Micro-engineered skeletal and cardiac muscle for Duchenne muscular dystrophy in vitro models
Year: 2012
Type: dissertation
Abstract: Duchenne muscular dystrophy (DMD) is the most common and severe genetic neuromuscular disorder affecting both skeletal and cardiac muscle functionality. More than twenty years have passed since the identification of the dystrophin gene, which mutations cause the disease. Many progresses have been made in understanding the pathogenesis and different experimental strategies has been tested both in vitro, on bench-top cell cultures, and in vivo, on different animal models. So far, despite some promising outcomes coming from recent clinical trials, this has not resulted in an effective and definitive cure significantly altering the relentless progression of this disease, which has still a 100% mortality rate. In this context, the aim of this PhD thesis is the development of micro-engineered human skeletal and cardiac muscles representing in vitro models of DMD patient tissues useful for testing therapeutic approaches aimed at restoring a proper dystrophin expression. The strategy applied for the obtainment of such human in vitro models rely on the application of micro-scale technologies for reproducing in vitro the main physiological cues that guide differentiation and allow functionality of skeletal and cardiac muscles in vivo. In particular, the mechanical properties of the cell micro-environment and the topologic organization over the cell culture were optimized for both skeletal and cardiac muscle. Such micro-scale technologies were coupled with an appropriate human cell source. Human primary myoblasts from biopsies of DMD patient were used for skeletal muscle engineering, while DMD patient-specific cardiomyocytes were differentiated from human pluripotent stem (hiPS) cells for modeling the cardiac muscle. Both the obtained DMD in vitro models were validated for testing the ability of different therapeutic approaches in restoring dystrophin expression. In particular, three different myogenic cell types were tested on the engineered DMD skeletal muscle, while dystrophin expression restoration by a human artificial chromosome carrying the full-length genomic dystrophin sequence was tested on hiPS cells-derived cardiomyocytes. In these perspectives, the developed human in vitro models can represent a useful platform for performing preliminary or pre-clinical tests of different therapeutic strategy for DMD. In addition, they can be used as complementary tool in a clinical trial, for test different batches of cells before using them on patients.
Author: Susi Zatti
Language: English
Subfield: Molecular Biology
Domain: Life Sciences
Open Access status: green
OpenAlex ID: W2399983424