Genome-wide analysis of PDX1 target genes in human pancreatic progenitors

Objective: Homozygous loss-of-function mutations in the gene coding for the homeobox transcription factor (TF) PDX1 leads to pancreatic agenesis, whereas heterozygous mutations can cause Maturity-Onset Diabetes of the Young 4 (MODY4). Although the function of Pdx1 is well studied in pre-clinical models during insulin-producing beta-cell development and homeostasis, it remains elusive how this TF controls human pancreas development by regulating a downstream transcriptional program. Also, comparative studies of PDX1 binding patterns in pancreatic progenitors and adult beta-cells have not been conducted so far. Furthermore, many studies reported the association between single nucleotide polymorphisms (SNPs) and T2DM, and it has been shown that islet enhancers are enriched in T2DM-associated SNPs. Whether regions, harboring T2DM-associated SNPs are PDX1 bound and active at the pancreatic progenitor stage has not been reported so far. Methods: In this study, we have generated a novel induced pluripotent stem cell (iPSC) line that efficiently differentiates into human pancreatic progenitors (PPs). Furthermore, PDX1 and H3K27ac chromatin immunoprecipitation sequencing (ChIP-seq) was used to identify PDX1 transcriptional targets and active enhancer and promoter regions. To address potential differences in the function of PDX1 during development and adulthood, we compared PDX1 binding profiles from PPs and adult islets. Moreover, combining ChIP-seq and GWAS meta-analysis data we identified T2DM-associated SNPs in PDX1 binding sites and active chromatin regions. Results: ChIP-seq for PDX1 revealed a total of 8088 PDX1-bound regions that map to 5664 genes in iPSC-derived PPs. The PDX1 target regions include important pancreatic TFs, such as PDX1 itself, RFX6, HNF1B, and ME1S1, which were activated during the differentiation process as revealed by the active chromatin mark H3K27ac and mRNA expression profiling, suggesting that auto-regulatory feedback regulation maintains PDX1 expression and initiates a pancreatic TF program. Remarkably, we identified several PDX1 target genes that have not been reported in the literature in human so far, including RFX3, required for ciliogenesis and endocrine differentiation in mouse, and the ligand of the Notch receptor DLL1, which is important for endocrine induction and tip-trunk patterning. The comparison of PDX1 profiles from PPs and adult human islets identified sets of stage-specific target genes, associated with early pancreas development and adult beta-cell function, respectively. Furthermore, we found an enrichment of T2DM-associated SNPs in active chromatin regions from iPSC-derived PPs. Two of these SNPs fall into PDX1 occupied sites that are located in the intronic regions of TCF7L2 and HNF1B. Both of these genes are key transcriptional regulators of endocrine induction and mutations in cis-regulatory regions predispose to diabetes. Conclusions: Our data provide stage-specific target genes of PDX1 during in vitro differentiation of stem cells into pancreatic progenitors that could be useful to identify pathways and molecular targets that predispose for diabetes. In addition, we show that T2DM-associated SNPs are enriched in active chromatin regions at the pancreatic progenitor stage, suggesting that the susceptibility to T2DM might originate from imperfect execution of a beta-cell developmental program

A psicoterapia analítica funcional na Alemanha : usos e aceitabilidade por terapeutas

Orientadora: Profª. Drª. Jocelaine Martins da SilveiraDissertação (mestrado) - Universidade Federal do Paraná, Setor de Ciências Humanas, Programa de Pós-Graduação em Psicologia. Defesa: Curitiba, 31/03/2017Inclui referências : f. 58-69Resumo: A Psicoterapia Analítica Funcional (FAP) é um modelo de psicoterapia com aproximadamente duas décadas de existência. Países sul-americanos e norteamericanos, como o Brasil, a Colômbia e os Estados Unidos apresentam grande número de publicações envolvendo a temática da FAP, o que demonstra haver boa aceitação dela por parte dos psicólogos na prática clínica. Por outro lado, na Europa existem registros de publicações apenas na Espanha. Em outros países existem apenas três registros de trabalhos publicados envolvendo a temática da FAP, sendo dois da Alemanha e um da Suíça. Esses dados sugerem que esta terapia ainda é desconhecida pela comunidade terapêutica destes países e consequentemente não é utilizada na prática clínica. Este estudo tem como objetivo investigar os usos e a forma de aceitação, por parte de terapeutas alemães, de técnicas referentes à FAP em intervenções no contexto clínico, para tal, procurar-se-à identificar com que frequência utilizam, na prática clínica, procedimentos similares às da FAP e, também, como avaliam os efeitos terapêuticos que resultam da aplicação destes procedimentos. Para tal, foram convidados a participar do estudo 1.586 terapeutas alemães, com conhecimento em fundamentos comportamentais, atuantes na área clínica. Os participantes responderam a um questionário denominado "A Psicoterapia Analítica Funcional na Alemanha - usos e aceitabilidade", contendo dez perguntas em escala Likert de quatro pontos e a duas questões de múltipla escolha. O programa Google Forms foi utilizado para a. Foi conduzido cálculo de desvio padrão, avaliando idade e tempo de experiência dos terapeutas. Foi conduzida uma análise descritiva e exploratória e análise fatorial multivariada. Os resultados indicaram que, no geral, os participantes aplicam no contexto clínico procedimentos que se assemelham às técnicas terapêuticas da FAP e os avaliam como terapêuticos. Palavras-Chaves: Psicoterapia Analítica Funcional, Psicologia na Alemanha, Terapias Contextuais, Terapia Comportamental, TransculturalidadeAbstract: The Functional Analytic Psychotherapy (FAP) is a model of psychotherapy with approximately two decades of existence. South and North American countries such as Brasil, Colombia and the United States present a great number of publications involving the thematic of FAP, which demonstrates a good acceptance of it by part of the psychologists in clinical practice. On the other hand, in Europe there are records of publications solely in Spain. In other countries there are only three records of papers published involving the thematic of FAP, two from Germany and one from Switzerland. This data suggests that this therapy is still unknown by the therapeutic community of these countries and therefore it isn't used in clinical practice. This study has the objective of studying the usages and the form of acceptance by german therapists to FAP related techniques in intervention in the clinical setting. To achieve this there will be and attempt on identifying the frequency used in clinical practice of procedures similar to those of FAP, as well as how the therapeutic effects that result from the application of these procedures are evaluated. Thus, 1,586 german clinical therapists with knowledge of behavioral fundaments were invited to participate in this study. The participants answered a questionaire denominated "Functional Analytic Psychotherapy in Germany - usages and acceptability", which contained ten questions in the Likert scale of four points and two multiple-choice questions. The program Google Forms was used for it. A standard deviation calculation was conducted, evaluating the age and experience time of the therapists. An exploratory and descriptive analysis was carried out, as well as a multivariate factor analysis. The results have indicated that, in general, the participants apply to the clinical context procedures that are similar to the therapeutic techniques of FAP and evaluate them as being therapeutic. Key-words: Functional Analytic Psychotherapy, Psychology in Germany, Contextual Therapies, Behavioral Therapy, Transculturatio

A psicoterapia analítica funcional na Alemanha : usos e aceitabilidade por terapeutas

Orientadora: Profª. Drª. Jocelaine Martins da SilveiraDissertação (mestrado) - Universidade Federal do Paraná, Setor de Ciências Humanas, Programa de Pós-Graduação em Psicologia. Defesa: Curitiba, 31/03/2017Inclui referências : f. 58-69Resumo: A Psicoterapia Analítica Funcional (FAP) é um modelo de psicoterapia com aproximadamente duas décadas de existência. Países sul-americanos e norteamericanos, como o Brasil, a Colômbia e os Estados Unidos apresentam grande número de publicações envolvendo a temática da FAP, o que demonstra haver boa aceitação dela por parte dos psicólogos na prática clínica. Por outro lado, na Europa existem registros de publicações apenas na Espanha. Em outros países existem apenas três registros de trabalhos publicados envolvendo a temática da FAP, sendo dois da Alemanha e um da Suíça. Esses dados sugerem que esta terapia ainda é desconhecida pela comunidade terapêutica destes países e consequentemente não é utilizada na prática clínica. Este estudo tem como objetivo investigar os usos e a forma de aceitação, por parte de terapeutas alemães, de técnicas referentes à FAP em intervenções no contexto clínico, para tal, procurar-se-à identificar com que frequência utilizam, na prática clínica, procedimentos similares às da FAP e, também, como avaliam os efeitos terapêuticos que resultam da aplicação destes procedimentos. Para tal, foram convidados a participar do estudo 1.586 terapeutas alemães, com conhecimento em fundamentos comportamentais, atuantes na área clínica. Os participantes responderam a um questionário denominado "A Psicoterapia Analítica Funcional na Alemanha - usos e aceitabilidade", contendo dez perguntas em escala Likert de quatro pontos e a duas questões de múltipla escolha. O programa Google Forms foi utilizado para a. Foi conduzido cálculo de desvio padrão, avaliando idade e tempo de experiência dos terapeutas. Foi conduzida uma análise descritiva e exploratória e análise fatorial multivariada. Os resultados indicaram que, no geral, os participantes aplicam no contexto clínico procedimentos que se assemelham às técnicas terapêuticas da FAP e os avaliam como terapêuticos. Palavras-Chaves: Psicoterapia Analítica Funcional, Psicologia na Alemanha, Terapias Contextuais, Terapia Comportamental, TransculturalidadeAbstract: The Functional Analytic Psychotherapy (FAP) is a model of psychotherapy with approximately two decades of existence. South and North American countries such as Brasil, Colombia and the United States present a great number of publications involving the thematic of FAP, which demonstrates a good acceptance of it by part of the psychologists in clinical practice. On the other hand, in Europe there are records of publications solely in Spain. In other countries there are only three records of papers published involving the thematic of FAP, two from Germany and one from Switzerland. This data suggests that this therapy is still unknown by the therapeutic community of these countries and therefore it isn't used in clinical practice. This study has the objective of studying the usages and the form of acceptance by german therapists to FAP related techniques in intervention in the clinical setting. To achieve this there will be and attempt on identifying the frequency used in clinical practice of procedures similar to those of FAP, as well as how the therapeutic effects that result from the application of these procedures are evaluated. Thus, 1,586 german clinical therapists with knowledge of behavioral fundaments were invited to participate in this study. The participants answered a questionaire denominated "Functional Analytic Psychotherapy in Germany - usages and acceptability", which contained ten questions in the Likert scale of four points and two multiple-choice questions. The program Google Forms was used for it. A standard deviation calculation was conducted, evaluating the age and experience time of the therapists. An exploratory and descriptive analysis was carried out, as well as a multivariate factor analysis. The results have indicated that, in general, the participants apply to the clinical context procedures that are similar to the therapeutic techniques of FAP and evaluate them as being therapeutic. Key-words: Functional Analytic Psychotherapy, Psychology in Germany, Contextual Therapies, Behavioral Therapy, Transculturatio

Generation of a human induced pluripotent stem cell (iPSC) line from a patient carrying a P33T mutation in the PDX1 gene

Homozygous loss-of-function mutations in the gene coding for the homeobox transcription factor PDX1 leads to pancreatic agenesis, whereas certain heterozygous point mutations are associated with Maturity-Onset Diabetes of the Young 4 (MODY4) and Type 2 Diabetes Mellitus (T2DM). To understand the pathomechanism of MODY4 and T2DM, we have generated iPSCs from a woman with a P33T heterozygous mutation in the transactivation domain of PDX1. The resulting PDX1 P33T iPSCs generated by episomal reprogramming are integration-free, have a normal karyotype and are pluripotent in vitro and in vivo. Taken together, this iPSC line will be useful to study diabetes pathomechanisms

Generation of a human induced pluripotent stem cell (iPSC) line from a patient with family history of diabetes carrying a C18R mutation in the PDX1 gene

Homozygous loss-of-function mutations in the gene coding for the homeobox transcription factor PDX1 leads to pancreatic agenesis, whereas certain heterozygous point mutations are associated with Maturity-Onset Diabetes of the Young 4 (MODY4) and Type 2 Diabetes Mellitus (T2DM). To understand the pathomechanism of MODY4 and T2DM, we have generated iPSCs from a woman with a C18R heterozygous mutation in the transactivation domain of PDX1. The resulting PDX1 C18R iPSCs generated by episomal reprogramming are integration-free, have a normal karyotype and are pluripotent in vitro and in vivo. Taken together, this iPSC line will be useful to study diabetes pathomechanisms