Cantilena, Bruno (2010) Identification of genes that regulate bone marrow stromal cell differentiation by RNA interference. [Tesi di dottorato] (Unpublished)


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Item Type: Tesi di dottorato
Resource language: English
Title: Identification of genes that regulate bone marrow stromal cell differentiation by RNA interference
Date: 30 January 2010
Number of Pages: 124
Institution: Università degli Studi di Napoli Federico II
Istituzioni (extra): CEINGE  Biotecnologie Avanzate
Department: CEINGE Biotecnologie avanzate
Scuola di dottorato: SEMM – European School of Molecular Medicine
Dottorato: PhD in Molecular Medicine (Molecular Oncology or Human Genetics)
Ciclo di dottorato: 21
Coordinatore del Corso di dottorato:
Karsenty, GerardUNSPECIFIED
Date: 30 January 2010
Number of Pages: 124
Keywords: Bone, osteoblast, mesenchymal stem cells, rna interference, differentiation
Settori scientifico-disciplinari del MIUR: Area 05 - Scienze biologiche > BIO/11 - Biologia molecolare
Area 05 - Scienze biologiche > BIO/13 - Biologia applicata
Additional information: Ciclo III/XXI, Curriculum Human Genetics
Date Deposited: 05 Feb 2010 16:16
Last Modified: 14 Jan 2015 12:11
DOI: 10.6092/UNINA/FEDOA/4318

Collection description

Mesenchymal stem cells (MSCs) can be isolated from bone marrow and can differentiate in a variety of cell types, including osteoblasts, adipocytes, chondrocytes, myoblasts, hepatocytes, and neural cells. For this reason, MSCs are very promising for the development of cell-based therapy for degenerative diseases. Understanding the mechanisms behind MSC cell fate determination is not easy, since the molecular processes that drive differentiation are complex and poorly understood. Even though in the recent years many improvements have been done, many mechanism need to be elucidated. We planned to investigate on differentiation mechanisms of MSC and, in particular, we focused our activities towards osteoblast differentiation; at this aim, we have silenced specific mRNAs using a mouse shRNA library in a 96 well plate-based screening strategy. With this methodology we identified sevral genes that are possible candidates for a role in osteoblast differentiation. Among the candidate genes, a consistent fraction was represented by genes whose function is still unknown: we focused our attention on a short number of these genes that we named ObI (Osteoblast inducer). In this thesis project we focused our experiments on a particular gene, that we named ObI-1. We also considered genes identified during the screening with a known function but an unknown role during the osteogenic differentiation process. Among these genes we focused our attention on Angiotensin receptor 1 (AT1R), whose role in osteogenic differentiation had been previously suggested. For both candidates that we analyzed for this project we confirmed the effects of silencing on osteogenic differentiation and we analyzed expression in tissues and in our cell culture system. Furthermore, we characterized function and mechanism of action in osteoblastogenesis for both genes.


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