A Muscle Hybrid Promoter as a Novel Tool for Gene Therapy

Piekarowicz, Katarzyna; Bertrand, Anne T.; Azibani, Fériel; Beuvin, Maud; Julien, Laura; Machowska, Magdalena; Bonne, Gisèle; Rzepecki, Ryszard

doi:10.1016/j.omtm.2019.09.001

Cited by 18 publications

(22 citation statements)

References 60 publications

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“…NCT02354781 [ 56 ]), has been replaced by muscle specific promoters, such as MHCK7 (clinical trial no. NCT03375164) [ 57 ], and muscle hybrid promoters [ 58 ], in order to restrict the expression of mini- or micro- dystrophin to skeletal muscle and heart tissues.…”

Section: Discussionmentioning

confidence: 99%

Effects of Mini-Dystrophin on Dystrophin-Deficient, Human Skeletal Muscle-Derived Cells

Meng

Counsell

Morgan

2020

IJMS

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Background: We are developing a novel therapy for Duchenne muscular dystrophy (DMD), involving the transplantation of autologous, skeletal muscle-derived stem cells that have been genetically corrected to express dystrophin. Dystrophin is normally expressed in activated satellite cells and in differentiated muscle fibres. However, in past preclinical validation studies, dystrophin transgenes have generally been driven by constitutive promoters that would be active at every stage of the myogenic differentiation process, including in proliferating muscle stem cells. It is not known whether artificial dystrophin expression would affect the properties of these cells. Aims: Our aims are to determine if mini-dystrophin expression affects the proliferation or myogenic differentiation of DMD skeletal muscle-derived cells. Methods: Skeletal muscle-derived cells from a DMD patient were transduced with lentivirus coding for mini-dystrophins (R3–R13 spectrin-like repeats (ΔR3R13) or hinge2 to spectrin-like repeats R23 (ΔH2R23)) with EGFP (enhanced green fluorescence protein) fused to the C-terminus, driven by a constitutive promoter, spleen focus-forming virus (SFFV). Transduced cells were purified on the basis of GFP expression. Their proliferation and myogenic differentiation were quantified by ethynyl deoxyuridine (EdU) incorporation and fusion index. Furthermore, dystrophin small interfering ribonucleic acids (siRNAs) were transfected to the cells to reverse the effects of the mini-dystrophin. Finally, a phospho-mitogen-activated protein kinase (MAPK) array assay was performed to investigate signalling pathway changes caused by dystrophin expression. Results: Cell proliferation was not affected in cells transduced with ΔR3R13, but was significantly increased in cells transduced with ΔH2R23. The fusion index of myotubes derived from both ΔR3R13- and ΔH2R23 -expressing cells was significantly compromised in comparison to myotubes derived from non-transduced cells. Dystrophin siRNA transfection restored the differentiation of ΔH2R23-expressing cells. The Erk1/2- signalling pathway is altered in cells transduced with mini-dystrophin constructs. Conclusions: Ectopic expression of dystrophin in cultured human skeletal muscle-derived cells may affect their proliferation and differentiation capacity. Caution should be taken when considering genetic correction of autologous stem cells to express dystrophin driven by a constitutive promoter.

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Section: Discussionmentioning

confidence: 99%

Effects of Mini-Dystrophin on Dystrophin-Deficient, Human Skeletal Muscle-Derived Cells

Meng

Counsell

Morgan

2020

IJMS

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“…Poorly conserved sequences are typically excluded from the design; the importance for the expression of the remaining promoter elements was verified by mutation analysis [ 29 ]. A similar approach is creating hybrid/ chimeric promoters by adding the enhancer elements of one gene to the promoter region of another gene [ 30 ]. The expression level and tissue specificity can be significantly improved by varying the copy number of the enhancers and individual TFBS and properly combining these sequences [ 31 ].…”

Section: Natural Promotersmentioning

confidence: 99%

Muscle-Specific Promoters for Gene Therapy

2021

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Many genetic diseases that are responsible for muscular disorders have been described to date. Gene replacement therapy is a state-of-the-art strategy used to treat such diseases. In this approach, the functional copy of a gene is delivered to the affected tissues using viral vectors. There is an urgent need for the design of short, regulatory sequences that would drive a high and robust expression of a therapeutic transgene in skeletal muscles, the diaphragm, and the heart, while exhibiting limited activity in non-target tissues. This review focuses on the development and improvement of muscle-specific promoters based on skeletal muscle -actin, muscle creatine kinase, and desmin genes, as well as other genes expressed in muscles. The current approaches used to engineer synthetic muscle-specific promoters are described. Other elements of the viral vectors that contribute to tissue-specific expression are also discussed. A special feature of this review is the presence of up-to-date information on the clinical and preclinical trials of gene therapy drug candidates that utilize muscle-specific promoters.

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“…Sk-CRM4, containing binding sites for the E2A, CEBP, LRF, MyoD and SREBP transcription factors, boosted transgene expression driven by Des or C5.12 promoters in heart and skeletal muscles (up to 400-fold), with a significant improvement of the mdx mouse phenotype [ 269 ]. Similarly, a 1030 bp modular muscle hybrid (MH) promoter composed of two enhancers (from the Des and Mck genes, respectively), a proximal promoter and an intron (modified from the Mck gene and core promoter) was more efficient than the Des promoter in skeletal and cardiac muscles, with limited expression in non-muscle tissues compared with the CMV promoter, showing a high potential for muscular gene therapy [ 270 ].…”

Section: Improvement Of the Therapeutic Toolboxmentioning

confidence: 99%

Of rAAV and Men: From Genetic Neuromuscular Disorder Efficacy and Toxicity Preclinical Studies to Clinical Trials and Back

Buscara

Gross

Danièle

2020

JPM

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Neuromuscular disorders are a large group of rare pathologies characterised by skeletal muscle atrophy and weakness, with the common involvement of respiratory and/or cardiac muscles. These diseases lead to life-long motor deficiencies and specific organ failures, and are, in their worst-case scenarios, life threatening. Amongst other causes, they can be genetically inherited through mutations in more than 500 different genes. In the last 20 years, specific pharmacological treatments have been approved for human usage. However, these “à-la-carte” therapies cover only a very small portion of the clinical needs and are often partially efficient in alleviating the symptoms of the disease, even less so in curing it. Recombinant adeno-associated virus vector-mediated gene transfer is a more general strategy that could be adapted for a large majority of these diseases and has proved very efficient in rescuing the symptoms in many neuropathological animal models. On this solid ground, several clinical trials are currently being conducted with the whole-body delivery of the therapeutic vectors. This review recapitulates the state-of-the-art tools for neuron and muscle-targeted gene therapy, and summarises the main findings of the spinal muscular atrophy (SMA), Duchenne muscular dystrophy (DMD) and X-linked myotubular myopathy (XLMTM) trials. Despite promising efficacy results, serious adverse events of various severities were observed in these trials. Possible leads for second-generation products are also discussed.

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A Muscle Hybrid Promoter as a Novel Tool for Gene Therapy

Cited by 18 publications

References 60 publications

Effects of Mini-Dystrophin on Dystrophin-Deficient, Human Skeletal Muscle-Derived Cells

Effects of Mini-Dystrophin on Dystrophin-Deficient, Human Skeletal Muscle-Derived Cells

Muscle-Specific Promoters for Gene Therapy

Of rAAV and Men: From Genetic Neuromuscular Disorder Efficacy and Toxicity Preclinical Studies to Clinical Trials and Back

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