Gene Modification and Three‐Dimensional Scaffolds as Novel Tools to Allow the Use of Postnatal Thymic Epithelial Cells for Thymus Regeneration Approaches

Abstract Defective functionality of thymic epithelial cells (TECs), due to genetic mutations or injuring causes, results in altered T‐cell development, leading to immunodeficiency or autoimmunity. These defects cannot be corrected by hematopoietic stem cell transplantation (HSCT), and thymus transpl...

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Autores principales:	Ileana Bortolomai, Monica Sandri, Elena Draghici, Elena Fontana, Elisabetta Campodoni, Genni Enza Marcovecchio, Francesca Ferrua, Laura Perani, Antonello Spinelli, Tamara Canu, Marco Catucci, Tiziano Di Tomaso, Lucia Sergi Sergi, Antonio Esposito, Angelo Lombardo, Luigi Naldini, Anna Tampieri, Georg A. Hollander, Anna Villa, Marita Bosticardo
Formato:	article
Lenguaje:	EN
Publicado:	Wiley 2019
Materias:	3D collagen scaffolds Lentiviral vector Thymic epithelial cells Thymic regeneration Medicine (General) R5-920 Cytology QH573-671
Acceso en línea:	https://doaj.org/article/84a5cdcf9f7d4b5e98536d3398bf01f3
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spelling	oai:doaj.org-article:84a5cdcf9f7d4b5e98536d3398bf01f32021-11-30T19:15:37ZGene Modification and Three‐Dimensional Scaffolds as Novel Tools to Allow the Use of Postnatal Thymic Epithelial Cells for Thymus Regeneration Approaches2157-65802157-656410.1002/sctm.18-0218https://doaj.org/article/84a5cdcf9f7d4b5e98536d3398bf01f32019-10-01T00:00:00Zhttps://doi.org/10.1002/sctm.18-0218https://doaj.org/toc/2157-6564https://doaj.org/toc/2157-6580Abstract Defective functionality of thymic epithelial cells (TECs), due to genetic mutations or injuring causes, results in altered T‐cell development, leading to immunodeficiency or autoimmunity. These defects cannot be corrected by hematopoietic stem cell transplantation (HSCT), and thymus transplantation has not yet been demonstrated to be fully curative. Here, we provide proof of principle of a novel approach toward thymic regeneration, involving the generation of thymic organoids obtained by seeding gene‐modified postnatal murine TECs into three‐dimensional (3D) collagen type I scaffolds mimicking the thymic ultrastructure. To this end, freshly isolated TECs were transduced with a lentiviral vector system, allowing for doxycycline‐induced Oct4 expression. Transient Oct4 expression promoted TECs expansion without drastically changing the cell lineage identity of adult TECs, which retain the expression of important molecules for thymus functionality such as Foxn1, Dll4, Dll1, and AIRE. Oct4‐expressing TECs (iOCT4 TEC) were able to grow into 3D collagen type I scaffolds both in vitro and in vivo, demonstrating that the collagen structure reproduced a 3D environment similar to the thymic extracellular matrix, perfectly recognized by TECs. In vivo results showed that thymic organoids transplanted subcutaneously in athymic nude mice were vascularized but failed to support thymopoiesis because of their limited in vivo persistence. These findings provide evidence that gene modification, in combination with the usage of 3D biomimetic scaffolds, may represent a novel approach allowing the use of postnatal TECs for thymic regeneration. Stem Cells Translational Medicine 2019;8:1107–1122Ileana BortolomaiMonica SandriElena DraghiciElena FontanaElisabetta CampodoniGenni Enza MarcovecchioFrancesca FerruaLaura PeraniAntonello SpinelliTamara CanuMarco CatucciTiziano Di TomasoLucia Sergi SergiAntonio EspositoAngelo LombardoLuigi NaldiniAnna TampieriGeorg A. HollanderAnna VillaMarita BosticardoWileyarticle3D collagen scaffoldsLentiviral vectorThymic epithelial cellsThymic regenerationMedicine (General)R5-920CytologyQH573-671ENStem Cells Translational Medicine, Vol 8, Iss 10, Pp 1107-1122 (2019)
institution	DOAJ
collection	DOAJ
language	EN
topic	3D collagen scaffolds Lentiviral vector Thymic epithelial cells Thymic regeneration Medicine (General) R5-920 Cytology QH573-671
spellingShingle	3D collagen scaffolds Lentiviral vector Thymic epithelial cells Thymic regeneration Medicine (General) R5-920 Cytology QH573-671 Ileana Bortolomai Monica Sandri Elena Draghici Elena Fontana Elisabetta Campodoni Genni Enza Marcovecchio Francesca Ferrua Laura Perani Antonello Spinelli Tamara Canu Marco Catucci Tiziano Di Tomaso Lucia Sergi Sergi Antonio Esposito Angelo Lombardo Luigi Naldini Anna Tampieri Georg A. Hollander Anna Villa Marita Bosticardo Gene Modification and Three‐Dimensional Scaffolds as Novel Tools to Allow the Use of Postnatal Thymic Epithelial Cells for Thymus Regeneration Approaches
description	Abstract Defective functionality of thymic epithelial cells (TECs), due to genetic mutations or injuring causes, results in altered T‐cell development, leading to immunodeficiency or autoimmunity. These defects cannot be corrected by hematopoietic stem cell transplantation (HSCT), and thymus transplantation has not yet been demonstrated to be fully curative. Here, we provide proof of principle of a novel approach toward thymic regeneration, involving the generation of thymic organoids obtained by seeding gene‐modified postnatal murine TECs into three‐dimensional (3D) collagen type I scaffolds mimicking the thymic ultrastructure. To this end, freshly isolated TECs were transduced with a lentiviral vector system, allowing for doxycycline‐induced Oct4 expression. Transient Oct4 expression promoted TECs expansion without drastically changing the cell lineage identity of adult TECs, which retain the expression of important molecules for thymus functionality such as Foxn1, Dll4, Dll1, and AIRE. Oct4‐expressing TECs (iOCT4 TEC) were able to grow into 3D collagen type I scaffolds both in vitro and in vivo, demonstrating that the collagen structure reproduced a 3D environment similar to the thymic extracellular matrix, perfectly recognized by TECs. In vivo results showed that thymic organoids transplanted subcutaneously in athymic nude mice were vascularized but failed to support thymopoiesis because of their limited in vivo persistence. These findings provide evidence that gene modification, in combination with the usage of 3D biomimetic scaffolds, may represent a novel approach allowing the use of postnatal TECs for thymic regeneration. Stem Cells Translational Medicine 2019;8:1107–1122
format	article
author	Ileana Bortolomai Monica Sandri Elena Draghici Elena Fontana Elisabetta Campodoni Genni Enza Marcovecchio Francesca Ferrua Laura Perani Antonello Spinelli Tamara Canu Marco Catucci Tiziano Di Tomaso Lucia Sergi Sergi Antonio Esposito Angelo Lombardo Luigi Naldini Anna Tampieri Georg A. Hollander Anna Villa Marita Bosticardo
author_facet	Ileana Bortolomai Monica Sandri Elena Draghici Elena Fontana Elisabetta Campodoni Genni Enza Marcovecchio Francesca Ferrua Laura Perani Antonello Spinelli Tamara Canu Marco Catucci Tiziano Di Tomaso Lucia Sergi Sergi Antonio Esposito Angelo Lombardo Luigi Naldini Anna Tampieri Georg A. Hollander Anna Villa Marita Bosticardo
author_sort	Ileana Bortolomai
title	Gene Modification and Three‐Dimensional Scaffolds as Novel Tools to Allow the Use of Postnatal Thymic Epithelial Cells for Thymus Regeneration Approaches
title_short	Gene Modification and Three‐Dimensional Scaffolds as Novel Tools to Allow the Use of Postnatal Thymic Epithelial Cells for Thymus Regeneration Approaches
title_full	Gene Modification and Three‐Dimensional Scaffolds as Novel Tools to Allow the Use of Postnatal Thymic Epithelial Cells for Thymus Regeneration Approaches
title_fullStr	Gene Modification and Three‐Dimensional Scaffolds as Novel Tools to Allow the Use of Postnatal Thymic Epithelial Cells for Thymus Regeneration Approaches
title_full_unstemmed	Gene Modification and Three‐Dimensional Scaffolds as Novel Tools to Allow the Use of Postnatal Thymic Epithelial Cells for Thymus Regeneration Approaches
title_sort	gene modification and three‐dimensional scaffolds as novel tools to allow the use of postnatal thymic epithelial cells for thymus regeneration approaches
publisher	Wiley
publishDate	2019
url	https://doaj.org/article/84a5cdcf9f7d4b5e98536d3398bf01f3
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Gene Modification and Three‐Dimensional Scaffolds as Novel Tools to Allow the Use of Postnatal Thymic Epithelial Cells for Thymus Regeneration Approaches

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