Study of the genotype-phenotype correlation in fibroblasts of patients with mutations in COQ4

Motivation: Coenzyme Q10 (CoQ10), or ubiquinone, has a crucial role in the energetic metabolism due to its redox capacity in the electron transport chain (ETC), where it shuttles electrons from complex I or II to complex III. Lack of this essential component leads to mitochondrial disorders characterized by a rare condition with a huge spectrum of different phenotypes and different genetic mutations. To date, specific genotype-phenotype correlations do not exist because the link between specific genetic defects and phenotypes is unclear. In this way, the diagnosis and treatment of this patients is so complicated. The diagnosis on time is extremely important to start the treatment in order to avoid the fulminant course of the disease with an irreversible damage and a fatal outcome. In a previous study reported in our lab it was described that CoQ10-deficient fibroblasts (independently from the etiology) showed a common transcriptomic profile. The expression of certain genes was modified in the same way, that it to say, they were always increased or decreased. COQ4 is one of the genes involved in CoQ10 biosynthesis. It has been also demonstrated that COQ4 mutations are responsible for early-onset mitochondrial diseases with heterogeneous clinical presentations and associated with CoQ10 deficiency. The aim of this work is to find a possible correlation between different COQ4 mutations, the pathological phenotype, the clinical severity of the disease and the level of markers genes expression.Methods: To achieve this goal, we have used two fibroblast cell lines as control, and four mutant fibroblast cell lines from patients with different COQ4 mutations. We have performed different genetic and biochemical assays such as analysis of the expression profile by microarray, Seahorse or flow cytometry.Results: Fibroblasts from subjects with COQ4 mutations show similar profiles between them, and at the same time, they show a different profile when we compared them to control fibroblasts. In adittion, a more differentiated profile is observed according as the severity of the symptoms increases. Taken together, these results suggest a correlation between the phenotype and the clinical severity.Conclusions: Cells with COQ4 mutations look for an adaptative biological response to face the mithocondrial damage due to the CoQ10 deficiency. And interestingly, our results suggest an association between genothype and phenotype in these patients

Neurodegeneration and astrogliosis in the entorhinal cortex in Alzheimer’s disease: Stereological layer-specific assessment and proteomic analysis

Introduction: The entorhinal cortex is among the earliest areas involved in Alzheimer’s disease. Volume reduction and neural loss in this area have been widely reported. Human entorhinal cortex atrophy is, in part, due to neural loss, but microglial and/or astroglial involvement in the different layers remains unclear. Additionally, -omic approaches in the human entorhinal cortex are scarce. Methods: Herein, stereological layer-specific and proteomic analyses were carried out in the human brain. Results: Neurodegeneration, microglial reduction, and astrogliosis have been demonstrated, and proteomic data have revealed relationships with up- (S100A6, PPP1R1B, BAG3, and PRDX6) and downregulated (GSK3B, SYN1, DLG4, and RAB3A) proteins. Namely, clusters of these proteins were related to synaptic, neuroinflammatory, and oxidative stress processes. Discussion: Differential layer involvement among neural and glial populations determined by proteinopathies and identified proteins related to neurodegeneration and astrogliosis could explain how the cortical circuitry facilitates pathological spreading within the medial temporal lobe.Introducción La corteza entorrinal se encuentra entre las primeras áreas involucradas en la enfermedad de Alzheimer. La reducción de volumen y la pérdida neural en esta área han sido ampliamente reportadas. La atrofia de la corteza entorrinal humana se debe, en parte, a la pérdida neural, pero la participación microglial y/o astroglial en las diferentes capas sigue sin estar clara. Además, los enfoques -ómicos en la corteza entorrinal humana son escasos. Métodos En este documento, se llevaron a cabo análisis proteómicos y específicos de capa estereológica en el cerebro humano. Resultados Se ha demostrado la neurodegeneración, la reducción microglial y la astrogliosis, y los datos proteómicos han revelado relaciones con proteínas reguladas al alza (S100A6, PPP1R1B, BAG3 y PRDX6) y a la baja (GSK3B, SYN1, DLG4 y RAB3A). Es decir, los grupos de estas proteínas estaban relacionados con procesos de estrés oxidativo, neuroinflamatorio y sináptico. Discusión La participación de capas diferenciales entre poblaciones neurales y gliales determinada por proteinopatías y proteínas identificadas relacionadas con la neurodegeneración y la astrogliosis podría explicar cómo el circuito cortical facilita la propagación patológica dentro del lóbulo temporal medial

Anosmin 1 interacts with the prokineticin receptor 2 in vitro indicating a molecular link between both proteins in the pathogenesis of kallmann syndrome

Sexual maturation and olfactory bulb defects found in prokineticin 2 (Pk2) and prokineticin receptor 2 (Pkr2) mutant mice resembling the phenotypic characteristics of Kallmann syndrome (KS), gave rise to the question of whether these genes would have a role in KS pathogenesis. Later, mutations in both genes were identified in patients suffering from KS. The gene responsible for the Xlinked form of KS, ANOS1, encodes the ECM protein anosmin 1. Among other functions, anosmin 1 can regulate the activity of FGFR1, encoded by one of the genes involved in the autosomal transmission of KS. Therefore, it has been proposed that anosmin 1 could interact with PKR2 to modulate its activity. We present the first evidence supporting this hypothesis and report the interaction of full-length anosmin 1 with three extracellular domains of PKR2. A truncated anosmin 1 protein comprising the first three domains of the protein interacts with the second extracellular loop of PKR2, involved in PK2 binding. Finally, last three FnIII repeats of anosmin 1 also interacted with the PKR2 domains that interacted with full-length anosmin 1. Our data represent a molecular link between two of the genes involved in KS pathogenesis.This research was supported with grants from the Gobierno de Castilla-La Mancha, Spain (PI2009/29) and Fundación Salud 2000, Spain (Merck-Serono Research Grant 2013) to PFE. VMB was a PhD student hired by Gobierno de Castilla-La Mancha (MOV2007-JI/19). PFE was a researcher hired by SESCAM, Gobierno de Castilla-La Mancha, Spain and with funds from the Spanish Ministerio de Economía, Innovación y Competitividad MINECO (ADE10-0010) granted to Fernando de Castro, Neurobiología del Desarrollo group, Hospital Nacional de Parapléjicos (Toledo, Spain).Peer reviewe