization probes regions. Conclusion: Our results indicate that CNVs of the gene coding regions analyzed here are not a common molecular cause of Hirschsprung disease. However, further studies are required to determine the presence of CNVs affecting non-coding regulatory regions, as well as other candidate genes.
Novel MLPA procedure using self-designed probes allows comprehensive analysis for CNVs of the genes involved in Hirschsprung disease
(Springer Nature, 2010-05-11) Sánchez-Mejías, Avencia; Núñez-Torres, Rocío; Fernández, Raquel M.; Antiñolo Gil, Guillermo; Borrego, Salud; Cirugía; Junta de Andalucía; Fondo de Investigacion Sanitaria, España
Background: Hirschsprung disease is characterized by the absence of intramural ganglion cells in the enteric plexuses, due to a fail during enteric nervous system formation. Hirschsprung has a complex genetic aetiology and mutations in several genes have been related to the disease. There is a clear predominance of missense/nonsense mutations in these genes whereas copy number variations (CNVs) have been seldom described, probably due to the limitations of conventional techniques usually employed for mutational analysis. In this study, we have looked for CNVs in some of the genes related to Hirschsprung (EDNRB, GFRA1, NRTN and PHOX2B) using the Multiple Ligation-dependent Probe Amplification (MLPA) approach. Methods: CNVs screening was performed in 208 HSCR patients using a self-designed set of MLPA probes, covering the coding region of those genes. Results: A deletion comprising the first 4 exons in GFRA1 gene was detected in 2 sporadic HSCR patients and in silico approaches have shown that the critical translation initiation signal in the mutant gene was abolished. In this study, we have been able to validate the reliability of this technique for CNVs screening in HSCR. Conclusions: The implemented MLPA based technique presented here allows CNV analysis of genes involved in HSCR that have not been not previously evaluated. Our results indicate that CNVs could be implicated in the pathogenesis of HSCR, although they seem to be an uncommon molecular cause of HSCR.
Novel association of severe neonatal encephalopathy and Hirschsprung disease in a male with a duplication at the Xq28 region
(Springer Nature, 2010-09-22) Fernández, Raquel M.; Núñez-Torres, Rocío; González-Meneses López, Juan; Antiñolo Gil, Guillermo; Borrego, Salud; Cirugía; Junta de Andalucía; Fondo de Investigacion Sanitaria, España
Background: Hirschsprung disease (HSCR) is a neurocristopathy characterized by the absence of parasympathetic intrinsic ganglion cells in the submucosal and myenteric plexuses along a variable portion of the intestinal tract. In approximately 18% of the cases HSCR also presents with multiple congenital anomalies including recognized syndromes. Methods: A combination of MLPA and microarray data analysis have been undertaken to refine a duplication at the Xq28 region. Results: In this study we present a new clinical association of severe neonatal encephalopathy (Lubs syndrome) and HSCR, in a male patient carrying a duplication at the Xq28 region which encompasses the MECP2 and L1CAM genes. Conclusions: While the encephalopathy has been traditionally attributed to the MECP2 gene duplication in patients with Lubs syndrome, here we propose that the enteric phenotype in our patient might be due to the dosage variation of the L1CAM protein, together with additional molecular events not identified yet. This would be in agreement with the hypothesis previously forwarded that mutations in L1CAM may be involved in HSCR development in association with a predisposing genetic background.
Comprehensive analysis of RET common and rare variants in a series of Spanish Hirschsprung patients confirms a synergistic effect of both kinds of events
(Springer Nature, 2011) Núñez-Torres, Rocío; Fernández, Raquel M.; Acosta, Manuel Jesús; Enguix Riego, María del Valle; Marbá, Martina; Agustín, Juan Carlos de; Castaño, Luis; Antiñolo Gil, Guillermo; Borrego, Salud; Cirugía; Junta de Andalucía; Fondo de Investigacion Sanitaria, España
Background: RET is the major gene associated to Hirschsprung disease (HSCR) with differential contributions of its rare and common, coding and noncoding mutations to the multifactorial nature of this pathology. In the present study, we have performed a comprehensive study of our HSCR series evaluating the involvement of both RET rare variants (RVs) and common variants (CVs) in the context of the disease. Methods: RET mutational screening was performed by dHPLC and direct sequencing for the identification of RVs. In addition Taqman technology was applied for the genotyping of 3 RET CVs previously associated to HSCR, including a variant lying in an enhancer domain within RET intron 1 (rs2435357). Statistical analyses were performed using the SPSS v.17.0 to analyze the distribution of the variants. Results: Our results confirm the strongest association to HSCR for the "enhancer" variant, and demonstrate a significantly higher impact of it in male versus female patients. Integration of the RET RVs and CVs analysis showed that in 91.66% of cases with both kinds of mutational events, the enhancer allele is in trans with the allele bearing the RET RV. Conclusions: A gender effect exists on both the transmission and distribution of rare coding and common HSCR causing mutations. In addition, these RET CVs and RVs seem to act in a synergistic way leading to HSCR phenotype.
Comprehensive Analysis of NRG1 Common and Rare Variants in Hirschsprung Patients
(PLOS, 2012-05-04) Luzón Toro, B.; Torroglosa, Ana; Núñez-Torres, Rocío; Enguix Riego, María del Valle; Fernández, Raquel María; Agustín, Juan Carlos de; Antiñolo Gil, Guillermo; Borrego, Salud; Cirugía; Instituto de Salud Carlos III; Junta de Andalucía; Ministerio de Innovación, Ciencia y Empresa. España
Hirschsprung disease (HSCR, OMIM 142623) is a developmental disorder characterized by the absence of ganglion cells along variable lengths of the distal gastrointestinal tract, which results in tonic contraction of the aganglionic gut segment and functional intestinal obstruction. The RET proto-oncogene is the major gene for HSCR with differential contributions of its rare and common, coding and noncoding mutations to the multifactorial nature of this pathology. Many other genes have been described to be associated with the pathology, as NRG1 gene (8p12), encoding neuregulin 1, which is implicated in the development of the enteric nervous system (ENS), and seems to contribute by both common and rare variants. Here we present the results of a comprehensive analysis of the NRG1 gene in the context of the disease in a series of 207 Spanish HSCR patients, by both mutational screening of its coding sequence and evaluation of 3 common tag SNPs as low penetrance susceptibility factors, finding some potentially damaging variants which we have functionally characterized. All of them were found to be associated with a significant reduction of the normal NRG1 protein levels. The fact that those mutations analyzed alter NRG1 protein would suggest that they would be related with HSCR disease not only in Chinese but also in a Caucasian population, which reinforces the implication of NRG1 gene in this pathology.
Subretinal Transplant of Induced Pluripotent Stem Cell-Derived Retinal Pigment Epithelium on Nanostructured Fibrin-Agarose
(SAGE Publications, 2019-11-04) García Delgado, Ana Belén; De la Cerda, Berta; Alba Amador, Julia; Valdés Sánchez, Maria Lourdes; Fernández Muñoz, Beatriz; Relimpio López, Isabel; Rodríguez de la Rúa Franch, Enrique; Díaz Corrales, Francisco J.; Farmacia y Tecnología Farmacéutica; Cirugía; Junta de Andalucía; European Commission (EC). Fondo Europeo de Desarrollo Regional (FEDER)
Damage to the retinal pigment epithelium (RPE) in age‐related macular degeneration (AMD) and other diseases results in photoreceptor cell death and blindness. Replacement of RPE is therefore being explored as a therapy for several retinal diseases. To move towards a future personalized autologous transplant approach, we have prepared a biocompatible implant using RPE derived from induced pluripotent stem cells (iPSC) reprogrammed from a healthy donor´s monocytes. The correct positioning of the polarized RPE is essential to fulfill its role and protect photoreceptors from degeneration. Hence, we have used a biocompatible hydrogel matrix of fibrin and agarose (FAH) that allows the surgical placement of an RPE sheet in the subretinal space, keeping its functional orientation. Our aim was to demonstrate safety and viability of the transplant in preclinical models. Pigs were used to test the feasibility of a regular vitreo‐retinal surgery. Our results show that this implant is suitable for subretinal transplantation allowing human RPE cells to survive and maintain their phenotype and orientation without any local or systemic adverse events. The ability to transplant the iPSC‐derived RPE sheet in its natural orientation will surely increase the chance to obtain a therapeutic effect in future translational studies.
Contributions of PHOX2B in the Pathogenesis of Hirschsprung Disease
(PLOS, 2013-01) Fernández, Raquel María; Mathieu, Yves; Luzón-Toro, Berta; Núñez-Torres, Rocío; González-Meneses, Antonio; Antiñolo Gil, Guillermo; Amiel, Jeanne; Borrego, Salud; Cirugía
Hirschsprung disease (HSCR) is a congenital malformation of the hindgut resulting from a disruption of neural crest cell migration during embryonic development. It has a complex genetic aetiology with several genes involved in its pathogenesis. PHOX2B plays a key function in the development of neural crest derivatives, and heterozygous mutations cause a complex dysautonomia associating HSCR, Congenital Central Hypoventilation Syndrome (CCHS) and neuroblastoma (NB) in various combinations. In order to determine the role of PHOX2B in isolated HSCR, we performed a mutational screening in a cohort of 207 Spanish HSCR patients. Our most relevant finding has been the identification of a de novo and novel deletion (c.393_410del18) in a patient with HSCR. Results of in silico and functional assays support its pathogenic effect related to HSCR. Therefore our results support that PHOX2B loss-of-function is a rare cause of HSCR phenotype.
The role of the interactome in the maintenance of deleterious variability in human populations
(Springer Nature, 2014) García-Alonso, Luz; Jiménez-Almazán, Jorge; Carbonell-Caballero, José; Vela-Boza, Alicia; Santoyo-López, Javier; Antiñolo Gil, Guillermo; Dopazo, Joaquín; Cirugía; Bull through the Bull Chair in Computational Genomics; Conselleria de Educación, Cultura y Universidades. Generalitat Valenciana; CIBER de Enfermedades Raras (CIBERER); Instituto de Salud Carlos III; National Institute of Bioinformatics; Ministerio de Economía y Competitividad (MINECO). España
Recent genomic projects have revealed the existence of an unexpectedly large amount of deleterious variability in the human genome. Several hypotheses have been proposed to explain such an apparently high mutational load. However, the mechanisms by which deleterious mutations in some genes cause a pathological effect but are apparently innocuous in other genes remain largely unknown. This study searched for deleterious variants in the 1,000 genomes populations, as well as in a newly sequenced population of 252 healthy Spanish individuals. In addition, variants causative of monogenic diseases and somatic variants from 41 chronic lymphocytic leukaemia patients were analysed. The deleterious variants found were analysed in the context of the interactome to understand the role of network topology in the maintenance of the observed mutational load. Our results suggest that one of the mechanisms whereby the effect of these deleterious variants on the phenotype is suppressed could be related to the configuration of the protein interaction network. Most of the deleterious variants observed in healthy individuals are concentrated in peripheral regions of the interactome, in combinations that preserve their connectivity, and have a marginal effect on interactome integrity. On the contrary, likely pathogenic cancer somatic deleterious variants tend to occur in internal regions of the interactome, often with associated structural consequences. Finally, variants causative of monogenic diseases seem to occupy an intermediate position. Our observations suggest that the real pathological potential of a variant might be more a systems property rather than an intrinsic property of individual proteins.
Integrated analysis of whole-exome sequencing and transcriptome profiling in males with autism spectrum disorders
(Springer Nature, 2015) Codina-Solà, Marta; Rodríguez-Santiago, Benjamín; Homs, Aïda; Santoyo, Javier; Rigau, Maria; Aznar-Laín, Gemma; Antiñolo Gil, Guillermo; Cuscó, Ivon; Cirugía; Programa Nacional de Proyectos de investigación Aplicada I + D + i; Subprograma de actuaciones Científicas y Tecnológicas en Parques Científicos y Tecnológicos (ACTEPARQ); FEDER; Fundación Alicia Koplowitz; Generalitat de Catalunya; Ministerio de Sanidad. España
Background: Autism spectrum disorders (ASD) are a group of neurodevelopmental disorders with high heritability. Recent findings support a highly heterogeneous and complex genetic etiology including rare de novo and inherited mutations or chromosomal rearrangements as well as double or multiple hits. Methods: We performed whole-exome sequencing (WES) and blood cell transcriptome by RNAseq in a subset of male patients with idiopathic ASD (n = 36) in order to identify causative genes, transcriptomic alterations, and susceptibility variants. Results: We detected likely monogenic causes in seven cases: five de novo (SCN2A, MED13L, KCNV1, CUL3, and PTEN) and two inherited X-linked variants (MAOA and CDKL5). Transcriptomic analyses allowed the identification of intronic causative mutations missed by the usual filtering of WES and revealed functional consequences of some rare mutations. These included aberrant transcripts (PTEN, POLR3C), deregulated expression in 1.7% of mutated genes (that is, SEMA6B, MECP2, ANK3, CREBBP), allele-specific expression (FUS, MTOR, TAF1C), and non-sense-mediated decay (RIT1, ALG9). The analysis of rare inherited variants showed enrichment in relevant pathways such as the PI3K-Akt signaling and the axon guidance. Conclusions: Integrative analysis of WES and blood RNAseq data has proven to be an efficient strategy to identify likely monogenic forms of ASD (19% in our cohort), as well as additional rare inherited mutations that can contribute to ASD risk in a multifactorial manner. Blood transcriptomic data, besides validating 88% of expressed variants, allowed the identification of missed intronic mutations and revealed functional correlations of genetic variants, including changes in splicing, expression levels, and allelic expression.
Identification of epistatic interactions through genome-wide association studies in sporadic medullary and juvenile papillary thyroid carcinomas
(Springer Nature, 2015) Luzón-Toro, Berta; Bleda, Marta; Navarro, Elena; García-Alonso, Luz; Ruiz Ferrer, Macarena; Medina, Ignacio; Martín Sánchez, Marta; Antiñolo Gil, Guillermo; Borrego, Salud; Cirugía; CIBERER; European Regional Development Funds (ERDF); la Marato Foundation TV3; Regional Ministry of Education of the Valencia Community; Junta de Andalucía
Background: The molecular mechanisms leading to sporadic medullary thyroid carcinoma (sMTC) and juvenile papillary thyroid carcinoma (PTC), two rare tumours of the thyroid gland, remain poorly understood. Genetic studies on thyroid carcinomas have been conducted, although just a few loci have been systematically associated. Given the difficulties to obtain single-loci associations, this work expands its scope to the study of epistatic interactions that could help to understand the genetic architecture of complex diseases and explain new heritable components of genetic risk. Methods: We carried out the first screening for epistasis by Multifactor-Dimensionality Reduction (MDR) in genome-wide association study (GWAS) on sMTC and juvenile PTC, to identify the potential simultaneous involvement of pairs of variants in the disease. Results: We have identified two significant epistatic gene interactions in sMTC (CHFR-AC016582.2 and C8orf37-RNU1-55P) and three in juvenile PTC (RP11-648k4.2-DIO1, RP11-648k4.2-DMGDH and RP11-648k4.2-LOXL1). Interestingly, each interacting gene pair included a non-coding RNA, providing thus support to the relevance that these elements are increasingly gaining to explain carcinoma development and progression. Conclusions: Overall, this study contributes to the understanding of the genetic basis of thyroid carcinoma susceptibility in two different case scenarios such as sMTC and juvenile PTC.
Exome sequencing reveals a high genetic heterogeneity on familial Hirschsprung disease
(Springer Nature, 2015) Luzón-Toro, Berta; Gui, Hongsheng; Ruiz Ferrer, Macarena; Sze-Man Tang, Clara; Fernández, Raquel M.; Sham, Pak-Chung; Antiñolo Gil, Guillermo; Borrego, Salud; Cirugía
Hirschsprung disease (HSCR; OMIM 142623) is a developmental disorder characterized by aganglionosis along variable lengths of the distal gastrointestinal tract, which results in intestinal obstruction. Interactions among known HSCR genes and/or unknown disease susceptibility loci lead to variable severity of phenotype. Neither linkage nor genome-wide association studies have efficiently contributed to completely dissect the genetic pathways underlying this complex genetic disorder. We have performed whole exome sequencing of 16 HSCR patients from 8 unrelated families with SOLID platform. Variants shared by affected relatives were validated by Sanger sequencing. We searched for genes recurrently mutated across families. Only variations in the FAT3 gene were significantly enriched in five families. Within-family analysis identified compound heterozygotes for AHNAK and several genes (N = 23) with heterozygous variants that co-segregated with the phenotype. Network and pathway analyses facilitated the discovery of polygenic inheritance involving FAT3, HSCR known genes and their gene partners. Altogether, our approach has facilitated the detection of more than one damaging variant in biologically plausible genes that could jointly contribute to the phenotype. Our data may contribute to the understanding of the complex interactions that occur during enteric nervous system development and the etiopathology of familial HSCR.
Author Correction: Genetic landscape of 6089 inherited retinal dystrophies affected cases in Spain and their therapeutic and extended epidemiological implications
(Springer Nature, 2021) Perea-Romero, Irene; Gordo, Gema; Iancu, Ionut F.; Del Pozo-Valero, Marta; Almoguera, Berta; Blanco-Kelly, Fiona; Antiñolo Gil, Guillermo; Ayuso, Carmen; Cirugía
Correction to: Scientific Reports https://doi.org/10.1038/s41598-021-81093-y, published online 15 January 2021.

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