Therapeutic Potential of RNAi Through Endocytotic Methods

The ability to manipulate gene expression promises to be an important tool for the management of infectious diseases and genetic disorders. However, a major limitation to effective delivery of therapeutic RNA to living cells is the cellular toxicity of conventional techniques. Team PANACEA’s research objective was to create new reagents based on a novel small-molecule delivery system that uses a modular recombinant protein vehicle consisting of a specific ligand coupled to a Hepatitis B Virus-derived RNA binding domain (HBV-RBD). Two such recombinant delivery proteins were developed: one composed of Interleukin-8, the other consisting of the Machupo Virus GP1 protein. The ability of these proteins to deliver RNA to cells were then tested. The non-toxic nature of this technology has the potential to overcome limitations of current methods and could provide a platform for the expansion of personalized medicine.

Aplicação de células estaminais dentárias na Medicina e na Medicina Dentária

A temática relativa às células estaminais inicia-se na década de 60 com a descoberta da primeira fonte viável destas células: a medula óssea. Diversos estudos permitiram definir a sua função de renovação tecidular e regeneração pós-dano, assim como a sua caraterização num grupo heterogéneo de células indiferenciadas, clonogénicas, definidas pela capacidade de auto-renovação e diferenciação em células maduras. Nos últimos anos, estas células ganharam popularidade face à alternativa terapêutica que representam para muitas doenças, tais como: diabetes, anomalias congénitas, danos do tecido nervoso, Parkinson, Alzheimer e outras alterações degenerativas, exposições pulpares, defeitos periodontais e perda do órgão dentário. Apesar do seu potencial terapêutico, apresentam vários efeitos adversos, especialmente em relação ao seu envolvimento direto (via transformação maligna das MSCs) e indireto (via efeito modulatório das MSCs) no desenvolvimento do cancro. Preconiza-se o seu uso no âmbito da Engenharia Tecidular, introduzindo o processo de regeneração tecidular através da utilização combinada de biomateriais e mediadores biológicos, a fim de proporcionar novas ferramentas para a medicina regenerativa. Mais tarde, tornou-se possível identificar cinco populações de células estaminais de origem dentária (DPSCs, SHEDs, DFPCs, SCAPs e PDLCs) que, para além da sua multipotência e capacidade de diferenciação, constituem fontes acessíveis para recolha. O isolamento destas células constitui ainda uma prática relativamente recente, na qual se torna preponderante isolar células com fenótipo pré-determinado e cultivá-las em meios de cultura adequados. Estudos comprovam que o método de isolamento e as condições de cultura utilizados podem dar origem a diferentes linhas celulares. A conservação é uma prática baseada na convicção de que a medicina regenerativa é o caminho mais promissor para o desenvolvimento da medicina personalizada. Informação adicional relativa à terapia com células estaminais é ainda necessária. Esta utiliza princípios de biomimética altamente desejáveis, pelo que os resultados obtidos têm vindo a despoletar grandes expetativas e a sua implementação na Engenharia Tecidular apresenta-se promissora.

Proteomics in the assessment of the therapeutic response of antineoplastic drugs: strategies and practical applications

Uncovering mechanisms of unknown pathological mechanisms and body response to applied medication are the drive forces toward personalized medicine. In this post-genomic era, all eyes are tuned to proteomic field, searching for the answers and explanations by investigating the final physiological functional units – proteins and their proteoforms. Development of cutting-edge mass spectrometric technologies and powerful bioinformatics tools, allowed life-science community mining of disease-specific proteins as biomarkers, which are often hidden by high complexity of the samples and/or small abundance. Nowadays, there are several proteomics-based approaches to study the proteome. This chapter focuses on gold standard proteomics strategies and related issues towards candidate biomarker discovery, which may have diagnostic/prognostic as well as mechanistic utility.

NGS y Metagenómica

The Next Generation Sequencing (NGS) allows to sequence the whole genome of an organism, compared to Maxam and Gilbert and Sanger sequencing that only allow to sequence, hardly, a single gene. Removing the separation of DNA fragments by electrophoresis, and the development of techniques that let the parallelization (analysing simultaneously several DNA fragments) have been crucial for the improvements of this process. The new companies in this ambit, Roche and Illumina, bet for different protocols to achieve these goals. Illumina bets for the sequencing by synthesis (SBS), requiring the library preparation and the use of adapters. Likewise, Illumina has replaced Roche because its lower rate of misincorporation, making it ideal for studies of genetic variability, transcriptomic, epigenomic, and metagenomic, in which this study will focus. However, it is noteworthy that the last progress in sequencing is carried out by the third generation sequencing, using nanotechnology to design small sequencers that sequence the whole genome of an organism quickly and inexpensively. Moreover, they provide more reliable data than current systems because they sequence a single molecule, solving the problem of synchronisation. In this way, PacBio and Nanopore allow a great progress in diagnostic and personalized medicine. Metagenomics provide to make a qualitative and quantitative analysis of the various species present in a sample. The main advantage of this technique is the no necessary isolation and growth of the species, allowing the analysis of nonculturable species. The Illumina protocol studies the variable regions of the 16S rRNA gene, which contains variable and not variables regions providing a phylogenetic classification. Therefore, metagenomics is a topic of interest to know the biodiversity of complex ecosystems and to study the microbiome of patients given the high involvement with certain microbial profiles on the condition of certain metabolic diseases.

Eficacia de terapias biológicas de tipo anti-angiogénico en el cáncer colorrectal metastásico revisión sistemática de la literatura

Introducción: El cáncer colorrectal (CCR) se encuentra entre los 5 tipos de cáncer con mayor incidencia a nivel mundial. Alrededor del 20% de los casos son diagnosticados en estadios metastásico, donde el tratamiento inicialmente era quimioterapia con una supervivencia global a 5 años de 12 a 14 meses. Es así que se investiga el papel de la angiogénesis tumoral, orientado al desarrollo de terapias, implementando su uso en estadios avanzados. Metodología: Se realizó una búsqueda sistemática en las bases de datos Embase, PubMed, SciELO y LILIACS con términos estandarizados a través de la herramienta MeSH y DECS bajo los lineamientos establecidos en las guías de revisiones sistemáticas y meta-análisis (Manual Cochrane). Se tomaron estudios clínicos aleatorizados controlados con pacientes con CCR metastásico, que hayan recibido quimioterapia sola o combinada con terapias antiangiogénicas, publicados en inglés y español entre el 2003 y 2013. Resultados: 6 artículos cumplieron con criterios de inclusión. Estos reportaron 15.8 meses en promedio de supervivencia global en el tratamiento de quimioterapia asociada a terapias biológicas frente a 14.4 meses con solo quimioterapia. Los eventos adversos de tipo vascular aumentaron más en el grupo de antiangiogénicos, reportando muertes debidas a perforaciones intestinales. Conclusiones: Los regímenes de quimioterapia asociadas a terapias antiangiogénicas brindan una mayor supervivencia global y libre de progresión, al igual que mayor número de tasas de respuesta. Son terapias con eventos adversos importantes pero que deberá seleccionarse bien al paciente para disminuir su riesgo de eventos. Palabras claves: Cáncer colorrectal metastásico, terapia anti-angiogénica, quimioterapia en segunda línea, receptor del factor de crecimiento de endotelio vascular, supervivencia global.

Revisión sistemática utilidad de asa y calcio con o sin vitamina d en la prevención de cáncer colorrectal y la recurrencia de pólipos adenomatosos en paciente con riesgo de padecer esta patología

Introducción: El cáncer colorrectal es una patología con alto impacto en la salud pública, debido a su prevalencia, incidencia, severidad, costo e impacto en la salud mental y física del individuo y la familia. Ensayos clínicos realizados en pacientes con antecedente de infarto al miocardio que consumían ácido acetil salicílico (asa), calcio con y sin vitamina D, mostraron asociación entre el consumo de estos medicamentos y disminución en la incidencia en cáncer colorrectal y pólipos adenomatosos. Objetivo: Evaluar la literatura sobre el uso de asa, calcio con y sin vitamina D con relación a su impacto en la prevención del cáncer colorrectal y pólipos adenomatosos. Métodos: Se realizó revisión sistemática buscando ensayos clínicos realizados en pacientes con factores de riesgo para cáncer colorrectal y pólipos adenomatosos que usaron asa, calcio con y sin vitamina D fueron incluidos. Resultados: se escogieron 105 para la revisión sistemática. Conclusiones: Es necesario desarrollar más estudios que lleven a evaluar el efecto protector de la aspirina, calcio y vitamina D. En los artículos revisados la aspirina a dosis de 81 a 325 mg día se correlaciona con reducción de riesgo de aparición de CRC aunque la dosis ideal, el tiempo de inicio y la duración de la ingesta continua no son claros. Hacen falta estudios que comparen poblaciones con ingesta de asa a diferentes dosis.

Factores asociados a respuesta de abatacept subcutaneo en artritis reumatoide

INTRODUCCION Dado que la artritis reumatoide es la artropatía inflamatoria más frecuente en el mundo, siendo altamente discapacitante y causando gran impacto de alto costo, se busca ofrecer al paciente opciones terapéuticas y calidad de vida a través del establecimiento de un tratamiento oportuno y eficaz, teniendo presentes aquellos predictores de respuesta previo a instaurar determinada terapia. Existen pocos estudios que permitan establecer aquellos factores de adecuada respuesta para inicio de terapia biológica con abatacept, por lo cual en este estudio se busca determinar cuáles son esos posibles factores. METODOLOGIA Estudio analítico de tipo corte transversal de 94 pacientes con diagnóstico de AR, evaluados para determinar las posibles variables que influyen en la respuesta a terapia biológica con abatacept. Se incluyeron 67 de los 94 pacientes al modelo de regresión logística, que son aquellos pacientes en que fue posible medir la respuesta al tratamiento (respuesta EULAR) a través de la determinación del DAS 28 y así discriminar en dos grupos de comparación (respuesta y no respuesta). DISCUSION DE RESULTADOS La presencia de alta actividad de la enfermedad al inicio de la terapia biológica, aumenta la probabilidad de respuesta al tratamiento respecto al grupo con baja/moderada actividad de la enfermedad; OR 4,19 - IC 95%(1,18 – 14.9), (p 0,027). La ausencia de erosiones óseas aumenta la probabilidad de presentar adecuada respuesta a la terapia biológica respecto aquellos con erosiones, con un OR 3,1 (1,01-9,55), (p 0,048). Niveles de VSG y presencia de manifestaciones extra-articulares son otros datos de interés encontrados en el análisis bivariado. Respecto a las variables o características como predictores de respuesta al tratamiento con abatacept, se encuentran estudios que corroboran los hallazgos de este estudio, respecto al alto puntaje del DAS 28 al inicio de la terapia (9, 12). CONCLUSIONES Existen distintas variables que determinan la respuesta a los diferentes biológicos para manejo de AR. Es imprescindible evaluar dichos factores de manera individual con el fin de lograr de manera efectiva el control de la enfermedad y así mejorar la calidad de vida del individuo (medicina personalizada). Existen variables tales como la alta actividad de la enfermedad y la ausencia de erosiones como predictores de respuesta en la terapia con abatacept.

Personalized cancer medicine: molecular diagnostics, predictive biomarkers, and drug resistance.

The progressive elucidation of the molecular pathogenesis of cancer has fueled the rational development of targeted drugs for patient populations stratified by genetic characteristics. Here we discuss general challenges relating to molecular diagnostics and describe predictive biomarkers for personalized cancer medicine. We also highlight resistance mechanisms for epidermal growth factor receptor (EGFR) kinase inhibitors in lung cancer. We envisage a future requiring the use of longitudinal genome sequencing and other omics technologies alongside combinatorial treatment to overcome cellular and molecular heterogeneity and prevent resistance caused by clonal evolution.

Infection systems biology:from reactive to proactive (P4) medicine

This short review establishes the conceptual bases and discusses the principal aspects of P4-shorthand for predictive, preventive, personalized and participatory medicine-medicine, in the framework of infectious diseases. P4 medicine is a new way to approach medical care; instead of acting when the patient is sick, physicians will be able to detect early warnings of disease to take early action. Furthermore, people might even be able to adjust their lifestyles to prevent disease. P4 medicine is fuelled by systems approaches to disease, including methods for personalized genome sequencing and new computational techniques for building dynamic disease predictive networks from massive amounts of data from a variety of OMICs. An excellent example of the effectiveness of the P4 medicine approach is the change in cancer treatments. Emphasis is placed on early detection, followed by genotyping of the patient to use the most adequate treatment according to the genetic background. Cardiovascular diseases and perhaps even neurodegenerative disorders will be the next targets for P4 medicine. The application of P4 medicine to infectious diseases is still in its infancy, but is a promising field that will provide much benefit to both the patients and the health-care system.

Targeting the undruggable: immunotherapy meets personalized oncology in the genomic era.

Owing to recent advances in genomic technologies, personalized oncology is poised to fundamentally alter cancer therapy. In this paradigm, the mutational and transcriptional profiles of tumors are assessed, and personalized treatments are designed based on the specific molecular abnormalities relevant to each patient's cancer. To date, such approaches have yielded impressive clinical responses in some patients. However, a major limitation of this strategy has also been revealed: the vast majority of tumor mutations are not targetable by current pharmacological approaches. Immunotherapy offers a promising alternative to exploit tumor mutations as targets for clinical intervention. Mutated proteins can give rise to novel antigens (called neoantigens) that are recognized with high specificity by patient T cells. Indeed, neoantigen-specific T cells have been shown to underlie clinical responses to many standard treatments and immunotherapeutic interventions. Moreover, studies in mouse models targeting neoantigens, and early results from clinical trials, have established proof of concept for personalized immunotherapies targeting next-generation sequencing identified neoantigens. Here, we review basic immunological principles related to T-cell recognition of neoantigens, and we examine recent studies that use genomic data to design personalized immunotherapies. We discuss the opportunities and challenges that lie ahead on the road to improving patient outcomes by incorporating immunotherapy into the paradigm of personalized oncology.

Personalized therapy in pain management: where do we stand?

Genomic variations influencing response to pharmacotherapy of pain are currently under investigation. Drug-metabolizing enzymes represent a major target of ongoing research in order to identify associations between an individual's drug response and genetic profile. Polymorphisms of the cytochrome P450 enzymes (CYP2D6) influence metabolism of codeine, tramadol, hydrocodone, oxycodone and tricyclic antidepressants. Blood concentrations of some NSAIDs depend on CYP2C9 and/or CYP2C8 activity. Genomic variants of these genes associate well with NSAIDs' side effect profile. Other candidate genes, such as those encoding (opioid) receptors, transporters and other molecules important for pharmacotherapy in pain management, are discussed; however, study results are often equivocal. Besides genetic variants, further variables, for example, age, disease, comorbidity, concomitant medication, organ function as well as patients' compliance, may have an impact on pharmacotherapy and need to be addressed when pain therapists prescribe medication. Although pharmacogenetics as a diagnostic tool has the potential to improve patient therapy, well-designed studies are needed to demonstrate superiority to conventional dosing regimes.

Systems medicine and integrated care to combat chronic noncommunicable diseases

We propose an innovative, integrated, cost-effective health system to combat major non-communicable diseases (NCDs), including cardiovascular, chronic respiratory, metabolic, rheumatologic and neurologic disorders and cancers, which together are the predominant health problem of the 21st century. This proposed holistic strategy involves comprehensive patient-centered integrated care and multi-scale, multi-modal and multi-level systems approaches to tackle NCDs as a common group of diseases. Rather than studying each disease individually, it will take into account their intertwined gene-environment, socio-economic interactions and co-morbidities that lead to individual-specific complex phenotypes. It will implement a road map for predictive, preventive, personalized and participatory (P4) medicine based on a robust and extensive knowledge management infrastructure that contains individual patient information. It will be supported by strategic partnerships involving all stakeholders, including general practitioners associated with patient-centered care. This systems medicine strategy, which will take a holistic approach to disease, is designed to allow the results to be used globally, taking into account the needs and specificities of local economies and health systems.

Adaptive Algorithms for Personalized Diabetes Treatment

Dynamic systems, especially in real-life applications, are often determined by inter-/intra-variability, uncertainties and time-varying components. Physiological systems are probably the most representative example in which population variability, vital signal measurement noise and uncertain dynamics render their explicit representation and optimization a rather difficult task. Systems characterized by such challenges often require the use of adaptive algorithmic solutions able to perform an iterative structural and/or parametrical update process towards optimized behavior. Adaptive optimization presents the advantages of (i) individualization through learning of basic system characteristics, (ii) ability to follow time-varying dynamics and (iii) low computational cost. In this chapter, the use of online adaptive algorithms is investigated in two basic research areas related to diabetes management: (i) real-time glucose regulation and (ii) real-time prediction of hypo-/hyperglycemia. The applicability of these methods is illustrated through the design and development of an adaptive glucose control algorithm based on reinforcement learning and optimal control and an adaptive, personalized early-warning system for the recognition and alarm generation against hypo- and hyperglycemic events.