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

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.

Study Overview

• Status: Completed
• Conditions: Kabuki Syndrome 1
• Intervention / Treatment: 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 Type: Observational
• Enrollment (Actual): 8

Contacts and Locations

Study Locations

• France
  • Herault
    • Montpellier, Herault, France, 34295
      • Arnaud de villeneuve Hospital

Participation Criteria

Eligibility Criteria

• Ages Eligible for Study: 4 years and older (Child, Adult, Older Adult)
• Accepts Healthy Volunteers: No
• Genders Eligible for Study: All

• Sampling Method: Non-Probability Sample

Study Population

Pediatric population with KS and healthy adult (unaffected sex match parents of patients)

Description

Inclusion Criteria:

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

Exclusion Criteria:

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

Study Plan

How is the study designed?

Design Details

• Observational Models: Cohort
• Time Perspectives: Prospective

Number of groups / cohorts

1

Cohorts and Interventions

Group / Cohort	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

What is the study measuring?

Primary Outcome Measures

Outcome Measure	Measure Description	Time Frame
success rate of fibroblasts culture	The aim is to establish a model of KS disease from fibroblasts from patients	one year maximum after inclusion

Secondary Outcome Measures

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

Collaborators and Investigators

• Sponsor: University Hospital, Montpellier
• Collaborators: Association Française contre les Myopathies Telethon
• Investigators: Principal Investigator: Alessio Zippo, PhD, Center of Integrative Biology

Study record dates

Study Major Dates

• Study Start (Actual): September 28, 2020
• Primary Completion (Actual): November 27, 2020
• Study Completion (Actual): November 27, 2020

Study Registration Dates

• First Submitted: February 22, 2019
• First Submitted That Met QC Criteria: February 25, 2019
• First Posted (Actual): February 27, 2019

Study Record Updates

• Last Update Posted (Actual): December 1, 2020
• Last Update Submitted That Met QC Criteria: November 30, 2020
• Last Verified: November 1, 2020

More Information

Terms related to this study

• Keywords: Kabuki syndrome CRIPSR/Cas9 KMT2D
• Additional Relevant MeSH Terms: Pathologic Processes; Disease; Congenital Abnormalities; Otorhinolaryngologic Diseases; Labyrinth Diseases; Ear Diseases; Syndrome; Hematologic Diseases; Vestibular Diseases; Genetic Diseases, Inborn; Abnormalities, Multiple
• Other Study ID Numbers: RECHMPL18_0242

Drug and device information, study documents

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