Mucopolysaccharidosis type I (or Hurler syndrome) is a rare genetic disorder, caused by mutations in the idua gene, resulting in the deficiency of α-L-iduronidase enzyme activity and intra-cellular accumulation of glycosaminoglycans. The aim of the present project was focused on the isolation and characterization of two different stem cell populations, multipotent and pluripotent, derived from patients affected by Hurler syndrome. We aim to use Mesenchymal Stem Cells (MSCs) and induced Pluripotent Stem Cells (iPSCs) as a tool to explore still unknown disease mechanisms involved in the genetic metabolic disorder of our interest. Our recently published study focused on the characterization of MSCs isolated from bone marrow of Hurler patients. The MSCs were characterized for their expansion rate, phenotype, telomerase activity, IDUA activity and differentiation capacity towards adipocytes, osteoblasts, chondrocytes and smooth muscle cells in vitro. Interestingly, affected MSCs displayed increased capacity to support osteoclastogenesis according to the upregulation of the RANKL/RANK/OPG molecular pathway in Hurler MSCs. The second study describes the isolation of iPSCs from fibroblasts of Hurler patients. The generated cell lines were fully characterized for their pluripotency markers, gene expression profile, viral copy number integration and differentiation potential both in vitro and in vivo. As a proof of principle, we are attempting to gene correct patient-derived iPSCs with an alternative and safer method than viral vectors, using a Zinc Finger Nucleases-mediated approach for gene targeting of pluripotent cells.

(2012). Exploring hurler syndrome through the study of disease-specific multipotent and pluripotent stem cells. (Tesi di dottorato, Università degli Studi di Milano-Bicocca, 2012).

Exploring hurler syndrome through the study of disease-specific multipotent and pluripotent stem cells

GATTO, FRANCESCA
2012

Abstract

Mucopolysaccharidosis type I (or Hurler syndrome) is a rare genetic disorder, caused by mutations in the idua gene, resulting in the deficiency of α-L-iduronidase enzyme activity and intra-cellular accumulation of glycosaminoglycans. The aim of the present project was focused on the isolation and characterization of two different stem cell populations, multipotent and pluripotent, derived from patients affected by Hurler syndrome. We aim to use Mesenchymal Stem Cells (MSCs) and induced Pluripotent Stem Cells (iPSCs) as a tool to explore still unknown disease mechanisms involved in the genetic metabolic disorder of our interest. Our recently published study focused on the characterization of MSCs isolated from bone marrow of Hurler patients. The MSCs were characterized for their expansion rate, phenotype, telomerase activity, IDUA activity and differentiation capacity towards adipocytes, osteoblasts, chondrocytes and smooth muscle cells in vitro. Interestingly, affected MSCs displayed increased capacity to support osteoclastogenesis according to the upregulation of the RANKL/RANK/OPG molecular pathway in Hurler MSCs. The second study describes the isolation of iPSCs from fibroblasts of Hurler patients. The generated cell lines were fully characterized for their pluripotency markers, gene expression profile, viral copy number integration and differentiation potential both in vitro and in vivo. As a proof of principle, we are attempting to gene correct patient-derived iPSCs with an alternative and safer method than viral vectors, using a Zinc Finger Nucleases-mediated approach for gene targeting of pluripotent cells.
Campo DC Valore Lingua
dc.authority.academicField2000 MED/38 - PEDIATRIA GENERALE E SPECIALISTICA en
dc.authority.advisor BIONDI, ANDREA en
dc.authority.people GATTO, FRANCESCA en
dc.authority.phdCourse MEDICINA TRASLAZIONALE E MOLECOLARE (DIMET) - 45R en
dc.authority.phdSchool Scuola di Dottorato in Medicina Traslazionale e Molecolare en
dc.collection.id.s e39773c1-7ce8-35a3-e053-3a05fe0aac26 *
dc.collection.name 07 - Tesi di dottorato Bicocca post 2009 *
dc.coverage.academiccycle 24 en
dc.coverage.academicyear 2010/2011 en
dc.date.accessioned 2012/04/05 15:38:33 -
dc.date.available 2012/04/05 15:38:33 -
dc.date.issued 2012-03-26 -
dc.description.abstract Mucopolysaccharidosis type I (or Hurler syndrome) is a rare genetic disorder, caused by mutations in the idua gene, resulting in the deficiency of α-L-iduronidase enzyme activity and intra-cellular accumulation of glycosaminoglycans. The aim of the present project was focused on the isolation and characterization of two different stem cell populations, multipotent and pluripotent, derived from patients affected by Hurler syndrome. We aim to use Mesenchymal Stem Cells (MSCs) and induced Pluripotent Stem Cells (iPSCs) as a tool to explore still unknown disease mechanisms involved in the genetic metabolic disorder of our interest. Our recently published study focused on the characterization of MSCs isolated from bone marrow of Hurler patients. The MSCs were characterized for their expansion rate, phenotype, telomerase activity, IDUA activity and differentiation capacity towards adipocytes, osteoblasts, chondrocytes and smooth muscle cells in vitro. Interestingly, affected MSCs displayed increased capacity to support osteoclastogenesis according to the upregulation of the RANKL/RANK/OPG molecular pathway in Hurler MSCs. The second study describes the isolation of iPSCs from fibroblasts of Hurler patients. The generated cell lines were fully characterized for their pluripotency markers, gene expression profile, viral copy number integration and differentiation potential both in vitro and in vivo. As a proof of principle, we are attempting to gene correct patient-derived iPSCs with an alternative and safer method than viral vectors, using a Zinc Finger Nucleases-mediated approach for gene targeting of pluripotent cells. -
dc.description.allpeople Gatto, F -
dc.description.allpeopleoriginal Gatto, -
dc.description.codicestruttura 1692 it
dc.description.doctoreuropaeus No en
dc.description.fulltext open en
dc.description.fulltextoriginal open en
dc.description.numberofauthors 1 -
dc.identifier.citation (2012). Exploring hurler syndrome through the study of disease-specific multipotent and pluripotent stem cells. (Tesi di dottorato, Università degli Studi di Milano-Bicocca, 2012). en
dc.identifier.uri http://hdl.handle.net/10281/29890 -
dc.language.iso eng en
dc.publisher.country Italy -
dc.publisher.name Università degli Studi di Milano-Bicocca -
dc.relation.alleditors SERAFINI, MARTA en
dc.subject.keywords GAGs, Hurler syndrome, bone marrow stromal cells, induced Pluripotent Stem Cells -
dc.subject.singlekeyword GAGs *
dc.subject.singlekeyword Hurler syndrome *
dc.subject.singlekeyword bone marrow stromal cells *
dc.subject.singlekeyword induced Pluripotent Stem Cells *
dc.title Exploring hurler syndrome through the study of disease-specific multipotent and pluripotent stem cells en
dc.type Tesi di dottorato -
dc.type.driver info:eu-repo/semantics/doctoralThesis -
dc.type.full Pubblicazioni::07 - Tesi di dottorato Bicocca post 2009 it
dc.type.miur -2.0 -
iris.bncf.datainvio 2024/11/25 18:17:44 *
iris.bncf.handle 20.500.14242/170009 *
iris.bncf.nbn URN:NBN:IT:UNIMIB-170009 *
iris.bncf.stato 2 *
iris.bncf.uuid 82d87c53-43b6-471a-ad12-2160427ace88 *
iris.orcid.lastModifiedDate 2023/12/21 12:08:02 *
iris.orcid.lastModifiedMillisecond 1703156882853 *
iris.sitodocente.maxattempts 1 -
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