Rejuvenation by cell reprogramming: A new horizon in gerontology

The discovery of animal cloning and subsequent development of cell reprogramming technology were quantum leaps as they led to the achievement of rejuvenation by cell reprogramming and the emerging view that aging is a reversible epigenetic process. Here, we will first summarize the experimental achievements over the last 7 years in cell and animal rejuvenation. Then, a comparison will be made between the principles of the cumulative DNA damage theory of aging and the basic facts underlying the epigenetic model of aging, including Horvath's epigenetic clock. The third part will apply both models to two natural processes, namely, the setting of the aging clock in the mammalian zygote and the changes in the aging clock along successive generations in mammals. The first study demonstrating that skin fibroblasts from healthy centenarians can be rejuvenated by cell reprogramming was published in 2011 and will be discussed in some detail. Other cell rejuvenation studies in old humans and rodents published afterwards will be very briefly mentioned. The only in vivo study reporting that a number of organs of old progeric mice can be rejuvenated by cyclic partial reprogramming will also be described in some detail. The cumulative DNA damage theory of aging postulates that as an animal ages, toxic reactive oxygen species generated as byproducts of the mitochondria during respiration induce a random and progressive damage in genes thus leading cells to a progressive functional decline. The epigenetic model of aging postulates that there are epigenetic marks of aging that increase with age, leading to a progressive derepression of DNA which in turn causes deregulated expression of genes that disrupt cell function. The cumulative DNA damage model of aging fails to explain the resetting of the aging clock at the time of conception as well as the continued vitality of species as millenia go by. In contrast, the epigenetic model of aging straightforwardly explains both biologic phenomena. A plausible initial application of rejuvenation in vivo would be preventing adult individuals from aging thus eliminating a major risk factor for end of life pathologies. Further, it may allow the gradual achievement of whole body rejuvenation.Fil: Goya, Rodolfo Gustavo. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Investigaciones Bioquímicas de La Plata "Prof. Dr. Rodolfo R. Brenner". Universidad Nacional de la Plata. Facultad de Ciencias Médicas. Instituto de Investigaciones Bioquímicas de La Plata "Prof. Dr. Rodolfo R. Brenner"; ArgentinaFil: Lehmann, Marianne. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Investigaciones Bioquímicas de La Plata "Prof. Dr. Rodolfo R. Brenner". Universidad Nacional de la Plata. Facultad de Ciencias Médicas. Instituto de Investigaciones Bioquímicas de La Plata "Prof. Dr. Rodolfo R. Brenner"; ArgentinaFil: Chiavellini, Priscila. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Investigaciones Bioquímicas de La Plata "Prof. Dr. Rodolfo R. Brenner". Universidad Nacional de la Plata. Facultad de Ciencias Médicas. Instituto de Investigaciones Bioquímicas de La Plata "Prof. Dr. Rodolfo R. Brenner"; ArgentinaFil: Canatelli Mallat, Martina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Investigaciones Bioquímicas de La Plata "Prof. Dr. Rodolfo R. Brenner". Universidad Nacional de la Plata. Facultad de Ciencias Médicas. Instituto de Investigaciones Bioquímicas de La Plata "Prof. Dr. Rodolfo R. Brenner"; ArgentinaFil: Hereñú, Claudia Beatriz. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Farmacología Experimental de Córdoba. Universidad Nacional de Córdoba. Facultad de Ciencias Químicas. Instituto de Farmacología Experimental de Córdoba; ArgentinaFil: Brown, Oscar Alfredo. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Investigaciones Bioquímicas de La Plata "Prof. Dr. Rodolfo R. Brenner". Universidad Nacional de la Plata. Facultad de Ciencias Médicas. Instituto de Investigaciones Bioquímicas de La Plata "Prof. Dr. Rodolfo R. Brenner"; Argentin

Traces of the (m)other: deconstructing hegemonic historical narrative in Teat(r)o Oficina Uzyna Uzona's Os Sertões

This article focuses on the way in which renowned São Paulo-based theatre company Teat(r)o Oficina Uzyna Uzona deconstructs hegemonic historical narrative in their 2000 - 2007 25 hour-long production of Euclides da Cunha’s seminal Brazilian novel Os sertões (“Rebellion in the Backlands”), an account of the War of Canudos (1896-1897), the first major act of State terrorism carried out by the nascent Brazilian Federal Government on the country’s subaltern population. The Teat(r)o Oficina’s epic adaptation fuses events from the colonial period, the military dictatorship and contemporary 21st Century São Paulo to relate the repetitive cycles of misappropriation, oppression and resistance that have characterized the history of Brazil and its people over the centuries. However, any fatalistic view of victimhood as an essential aspect of Brazilian subjectivity is radically challenged by the vibrant, rhythmic, material impact of the theatrical super-signs underpinning the performance text. Drawing on Julia Kristeva’s notion of the semiotic - the pre-linguistic, illogical, rhythmical materialism of language intimately related to a primordial relationship with the abject mother – I shall suggest that it is the rhythmic, libidinal force of the performance and its extensive use of the cultural manifestations of Brazil’s subaltern population that imbues Os Sertões with the silent presence-as-absence of the abject Brazilian (M)Other – the Black, Indigenous and Mestiza matriarchal line whose alternative discourse is often barred from hegemonic accounts of Brazilian historiography. Her silent heritage is embodied on stage by the members of the Oficina, who reclaim an alienating national heritage for themselves by transforming the often tragic tale of Brazil’s past into a joyous celebration of tenacious vitality

Masculinising testosterone treatment and effects on preclinical cardiovascular disease, muscle strength and power, aggression, physical fitness and respiratory function in transgender men : protocol for a 10-year, prospective, observational cohort study in Denmark at the Body Identity Clinic (BIC)

Introduction The number of individuals with gender dysphoria seeking gender-affirming treatment is increasing. The short-term and long-term effects of masculinising treatment with testosterone are debated as serum testosterone increases up to 20-fold compared with cisgender women. We will investigate short-term and long-term effects of masculinising testosterone treatment on preclinical and clinical coronary disease, muscle strength and power, oxygen consumption (VO2) max, cardiac and respiratory function and quality of life including aggression in transgender men. Methods and analyses Prospective, single-centre, observational cohort study at the Body Identity Clinic (BIC), Odense University Hospital, Denmark. Investigations are performed at inclusion and following 1, 3, 5 and 10 years of testosterone therapy. Non-calcified coronary plaque volume and calcium score are estimated by coronary CT angiography. CT is only performed at inclusion and following 1 and 10 years. Upper body muscle strength and power are measured by a 'low row' weight stack resisted exercise machine. Evaluation of aggression and quality of life is assessed by questionnaires, VO2 max is estimated by maximal testing on bike ergometer, and cardiac and respiratory functions are measured by echocardiography and spirometry, respectively. Markers of cardiovascular risk and inflammation and also cortisol and cortisone are assessed in blood, diurnal urine and/or hair samples. Our cohort (BIC), including dropouts, will be an embedded subcohort in a future national registry study in all individuals with gender dysphoria and controls. Data are available on International Statistical Classification of Diseases and Related Health Problems 10(th) version diagnostic codes, prescriptions, socioeconomics and causes of death. Ethics and dissemination The Regional Committee on Health Research Ethics for Southern Denmark (S-20190108) and the Danish Data Protection Agency (19/27572) approved the study. Signed informed consent will be obtained from all participants. All findings will be published in peer-reviewed journals or at scientific conferences

A hierarchical model for the control of epigenetic aging in mammals

Regulatory mechanisms range from a single level of control in simple metazoans to multi-level hierarchical control networks in higher animals. Organismal regulation encompasses homeostatic and circadian networks that are interconnected, with no documented exceptions. The epigenetic clock is a highly accurate biomarker of age in humans, defined by a mathematical algorithm based on the methylation of a subset of age-related CpG sites on DNA. Experimental evidence suggests the existence of an underlying regulatory mechanism. By analogy with other integrative systems as the neuroendocrine-immune network and the circadian clocks, a hierarchical organization in the control of the ticking rate of the epigenetic clock is hypothesized here. The hierarchical organization of the neuroendocrine, immune and circadian systems is briefly reviewed. This is followed by a brief review of the epigenetic clock at cell level. Finally, different lines of indirect evidence, consistent with the existence of a central pacemaker controlling the ticking rate of the epigenetic clock at organismal level are discussed. The concluding remarks put the hierarchical model proposed for the control of the clock into an evolutionary perspective. Within this perspective, the present hypothesis is intended as a conceptual outline based on designs consistently favored by evolution in higher animals.Instituto de Investigaciones Bioquímicas de La Plat

Construcción de un adenovirus recombinante regulable portador de los genes de pluripotencia y de gen GFP para implementar en rejuvenecimiento celular mediante reprogramación celular parcial

La reprogramación celular convencional involucra convertir un linaje celular somático a células madre pluripotentes inducidas (iPSC), las cuales pueden ser posteriormente rediferenciadas a tipos celulares específicos. De manera alternativa, la reprogramación celular parcial convierte células somáticas en otros tipos celulares por expresión transitoria de genes de pluripotencia, procedimiento que genera intermediarios celulares pluripotentes que retienen la identidad celular original, pero que están rejuvenecidos y responden a cócteles apropiados de factores de diferenciación específicos. En este contexto, el rejuvenecimiento por reprogramación celular parcial constituye un campo de investigación emergente. Para su implementación, los sistemas de expresión regulables policistrónicos, generalmente integrativos, son ampliamente utilizados. Con el mismo propósito, hemos construido un adenovirus recombinante (no integrativo) regulable de alta capacidad que contiene el gen de la proteína fluorescente verde (GFP) y los genes oct4, sox2, klf4 y c-myc (OSKM, o genes de pluripotencia). Estos genes OSKM se encuentran ensamblados como un tándem bicistrónico (STEMCCA) bajo el control de un promotor bidireccional regulable por sistema Tet-Off que controla también la expresión de la GFP. Separadamente, un segundo cassette expresa constitutivamente la proteína regulatoria tTA. En este trabajo se describe la generación de dicho adenovector dependiente de virus auxiliar y la caracterización del mismo mediante diversas técnicas. Este adenovector constituye una herramienta promisoria para la implementación de reprogramación parcial no integrativa.Eje: Salud humanaInstituto de Investigaciones Bioquímicas de La Plat

Generación de cardiomiocitos humanos a partir de células madre pluripotentes inducidas (iPSCs) generadas a partir de eritoblastos de sangre periférica

Objetivos del trabajo: 1) Generar y caracterizar líneas de iPSCs a partir de eritoblastos de sangre periférica humana de pacientes con Síndrome QT largo e individuos sanos control, por medio de la transducción con vectores virales conteniendo las regiones codificantes de los genes de los factores de transcripción OCT3/4, SOX2, KLF4 y c-MYC. 2) Diferenciar dichas líneas de iPSCs a cardiomiocitos con la finalidad futura de modelar in vitro esta cardiopatía y realizar estudios comparativos a nivel morfológico y funcional entre las células derivadas de individuos sanos y pacientes, así como testear posibles drogas.Facultad de Ciencias Médica

Generación de cardiomiocitos humanos a partir de células madre pluripotentes inducidas (iPSCs) generadas a partir de eritoblastos de sangre periférica

Objetivos del trabajo: 1) Generar y caracterizar líneas de iPSCs a partir de eritoblastos de sangre periférica humana de pacientes con Síndrome QT largo e individuos sanos control, por medio de la transducción con vectores virales conteniendo las regiones codificantes de los genes de los factores de transcripción OCT3/4, SOX2, KLF4 y c-MYC. 2) Diferenciar dichas líneas de iPSCs a cardiomiocitos con la finalidad futura de modelar in vitro esta cardiopatía y realizar estudios comparativos a nivel morfológico y funcional entre las células derivadas de individuos sanos y pacientes, así como testear posibles drogas.Facultad de Ciencias Médica

Generación de cardiomiocitos humanos a partir de células madre pluripotentes inducidas (iPSCs) generadas a partir de eritoblastos de sangre periférica

Objetivos del trabajo: 1) Generar y caracterizar líneas de iPSCs a partir de eritoblastos de sangre periférica humana de pacientes con Síndrome QT largo e individuos sanos control, por medio de la transducción con vectores virales conteniendo las regiones codificantes de los genes de los factores de transcripción OCT3/4, SOX2, KLF4 y c-MYC. 2) Diferenciar dichas líneas de iPSCs a cardiomiocitos con la finalidad futura de modelar in vitro esta cardiopatía y realizar estudios comparativos a nivel morfológico y funcional entre las células derivadas de individuos sanos y pacientes, así como testear posibles drogas.Facultad de Ciencias Médica