Custom-designed gene editing of induced epidermal stem cells for gene therapy of genetic diseases of squamous epithelia (HOLO-GT)


PROJECT DESCRIPTION

Gene therapy for inherited skin disorders

Epidermolysis bullosa (EB) is a group of rare inherited skin disorders associated with fragile, blistering skin. EB diseases result from mutations in a variety of genes responsible for intraepidermal cohesion including those coding for keratin, integrin and laminin. Scientists of the EU-funded Holo-GT project have previously demonstrated the efficacy of a combinatorial and gene therapy approach for EB. However, given the dominant inheritance pattern of many EB conditions, gene addition is unsuitable. The Holo-GT consortium aims to develop a gene editing strategy as well as an approach for replenishing skin stem cells that are often lost in patients with EB.

Project Objectives

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).

PROJECT INFORMATION

Grant agreement ID: 101019289


DOI
10.3030/101019289

Start date: 1 September 2021 – End date: 31 August 2026

Funded under

  • EXCELLENT SCIENCE – European Research Council (ERC)

Total cost: € 2 462 375,00

EU contribution: € 2 462 375,00

Coordinated by:

UNIVERSITA DEGLI STUDI DI MODENA E REGGIO EMILIA – Italy

Open Access Publications

  • De Rosa L, Enzo E, Palamenghi M, Sercia L, De Luca M. Stairways to Advanced Therapies for Epidermolysis Bullosa. Cold Spring Harb Perspect Biol. 2023 Apr 3;15(4):a041229. doi: 10.1101/cshperspect.a041229. PMID: 36167646; PMCID: PMC10071437.
  • De Rosa L, De Luca M. The joint battle to tackle epidermolysis bullosa through gene therapy. Trends Mol Med. 2022 Jul;28(7):533-535. doi: 10.1016/j.molmed.2022.05.001. Epub 2022 May 19. PMID: 35599142.
  • Kueckelhaus M, Rothoeft T, De Rosa L, Yeni B, Ohmann T, Maier C, Eitner L, Metze D, Losi L, Secone Seconetti A, De Luca M, Hirsch T. Transgenic Epidermal Cultures for Junctional Epidermolysis Bullosa – 5-Year Outcomes. N Engl J Med. 2021 Dec 9;385(24):2264-2270. doi: 10.1056/NEJMoa2108544. PMID: 34881838.

Scientific Dissemination

International Meetings

  • EMBO Regeneration, 26-30 September 2022, Barcelona, Spain, Prof. Michele De Luca (remote)
  • ISSCR/ASGCT 2022 Madison Symposium, “Emerging Therapies at the Intersection of Genetic and Cellular Technologies”, 22-23 September 2022, Prof. Michele De Luca (remote)
  • Azienda Ospedaliero-Universitaria Città della Salute e della Scienza di Torino, 19th European Burns Association Congress (EBA), 7-10 September 2022, Torino, Italy, Prof. Michele De Luca
  • Mendel Genetics Conference 2022, 20-23 July 2022, Brno, Czech Republic, Prof. Michele De Luca (remote)
  • The Francis Crick Institute/UCL, Conference on “Stemness, Regeneration and Immunity: from development to therapy”, 20-21 June 2022, London, UK, Prof. Michele De Luca
  • 1st World Congress on Rare Skin Diseases 2022, 7-9 June 2022, Paris, Dr Laura De Rosa
  • NSC-Reconstruct Annual Meeting, 5-7 April 2022, Bellagio, Italia, Prof. Michele De Luca
  • SkinTERM Annual Meeting 2022, 26-27 March 2022, Prof. Michele De Luca (remote)
  • UC San Diego Stem Cell Symposium, 10 febbraio 2022, La Jolla, California, Prof. Michele De Luca (remote)
  • ISSCR Shanghai International Symposium, 12-14 January 2022, Shanghai, China, Prof. Michele De Luca (remote)
  • 2021 Virtual Ottawa “Till & McCulloch Meeting” (TMM), November 15-17, 2021, Prof. Michele De Luca (remote)
  • National Foundation for Ectodermal Dysplasias, NFED 2021 Research Conference, October 21-24, 2021, Prof. Michele De Luca (remote)
  • 14th World Congress of Paediatric Dermatology, 22-25 September 2021, Edinburgh, UK, Prof. Michele De Luca
  • Nano-Innovation 2021, 22 September 2021, Prof. Michele De Luca (remote)
  • DEBRA International Congress 2021, 16-19 September 2021, Prof. Michele De Luca (remote)

Workshop & Seminars

  • Institut Pasteur, Revive ‘Advances in Stem Cell Biology’ Course, 27 June 2023, Paris, Prof. Michele De Luca
  • ERN Workshop “Epidermolysis bullosa: from genes to translation into therapies”, 23-24 March 2023, Freiburg, Germany, Dr Laura De Rosa
  • Accademia Medica Roma, Seminario, 16 March 2023, Roma, Prof. Michele De Luca
  • SIGU (Società Italiana di Genetica Umana), Webinar “Medical Genetics Academy SIGU 2022”, 13 October 2022, Prof. Michele De Luca (remote)
  • Hydra 2022, XV European Summer School on Stem Cell Biology and Regenerative Medicine, 13-20 September 2022, Hydra, Greece, Prof. Michele De Luca
  • Institut Pasteur, 9th Advances in Stem Cell Biology Course 2021-2022, 28 June 2022, Prof. Michele De Luca (remote)
  • King’s College London, Randall Seminar Series, 14 June 2022, Prof. Michele De Luca (remote)
  • EXPOSANITA’, Workshop Clust-ER Health, Industrie della Salute e del Benessere,  “Prodotti medicinali per le terapie avanzate: prospettive di sviluppo e industrializzazione”, 12 May 2022, Bologna, Italy, Prof. Michele De Luca
  • Workshop on “Stem Cells and Their Promise for Regenerative Medicine”, 5-6 May 2022, Vaticano, Prof. De Luca (remote)
  • Accademia Medica di Roma, Conferenza “Epiteli transgenici: dal laboratorio alla clinica (e ritorno)”, 7 April 2022, Prof. Michele De Luca
  • Helmholtz Zentrum Munchen, HDC Hybrid Seminar Series, 15 February 2022, Prof. Michele de Luca (remote)
  • Unimore, Lettura Magistrale, XX Modulo del Master in Wound-Care, 28 January 2022, Prof. Michele De Luca (remote)
  • Università degli Studi di Torino, Master II Livello “Cellule Staminali in Medicina Rigenerativa e Management della Cell Factory” – Secondo Ciclo, 13 January 2022, Michele De Luca (remote)
  • Utrecht University, PhD Programme on “Cancer Stem Cells and Developmental Biology” (CSnD) and Regenerative Medicine, 14 December 2021, Prof. Michele De Luca (remote)
  • Osservatorio Terapie Avanzate (OTA), Il Valore delle Terapie Avanzate in Italia, Presentazione Consensus “Proposta di misure per la determinazione del valore delle Terapie Avanzate in Italia”, 17 November 2021, Prof. Michele De Luca (remote)
  • Università di Torino, Workshop del Master di II° Livello “Cellule Staminali in Medicina Rigenerativa e Management della Cell Factory”, sessione “La manipolazione genetica ex vivo – ri-espressione genica”, 18 September 2021, Prof. De Luca (remote)
  • Sorbonne University, 15 September 2021, Prof. Michele De Luca (remote)

Communication

Public Engagement

  • R2B, Research to Business, Bologna, 8-9 June 2023, Dr Eleonora Maurizi e dr.ssa Maria Pia Polito
  • Le ali di Camilla, EB Days, 29-30 April 2023, Modena, Prof. Michele De Luca
  • Accademia Medica di Roma, Conferenza AMR, 16 March 2023, Roma, Prof. Michele De Luca
  • Unione Italiana Ittiosi, Convegno Italiano UNITI 2022, 1 October 2022, Palermo, Prof. Michele De Luca, Dr Laura De Rosa, Dr Laura Sercia e Dr Stefania Bettinelli
  • Comune di Casina, I martedì della cantoniera, 2 August 2022, Casina (RE), Prof. Michele De Luca e dr.ssa Stefania Bettinelli
  • Le ali di Camilla, EB Day, 21 May 2022, Modena, Prof. Michele De Luca
  • Festa della Scienza, XIII Edizione, 12-14 May 2022, Salento, Prof. Michele De Luca
  • XI edizione Festa di Scienza e di Filosofia-Virtute e Canoscenza, “Riprendiamo il cammino. La Scienza. Il nuovo sviluppo. Il libero pensiero”, 21-24 April 2022, Foligno (PG), Prof. Michele De Luca
  • Associazione Luca Coscioni, Progetto “ScolarMente” – La ricerca sulle Cellule Staminali, 11 March 2022, Prof. Michele De Luca (remote)
  • Expo Dubai 2020, Personalized Medicine: New tools and active polices for prevention, diagnosis and therapy, 27-28 January 2022, Dubai, UAE, Dr Stefania Bettinelli
  • Scienza: Ultima Frontiera a Brugherio, Edizione 2021, 5 November 2021, Prof. Michele De Luca
  • B.T. Expò, Workshop “L’innovazione nel trattamento e nella cura di patologie: nuovi approcci per la medicina del futuro”, 5 October 2021, Modena, Dott.ssa Stefania Bettinelli
  • European Biotech Week, HoloTour – Alla scoperta di Holostem e delle Terapie avanzate a Modena, Modena, 1 October 2021, Dr Stefania Bettinelli (remote)

Coordinator

UNIVERSITA DEGLI STUDI DI MODENA E REGGIO EMILIA

Net EU contribution € 2 118 125,00 – Other funding € 0,00

Address

Via Università 4
41121 Modena
 Italy 

Beneficiaries (3)


UNIVERSITA DEGLI STUDI DI MODENA E REGGIO EMILIA – Italy

Net EU contribution: € 2 118 125,00

THIRD PARTY

HOLOSTEM TERAPIE AVANZATE SRL – Italy

Net EU contribution: € 125.000,00


UNIVERSITA DEGLI STUDI DI ROMA LA SAPIENZA – Italy

Net EU contribution: € 219.250,00