LAMA2 Genetic Correction

Ex Vivo Genetic Correction of LAMA2 Mutation(s) in Myogenic Stem Cells of Patients with Merosin-deficient Congenital Muscle Dystrophy Type 1a (MDC1a)

Merosin-deficient congenital muscle dystrophy type 1a (MDC1a), or LAMA2 muscular dystrophy (LAMA2-MD) is a severe autosomal recessive form of muscular dystrophy that is caused by homozygous or compound heterozygous mutations in the laminin alpha 2 (LAMA-2) gene. Many different LAMA-2 mutations have been reported. In most cases, MDC1a is diagnosed within the first year of life, and is characterized by hypotonia, delayed motor development and white matter abnormalities. Currently, no efficient treatment is available for this patient group. Generally, MDC1a patients with mutations causing a premature stop codon are most severely affected (early onset LAMA2-MD) and patients with missense mutations are generally affected more mild affected and more late-onset (late onset LAMA2-MD). However, large variation in disease severity and clinical course is observed, even between individuals with the same mutation, e.g. the LAMA2 c.5562+5G>C mutation, which is frequently observed in Dutch MDC1a patients. This study aims to isolate and culture fibroblasts and myogenic stem cells called mesoangioblasts from the skin and muscle biopsies of adult LAMA2 mutation carriers to explore if genetic correction of LAMA2 mutations using CRISPR-Cas9 can be achieved and subsequently assess the effect in vitro, as a first step towards therapy development.

Study Overview

• Status: Completed
• Conditions: MDC1A

• Study Type: Observational
• Enrollment (Actual): 7

Participation Criteria

Eligibility Criteria

• Ages Eligible for Study: Adult; Older Adult
• Accepts Healthy Volunteers: Yes

• Sampling Method: Non-Probability Sample

Study Population

The study will only include patients/controls living in The Netherlands

Description

Inclusion Criteria:

• LAMA2 mutation carriers:
• Age >18 years
• Heterozygous or homozygous LAMA2 c.5562+5G>C mutation
• Written informed consent

Controls:

• Written informed consent
• Age >18 years
• No muscular dystrophy or other disease known to affect muscle morphology or function

Exclusion Criteria:

• MDC1a patients and controls:
• No informed consent
• Use of anti-coagulants, anti-thrombotics and other medication influencing coagulation
• Have a weekly alcohol intake of ≥ 35 units (men) or ≥ 24 units (women)
• Current history of drug abuse
• A history of strokes
• Significant concurrent illness
• Ongoing participation in other clinical trials
• Major surgery within 4 weeks of the visit
• Pregnant or lactating women
• Patients unable and/or unwilling to comply with treatment and study instructions
• Any other factor that in the opinion of the investigator excludes the patient from the study

Study Plan

How is the study designed?

What is the study measuring?

Primary Outcome Measures

Outcome Measure	Measure Description	Time Frame
Compare RNA transcription of corrected and not corrected DNA	qPCR	1 day
Assess effect LAMA2 mutations on muscle protein quantity (amount of LAMA2)	LAMA2 immunostaining	1 day
Assess effect LAMA2 mutations on muscle protein quality (localization of LAMA2)	LAMA2 immunostaining	1 day
Assess effect LAMA2 mutations on muscle protein quantity (LAMA2 overall levels)	LAMA2 western blot	1 day
Assess effect LAMA2 mutations on muscle protein quality (fibrosis)	LAMA2 HE staining	1 day

Secondary Outcome Measures

Outcome Measure	Measure Description	Time Frame
Verification of the LAMA2 mutations or exclusion of LAMA2 mutations in controls	DNA analysis	1 day
Blood markers to assess level of muscle inflammation, damage and regeneration in MDC1a patients: CK	ELISA assay	1 day
Blood markers to assess level of muscle inflammation, damage and regeneration in MDC1a patients: TNFa	ELISA assay	1 day
Blood markers to assess level of muscle inflammation, damage and regeneration in MDC1a patients: IL-6	ELISA assay	1 day
Blood markers to assess level of muscle inflammation, damage and regeneration in MDC1a patients: SDF1	ELISA assay	1 day

Collaborators and Investigators

• Sponsor: Maastricht University

Study record dates

Study Major Dates

• Study Start (Actual): July 1, 2020
• Primary Completion (Actual): July 18, 2024
• Study Completion (Actual): July 18, 2024

Study Registration Dates

• First Submitted: January 11, 2022
• First Submitted That Met QC Criteria: August 30, 2024
• First Posted (Actual): September 3, 2024

Study Record Updates

• Last Update Posted (Actual): September 3, 2024
• Last Update Submitted That Met QC Criteria: August 30, 2024
• Last Verified: July 1, 2024

More Information

Terms related to this study

• Keywords: Muscular Dystrophy; MDC1A; LAMA2; Mesoangioblasts
• Other Study ID Numbers: MABS03

Plan for Individual participant data (IPD)

• Plan to Share Individual Participant Data (IPD)?: NO

Drug and device information, study documents

• Studies a U.S. FDA-regulated drug product: No
• Studies a U.S. FDA-regulated device product: No
• product manufactured in and exported from the U.S.: No