Ex Vivo Gene Therapy Clinical Trial for RDEB Using Genetically Corrected Autologous Skin Equivalent Grafts (EBGraft)

Phase I/II ex Vivo Gene Therapy Clinical Trial for RDEB Using Autologous Skin Equivalent Grafts Genetically Corrected With a COL7A1-encoding SIN Retroviral Vector

This phase I/II clinical trial aims to treat 3 adult subjects with Recessive Dystrophic Epidermolysis Bullosa, expressing residual C7 levels, by genetically corrected autologous skin equivalent grafts on selected areas (up to 300 cm2).

Study Overview

• Status: Active, not recruiting
• Conditions: Epidermolysis Bullosa Dystrophica, Recessive
• Intervention / Treatment: Biological: COL7A1-SIN retroviral vector engineered autologous tissue-engineered skin

Detailed Description

Recessive Dystrophic Epidermolysis Bullosa (RDEB) is a severe orphan genetic disease responsible for skin and mucosal detachments due to a loss of adhesion of the epidermis to the underlying dermis. The disease is caused by loss of function mutations of the COL7A1 encoding type VII collagen (C7) which forms anchoring fibers, which are essential structures for dermal-epidermal adherence. Current treatments are only symptomatic and do not effectively treat or prevent the occurrence of cutaneous and mucosal detachments responsible for local and systemic complications that threaten the vital prognosis.

EBGRAFT is a prospective open-label international monocentric phase I/II clinical trial. It aims to treat 3 adult subjects with RDEB, expressing residual C7 levels, by genetically corrected autologous skin equivalent grafts.

The skin equivalent consists of keratinocytes and fibroblasts from the patient, genetically corrected ex vivo with a secure Self INactivating (SIN) retroviral vector expressing the COL7A1 cDNA under the control of the ubiquitous human promoter EF1a.

Each patient will be grafted sequentially at Necker Hospital in Paris using autologous genetically corrected skin equivalents of approximately 300 cm2 (up to 6 grafts of 50 cm2 each).

The main objective is to evaluate the safety of autologous skin equivalent grafts genetically corrected with a SIN COL7A1 retroviral vector (RV) in adults with RDEB.

The secondary objectives are:

1. To evaluate the efficacy of transplanting autologous skin equivalent genetically corrected with RV SIN COL7A1 in adults with RDEB.
2. To evaluate the immune response against recombinant type VII collagen (C7).

This clinical trial should evaluate whether the grafting of these genetically corrected autologous skin equivalents is well tolerated and whether they restore normal dermal-epidermal adherence of the grafted areas. The proposed treatment aims to obtain a permanent correction of the grafted areas, allowing skin healing and reducing pain. It has the potential to reduce itching, to prevent the occurrence of blisters and skin detachments, reduce the risk of infections, the duration and cost of care and also the risk of development of squamous cell carcinomas in the grafted areas.

• Study Type: Interventional
• Enrollment (Anticipated): 3
• Phase: Phase 2; Phase 1

Contacts and Locations

Study Locations

• France
  • Paris, France, 75743
    • Institut Imagine Necker Hospital

Participation Criteria

Eligibility Criteria

• Ages Eligible for Study: 18 years and older (Adult, Older Adult)
• Accepts Healthy Volunteers: No
• Genders Eligible for Study: All

Description

Inclusion Criteria:

1. Clinical and molecular diagnosis of RDEB with confirmed bi-allelic COL7A1 mutations
2. Reduced staining of C7 on skin biopsy, measured by immunofluorescence microscopy (IF)
3. A reduced number of/or morphologically abnormal anchoring fibrils confirmed by TEM
4. Detection of non-collagenous-1 domain (NC-1) of C7 on skin biopsy, measured by immunofluorescence microscopy (IF) and/or Western blot (WB) analysis
5. Presence of ≥100cm2 of blistered and/or erosive skin areas including chronic wounds suitable for skin grafting
6. Ability to undergo anaesthesia for skin grafting procedures
7. Subjects aged 18 years, willing and able to give informed consent

Exclusion Criteria:

1. Recipients of other investigational medicinal products within 6 months prior to enrolment into this study
2. Past medical history of biopsy proven skin malignancy
3. Immunotherapy including oral corticosteroids (Prednisolone >1mg/kg) for more than one week (intranasal and topical preparations are permitted) or chemotherapy within 60 days of enrolment into this study
4. Known allergy to any of the constituents of the investigational medicinal product (IMP) including Penicillin

5. Subjects with BOTH:

   • positive serum antibodies to C7 confirmed by ELISA and
   • positive IIF with binding to the base of salt split skin and/or
   • positive Western blot
6. Positive results for HIV, Hepatitis BsAg, Hepatitis BcAb, Hepatitis C IgG, HTLV1&2 or Syphilis serology
7. Clinically significant medical, psychological or laboratory abnormalities limiting the ability of the subject to travel to the trial site(s) and to undergo grafting and follow-up procedures, as determined by the Investigator
8. Absence of adequate social support
9. Subjects who are pregnant, breast-feeding or of child-bearing potential who are neither abstinent nor practicing an acceptable means of contraception when this is in line with the usual and preferred lifestyle of the subject, as determined by the Investigator, for the duration of the trial

Study Plan

How is the study designed?

Design Details

• Primary Purpose: Treatment
• Allocation: N/A
• Interventional Model: Single Group Assignment
• Masking: None (Open Label)

Number of Arms

1

Arms and Interventions

Participant Group / Arm	Intervention / Treatment
Experimental: Autologous genetically modified tissue-engineered skin graft; Graft of SIN RV-mediated COL7A1 gene-modified autologous skin equivalent	Biological: COL7A1-SIN retroviral vector engineered autologous tissue-engineered skin; Graft of SIN RV-mediated COL7A1 gene-modified autologous skin equivalent

What is the study measuring?

Primary Outcome Measures

Outcome Measure	Measure Description	Time Frame
Safety of grafting SIN RV-mediated COL7A1 gene-modified autologous skin equivalent: Adverse Events (AE), Serious Adverse Events (SAEs), Adverse Reactions (ARs) and Serious Adverse Reactions (SARs)	The primary objective is to evaluate the safety of autologous autologous skin equivalent grafts genetically corrected with a SIN COL7A1 retroviral vector (RV) in adults with RDEB Primary Endpoints: Record of Adverse Events (AE), Serious Adverse Events (SAEs), Adverse Reactions (ARs) and Serious Adverse Reactions (SARs).	Month 12 post grafting.

Secondary Outcome Measures

Outcome Measure	Measure Description	Time Frame
Change in C7 protein expression	Skin biopsy analysis of grafted skin compared to baseline for : C7 protein expression by immunofluorescence microscopy (IF) using several specific antibodies	Month 1, Month 3, Month 6, Month 12 post grafting.
Change in anchoring fibrils number	Count of anchoring fibril (AF) at the dermal-epidermal junction (DEJ) by transmission electron microscopy (TEM) will be performed on skin biopsies and	Month 1, Month 3, Month 6, Month 12 post grafting.
Change in scar quality: Vancouver Scar Scale (VSS)	Scar quality will be measured using the Vancouver Scar Scale (VSS) and compared to baseline at several time points. Minimum value=0, Maximum value=12. Lower value is better.	Month 1, Month 2, Month 3, Month 6, Month 12 post grafting.
Changes in blister number over the grafted skin	Blister formation will be monitored and counted by a skilled dermatologist and compared to baseline at several time points.	Month 1, Month 2, Month 3, Month 6, Month 12 post grafting.
Changes in clinical appearance of grafted skin	Clinical appearance of the grafted skin areas will be assessed by 3D photographic reconstruction using a CANFIELD Vectra H1 device and compared to baseline at several time points.	Month 1, Month 2, Month 3, Month 6, Month 12 post grafting.
Changes in pruritus of grafted skin	Pruritus will be measured by the 5D pruritus score and compared to baseline at several time points. Minimum value=4, Maximum value=35. Lower value is better.	Month 1, Month 2, Month 3, Month 6, Month 12 post grafting.
Change in Quality of life: QOLEB questionnaire (Quality of Life for Epidermolysis Bullosa)	Quality of life will be measured using the specific QOLEB questionnaire (Quality of Life for Epidermolysis Bullosa) and compared to baseline at several time points. Minimum value=0, Maximum value=51. Lower value is better.	Month 1, Month 2, Month 3, Month 6, Month 12 post grafting.
Change in Birmingham Epidermolysis Score (BEBS)	Change in disease severity will be assessed by the Birmingham Epidermolysis Score (BEBS). Score will be compared to baseline at several timepoints. Minimum value=0, Maximum value=100. Lower value is better.	Month 1, Month 2, Month 3, Month 6, Month 12 post grafting.
Change in Epidermolysis Bullosa Disease Activity and Scarring Index (EBDASI) - Activity	Change in disease severity will be assessed by the Epidermolysis Bullosa Disease Activity and Scarring Index (EBDASI). Score will be compared to baseline at several timepoints. Score measure activity (minimum value=0, maximum value=276). Lower value is better.	Month 1, Month 2, Month 3, Month 6, Month 12 post grafting.
Change in Epidermolysis Bullosa Disease Activity and Scarring Index (EBDASI) - Damage	Change in disease severity will be assessed by the Epidermolysis Bullosa Disease Activity and Scarring Index (EBDASI). Score will be compared to baseline at several timepoints. Score measure damage (minimum value=0, maximum value=230). Lower value is better.	Month 1, Month 2, Month 3, Month 6, Month 12 post grafting.
Change in instrument for Scoring Clinical Outcomes of Research for Epidermolysis Bullosa (iscorEB).	Change in disease severity will be assessed by the instrument for Scoring Clinical Outcomes of Research for Epidermolysis Bullosa (iscorEB). Score will be compared to baseline at several timepoints. Minimum value=0, Maximum value=120. Lower value is better.	Month 1, Month 2, Month 3, Month 6, Month 12 post grafting.
Evaluation of the humoral immune response against recombinant C7	Detection of circulating anti-C7 antibodies in patient blood by ELISA and/or indirect immunofluorescence (IIF) on split skin at several time points	Month 1, Month 6, Month 12 post grafting.
Evaluation of the cytotoxic immune response against recombinant C7	Detection of T-cell responses to the full length C7 in patient blood by ELISPOT assay.	Month 1, Month 6, Month 12 post grafting.

Collaborators and Investigators

• Sponsor: Institut National de la Santé Et de la Recherche Médicale, France

Publications and helpful links

General Publications

• Gaucher S, Lwin SM, Titeux M, Abdul-Wahab A, Pironon N, Izmiryan A, Miskinyte S, Ganier C, Duchatelet S, Mellerio JE, Bourrat E, McGrath JA, Hovnanian A. EBGene trial: patient preselection outcomes for the European GENEGRAFT ex vivo phase I/II gene therapy trial for recessive dystrophic epidermolysis bullosa. Br J Dermatol. 2020 Mar;182(3):794-797. doi: 10.1111/bjd.18559. Epub 2019 Nov 27. No abstract available.
• Titeux M, Pendaries V, Zanta-Boussif MA, Decha A, Pironon N, Tonasso L, Mejia JE, Brice A, Danos O, Hovnanian A. SIN retroviral vectors expressing COL7A1 under human promoters for ex vivo gene therapy of recessive dystrophic epidermolysis bullosa. Mol Ther. 2010 Aug;18(8):1509-18. doi: 10.1038/mt.2010.91. Epub 2010 May 18.
• Hennig K, Raasch L, Kolbe C, Weidner S, Leisegang M, Uckert W, Titeux M, Hovnanian A, Kuehlcke K, Loew R. HEK293-based production platform for gamma-retroviral (self-inactivating) vectors: application for safe and efficient transfer of COL7A1 cDNA. Hum Gene Ther Clin Dev. 2014 Dec;25(4):218-28. doi: 10.1089/humc.2014.083.

Study record dates

Study Major Dates

• Study Start (Actual): January 10, 2020
• Primary Completion (Anticipated): June 9, 2027
• Study Completion (Anticipated): June 9, 2027

Study Registration Dates

• First Submitted: November 25, 2019
• First Submitted That Met QC Criteria: December 4, 2019
• First Posted (Actual): December 5, 2019

Study Record Updates

• Last Update Posted (Actual): October 19, 2021
• Last Update Submitted That Met QC Criteria: October 18, 2021
• Last Verified: October 1, 2021

More Information

Terms related to this study

• Keywords: COL7A1; Genetic Therapy; Autologous Skin Graft; SIN retroviral vector; Type VII collagen
• Additional Relevant MeSH Terms: Skin Diseases; Congenital Abnormalities; Genetic Diseases, Inborn; Connective Tissue Diseases; Skin Diseases, Genetic; Skin Diseases, Vesiculobullous; Skin Abnormalities; Collagen Diseases; Epidermolysis Bullosa; Epidermolysis Bullosa Dystrophica
• Other Study ID Numbers: C12-48; 2016-002790-35 (EudraCT Number)

Drug and device information, study documents

• Studies a U.S. FDA-regulated drug product: No
• Studies a U.S. FDA-regulated device product: No