Biology subjects

Rodriguez-Rovira, I.

Publications and source records attributed to Rodriguez-Rovira, I..

7 recordsLinked to original sources

bioRxiv2026

Integrated luminescence and phenotypic profiling for drug discovery in a zebrafish model of Marfan syndrome

BackgroundMarfan syndrome (MFS) is a life-threatening heritable connective tissue disorder caused by pathogenic variants in fibrillin-1, characterized by progressive cardiovascular disease. Current medical therapies slow disease progression but do not prevent major complications, underscoring the need for new treatment strategies and unbiased discovery approaches. MethodsWe used a zebrafish model of MFS lacking fibrillin-3 (fbn3-/-), which recapitulates key cardiovascular phenotypes including cardiac stress, valvular defects, arrhythmia, and aortic dilation. To enable sensitive, quantitative assessment of cardiac stress, we generated a novel transgenic zebrafish reporter expressing secreted nanoluciferase under control of the stress-responsive nppb promoter. This reporter was combined with morphological phenotyping and bulbus arteriosus (BA) imaging. We evaluated standard MFS therapies, targeted modulators of TGF-{beta} signaling, and performed an unbiased high-throughput drug screen of over 1 500 clinically approved compounds across multiple developmental treatment windows. Resultsfbn3-/- larvae exhibited markedly elevated nppb activity that correlated with phenotypic severity and peaked during stages of highest mortality. The nanoluciferase reporter provided a [~]1 000-fold dynamic range, substantially outperforming Firefly luciferase-based assays. Pharmacological inhibition of TGF-{beta} signaling produced transient or deleterious effects, while {beta}-blockers, losartan, and allopurinol failed to consistently improve cardiac stress, pericardial edema, or BA dilation. The unbiased high-throughput drug screen identified a small number of primary and secondary hits; however, none demonstrated reproducible phenotypic rescue upon rigorous multi-dose, multi-time window validation. ConclusionsThis study establishes a sensitive zebrafish-based platform for early, quantitative assessment of cardiovascular stress in MFS. Our findings highlight the limited efficacy of current therapies, the context-dependent nature of TGF-{beta} modulation, and the biological complexity underlying MFS pathogenesis. Although no definitive therapeutic candidates were identified, this work lays a robust foundation for expanded unbiased discovery efforts aimed at identifying disease-modifying interventions for MFS.

pharmacology and toxicology↗

bioRxiv2025

Impact of the maternal environment on cardiovascular features of the offspring in a mouse model of Marfan syndrome

BackgroundMarfan syndrome (MFS) is a systemic disorder of the connective tissue caused by heterozygous mutations in the FBN1 gene, which encodes fibrillin 1, a glycoprotein that constitutes elastic fibers. MFS does not exhibit sexual dimorphism regarding prevalence; however, it remains unknown whether the paternal or maternal inheritance of the FBN1 mutation affects the cardiovascular pathology of the offspring. In this study, we aimed to determine the impact of the parental origin of the FBN1 mutation on the cardiovascular manifestations of the offspring using the Fbn1C1041G/+ mouse model of MFS. Methods and ResultsFour experimental groups were generated by crossing wild-type (WT) and MFS mice to obtain WT and MFS offspring from either a paternal (MFS-P) or maternal (MFS-M) MFS parent. Cardiovascular phenotyping of offspring was performed from childhood to adulthood (from one to six months of age), including echocardiography, tail-cuff plethysmography, histopathology, and canonical (pSmad2) and non-canonical (pERK) TGF-{beta} signaling activity in the aortic tissue. At one month of age, both WT and MFS offspring from MFS-M presented lower body weight than those from MFS-P. However, with age, MFS-M offspring became persistently and significantly overweight. Both MFS-P and MFS-M offspring exhibited a significantly increased aortic root diameter compared with WT offspring; however, this enlargement appeared earlier in MFS-M than in MFS-P offspring. These parental and age-related differences in aortic root diameter were accompanied by increased canonical and non-canonical TGF-{beta} signaling. The cardiac ejection fraction was reduced at early ages in both WT and MFS offspring from MFS-M compared with MFS-P, with the difference persisting only in MFS-M offspring at adulthood. Systolic blood pressure was initially lower in MFS-M offspring across both genotypes. However, it progressively increased, resulting in elevated levels in both WT and MFS offspring from MFS-M by six months of age. ConclusionOur results indicate the existence of a gestational maternal MFS environmental factor with an early impact on the aorta and heart of MFS offspring. In adulthood, this becomes normalized in the aorta but not in the heart.

pathology↗

bioRxiv2025

Allopurinol improves cardiovascular phenotypes of a mouse model for Williams-Beuren syndrome reducing redox stress

Cardiovascular disease represents the primary cause of morbidity in Williams-Beuren syndrome (WBS), a neurodevelopmental disorder resulting from a hemizygous deletion of 26-28 genes on chromosome 7q11.23. The clinical phenotype includes systemic hypertension, cardiac dysfunction, and progressive vascular remodelling, features that are further exacerbated by increased oxidative stress. Given the early onset and progression of these cardiovascular manifestations, patients often require long-term pharmacological management from childhood. This raises important concerns about cumulative drug toxicity, dosing strategies, and the long-term safety of therapeutic interventions in pediatric populations. Using the Complete deletion (CD) mouse model of WBS, this study aims to evaluate whether pediatric-equivalent doses of Allopurinol (ALO) (20mg/kg/day equivalent to 1,66mg/kg/day in humans) a highly specific xanthine oxidoreductase (XOR) inhibitor, and Losartan (LOS) (12,5 mg/kg/day equivalent to 1 mg/kg/day in humans) an angiotensin II type1 receptor antagonist, will exert beneficial effects on the cardiovascular phenotype of CD mice. To enable comparative analysis, the pharmacological effects of ALO and LOS were evaluated against both untreated CD mice and wild-type (WT) controls. In CD mice, both treatments significantly reduced systemic blood pressure; however, their effects on cardiac outcomes were different. While treatment with ALO improved cardiac pathology, treatment with LOS did not. ALO likely acts by reducing XOR protein levels, reactive oxygen species (ROS) production and NRLP3 activation. These findings support a therapeutic model in which minimal doses of ALO offers broader cardiovascular protection in WBS.

genetics↗

bioRxiv2025

Differential contribution of elastin and fibrillin-1 to the cardiovascular phenotype of a double heterozygous Marfan and Williams-Beuren syndrome mouse model

Marfan syndrome (MFS) and Williams-Beuren syndrome (WBS) are two genetic diseases of connective tissue caused respectively by mutations in the fibrillin-1 gene (FBN1) and hemizygous loss of the elastin gene (ELN) from an allelic chromosomic deletion. Their respective vascular manifestations are opposed, resulting in thoracic aortic aneurysm in MFS and supravalvular aortic stenosis in WBS. To investigate the interdependence of both essential molecular constituents of elastic fibers, we have generated a double heterozygous mouse model by crossing an MFS female (Fbn1C1041G/+) with a Complete Deletion (CD)-WBS male. We evaluated blood pressure, cardiac and aortic phenotypes, and some physical and cognitive functions in the offspring. Double heterozygous mice (CDMFS) presented the characteristic CD altered behavior. CDMFS mice developed an aneurysm, which progressed over age indistinguishably from MFS mice; CDMFS aortic wall showed a thicker tunica media aorta layer and thinner elastic fibers in accordance with CD mice. The characteristic MFS reduction in vascular smooth muscle cell density is not observed in the CDMFS aorta. CDMFS mice develop the high blood pressure observed in CD animals which does not occur in MFS. Ejection fraction was significantly reduced in MFS, CD, and CDMFS mice compared with WT littermates. Cardiac redox stress markers increased only in CD mice, but not in CDMFS animals. Metalloproteinase-2 protein levels increased only in MFS hearts and partially reduced in CDMFS ones. In conclusion, CDMFS mice exhibited the characteristic phenotypic manifestations of each syndrome, along with the pathological outcomes associated with each elastic fiber component and the respective disease.

genetics↗

bioRxiv2023

HYPERURICAEMIA DOES NOT INTERFERE WITH AORTOPATHY IN A MURINE MODEL OF MARFAN SYNDROME

Redox stress is involved in the aortic aneurysm pathogenesis in Marfan syndrome (MFS). We recently reported that allopurinol, a recognised inhibitor of XOR, inhibited aortopathy in a mouse model of MFS acting as an antioxidant. Hyperuricaemia is ambiguously associated with cardiovascular injuries as uric acid (UA), having antioxidant or pro-oxidant properties depending on the concentration and accumulation site. We aimed to evaluate whether hyperuricaemia causes harm or relief in MFS aortopathy and cardiopathy pathogenesis. Two-month-old male wild-type (WT) and MFS mice (Fbn1C1041G/+) were injected intraperitoneally for several weeks with potassium oxonate (PO), an inhibitor of uricase, an enzyme that catabolises UA to allantoin. Plasma UA and allantoin levels were measured via several techniques, aortic root diameter and cardiac parameters by ultrasound imaging, aortic wall structure by histopathology, and pNRF2 levels by immunofluorescence. PO induced a significant increase in UA in blood plasma both in WT and MFS mice, reaching a peak at three and four months of age but decaying at six months. Hyperuricaemic MFS mice showed no change in the characteristic aortic aneurysm progression or aortic wall disarray evidenced by large elastic laminae ruptures. There were no changes in cardiac parameters or the redox stress-induced nuclear translocation of pNRF2 in the aortic tunica media. Altogether, the results suggest that hyperuricaemia interferes neither with aortopathy nor cardiopathy in MFS mice.

genetics↗

bioRxiv2022

Curcumin combined with verapamil improve cardiovascular phenotype of a Williams-Beuren Syndrome mice model reducing oxidative stress

Williams-Beuren syndrome (WBS) is a rare neurodevelopmental disorder with hallmarks in the cardiovascular manifestations and no well-defined therapeutic strategies. We investigated the progression of cardiovascular phenotype present in CD (complete deletion) mice, a murine model of WBS, after chronic treatment with curcumin and verapamil, both compounds with positive effects on related pathologies. Treatment was administered orally dissolved in drinking water. After measuring "in vivo" systolic blood pressure, aortic and left ventricular myocardium were histological and molecular analysed to determine the effects of treatment and its underlying mechanism in CD mice. We observed upregulated xanthine oxidoreductase (XOR) expression in both aortic and left ventricular myocardium of CD mice. This overexpression is concomitant with increased levels of nitrated proteins as example of byproduct-mediated oxidative stress damage, indicating of XOR-generated oxidative stress impact on the pathophysiology of cardiovascular manifestations in WBS, and suggesting that the inhibition of XOR and/or oxidative stress damage could help to ameliorate the severe cardiovascular injuries presented in WBS.

genetics↗

bioRxiv2021

ALLOPURINOL BLOCKS THE FORMATION AND PROGRESSION OF AORTIC ANEURYSM IN A MOUSE MODEL OF MARFAN SYNDROME ACTING AS SCAVENGER OF REACTIVE OXYGEN SPECIES

BackgroundIncreasing evidence indicates that redox stress participates in MFS aortopathy, though its mechanistic contribution is little known. We reported elevated reactive oxygen species (ROS) formation and NADPH oxidase NOX4 upregulation in MFS patients and mouse aortae. Here we address the contribution of xanthine oxidoreductase (XOR), which catabolizes purines into uric acid and ROS in MFS aortopathy. Methods and ResultsIn aortic samples from MFS patients, XOR protein expression, revealed by immunohistochemistry, increased in both the tunicae intima and media of the dilated zone. In MFS mice (Fbn1C1041G/+), aortic XOR mRNA transcripts and enzymatic activity of the oxidase form (XO) were augmented in the aorta of 3-month-old mice but not in older animals. The administration of the XOR inhibitor allopurinol (ALO) halted the progression of aortic root aneurysm in MFS mice. ALO administrated before the onset of the aneurysm prevented its subsequent development. ALO also inhibited MFS-associated endothelial dysfunction as well as elastic fiber fragmentation, fibrotic collagen remodeling, nuclear translocation of pNRF2 and increased 3-nitrotyrosine levels all occurring in the tunica media. ALO reduced the MFS-associated large aortic production of H2O2, and NOX4 and MMP2 transcriptional overexpression. ConclusionsAllopurinol interferes in aortic aneurysm progression acting as a potent antioxidant. This study strengthens the concept that redox stress is an important determinant of aortic aneurysm formation and progression in MFS and warrants the evaluation of ALO therapy in MFS patients. HIGHLIGHTSO_LIXanthine oxidoreductase (XOR) is upregulated in the aortic aneurysm of Marfan syndrome (MFS) both in patients and young mice. C_LIO_LIAllopurinol halts the formation and progression of aortic aneurysm in MFS mice C_LIO_LIAllopurinol reduces a variety of oxidative stress-associated molecular reactions. C_LIO_LIAllopurinol prevents MFS endothelial-dependent vasodilator dysfunction. C_LIO_LIThe antioxidant action of allopurinol suggests its repositioning for pharmacological use in MFS aortopathy. C_LI GRAPHICAL ABSTRACT O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=146 SRC="FIGDIR/small/464182v5_ufig1.gif" ALT="Figure 1"> View larger version (30K): org.highwire.dtl.DTLVardef@1e6f784org.highwire.dtl.DTLVardef@84075eorg.highwire.dtl.DTLVardef@1ffcb5borg.highwire.dtl.DTLVardef@800503_HPS_FORMAT_FIGEXP M_FIG C_FIG

pathology↗