A scalable, clinically severe pig model for Duchenne muscular dystrophy

Abstract

Large animal models for Duchenne muscular dystrophy (DMD) are crucial for evaluation of diagnostic procedures and treatment strategies. Pigs cloned from male cells lacking DMD exon 52 (DMDΔ52) resemble molecular, clinical and pathological hallmarks of DMD, but die before sexual maturity and cannot be propagated by breeding. Therefore, we generated female DMD+/- carriers. A single founder animal had 11 litters with 29 DMDY/-, 34 DMD+/- as well as 36 male and 29 female wild-type offspring. Breeding with F1 and F2 DMD+/- carriers resulted in additional 114 DMDY/- piglets. With intensive neonatal management, the majority survived for 3-4 months, providing statistically relevant cohorts for experimental studies. Pathological investigations and proteome studies of skeletal muscles and myocardium confirmed the resemblance of human disease mechanisms. Importantly, DMDY/- pigs reveal progressive myocardial fibrosis and increased expression of connexin-43, associated with significantly reduced left ventricular ejection fraction already at age 3 months. Furthermore, behavioral tests provided evidence for impaired cognitive ability. Our breeding cohort of DMDΔ52 pigs and standardized tissue repositories provide important resources for studying DMD disease mechanisms and for testing novel treatment strategies.

Cardiac Macrophages and Their Effects on Arrhythmogenesis

“…Furthermore, the porcine immune system is very similar to the human immune system, especially compared to mice ( Mair et al, 2014 ; Pabst, 2020 ). Clinical routine techniques and equipment used for humans can easily be applied to pigs, a large number of disease models have been established and well characterized ( Schuttler et al, 2022 ), and targeted genetic modification has become feasible with recent technological advances ( Park et al, 2015 ; Langin et al, 2018 ; Renner et al, 2020 ; Riedel et al, 2020 ; Stirm et al, 2021 ). Moreover, pigs present action potentials and electrophysiological properties similar to humans as they express the same major ion currents (except I to ) ( Kaese et al, 2013 ; Piktel and Wilson, 2019 ).…”

Human muscle in gene edited pigs for treatment of volumetric muscle loss

“…The establishment and importance of pig models in biomedical research is clear ( Lunney et al, 2021 ). While the use of the pig for the study of muscular dystrophies ( Duque et al, 2015 ; Stirm et al, 2021 ) is established, its use for traumatic applications is more recent. The generation of chimeric animals represents an emerging field that holds remarkable potential for the treatment of VML injuries as well as the opportunity for the development of unique clinically based disease models.…”

Isolation and Characterization of Primary DMD Pig Muscle Cells as an In Vitro Model for Preclinical Research on Duchenne Muscular Dystrophy

“…The generation and characterization of the DMD pig have been published elsewhere [ 12 , 19 ]. In short, a German Landrace and Swabian-Hall mix sow with a heterozygous DMD exon 52 deletion ( DMD Δ52) was interbred with wild-type boars, resulting in the following offspring: male DMD Δ52 pigs (DMD Y/− or DMD pig), heterozygous DMD Δ52 carrier pigs ( DMD +/− ), and male and female wild-type pigs (WT).…”