Combined cell transplantation as a therapeutic approach for dysferlinopathies and transversal study to evaluate the frequency of carriers of dysferlinopathy in Caucasian population using a monocyte’s test to detect dysferlin expression

Publications

Genetic and epigenetic determinants of low dysferlin expression in monocytes (2014)

Grant Duration
01/12 – 12/12

Project 1

Cell transplantation as a therapy for dysferlinopathy

So far, there is no effective treatment for dysferlinopathies. Preliminary studies from our group have shown some functional improvement in A/J mice treated with bone marrow transplantation. Dr Diaz, a neurologist in our research group, demonstrated that transfer of mesoangioblasts into dysferlin deficient mice can restore dysferlin expression in muscle after one month and in vitro experiments showed that isolated fibers from mesoangioblast treated mice improved their ability to repair sarcolemma after injury. We are interested in comparing the efficiency of these two treatments alone or in combination and analyze their efficacy 5 months after the beginning of the treatment. We will study dysferlin expression in skeletal muscle and peripheral blood and perform functional studies i.e. electromyography and gait assessment.

Project 2

Estimating the frequency of dysferlinopathy

Currently, we can only estimate the frequency of dysferlinopathy in the global population. After 8 years of performing dysferlin expression analysis in peripheral blood monocytes we have observed an incidence of subjects with reduced levels of dysferlin higher than expected in our local “controls” (volunteers with no clinical symptoms). Our goal is to determine the carrier frequency in the general population and whether it is consistent with the frequency with which we see affected individuals. Analysis of the frequency of dysferlin mutation in a sample population could provide clues as to the true likelihood of a child inheriting dysferlinopathy.

By Eduard Gallardo, PhD, Institut de Recerca de l’Hospital de la Santa Creu i Sant Pau|2025-05-06T22:05:43+00:00January 1st, 2013|Past Projects|Comments Off

About the Author: Eduard Gallardo, PhD, Institut de Recerca de l’Hospital de la Santa Creu i Sant Pau

Dr. Eduard Gallardo is a Senior Researcher at the Institut de Recerca de l’Hospital de la Santa Creu i Sant Pau in Barcelona. Dr. Gallardo is a biologist who holds a PhD in Inflammatory Myopathies. He was among the first to characterize the inflammatory findings in dysferlinopathies. The Neuromuscular Diseases group in Barcelona is focused in immune mediated diseases of peripheral nerve and muscle and on muscular dystrophies. They have a special interest in dysferlinopathies and they have been studying them since the cloning of the gene in 1998.

Bla/J mice (B6.A-Dysf^prmd/GeneJ)

Availability:
The Jackson Laboratory
Stock number: 012767
Strain information: http://jaxmice.jax.org/strain/012767.html
Phone: 800-422-MICE (800-422-6423)
610 Main Street, Bar Harbor, Maine 04609 USA

Please note that this strain has been cryopreserved by the Jackson Laboratories, but the Jain Foundation maintains a private colony of these mice for public use. To obtain these mice, please contact the Jain Foundation:

Jain Foundation Inc.
9706 Fourth Ave NE, Suite 101
Seattle, Washington 98115
Phone: 425-882-1492
Fax: 425-882-1050

Email: admin@jain-foundation.org

Development:
In these mice the progressive muscular dystrophy (prmd) allele from the A/J inbred strain is crossed onto the C57BL/6 genetic background. The cross was performed in the laboratory of Dr. Isabelle Richard at Genethon and the backcross generation reached N8. In collaboration with the Jain Foundation, Dr. Richard donated the strain to The Jackson Laboratory in 2010. Upon arrival, mice were bred to C57BL/6J for at least 2 generations to establish the colony.

Mutation:
An ETn retrotransposon (5-6kb) is inserted in intron 4 of the dysferlin gene.

Symptoms:
Disease onset is observed by 2 months and is characterized by the presence of centronucleated fibers and areas of inflammation. As seen with the original background A/J, mice homozygous for the prmd allele on the C57BL/6J background display an increasing number of centronucleated fibers and impairment in the majority of muscles by 4 months of age. In order of severity, the most affected muscles are psoas, quadriceps femoris, tibialis anterior, and gastrocnemius. Mice exhibit a decreased membrane repair capacity following laser wounding experiments. In an open space assay, mice cover less distance and are less active than wild-type. Mice that are homozygous for this allele are viable, fertile and normal in size.

Comparison with other disease strains:
Disease progression is similar to A/J: slightly slower than in SJL/J, Dysf-/- (Campbell), Dysf-/- (Brown), and C57BL/10.SJL mice. As in both SJL/J and Dysf-/- (Brown) mice, proximal muscles are more severely affected than distal muscles. As in Dysf-/- (Brown) mice, abdominal muscles are also affected.

Control strain(s):
C57BL/6 (for homozygotes); littermates (for heterozygotes).

References:
Lostal W; Bartoli M; Bourg N; Roudaut C; Bentaib A; Miyake K; Guerchet N; Fougerousse F; McNeil P; Richard I. 2010. Efficient recovery of dysferlin deficiency by dual adeno-associated vector-mediated gene transfer. Hum Mol Genet 19(10):1897-907.

PMID: 20154340

Below is a quick summary of the inclusion/exclusion criteria. Please review this information carefully prior to making any decision regarding participating.

Inclusion Criteria:

Must be Non-ambulant (cannot walk 10 meters in ≤ 30 sec) and age 18 years or older
Established mutations of the dysferlin gene on both alleles
Impaired muscle function but with sufficient muscle preservation to ensure muscle transfection based on magnetic resonance image of the EDB showing sufficient muscle preservation to permit transfection
Willingness of sexually active subjects with reproductive capacity to practice reliable method of contraception (If appropriate), during the first six months after gene transfer (females) or until two negative sperm samples are obtained post gene transfer (males).

Exclusion Criteria:

Active viral infection based on clinical observations or serological evidence of HIV, or Hepatitis A, B or C infection
The presence of dysferlin mutations without weakness or loss of function
Symptoms or signs of cardiomyopathy, including:
Dyspnea on exertion, pedal edema, shortness of breath upon lying flat, or rales at the base of the lungs
Echocardiogram with ejection fraction below 40%
Diagnosis of (or ongoing treatment for) an autoimmune disease
Persistent leukopenia or leukocytosis (WBC ≤ 3.5 K/µL or ≥ 20.0 K/µL) or an absolute neutrophil count < 1.5K/µL
Concomitant illness or requirement for chronic drug treatment that in the opinion of the PI creates unnecessary risks for gene transfer
Pregnancy
AAVrh74 or AAV8 binding antibody titers > 1:50 as determined by ELISA immunoassay
Abnormal laboratory values in the clinically significant range in the table below, based upon normal values in the Nationwide Children's Hospital Laboratory: GGT, Total Bilirubin, Cystatine, Hemoglobin, White Blood Cells
This first phase of the trial will be small and will only evaluate safety; patients enrolled are not expected to show improvement in their symptoms. If the initial trial is successful, a Phase 2 trial will be scheduled, which will look for efficacy (improvement in symptoms) as well as safety. Each phase would include more patients. So, it is a process and we are pacing ourselves as we support this major milestone in the field of therapeutic intervention for people suffering from dysferlinopathy.