Limb Girdle Muscular Dystrophy (LGMD) Gene Therapy
Publications


LGMD2D [alpha-sarcoglycan deficiency (SGCA)]

Lack of toxicity of alpha-sarcoglycan overexpression supports clinical gene transfer trial in LGMD2D.

L. R. Rodino-KlapacJ-S LeeR. C. MulliganK. R. ClarkJ. R. Mendell

This was a preclinical study to evaluate safety and efficacy AAV1.SGCA intramuscular gene delivery comparing muscle specific versus a constitutive promoter. This study was pivotal for the approval of a Phase I intramuscular safety trial of AAV1.tMCK.SGCA (IND# 13434)

Limb‐girdle muscular dystrophy type 2D gene therapy restores α‐sarcoglycan and associated proteins.

Mendell, JR, Rodino-Klapac, LR, Rosales-Quintero, X, Kota, J, Coley, BD, Galloway, G, Craenen, JM, Lewis, S, Malik, V, Shilling, CJ, Byrne, B, Conlon, T, Campbell, KJ, Bremer, WG, Viollet, L, Walker, CM, Sahenk, Z, Clark, KR

This was the first publication under IND# 13434 (Phase I intramuscular trial of AAV1.tMCK.SGCA to the extensor digitorum brevis (EDB) muscle in LGMD2D patients – double blind placebo controlled. This first publication describes safety and efficacy in the first three subjects. Demonstrated evidence of safety, SGCA gene expression and improved muscle fiber size.

Sustained alpha‐sarcoglycan gene expression after gene transfer in limb‐girdle muscular dystrophy, type 2D.

Mendell, JR, Rodino-Klapac, LR, Rosales-Quintero, X, Kota, J, Coley, BD, Galloway, JM, Lewis, S, Malik, V, Shilling, CJ, Byrne, B, Conlon, T, Campbell, KJ, Bremer, WG, Taylor, LE, Flanigan, KM, Kota, J, Sahenk, Z, Walker, CM, Clark, KR.

This was the second publication under IND# 13434 (Phase I intramuscular trial of AAV1.tMCK.SGCA to the extensor digitorum brevis (EDB) muscle in LGMD2D patients – double blind placebo controlled. This second publication describes safety and efficacy in subjects 4-6 and overall study comparisons. Demonstrated evidence of safety, SGCA gene expression and improved muscle fiber size. Biopsies were done at 6 months in subjects 4-6 to demonstrated SGCA expression was sustained and not diminished compared to biopsies done at 6 or 12 weeks.


LGMD2E [beta-sarcoglycan deficiency (SGCB)]

β-Sarcoglycan Gene Transfer Decreases Fibrosis and Restores Force in LGMD2E Mice.

Pozsgai, ER, Griffin, DA, Heller, KN, Mendell, JR and Rodino-Klapac, LR.

This was a preclinical study performed in beta-sarcoglycan deficient mice to show safety and efficacy following intramuscular injection and isolated limb perfusion (ILP) of scAAVrh74.tMCK.SGCB. This publication demonstrated of robust gene expression, decreased fibrosis and correction of force deficits.

Systemic AAV-Mediated β-Sarcoglycan Delivery Targeting Cardiac and Skeletal Muscle Ameliorates Histological and Functional Deficits in LGMD2E Mice.

Pozsgai, ER, Griffin, DA, Heller, KN, Mendell, JR and Rodino-Klapac, LR.

This follow on study demonstrated safety and efficacy of intravenous delivery of scAAVrh74.MHCK7.SGCB. The promoter was changed from tMCK to MHCK7 to allow for expression in cardiac muscle in addition to skeletal muscle. This publication demonstrated 98.1% of fibers expressed SGCB across all skeletal muscles and >99% in the heart. Fibrosis was reversed, force was normalized, creatine kinase (CK) levels were normalized, kyphosis was improved, and overall activity was improved. This study was pivotal for the approval of a Phase I/IIa intravenous trial in LGMD2E patients (IND# 17060).


LGMD2B [dysferlin deficiency]

Homologous Recombination Mediates Functional Recovery of Dysferlin Deficiency Following AAV5 Gene Transfer.

Grose WG, Clark KR, Griffin DG, Shontz K, Malik V, Montgomery CL, Janssen, PM, Brown RJ, Mendell JR, Rodino-Klapac LR.

This proof of principle preclinical study demonstrated full-length dysferlin could be assembled via homologous recombination of AAV5 encapsidated vector fragments when delivered to Dysf-/- mice. Dysferlin expression improved histology of the muscle and the ability of Dysf-/- muscle to repair itself following injury.

Dysferlin Restoration using Overlapping AAV Vectors Restores Function in Animal Models of Dysferlinopathies. 

Sondergaard, PC, Griffin, DA, Pozsgai, ER, Johnson, RW, Grose, WE, Heller, KN, Shontz, KM, Montgomery, CL, Liu, J, Clark, KR, Sahenk, Z, Mendell, JR, and Rodino-Klapac, LR.

This preclinical study demonstrated full-length dysferlin can be delivered to muscle using two discrete vectors packaged using AAVrh74 that harbor a 1kb region of overlap as a substrate for recombination. The study demonstrated efficacy or intramuscular delivery, isolated limb perfusion, and systemic delivery. Efficacy included histological improvement, rescue of diaphragm force and membrane repair deficits. Safety was further confirmed in non-human primates. This study was pivotal for approval of a Phase I Intramuscular safety trial in LGMD2B patients under IND#16581.

Systemic Delivery of Dysferlin Overlap Vectors Provides Long-Term Functional Improvement for Dysferlinopathy.

Potter, RA, Griffin, DA, Sondergaard, PC, Johnson, RW, Pozsgai, ER, Heller, KN, Peterson, EL, Lehtimaki, K, Windish, H, Albrecht, D, Mendell, JR, and Rodino-Klapac, LR.

This is a follow-up pre-clinical study to demonstrated long-term benefit following intravenous (systemic) delivery of AAVrh74.MHCK7.DYSF.DV. Expression was sustained for the length of the study (2 years). MRI/MRS measurements were completed in the most affected muscles in the BlaJ dysferlin deficient mouse– the gluteus muscles. AAVrh74.MHCK7.DYSF.DV preserved these muscles for at least 15 months –study endpoint. Primates were also studied and confirmed safety/no toxicity. This study further validated systemic delivery as a viable method for treatment of LGMD2B.


LGMD2L [anoctamin 5 deficiency (ANO5)]

Defective membrane fusion and repair in anoctamin5-deficient muscular dystrophy.

Griffin, DA, Johnson, RW, Whitlock, JM, Pozsgai, ER, Heller, KH, Grose, WE, Arnold, WD, Sahenk, Z; Hartzell HC and Rodino-Klapac, LR.

Generation of first mouse model of LGMD2L. Proof of principle for delivery of AAV.ANO5 and improvement in membrane repair.


Additional Articles Related to the Use of AAVrh74 and Vascular Delivery

A translational approach for limb vascular delivery of the micro-dystrophin gene without high volume or high pressure for treatment of Duchenne muscular dystrophy.

Rodino-Klapac, LR, Janssen, PML, Montgomery, CL, Coley, BD, Chicoine, LG, Clark, KR and Mendell, JR.

Comparison of AAV serotypes 1, 6, and rh74 by intramuscular and vascular delivery.  Rationale for choice of AAVrh74

Persistent expression of FLAG tagged micro-dystrophin in non-human primates with intramuscular and vascular delivery.

Rodino-Klapac, LR, Montgomery, CL, Bremer, WG,  Shontz, KM, Malik,V, Davis, N, Sprinkle, S, Campbell, KJ,  Sahenk, Z, Clark, KR, Walker, CM, Mendell, JR, Chicoine, LG.

Safety and Efficacy of AAVrh74 by vascular delivery in non-human primates.

Plasmapheresis eliminates the negative impact of AAV antibodies on micro-dystrophin gene expression following vascular delivery.

Chicoine, LG, Montgomery, CL, Bremer, WG, Shontz, KM, Griffin, DA, Heller, KN, Lewis, S, Malik, V, Shilling, CJ, Campbell, KJ, Preston, TJ, Coley, BD, Martin, PT, Walker, CM, Clark, KR, Sahenk, Z, Mendell, JR, and Rodino-Klapac, LR.

Safety and Efficacy of AAVrh74 by vascular delivery in non-human primates.  Biodistribution of rh74 following vascular delivery. Plasmapheresis can effectively remove AAV antibodies and enhance gene expression.


Evidence of Safety and Efficacy following Systemic Delivery using high doses of AAV

Single-Dose Gene-Replacement Therapy for Spinal Muscular Atrophy.

Mendell JR, Al-Zaidy S, Shell R, Arnold WD, Rodino-Klapac LR, Prior TW, Lowes L, Alfano L, Berry K, Church K, Kissel JT, Nagendran S, L’Italien J, Sproule DM, Wells C, Cardenas JA, Heitzer MD, Kaspar A, Corcoran S, Braun L, Likhite S, Miranda C, Meyer K, Foust KD, Burghes AHM, Kaspar BK.

Demonstrated Safety and Efficacy of AAV9.SMN for the treatment of Spinal Muscular Atrophy Type 1 in Phase I/IIa study.  15/15 patients were alive and event-free at 20 months of age, as compared with a rate of survival of 8% in a historical cohort and high dose cohort (n=12) demonstrated rapid improvement from baseline in CHOP INTEND score.


Proof of Principle for Newborn Screening for LGMD

Evidence-Based Path to Newborn Screening for Duchenne Muscular Dystrophy. 

Mendell, JR, Shilling, CJ, Leslie ND, Flanigan, KM, Al-Dahhak, R, Gastier-Foster, J, Kneile, K, Dunn, D, Duval, B, Alexander, A, Hamil, C, Mahmoud, M, Roush, K, Bird, L, Rankin, C, Lilly, H, Street, N, Chandrasekar, R and Weiss, RB.

This study demonstrated that patients with LGMD can be identified with newborn screening.  The study evaluated <37,000 newborn males with the intention of identifying boys with DMD at birth.  A two-tiered system was used by first testing for elevated creatine kinase (CK) levels on dried blood spots. Subjects with CK elevation of >2000 U/l were sent for DNA testing.  In ~65% of the cases, DMD mutations were identified. In the remaining subjects, mutations were identified for 3 LGMD subtypes (LGMD2E, LGMD2B, and LGMD2i). This study demonstrated that this method of newborn screening is feasible for LGMD newborn screening and should also be applies to females as both genders are affected equally.