Epi-KAB: Exploiting Epigenome Editing in Kabuki Syndrome: a New Route Towards Gene Therapy for Rare Genetic Disorders

Sponsor

University Hospital, Montpellier (Other)

Overall Status

Completed

CT.gov ID

NCT03855631

Collaborator

Association Française contre les Myopathies Telethon (Other)

Enrollment

Location

Actual Duration (Months)

4.1

Patients Per Site Per Month

Study Details

Study Description

Brief Summary

Starting from isolating primary cells from affected patients, an in vitro disease model system for KS will be developed. Using alternative strategies to obtain patient-derived mesenchymal stem cells, an integrative approach will be adopted for defining both the transcriptional and epigenetic regulatory networks perturbed upon the loss of function of KMT2D. Combining the self-renewal potential of mesenchymal stem cells (MSCs) with CRISPR/Cas9 technology, an epigenome editing approach as therapeutic strategy to rescue the activity of MLL4 will be developed.

A step forward is expected towards the understanding of those the molecular mechanisms governing the aetiology of Kabuki Syndrome (KS) and that the proposed in vitro disease model will provide to the scientific community an experimental system to study the KS. Importantly, the aim is to define the molecular bases of KS and to develop a therapeutic strategy that could ameliorate some of the abnormalities associated with KS.

Condition or Disease	Intervention/Treatment	Phase
Kabuki Syndrome 1	Genetic: Intervention on primary cultured cells

Detailed Description

Main and secondary objectives :

The main objective is to study the pathological role of ML mutations in KS to facilitate the identification and characterization of therapeutic strategies to improve the symptoms of patients with KS. Due to the lack of treatment in the KS, the aim is to develop a model of this disease from fibroblasts from patients and reprogram them into mesenchymal stem cells. This approach will generate a KS-specific stem cell bio-bank, allowing the identification of common disturbances caused by the loss of function (LoF) of KMT2D. The impact of the KMT2D LoF on transcription but also the epigenetic mechanisms activated during MSC differentiation will be described. Finally, the therapeutic potential of an epigenome "editing" approach to increase the expression of the wild KMT2D allele to restore the functional activity of MLL4 in treated MSCs will be evaluated.

The secondary objective is to evaluate the ability to return to normal after CRISPR/Case9 gene therapy treatment on patients' cells

Methodology (design study, population, description of groups (if applicable), inclusion criteria, non-inclusion, main and secondary judgment criteria, number of subjects to be included, statistical analysis...) :

Starting from primary cells isolated from affected patients, an in vitro model of the disease will be developed. An integrative but alternative strategy approach to obtain mesenchymal stem cells derived from patients to define normal and abnormal transcription and epigenetic circuits during KMT2D LoF will be adopted. By combining the autonomous renewal potential of MSCs with CRISPR/Cas9 technology, an approach to "editing" the epigenome for therapeutic purposes to restore MLL4 activity will be developed.

This project will represent a step forward in understanding the molecular mechanisms responsible for KS. The in vitro model will provide the scientific community with an experimental system to study KS. Finally, the aim is to define the molecular basis of KS and develop a therapeutic strategy to improve certain symptoms in patients with KS.

Process (number of visits, duration of inclusions, duration of follow-up):

8 people will be included: 4 patients with Kabuki syndrome authenticated by the molecular genetics study of the KMT2D gene.

4 parents of the same sex as the patient

Feasibility :

In the clinical genetics department of the Montpellier University Hospital, more than 40 patients with Kabuki syndrome are followed.

Impacts / prospects :

The identification of the consequences of mutations in the KMT2D gene on epigenetic mark changes and cellular structural changes as well as the attempt at gene correction by CRISPR/Cas9 will provide a better understanding of the disease (and the genes that are deregulated by post-KMT2D epigenetic changes) and will also open a promising path for gene therapy by CRISPR/Cas9 method.

Study Design

Study Type:

Observational

Actual Enrollment :

8 participants

Observational Model:

Cohort

Time Perspective:

Prospective

Official Title:

Exploiting Epigenome Editing in Kabuki Syndrome: a New Route Towards Gene Therapy for Rare Genetic Disorders

Actual Study Start Date :

Sep 28, 2020

Actual Primary Completion Date :

Nov 27, 2020

Actual Study Completion Date :

Nov 27, 2020

Arms and Interventions

Arm	Intervention/Treatment
Kabuki syndrome/ unaffected parents Intervention on primary cultured cells from 4 patients with KS and sex match parents	Genetic: Intervention on primary cultured cells The intervention includes primary cultured cells, reprograming them into mesenchymal stem cells and CRISPR/Case9 gene therapy treatment on patients' cells

Arm

Intervention/Treatment

Kabuki syndrome/ unaffected parents

Intervention on primary cultured cells from 4 patients with KS and sex match parents

Genetic: Intervention on primary cultured cells

The intervention includes primary cultured cells, reprograming them into mesenchymal stem cells and CRISPR/Case9 gene therapy treatment on patients' cells

Outcome Measures

Primary Outcome Measures

success rate of fibroblasts culture [one year maximum after inclusion]
The aim is to establish a model of KS disease from fibroblasts from patients

Secondary Outcome Measures

success rate of turning fibroblasts into mesenchymal stem cells [one year maximum after inclusion]
The aim is to determine the success rate of turning fibroblasts into mesenchymal stem cells
success rate of CRISPR/Case9 gene therapy treatment on patients' cells [one year maximum after inclusion]
The aim is to evaluate the ability to return to normal after CRISPR/Case9 gene therapy treatment on patients' cells

Eligibility Criteria

Criteria

Ages Eligible for Study:

6 Years and Older

Sexes Eligible for Study:

All

Accepts Healthy Volunteers:

Inclusion Criteria:

For the patient = to have a Kabuki syndrome authenticated by molecular genetics (proof of mutation in the KMT2D gene)
For parents = having the same sex as your child
To be affiliated to a French social security system
Authorize the participation of the study

Exclusion Criteria:

Not be affiliated to a social security scheme (CMU is included)
Existence of a significant coagulation ruble (especially thrombocytopenic purpura in Kabuki patients with platelet counts < 20,000 Units).
Genetic skin disease responsible for poor healing
Refusal to participate in the child's and/or parent's study