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- Ear wound regeneration in the African spiny mouse Acomys cahirinusPublication . Santos Matias, Dino; Rita, Ana Martins; Casanellas, Ignasi; Ova, Adelia Brito; Araújo, Inês; Power, Deborah; Tiscornia, GustavoWhile regeneration occurs in a number of taxonomic groups across the Metazoa, there are very few reports of regeneration in mammals, which generally respond to wounding with fibrotic scarring rather than regeneration. A recent report described skin shedding, skin regeneration and extensive ear punch closure in two rodent species, Acomys kempi and Acomys percivali. We examined these striking results by testing the capacity for regeneration of a third species, Acomys cahirinus, and found a remarkable capacity to repair full thickness circular punches in the ear pinna. Four-millimeter-diameter wounds closed completely in 2 months in 100% of ear punches tested. Histology showed extensive formation of elastic cartilage, adipose tissue, dermis, epidermis and abundant hair follicles in the repaired region. Furthermore, we demonstrated abundant angiogenesis and unequivocal presence of both muscle and nerve fibers in the reconstituted region; in contrast, similar wounds in C57BL/6 mice simply healed the borders of the cut by fibrotic scarring. Our results confirm the regenerative capabilities of Acomys, and suggest this model merits further attention.
- Coenzyme Q10 partially restores pathological alterations in a macrophage model of Gaucher diseasePublication . Mata, Mario de la; Cotán, David; Oropesa-Ávila, Manuel; Villanueva-Paz, Marina; Lavera, Isabel de; Álvarez-Córdoba, Mónica; Luzón-Hidalgo, Raquel; Suárez-Rivero, Juan M.; Tiscornia, Gustavo; Sánchez-Alcázar, José A.Background Gaucher disease (GD) is caused by mutations in the GBA1 gene which encodes lysosomal β-glucocerebrosidase (GCase). In GD, partial or complete loss of GCase activity causes the accumulation of the glycolipids glucosylceramide (GlcCer) and glucosylsphingosine in the lysosomes of macrophages. In this manuscript, we investigated the effects of glycolipids accumulation on lysosomal and mitochondrial function, inflammasome activation and efferocytosis capacity in a THP-1 macrophage model of Gaucher disease. In addition, the beneficial effects of coenzyme Q10 (CoQ) supplementation on cellular alterations were evaluated. Chemically-induced Gaucher macrophages were developed by differentiateing THP-1 monocytes to macrophages by treatment with phorbol 12-myristate 13-acetate (PMA) and then inhibiting intracellular GCase with conduritol B-epoxide (CBE), a specific irreversible inhibitor of GCase activity, and supplementing the medium with exogenous GlcCer. This cell model accumulated up to 16-fold more GlcCer compared with control THP-1 cells. Results Chemically-induced Gaucher macrophages showed impaired autophagy flux associated with mitochondrial dysfunction and increased oxidative stress, inflammasome activation and impaired efferocytosis. All abnormalities were partially restored by supplementation with CoQ. Conclusion These data suggest that targeting mitochondria function and oxidative stress by CoQ can ameliorate the pathological phenotype of Gaucher cells. Chemically-induced Gaucher macrophages provide cellular models that can be used to investigate disease pathogenesis and explore new therapeutics for GD.
- Coenzyme Q(10) partially restores pathological alterations in a macrophage model of Gaucher diseasePublication . de la Mata, Mario; Cotan, David; Oropesa-Avila, Manuel; Villanueva-Paz, Marina; de lavera, Isabel; Alvarez-Cordoba, Monica; Luzón-Hidalgo, Raquel; Suarez-Rivero, Juan M.; Tiscornia, Gustavo; Sanchez-Alcazar, José A.Background: Gaucher disease (GD) is caused by mutations in the GBA1 gene which encodes lysosomal beta-glucocerebrosidase (GCase). In GD, partial or complete loss of GCase activity causes the accumulation of the glycolipids glucosylceramide (GlcCer) and glucosylsphingosine in the lysosomes of macrophages. In this manuscript, we investigated the effects of glycolipids accumulation on lysosomal and mitochondrial function, inflammasome activation and efferocytosis capacity in a THP-1 macrophage model of Gaucher disease. In addition, the beneficial effects of coenzyme Q(10) (CoQ) supplementation on cellular alterations were evaluated. Chemically-induced Gaucher macrophages were developed by differentiateing THP-1 monocytes to macrophages by treatment with phorbol 12-myristate 13-acetate (PMA) and then inhibiting intracellular GCase with conduritol B-epoxide (CBE), a specific irreversible inhibitor of GCase activity, and supplementing the medium with exogenous GlcCer. This cell model accumulated up to 16 fold more GlcCer compared with control THP-1 cells. Results: Chemically-induced Gaucher macrophages showed impaired autophagy flux associated with mitochondrial dysfunction and increased oxidative stress, inflammasome activation and impaired efferocytosis. All abnormalities were partially restored by supplementation with CoQ. Conclusion: These data suggest that targeting mitochondria function and oxidative stress by CoQ can ameliorate the pathological phenotype of Gaucher cells. Chemically-induced Gaucher macrophages provide cellular models that can be used to investigate disease pathogenesis and explore new therapeutics for GD.
- Coronal brain atlas in stereotaxic coordinates of the African spiny mouse, acomys cahirinusPublication . Vitorino, Marta; S, Simão; Moreira, João B.; Nogueira‐Rodrigues, Joana; Silva, Joana A; Sofia Lourenço, Ana; Vitor Fernandes, Dr; Sousa, Monica M.; Tiscornia, Gustavo; Araújo, Inês MariaThe African spiny mouse (Acomys cahirinus) is an emerging model of mammalian epimorphic regeneration that has aroused the interest of the scientific community in the last decade. To date, studies on brain repair have been hindered by the lack of knowledge on the neuroanatomy of this species. Here, we present a coronal brain atlas in stereotaxic coordinates, which allows for three-dimensional identification and localization of the brain structures of this species. The brain of 12-week-old spiny mice was mapped in stereotaxic coordinates using cresyl violet-stained brain sections obtained from coronal cryosectioning of the brain after transcardial perfusion with fixative. The atlas is presented in 42 plates representing sections spaced 240 mu m apart. Stereotaxic coordinates were validated using both a model of Parkinsonian lesion of the striatum with 6-hydroxydopamine and labeling of the corticospinal tract in the spiny mouse spinal cord using AAV1/2-GFP intracortical injections. This work presents a new tool in A. cahirinus neurobiology and opens new avenues of research for the investigation of the regenerative ability of A. cahirinus in models of brain disorders.
- Human oocyte meiotic maturation is associated with a specific profile of alternatively spliced transcript isoformsPublication . Cornet‐Bartolomé, David; Barragán, Montserrat; Zambelli, Filippo; Ferrer‐Vaquer, Anna; Tiscornia, Gustavo; Balcells, Susanna; Rodriguez, Amelia; Grinberg, Daniel; Vassena, RitaThe transition from a transcriptionally active state (GV) to a transcriptionally inactive state (mature MII oocytes) is required for the acquisition of oocyte developmental competence. We hypothesize that the expression of specific genes at the in vivo matured (MII) stage could be modulated by posttranscriptional mechanisms, particularly regulation of alternative splicing (AS). In this study, we examined the transcriptional activity of GV oocytes after ovarian stimulation followed by oocyte pick-up and the landscape of alternatively spliced isoforms in human MII oocytes. Individual oocytes were processed and analyzed for transcriptional activity (GV), gene expression (GV and MII), and AS signatures (GV and MII) on HTA 2.0 microarrays. Samples were grouped according to maturation stage, and then subgrouped according to women's age and antral follicular count (AFC); array results were validated by quantitative polymerase chain reaction. Differentially expressed genes between GV and MII oocytes clustered mainly in biological processes related to mitochondrial metabolism. Interestingly, 16 genes that were related to the regulation of transcription and mitochondrial translation showed differences in alternatively spliced isoform profiles despite not being differentially expressed between groups. Altogether, our results contribute to our understanding of the role of AS in oocyte developmental competence acquisition.
- Vaginal microbiota profile at the time of embryo transfer does not affect live birth rate in IVF cycles with donated oocytesPublication . Vergaro, Paula; Tiscornia, Gustavo; Barragan, Montserrat; Garcia, Desiree; Rodriguez, Amelia; Santalo, Josep; Vassena, RitaResearch question: What is the relationship between the vaginal microbiota profile at the time of embryo transfer and live birth rates in women undergoing IVF/intracytoplasmic sperm injection (ICSI) with donated oocytes? Design: One hundred and fifty Caucasian women receiving donated oocytes were prospectively included in the study from March 2017 to January 2018. Samples of vaginal fluid were taken immediately before transfer of a fresh single blastocyst and genomic DNA (gDNA) was extracted. Bacterial load as well as the presence of four lactobacilli (L. crispatus, L. gasseri, L. jensenii and L. iners) and species associated with bacterial vaginosis (Gardnerella vaginalis, Atopobium vaginae, Mycoplasma hominis and Prevotella spp. - here collectively termed BVB) were determined by quantitative polymerase chain reaction. Vaginal microbiota profiles for each patient were characterized and correlated with reproductive results. Results: Although bacterial load was variable, a majority of samples were dominated by a single species (80.7%, 121/150). Most samples (76.7%, 115/150) were dominated by Lactobacillus spp., while 23.3% (35/150) were dominated by bacteria associated with bacterial vaginosis. The distribution of microbiota profiles among women who achieved a live birth and women who did not was similar (P = 0.43). Interestingly, we found a significantly higher proportion of samples dominated by L. crispatus- in women achieving live birth compared with those who did not (P = 0.021)
- Trophoblast attachment to the endometrial epithelium elicits compartment-specific transcriptional waves in an in-vitro modelPublication . Vergaro, Paula; Tiscornia, Gustavo; Zambelli, Filippo; Rodríguez, Amelia; Santaló, Josep; Vassena, RitaImplantation is a major bottleneck in human reproduction (Polanski et al., 2014). The average implantation rate for an embryo ranges from 30% to 40% (Coughlan et al., 2014). Recurrent implantation failure (RIF) is estimated to occur in approximately 4% of IVF cycles (Koot et al., 2012), although estimates vary because there are several somewhat different definitions of RIF in the literature. Implantation of the blastocyst in the receptive endometrium is a sequential process involving apposition, attachment and invasion that precedes the establishment of pregnancy (Wang and Dey, 2006). Successful implantation requires embryo competence and endometrial receptivity, both of which are dynamic and highly regulated states (Wang and Dey, 2006). In addition to genetic disorders (which are a major cause of implantation failure and miscarriage), embryo competence, quality and ultimately developmental potential depend on the embryo achieving the correct regulatory, signalling and metabolic states (Fu et al., 2009; Hourvitz et al., 2006; Lundin et al., 2001; Simon and Laufer, 2012; Sjoblom et al., 2006). A key determinant of these embryonic states is their underlying transcriptional dynamics; for instance, waves of embryonic transcriptional activation direct early development and the symmetry breaking needed for cell fate specification (Shi et al., 2015; Vassena et al., 2011).
- Matrisomal components involved in regenerative wound healing in axolotl and acomys: implications for biomaterial developmentPublication . Avila-Martinez, Nancy; Gansevoort, Merel; Verbakel, Juul; Jayaprakash, Haarshaadri; Araujo, Ines; Vitorino, Marta; Tiscornia, Gustavo; van Kuppevelt, Toin H.; Daamen, Willeke F.Achieving regeneration in humans has been a long-standing goal of many researchers. Whereas amphibians like the axolotl (Ambystoma mexicanum) are capable of regenerating whole organs and even limbs, most mammals heal their wounds via fibrotic scarring. Recently, the African spiny mouse (Acomys sp.) has been shown to be injury resistant and capable of regenerating several tissue types. A major focal point of research with Acomys has been the identification of drivers of regeneration. In this search, the matrisome components related to the extracellular matrix (ECM) are often overlooked. In this review, we compare Acomys and axolotl skin wound healing and blastema-mediated regeneration by examining their wound healing responses and comparing the expression pattern of matrisome genes, including glycosaminoglycan (GAG) related genes. The goal of this review is to identify matrisome genes that are upregulated during regeneration and could be potential candidates for inclusion in pro-regenerative biomaterials. Research papers describing transcriptomic or proteomic coverage of either skin regeneration or blastema formation in Acomys and axolotl were selected. Matrisome and GAG related genes were extracted from each dataset and the resulting lists of genes were compared. In our analysis, we found several genes that were consistently upregulated, suggesting possible involvement in regenerative processes. Most of the components have been implicated in regulation of cell behavior, extracellular matrix remodeling and wound healing. Incorporation of such pro-regenerative factors into biomaterials may help to shift pro-fibrotic processes to regenerative responses in treated wounds.
- Induced pluripotent stem cell modeling of Gaucher's disease: what have we learned?Publication . Santos Matias, Dino; Tiscornia, GustavoGaucher's disease (GD) is the most frequently inherited lysosomal storage disease, presenting both visceral and neurologic symptoms. Mutations in acid beta-glucocerebrosidase disrupt the sphingolipid catabolic pathway promoting glucosylceramide (GlcCer) accumulation in lysosomes. Current treatment options are enzyme replacement therapy (ERT) and substrate reduction therapy (SRT). However, neither of these approaches is effective in treating the neurological aspect of the disease. The use of small pharmacological compounds that act as molecular chaperones is a promising approach that is still experimental. In recent years, an association between GD and Parkinson like synucleinopathies has been discovered. Since 1992, a number of mouse models of GD have been the developed and partially reproduce phenotype of the disease. More recently, the discovery of direct reprograming has allowed the derivation of induced pluripotent stem cells (iPSc) from fibroblasts obtained from GD patients. iPSc can be expanded indefinitely in vitro and differentiated to macrophages and neurons, the main relevant cell types involved in GD. In this work, we review iPSc models of GD and summarize what we have learned from this system.