Targeting Autophagy in Myogenic Cells to Counteract Muscle Aging and Neuromuscular Diseases: Novel Bioengineering Tools for Validating Pharmacological Candidates
Description: Several muscular dystrophies (MDs) do share with aging common features and underlying mechanisms impinging on muscle ability to regenerate and recover disease-induced damage. In both MDs and aging, muscle regeneration and autophagy display a decline in activity, thus impairing muscle homeostasis. In this project we propose to throw light on a major missing link in this context, concerning the impact of autophagy deregulation on muscle regenerative potential, by investigating the influence of aging and disease condition on the autophagic activity of the main myogenic cell populations involved in muscle regeneration, namely satellite cells (SCs), myoblasts (MBs) and muscle pericytes (MPs). Toward this aim, we will set up and exploit biomimetic 3D-printed culturing systems, in order to reproduce the physiological environment and mimic the biomechanical, structural and biochemical properties of young and aged skeletal muscle. Primary murine SCs and MBs will be cultured in young- and aged-like 3D scaffolds, and autophagy activity along with myogenic potential will be monitored in vitro. In parallel, primary SCs and MBs will be isolated from Ambra1flox/flox and Becn1flox/flox mice and, following transfection with Cre, used as a control of autophagy inhibition, based on the role of Ambra1 and Beclin1 in the induction of autophagy in different tissues. To fully exploit this novel in vitro system, young- and aged-like 3D scaffolds will be used as a culturing platform to validate autophagy modulating molecules, such as spermidine, resveratrol, rapamycin and analogues, able to enhance the autophagic activity of myogenic cells in aging and disease conditions. Towards this aim, primary SCs and MBs derived from wild-type and LmnaG609G progeria mouse (a model of enhanced aging) will be tested, in parallel with cells derived from Col6a1–/–, mdx and Lmna–/–, mouse models for Collagen VI-related myopathies [Ullrich congenital muscular dystrophy (UCMD), Bethlem myopathy (BM), myosclerosis myopathy (MM)], Duchenne muscular dystrophy (DMD), and Emery-Dreifuss muscular dystrophy (EDMD), respectively. In addition, we will use the same experimental setup to test a selection of positive drug candidates also in primary SCs, MBs and MPs derived from muscle biopsies of UCMD, BM, MM, DMD and EDMD patients. Finally, drug candidates able to enhance autophagy in disease-derived cells will be administered in vivo in progeria, Col6a1 null and mdx mice, thus testing the ability of the selected drugs to ameliorate muscle phenotype, by either counteracting muscle aging or recover/slow down disease progression. The achievement of the proposed aims will settle a novel cutting-edge bioengineered application in the field of muscle physiology and pathology. Moreover, they will pave the way for the development of new therapies targeting autophagy in myogenic cells and counteracting aging and disease in different muscle conditions.
Amount: €171,500
Funding type: research
Funder scheme: PRIN 2015
Investigators: Paolo Bonaldo, Stefano Maria Cannata, Paolo Bonaldo, Stefano Maria Cannata, Cesare Faldini, Giovanna Lattanzi, Cecilia Gelfi
Institutions: Università degli Studi di PADOVA, Università degli Studi di ROMA "Tor Vergata", Alma Mater Studiorum - Università di BOLOGNA, Consiglio Nazionale delle Ricerche, Università degli Studi di MILANO
Funded outputs count: 19
Funder award ID: 2015FBNB5Y
OpenAlex ID: G3151843983