Descripción del proyecto
Terapia génica contra las dermatosis hereditarias
La epidermólisis ampollosa (EA) es un grupo de dermatosis hereditarias raras asociadas a una piel frágil y ampollosa. Las enfermedades relacionadas con la EA son el resultado de mutaciones en una variedad de genes responsables de la cohesión intraepidérmica, incluidos los que codifican la queratina, la integrina y la laminina. Los científicos del proyecto Holo-GT, financiado con fondos europeos, ya han demostrado la eficacia de un enfoque genoterapéutico y combinatorio para tratar la EA. Sin embargo, dado el patrón de herencia dominante de muchas afecciones de la EA, la adición de genes es inadecuada. El consorcio de Holo-GT pretende desarrollar una estrategia de edición genética, así como un método para reponer las células madre de la piel que suelen perderse en los pacientes con EA.
Objetivo
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).
Ámbito científico
Palabras clave
Programa(s)
Régimen de financiación
ERC-ADG - Advanced GrantInstitución de acogida
41121 Modena
Italia