Limb Girdle Muscular Dystrophy (LGMD) Gene Therapy
Publications

Publications


LGMD2D [alpha-sarcoglycan deficiency (SGCA)]

Rodino-Klapac, LR, Lee, JS, Mulligan, RC, Clark, KR and Mendell, JR. (2008) Lack of toxicity of alpha-sarcoglycan overexpression supports clinical gene transfer trial in LGMD2D. Neurology 71:240-247. PMID: 18525034

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)

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. (2009) LGMD 2D gene therapy restores alpha-sarcoglycan and associated proteins. Annals of Neurology 66: 290-297. PMID:19798725

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.

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. (2010) Sustained alpha-sarcoglycan gene expression after gene transfer in Limb-Girdle Muscular Dystrophy, Type 2D. Annals of Neurology 68: 629-638. PMID:21031578

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.

Publications


LGMD2E [beta-sarcoglycan deficiency (SGCB)]

Pozsgai, ER, Griffin, DA, Heller, KN, Mendell, JR and Rodino-Klapac, LR. (2016). β-Sarcoglycan Gene Transfer Decreases Fibrosis and Restores Force in LGMD2E Mice. Gene Therapy. 23: 57-66. PMID: 26214262

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.

Pozsgai, ER, Griffin, DA, Heller, KN, Mendell, JR and Rodino-Klapac, LR. (2017) Systemic AAV-Mediated β-Sarcoglycan Delivery Targeting Cardiac and Skeletal Muscle Ameliorates Histological and Functional Deficits in LGMD2E Mice. Molecular Therapy. 25: 855-869. PMID: 28284983

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).

Publications


LGMD2B [dysferlin deficiency]

Grose WG, Clark KR, Griffin DG, Shontz K, Malik V, Montgomery CL, Janssen, PM, Brown RJ, Mendell JR, Rodino-Klapac LR. (2012) Homologous Recombination Mediates Functional Recovery of Dysferlin Deficiency Following AAV5 Gene Transfer. PLoS One. 7:e39233. PMID:22720081

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.

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. (2015) Dysferlin Restoration using Overlapping AAV Vectors Restores Function in Animal Models of Dysferlinopathies. Annals of Clinical and Translational Neurology2: 256-270. PMID: 25815352

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.

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. (2017) Systemic Delivery of Dysferlin Overlap Vectors Provides Long-Term Functional Improvement for Dysferlinopathy. Human Gene Therapy. In Press. PMID: 28707952

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.

Publications


LGMD2L [anoctamin 5 deficiency (ANO5)]

Griffin, DA, Johnson, RW, Whitlock, JM, Pozsgai, ER, Heller, KH, Grose, WE, Arnold, WD, Sahenk, Z; Hartzell HC and Rodino-Klapac, LR. (2016). Defective membrane fusion and repair in anoctamin5-deficient muscular dystrophy. Human Molecular Genetics 25: 1900-1911. PMID: 26911675

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

Publications


Additional Articles Related to the Use of AAVrh74 and Vascular Delivery

Rodino-Klapac, LR, Janssen, PML, Montgomery, CL, Coley, BD, Chicoine, LG, Clark, KR and Mendell, JR.  (2007) A translational approach for limb vascular delivery of the micro-dystrophin gene without high volume or high pressure for treatment of Duchenne muscular dystrophy.  Journal of Translational Medicine 5:45. PMID:17892583

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

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.  (2010) Persistent expression of FLAG tagged micro-dystrophin in non-human primates with intramuscular and vascular delivery.  Molecular Therapy. 18: 109-117. PMID:19904237

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

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. (2014). Plasmapheresis eliminates the negative impact of AAV antibodies on micro-dystrophin gene expression following vascular delivery.  Molecular Therapy. 22:338-347.  PMID: 24196577

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.

Publications


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

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. (2017). Single-Dose Gene-Replacement Therapy for Spinal Muscular Atrophy. New England Journal of Medicine 377: 1713-1722. PMID: 29091557

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.

Publications


Proof of Principle for Newborn Screening for LGMD

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. (2012). Evidence-Based Path to Newborn Screening for Duchenne Muscular Dystrophy. Annals of Neurology 71: 304-313. PMID: 22451200

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.