Projektbeschreibung
Gentherapie für vererbte Hautkrankheiten
Epidermolysis bullosa (EB) ist eine Gruppe seltener erblicher Hauterkrankungen, die Fragilität und Blasenbildung in der Haut verursachen. EB-Erkrankungen entstehen durch Mutationen in einer Reihe von Genen, die für den intraepidermalen Zusammenhalt verantwortlich sind. Dazu zählen solche, die für Keratin, Integrin und Laminin kodieren. Das wissenschaftliche Team hinter dem EU-finanzierten Projekt Holo-GT konnte bereits die Wirksamkeit eines kombinatorischen und gentherapeutischen Ansatzes für Epidermolysis bullosa nachweisen. Angesichts des dominanten Vererbungsmusters vieler EB-Erkrankungen ist die Zugabe von Genen hier jedoch ungeeignet. Das Holo-GT-Konsortium hat sich daher zum Ziel gesetzt, eine Technik zur Gen-Editierung sowie einen Ansatz zur Wiederherstellung von Hautstammzellen zu entwickeln, die bei Patientinnen und Patienten mit Epidermolysis bullosa häufig verloren gehen.
Ziel
Holo-GT will define innovative ex vivo gene therapy of dominant genetic skin disease, by using Epidermolysis Bullosa (EB) as a model system. EB is a group of dominantly or recessively inherited, devastating, incurable diseases marked by structural fragility of the integuments. The applicant has shown that combined ex vivo cell and gene therapy can cure the skin of recessive LAMB3-Junctional EB. This lifesaving procedure unveiled that the human epidermis is sustained solely by long-lived stem cells, detected as holoclone-forming cells, which generate transient progenitors referred to as meroclones and paraclones.
However, over 50% of EB are dominantly inherited, making gene addition unsuitable. Gene editing does not efficiently correct epidermal stem cells, which define a small proportion of clonogenic keratinocytes and cannot be prospectively isolated.
Holo-GT aims at designing a safe and efficacious protocol for dominant forms of EB through:
• Molecular characterization of the keratinocyte clonal types and pathways sustaining holoclone-forming cells by single-cell RNA profiling.
• Generation of induced holoclone-forming cells (iHolo) by transient expression of specific sets of transcription factors (identified in aim 1) able to reprogram progenitors into holoclone-forming cells.
• Development of custom-engineered CRISPR/Cas nucleases, based on selection-based directed evolution or structure-guided mutagenesis, able to fully distinguish wild type and mutant alleles.
• Using this newly developed CRISPR/Cas system, gene editing of iHolo prepared from keratinocyte cultures initiated from patients with dominant forms of EB.
Holo-GT could tackle not only dominant EBs but also other dominant genetic diseases of the epidermis and other squamous epithelia, as well as recessive forms of EB, particularly those characterized by the well-known depletion of stem cells, which could be rescued by the reprogramming of transient progenitors into holoclone-forming cells (iHolo).
Wissenschaftliches Gebiet
Schlüsselbegriffe
Programm/Programme
Thema/Themen
Finanzierungsplan
ERC-ADG - Advanced GrantGastgebende Einrichtung
41121 Modena
Italien