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Previous issue date: 2017-09-01 / Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior (CAPES) / A diferencia??o das c?lulas-tronco mesenquimais humanas (CTMh) em osteoblasto segue um programa espec?fico de express?o g?nica, comprometendo-se inicalmente sob a influ?ncia das vias Wnt (Wingless pathway) e das BMPs (bone morphogenetic protein) que direcionam a c?lula a sua transforma??o em osteoblasto. Entretanto, as vias ativadas especificamente durante a fase inicial da diferencia??o s?o ainda pouco exploradas. Assim, o objetivo deste trabalho foi caracterizar o perfil transcricional do in?cio do processo de diferencia??o de CTMh, obtidas da veia do cord?o umbilical, em osteoblasto, a partir da t?cnica de RNA-Seq. A partir dos dados de transcriptoma, foi produzida uma lista ordenada de genes funcionalmente associados, obtida pela me?dia da expressa?o ge?nica tomada sobre genes vizinhos nesta lista, facilitando, assim, a sua interpretac?a?o biolo?gica. Para o estudo de ontologia g?nica e delineamento do perfil de express?o durante a osteog?nese, os processos metab?licos e fun??es moleculares significativamente alterados durante o decorrer do processo de diferencia??o foram analisados em diversas ferramentas (REVIGO, GOrila, PANTHER, LNCipedia e NONCODE) e descritos. Durante a indu??o ? diferencia??o das CTMh em osteoblasto, foi observado o aumento da express?o de genes caracter?sticos do fen?tipo osteobl?stico j? a partir do primeiro dia de diferencia??o. Foram identificados RNAs n?o codificantes, consoante a evolu??o da diferencia??o, bem como genes envolvidos na forma??o de rafts de membrana, j? a partir do terceiro dia de diferencia??o. Durante o terceiro dia de indu??o, genes envolvidos na regula??o da diferencia??o celular e em outros processos biol?gicos que precedem a diferencia??o, como ades?o celular, sinaliza??o e resposta ? fatores qu?micos externos j? apresentaram aumento em sua express?o. O estudo da express?o g?nica durante estes tr?s primeiros dias revelou ainda a diminui??o na express?o de genes envolvidos em processos biol?gicos de manuten??o de metabolismo basal, degrada??o de RNA e organiza??o do citoesqueleto, indicando assim que as mudan?as celulares que levam a c?lula a entrar em diferencia??o podem ter origem durante os tr?s primeiros dias de tratamento osteog?nico. / Human mesenchymal stem cells (hMSC) differentiation into osteoblast follows a specific gene expression program, committing itself initially under Wnt and BMPs pathways influence then differentiating into osteoblasts. However, specifically activated pathways during the first three days of differentiation are still poorly understood. Considering next generation sequencing technologies efficiency, and the lack of characterization in early hMSC differentiation process, in this work we used Illumina RNA-Seq to investigate the changes in these cells transcriptome. We used ex-vivo cultures of two human umbilical cord veins. Data from the complete transcriptome were analyzed in Transcriptogramer for the production of an ordered list of functionally associated genes, obtained by the mean of the gene expression taken on neighboring genes in this list, thus facilitating their biological interpretation. To study gene ontology and gene expression profile during osteogenesis, the metabolic processes and molecular functions significantly altered during the course of the differentiation process were analyzed in several tools (REVIGO, GOrila, PANTHER, LNCipedia and NONCODE) and properly described. During hMSC differentiation, an increase in expression of osteoblastic phenotype genes was observed as soon as the first day of differentiation started. Noncoding RNAs were also identified, depending on the evolution of the differentiation process, as well as genes involved in the formation of membrane rafts, on the third day of differentiation. During the third day of induction, cell differentiation regulation genes and other important genes on biological processes that precede differentiation, such as cell adhesion, signaling and external chemical factors response, have already increased expression. The study of gene expression during these first three days also revealed the decrease in the expression of groups of genes crucial to basal metabolism maintenance, RNA degradation and cytoskeleton organization, thus indicating that the cellular changes that lead the cell into differentiation may originate during the first three days of osteogenic treatment.
| Identifer | oai:union.ndltd.org:IBICT/oai:repositorio.ufrn.br:123456789/24461 |
| Date | 01 September 2017 |
| Creators | Rocha, Ana Carolina Pereira |
| Contributors | 32398336468, Lima, Daniel Chaves de, 06756927418, Marques, Joana Cristina Medeiros Tavares, 05623250424, Lima, Jo?o Paulo Matos Santos, 79332021368, Medeiros, Silvia Regina Batistuzzo de |
| Publisher | PROGRAMA DE P?S-GRADUA??O EM BIOQU?MICA, UFRN, Brasil |
| Source Sets | IBICT Brazilian ETDs |
| Language | Portuguese |
| Detected Language | English |
| Type | info:eu-repo/semantics/publishedVersion, info:eu-repo/semantics/masterThesis |
| Source | reponame:Repositório Institucional da UFRN, instname:Universidade Federal do Rio Grande do Norte, instacron:UFRN |
| Rights | info:eu-repo/semantics/openAccess |
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