Repositorio Institucional Fleni
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Music Based Assessment for Cognitive Functions for Spanish-Speaking Adults With Acquired Brain Injury (ECMUS): A Pilot-Validation Study
No full textAcquired brain injury (ABI) commonly causes cognitive dysfunction that needs to be assessed and treated to maximize rehabilitation outcomes. Research suggests that music, emotion, and cognition are intimately linked, and that music can contribute to the assessment and treatment of cognitive functions of adults who have suffered from ABI. To this date, no standardized music based assessment tool exists to identify and measure cognitive functioning and mood states of Spanish-speaking persons with ABI at treatment intake and over time. The objective of this study was to develop such a scale and determine its psychometric properties in terms of internal consistency, reliability, and concurrent validity. The “Evaluación de la Cognición Musical para Adultos con Lesión Cerebral Adquirida” (in English “Music Based Assessment for Cognitive Functions of Adults with Acquired Brain Injury – ECMUS”) was developed and tested at a neurorehabilitation institute in Argentina. Twenty-four healthy adults and 20 adults with ABI were recruited and assessed with the ECMUS. Despite the limited number of participants, this preliminary psychometric examination shows promising results. The tool has an acceptable internal consistency, excellent test–retest and inter-rater reliability, and, depending on the subscale, weak to strong correlations to related, nonmusical constructs. Overall, this pilot study opens the possibility to further explore the inclusion of music in assessment procedures of Spanish-speaking individuals with ABI in rehabilitation settings. It is clinically derived and emphasizes evidence-based contributions of the field of music therapy to interdisciplinary assessment and treatment in rehabilitation settings.Fil: Pfeiffer, Camila Floreana. Fleni. Centro de Rehabilitación Adultos CR. Servicio de Musicoterapia; Argentina. Berklee College of Music; Estados Unidos.Fil: Hanser, Suzanne. Berklee College of Music; Estados Unidos.Fil: Russo, María Julieta. Fleni. Centro de Rehabilitación Adultos CR. Servicio de Musicoterapia; Argentina.Fil: Goyheneix, Mercedes. Fleni. Centro de Rehabilitación Adultos CR. Servicio de Musicoterapia; Argentina.Fil: Oliva, Miguel. Universidad Nacional de Tres de Febrero; Argentina.Fil: Codding, Peggy. Berklee College of Music; Estados Unidos.Fil: Allegri, Ricardo Francisco. Fleni. Departamento de Neurología. Servicio de Neurología Cognitiva, Neuropsicología y Neuropsiquiatría; Argentina
Editorial: Consequences of the COVID-19 Pandemic on Care for Neurological Conditions
No full textEditorial on the Research Topic Consequences of the COVID-19 Pandemic on Care for Neurological ConditionsFil: Matias-Guiu, Jordi A. Universidad Complutense de Madrid; España.Fil: Sung, Sheng-Feng. Ditmanson Medical Foundation Chia-Yi Christian Hospital; Taiwan.Fil: Hsieh, Cheng-Yang. Tainan Sin Lau Hospital; Taiwan.Fil: Nezu, Tomohisa. Hiroshima University Graduate School of Biomedical and Health Sciences; Japón.Fil: Porta-Etessam, Jesús. Universidad Complutense de Madrid; España.Fil: Allegri, Ricardo Francisco. Fleni. Departamento de Neurología. Servicio de Neurología Cognitiva, Neuropsicología y Neuropsiquiatría; Argentina
Obesity and the risk of Multiple Sclerosis. The role of Leptin
No full textObjective: To investigate the effects of leptin on different T-cell populations, in order to gain more insight into the link between leptin and obesity.
Methods: Three hundred and nine RRMS patients and 322 controls participated in a cross-sectional survey, to confirm whether excess weight/obesity in adolescence or early adulthood increased the risk of MS. Serum leptin levels were determined by ELISA. MBP83-102 , and MOG63-87 peptide-specific T cells lines were expanded from peripheral blood mononuclear cells. Leptin receptor expression was measured by RT-PCR and flow cytometry. Bcl-2, p-STAT3, pERK1/2, and p27kip1 expression were assayed using ELISA, and apoptosis induction was determined by Annexin V detection. Cytokines were assessed by ELISPOT and ELISA, and regulatory T cells (Tregs) by flow cytometry.
Results: Logistic regression analysis, showed excess weight at age 15, and obesity at 20 years of age increased MS risk (OR = 2.16, P = 0.01 and OR = 3.9, P = 0.01). Leptin levels correlated with BMI in both groups. The addition of Leptin increased autoreactive T-cell proliferation, reduced apoptosis induction, and promoted proinflammatory cytokine secretion. Obese patients produced more proinflammatory cytokines compared to overweight/normal/underweight subjects. Inverse correlation was found between leptin levels and circulating Treg cells (r = -0.97, P < 0.0001). Leptin inhibited Treg proliferation. Effects of leptin on CD4+ CD25- effector T cells were mediated by increased STAT3 and ERK1/2 phosphorylation, and down modulation of the cell cycle inhibitor P27kip1 . In contrast, leptin effects on Tregs resulted from decreased phosphorylation of ERK1/2 and upregulation of p27kip1 .
Interpretation: Leptin promotes autoreactive T-cell proliferation and proinflammatory cytokine secretion, but inhibits Treg-cell proliferation.Fil: Marrodán, Mariano. Fleni. Departamento de Neurología. Servicio de Neuroinmunología y Enfermedades Desmielinizantes; Argentina.Fil: Farez, Mauricio Franco. Fleni. Departamento de Neurología; Argentina.Fil: Correale, Jorge. Fleni. Departamento de Neurología. Servicio de Neuroinmunología y Enfermedades Desmielinizantes; Argentina.Fil: Balbuena Aguirre, María Eugenia. Hospital de Clínicas José de San Martín. Departamento de Neurología; Argentina
Immunosuppressive Amino-Acid Catabolizing Enzymes in Multiple Sclerosis
Full text linkMultiple sclerosis (MS) is a chronic inflammatory demyelinating disease that affects the central nervous system. Although the pathogenesis of MS is not yet fully elucidated, several evidences suggest that autoimmune processes mediated by Th1, Th17, and B cells play an important role in the development of the disease. Similar to other cells, immune cells need continuous access to amino acids (AA) in order to maintain basal metabolism and maintain vitality. When immune cells are activated by inflammation or antigenic signals, their demand for AA increases rapidly. Although AA deprivation itself may weaken the immune response under certain conditions, cells also have AA sensitive pathways that can activate intense alterations in cell metabolism based on changes in AA levels. Several data indicate that cells expressing enzymes that can degrade AA can regulate the functions of antigen-presenting cells and lymphocytes, revealing that the AA pathways are essential for controlling the function, and survival of immune cells, as well as immune cell gene expression. Basal AA catabolism may contribute to immune homeostasis and prevent autoimmunity, while increased AA catalytic activity may enhance immune suppression. In addition, there is increasing evidence that some downstream AA metabolites are important biological mediators of autoimmune response regulation. Two of the most important AA that modulate the immune response are L-Tryptophan (Trp) and L-Arginine (Arg). Tryptophan is catabolized through 2,3-dioxygenase (TDO) and indoleamine 2,3-dioxygenase (IDO) 1 and IDO2 enzymes, while three other enzymes catabolize Arg: inducible nitric oxide synthetase (iNOS), and two arginase isoforms (ARG1, ARG2). Genes encoding IDO, iNOS and ARG are induced by inflammatory cues such as cytokines, a key feature that distinguishes them from enzymes that catabolize other AA. Evidence suggests that AA catabolism is decreased in MS patients and that this decrease has functional consequences, increasing pro-inflammatory cytokines and decreasing Treg cell numbers. These effects are mediated by at least two distinct pathways involving serine/threonine kinases: the general control nonderepressible 2 kinase (GCN2K) pathway; and the mammalian target of rapamycin (mTOR) pathway. Similarly, IDO1-deficient mice showed exacerbation of experimental autoimmune encephalomyelitis (EAE), increased Th1 and Th17 cells, and decreased Treg cells. On the contrary, the administration of downstream Trp metabolite 3-HAA, inhibits Th1/Th17 effector cells and promotes Treg response by up-regulating TGF-β production by dendritic cells, thereby improving EAE. Collectively, these observations stand out the significance of AA catabolism in the regulation of the immune responses in MS patients. The molecules related to these pathways deserve further exploration as potential new therapeutic targets in MSFil: Correale, Jorge. Fleni. Departamento de Neurología. Servicio de Neuroinmunología y Enfermedades Desmielinizantes; Argentina
Limitations of the Unified Multiple System Atrophy Rating Scale as outcome measure for clinical trials and a roadmap for improvement
No full textPurpose
The unified multiple system atrophy (MSA) rating scale (UMSARS) was developed almost 20 years ago as a clinical rating scale to capture multiple aspects of the disease. With its widespread use, the shortcomings of the UMSARS as a clinical outcome assessment (COA) have become increasingly apparent. We here summarize the shortcomings of the scale, confirm some of its limitations with data from the Natural History Study of the Synucleinopathies (NHSS), and suggest a framework to develop and validate an improved COA to be used in future clinical trials of disease-modifying drugs in patients with MSA.
Methods
Expert consensus assessment of the limitations of the UMSARS and recommendations for the development and validation of a novel COA for MSA. We used UMSARS data from the ongoing NHSS (ClinicalTrials.gov: NCT01799915) to showcase some of these limitations.
Results
The UMSARS in general, and specific items in particular, have limitations to detect change resulting in a ceiling effect. Some items have specific limitations including unclear anchoring descriptions, lack of correlation with disease severity, susceptibility to improve with symptomatic therapies (e.g., orthostatic hypotension, constipation, and bladder dysfunction), and redundancy, among others.
Conclusions
Because of the limitations of the UMSARS, developing and validating an improved COA is a priority. The time is right for academic MSA clinicians together with industry, professional societies, and patient advocacy groups to develop and validate a new COA.Fil: Palma, José Alberto. New York University School of Medicine; Estados Unidos.Fil: Millar Vernetti, Patricio. New York University School of Medicine; Estados Unidos.Fil: Perez, Miguel A. New York University School of Medicine; Estados Unidos.Fil: Krismer, Florian. Medical University of Innsbruck; Austria.Fil: Seppi, Klaus. Medical University of Innsbruck; Austria.Fil: Fanciulli, Alessandra. Medical University of Innsbruck; Austria.Fil: Singer, Wolfgang. Mayo Clinic Rochester; Estados Unidos.Fil: Low, Phillip. Mayo Clinic Rochester; Estados Unidos.Fil: Biaggioni, Italo. Vanderbilt University; Estados Unidos.Fil: Norcliffe-Kaufmann, Lucy. New York University School of Medicine; Estados Unidos.Fil: Pellecchia, Maria Teresa. Salerno University; Italia.Fil: Martí, Maria José. Hospital Clinic Barcelona; España.Fil: Kim, Han-Joon. Seoul Medical University; Corea del Sur.Fil: Merello, Marcelo. Fleni. Departamento de Neurología. Servicio de Movimientos Anormales; Argentina.Fil: Stankovic, Iva. University of Belgrade; Serbia.Fil: Poewe, Werner. Medical University of Innsbruck; Austria.Fil: Betensky, Rebecca. New York University School of Global Public Health; Estados Unidos.Fil: Wenning, Gregor. Medical University of Innsbruck; Austria.Fil: Kaufmann, Horacio. New York University School of Medicine; Estados Unidos
Status of the neuromyelitis optica spectrum disorder in Latin America
No full textBackground
Neuromyelitis optica spectrum disorders (NMOSD) is an increasing diagnostic and therapeutic challenge in Latin America (LATAM). Despite the heterogeneity of this population, ethnic and socioeconomic commonalities exist, and epidemiologic studies from the region have had a limited geographic and population outreach. Identification of some aspects from the entire region are lacking.
Objectives
To determine ethnic, clinical characteristics, and utilization of diagnostic tools and types of therapy for patients with NMOSD in the entire Latin American region.
Methods
The Latin American Committee for Treatment and Research in MS (LACTRIMS) created an exploratory investigational survey addressed by Invitation to NMOSD Latin American experts identified through diverse sources. Data input closed after 30 days from the initial invitation. The questionnaire allowed use of absolute numbers or percentages. Multiple option responses covering 25 themes included definition of type of practice; number of NMOSD cases; ethnicity; utilization of the 2015 International Panel criteria for the diagnosis of Neuromyelitis optica (IPDN); clinical phenotypes; methodology utilized for determination of anti-Aquaporin-4 (anti AQP4) antibodies serological testing, and if this was performed locally or processed abroad; treatment of relapses, and long-term management were surveyed.
Results
We identified 62 investigators from 21 countries reporting information from 2154 patients (utilizing the IPDN criteria in 93.9% of cases), which were categorized in two geographical regions: North-Central, including the Caribbean (NCC), and South America (SA). Ethnic identification disclosed Mestizos 61.4% as the main group. The most common presenting symptoms were concomitant presence of optic neuritis and transverse myelitis in 31.8% (p=0.95); only optic neuritis in 31.4% (more common in SA), p<0.001); involvement of the area postrema occurred in 21.5% and brain stem in 8.3%, both were more frequent in the South American cases (p<0.001). Anti-AQP4 antibodies were positive in 63.9% and anti-Myelin Oligodendrocyte Glycoprotein (MOG) antibodies in 4.8% of total cases. The specific laboratorial method employed was not known by 23.8% of the investigators. Acute relapses were identified in 81.6% of cases, and were treated in 93.9% of them with intravenous steroids (IVS); 62.1% with plasma exchange (PE), and 40.9% with intravenous immunoglobulin-G (IVIG). Therapy was escalated in some cases due to suboptimal initial response. Respondents favored Rituximab as long-term therapy (86.3%), whereas azathioprine was also utilized on 81.8% of the cases, either agent used indistinctly by the investigators according to treatment accessibility or clinical judgement. There were no differences among the geographic regions.
Conclusions
This is the first study including all countries of LATAM and the largest cohort reported from a multinational specific world area. Ethnic distributions and phenotypic features of the disease in the region, challenges in access to diagnostic tools and therapy were identified. The Latin American neurological community should play a determinant role encouraging and advising local institutions and health officials in the availability of more sensitive and modern diagnostic methodology, in facilitating the the access to licensed medications for NMOSD, and addressing concerns on education, diagnosis and management of the disease in the community.Fil: Rivera, Victor M. Baylor College of Medicine; Estados Unidos.Fil: Hamuy, Fernando. Centro Nacional de Esclerosis Múltiple; Paraguay.Fil: Rivas, Verónica. Instituto Nacional de Neurología y Neurocirugía "Manuel Velasco Suarez"; México.Fil: Gracia, Fernando. Hospital Santo Tomas; Panamá.Fil: Rojas, Juan Ignacio. Centro de Esclerosis Múltiple de Buenos Aires; Argentina.Fil: Bernardi Bichuetti, Denis. Universidade Federal de Sao Paulo; Brasil.Fil: Villa, Andrés. Hospital Ramos Mejía; Argentina.Fil: Daccah Marques, Vanessa. Hospital das Clinicas da Faculdade de Medicina de Ribeirao Preto; Brasil.Fil: Soto, Arnoldo. Centro Medico Docente La Trinidad; Venezuela.Fil: Bertado, Brenda. Instituto Mexicano del Seguro Social; México.Fil: Trevino Frenk, Irene. lnstituto Nacional de Ciencias Médicas y Nutrición "Salvador Zubirán"; México.Fil: Galleguillos, Lorna. Clínica Alemana de Santiago; Chile.Fil: Quiñones, Jairo. Fundación Valle del Lili; Colombia.Fil: Ramírez, Deyanira A. Hospital Docente Padre Bellini; República Dominicana.Fil: Caparó-Zamalloa, César. lnstituto Nacional de Ciencias Neurológicas; Perú.Fil: Ciampi, Ethel. Pontificia Universidad Católica de Chile; Chile.Fil: Lana-Peixoto, Marco A. Universidade Federal de Minas Gerais; Brasil.Fil: Rodríguez, Emmanuel. Centro Médico Nacional "La Raza"; México.Fil: Zarco, Luis. Pontificia Universidad Javeriana; Colombia.Fil: Correale, Jorge. Fleni. Departamento de Neurología. Servicio de Neuroinmunología y Enfermedades Desmielinizantes; Argentina
Research priorities in multiple sclerosis in Latin America: A multi-stakeholder call to action to improve patients care: Research priorities in MS in LATAM
No full textAs human and economic resources are limited, especially in Latin America (LATAM), it is important to identify research priorities to improve multiple sclerosis (MS) patients care in the region. The objective was to generate a multidisciplinary consensus on research priorities in MS for patients care in LATAM by involving healthcare professionals and MS patient associations.
Methods
consensus was reached through a four-step modified Delphi method designed to identify and rate research priorities in MS in LATAM. The process consisted of two qualitative assessments, a general ranking phase and a consensus meeting followed by a more detailed ranking phase
Results
a total of 62 participants (35 neurologists, 4 nurses, 12 kinesiologists, 7 neuropsychologists and 4 patient association members) developed the process. At the final ranking stage following the consensus meeting, each participant provided their final rankings, and the top priority research questions were outlined. 11 research priorities were identified focusing on healthcare access, costs of the disease, physical and cognitive evaluation and rehabilitation, quality of life, symptoms management, prognostic factors, the need of MS care units and patient's management in emergencies like COVID-19.
Conclusion
this work establishes MS research priorities in LATAM from multiple perspectives. To pursue the actions suggested could launch the drive to obtain information that will help us to better understand the disease in our region and, especially, to better care for affected patients.Fil: Rojas, Juan I. Centro de Esclerosis Múltiple de Buenos Aires; Argentina.Fil: Carnero Contentti, Edgar. Hospital Alemán de Buenos Aires; Argentina.Fil: Abad, Patricio. Hospital Metropolitano de Quito; Ecuador.Fil: Aguayo, Adriana. Universidad de Guadalajara; México.Fil: Alonso, Ricardo. Hospital Ramos Mejía; Argentina.Fil: Bauer, Johana. Esclerosis Múltiple Argentina; Argentina.Fil: Becker, Jefferson. Pontifical Catholic University of Rio Grande do Sul (PUCRS); Brasil.Fil: Bustos, Andrea K. Fundación AlunCo; Argentina.Fil: Carcamo, Claudia A. Pontificia Universidad Católica de Chile; Chile.Fil: Carrá, Adriana. Hospital Británico Buenos Aires; Argentina.Fil: Correa Díaz, Edgar Patricio. Hospital de Especialidades Carlos Andrade Marín; Ecuador.Fil: Correale, Jorge. Fleni. Departamento de Neurología. Servicio de Neuroinmunología y Enfermedades Desmielinizantes; Argentina.Fil: Cristiano, Edgardo. Hospital Universitario CEMIC; Argentina.Fil: Diaz, Alejandro J. Instituto Guatemalteco de Seguridad Social; Guatemala.Fil: Fernandez Liguori, Nora. Hospital Universitario Sanatorio Guemes; Argentina.Fil: Flores, José. Instituto Nacional de Neurología y Neurocirugía; México.Fil: Fruns, Manuel. Clínica Las Condes; Chile.Fil: Ysrraelit, Maria Célica. Fleni. Departamento de Neurología. Servicio de Neuroinmunología y Enfermedades Desmielinizantes; Argentina
The commercial genetic testing landscape for Parkinson's disease
No full textIntroduction: There have been no specific guidelines regarding which genes should be tested in the clinical setting for Parkinson's disease (PD) or parkinsonism. We evaluated the types of clinical genetic testing offered for PD as the first step of our gene curation.
Methods: The National Institutes of Health (NIH) Genetic Testing Registry (GTR) was queried on 12/7/2020 to identify current commercial PD genetic test offerings by clinical laboratories, internationally.
Results: We identified 502 unique clinical genetic tests for PD, from 28 Clinical Laboratory Improvement Amendments (CLIA)-approved clinical laboratories. These included 11 diagnostic PD panels. The panels were notable for their differences in size, ranging from 5 to 62 genes. Five genes for variant query were included in all panels (SNCA, PRKN, PINK-1, PARK7 (DJ1), and LRRK2). Notably, the addition of the VPS35 and GBA genes was variable. Panel size differences stemmed from inclusion of genes linked to atypical parkinsonism and dystonia disorders, and genes in which the link to PD causation is controversial.
Conclusion: There is an urgent need for expert opinion regarding which genes should be included in a commercial laboratory multi-gene panel for PD.Fil: Cook, Lola. Indiana University School of Medicine; Estados Unidos.Fil: Schulze, Jeanine. Indiana University School of Medicine; Estados Unidos.Fil: Verbrugge, Jennifer. Indiana University School of Medicine; Estados Unidos.Fil: Beck, James C. Parkinson's Foundation; Estados Unidos.Fil: Marder, Karen S. Columbia University Medical Center; Estados Unidos.Fil: Saunders-Pullman, Rachel. Mount Sinai Beth Israel; Estados Unidos.Fil: Klein, Christine. University of Luebeck; Alemania.Fil: Naito, Anna. Parkinson's Foundation; Estados Unidos.Fil: Alcalay, Roy N. Columbia University Medical Center; Estados Unidos.Fil: Merello, Marcelo. Fleni. Departamento de Neurología. Servicio de Movimientos Anormales; Argentina
Multiple sclerosis and neuromyelitis optica spectrum disorders in Argentina: comparing baseline data from the Argentinean MS Registry (RelevarEM)
No full textThe objective of this study was to describe and compare the baseline epidemiological data of multiple sclerosis (MS) and neuromyelitis optica spectrum disorder (NMOSD) patients included in RelevarEM (Clinical Trials registry number NCT03375177).
Methods: RelevarEM is a longitudinal, strictly observational MS and NMOSD registry in Argentina. Epidemiological and comorbidity data from MS and NMOSD patients were described and compared. For comorbidities, the Charlson comorbidity index (CCI) was used to calculate the burden at entry. CCI was stratified in 0 and ≥ 1 and described for the entire cohort.
Results: A total of 1588 and 75 MS and NMOSD patients (respectively) were included. For MS patients, the mean age was 42 ± 7 years, female sex 65.3%, mean EDSS 2, and mean disease duration 8 ± 6 years. In NMOSD, the mean age was 40 ± 7 years, female sex 78.7%, mean disease duration 5 ± 3.5 years, and mean EDSS 2.5. The most frequent MS phenotype was RRMS in 82.4%. In MS, the CCI was 0 in 85.8.2% while ≥ 1 was in 14.2% of patients. Regarding phenotype stratification, CCI ≥ 1 was 3.9% in CIS, 13.5% in RRMS, 28.7% in SPMS, and 17.4% in PPMS (p < 0.001 between groups). In NMOSD, the CCI was 0 in 64% while ≥ 1 was in 36%. The MS/NMOSD ratio found was 21/1.
Conclusions: This is the first analysis of the longitudinal Argentinean registry of MS and NMOSD describing and comparing conditions that contributes to provide reliable real-world data in the country.Fil: Correale, Jorge. Fleni. Departamento de Neurología. Servicio de Neuroinmunología y Enfermedades Desmielinizantes; Argentina.Fil: Gaitán, María Inés. Fleni. Departamento de Neurología. Servicio de Neuroinmunología y Enfermedades Desmielinizantes; Argentina.Fil: Marrodán, Mariano. Fleni. Departamento de Neurología. Servicio de Neuroinmunología y Enfermedades Desmielinizantes; Argentina.Fil: Negrotto, Laura. Fleni. Departamento de Neurología. Servicio de Neuroinmunología y Enfermedades Desmielinizantes; Argentina.Fil: Ysrraelit, Maria. Fleni. Departamento de Neurología. Servicio de Neuroinmunología y Enfermedades Desmielinizantes; Argentina.Fil: Rojas, Juan Ignacio. Hospital Italiano de Buenos Aires. Servicio de Neurología; Argentina. Centro de Esclerosis Múltiple de Buenos Aires; Argentina.Fil: Alonso Serena, Marina. Hospital Italiano de Buenos Aires. Área de Investigación en Medicina Interna; Argentina.Fil: Garcea, Orlando. Universidad de Buenos Aires. Facultad de Medicina. Centro Universitario de Esclerosis Múltiple; Argentina. Hospital Dr. J. M. Ramos Mejía; Argentina.Fil: Patrucco, Liliana. Hospital Italiano de Buenos Aires. Servicio de Neurología; Argentina.Fil: Carrá, Adriana. Hospital Británico. Sección de Enfermedades Desmielinizantes; Argentina. Fundación Favaloro/INECO; Argentina.Fil: Vrech, Carlos. Sanatorio Allende. Departamento de Enfermedades desmielinizantes; Argentina.Fil: Pappolla, Agustín. Hospital Italiano de Buenos Aires. Servicio de Neurología; Argentina.Fil: Miguez, Jimena. Hospital Italiano de Buenos Aires. Servicio de Neurología; Argentina.Fil: Doldan, María L. Hospital Italiano de Buenos Aires. Servicio de Neurología; Argentina. Centro de Esclerosis Múltiple de Buenos Aires, Buenos Aires, Argentina.Fil: Silveira, Facundo. Hospital Italiano de Buenos Aires. Servicio de Neurología; Argentina.Fil: Alonso, Ricardo. Universidad de Buenos Aires. Facultad de Medicina. Centro Universitario de Esclerosis Múltiple; Argentina. Hospital Dr. J. M. Ramos Mejía; Argentina. Sanatorio Güemes; Argentina.Fil: Cohen, Leila. Universidad de Buenos Aires. Facultad de Medicina. Centro Universitario de Esclerosis Múltiple; Argentina. Hospital Dr. J. M. Ramos Mejía; Argentina.Fil: Pita, Cecilia. Universidad de Buenos Aires. Facultad de Medicina. Centro Universitario de Esclerosis Múltiple; Argentina. Hospital Dr. J. M. Ramos Mejía; Argentina.Fil: Silva, Berenice Anabel. Universidad de Buenos Aires. Facultad de Medicina. Centro Universitario de Esclerosis Múltiple; Argentina. Hospital Dr. J. M. Ramos Mejía; Argentina
Generation of myostatin edited horse embryos using CRISPR/Cas9 technology and somatic cell nuclear transfer
No full textThe application of new technologies for gene editing in horses may allow the generation of improved sportive individuals. Here, we aimed to knock out the myostatin gene (MSTN), a negative regulator of muscle mass development, using CRISPR/Cas9 and to generate edited embryos for the first time in horses. We nucleofected horse fetal fibroblasts with 1, 2 or 5 µg of 2 different gRNA/Cas9 plasmids targeting the first exon of MSTN. We observed that increasing plasmid concentrations improved mutation efficiency. The average efficiency was 63.6% for gRNA1 (14/22 edited clonal cell lines) and 96.2% for gRNA2 (25/26 edited clonal cell lines). Three clonal cell lines were chosen for embryo generation by somatic cell nuclear transfer: one with a monoallelic edition, one with biallelic heterozygous editions and one with a biallelic homozygous edition, which rendered edited blastocysts in each case. Both MSTN editions and off-targets were analyzed in the embryos. In conclusion, CRISPR/Cas9 proved an efficient method to edit the horse genome in a dose dependent manner with high specificity. Adapting this technology sport advantageous alleles could be generated, and a precision breeding program could be developed.Fil: Miriuka, Santiago Gabriel. Fleni. Laboratorio de Investigación Aplicada a las Neurociencias; Argentina. Consejo Nacional de Investigaciones Científicas y Tecnológicas; Argentina.Fil: Moro, Lucía Natalia. Fleni. Laboratorio de Investigación Aplicada a las Neurociencias; Argentina.Fil: Viale, Diego Luis. Consejo Nacional de Investigaciones Científicas y Tecnológicas; Argentina. Laboratorio de Neurología y Citogenética Molecular, CESyMA; Argentina.Fil: Bastón, Juan Ignacio. KHEIRON BIOTECH; Argentina.Fil: Arnold, Victoria. KHEIRON BIOTECH; Argentina.Fil: Suvá, Mariana. KHEIRON BIOTECH; Argentina.Fil: Wiedenmann, Elisabet. KHEIRON BIOTECH; Argentina.Fil: Olguín, Martín. KHEIRON BIOTECH; Argentina.Fil: Vichera, Gabriel. KHEIRON BIOTECH; Argentina