Rare diseases, less than 50-60/100,000 people, include neurological disorders and myopathies. There is a gap between basic research and therapy development and even if the number of orphan drugs is growing, the majority of rare diseases with Mendelian inheritance still need gene or cell therapy. Muscular dystrophies, MDs, are rare genetic myopathies characterized by progressive muscle weakness, muscle degeneration, and fibro-adipose substitution. The pathological phenotype of MDs is rarely linked only to the malfunction of a single gene, but rather derives from the alteration of multiple molecular networks. No definitive cure is available for MDs and significant effort is being placed to either correct the genetic defect and to stimulate skeletal muscle regeneration. Inflammation is fundamental to the regenerative process, but in several MDs, the continuous release of the myofiber content upon degeneration leads to sustained inflammation, which contributes to the pathogenesis of the disease. Moreover, several findings in MDs show there is not a single immune response and suggest that endogenous immunomodulators may contribute to diverse immune responses to distinct MDs. Standard treatment for MDs relies on corticosteroids, with side effects such as muscle weakness and atrophy. Therefore, the modulation of the immune response is a relevant therapeutic option for MDs, but we still need new strategies without or with minor side effects. Cell therapy, as mesenchymal stem cell (MSC) transplantation, could be a strategy for many rare diseases to improve tissue repairing/regenerating. In addition, MSCs mediate favorable effects through paracrine signaling, which stimulate tissue healing by its anti-inflammatory, anti-fibrotic and anti-angiogenic properties. In this project we will use immortalized myoblasts from different MDs, culturing them on a new 3D-bioprinted scaffold, recreating the muscle stem cell niche microenvironment; then we will set up co-cultures with immune cells and MSCs to study inflammation and its modulation by MSCs as a common feature among several MDs. We will focus on the investigation of inflammation markers and related pathways analyzing secretome and transcriptome, as well as the miRNA profile. This step is crucial in understanding both the role of inflammation in muscle disorders and the immunomodulatory action of MSCs to find possible common key actors and druggable targets. The use of a 3D-system will also enable drug repositioning and testing of multiple compounds, as chemical libraries and natural drugs, in a high-throughput manner. The reduction in costs and time expenditure assured by these strategies make them interesting, especially for those diseases for which the canonical drug discovery process would be long and expensive. The project results could be also considered as a proof-of-concept of an innovative tool in drug testing, not only for rare disease, overcoming the needs of animal experimentation.
Study of myopathy 3D models for anti-inflammatory treatment strategy
MONICA BORGATTI
2023
Abstract
Rare diseases, less than 50-60/100,000 people, include neurological disorders and myopathies. There is a gap between basic research and therapy development and even if the number of orphan drugs is growing, the majority of rare diseases with Mendelian inheritance still need gene or cell therapy. Muscular dystrophies, MDs, are rare genetic myopathies characterized by progressive muscle weakness, muscle degeneration, and fibro-adipose substitution. The pathological phenotype of MDs is rarely linked only to the malfunction of a single gene, but rather derives from the alteration of multiple molecular networks. No definitive cure is available for MDs and significant effort is being placed to either correct the genetic defect and to stimulate skeletal muscle regeneration. Inflammation is fundamental to the regenerative process, but in several MDs, the continuous release of the myofiber content upon degeneration leads to sustained inflammation, which contributes to the pathogenesis of the disease. Moreover, several findings in MDs show there is not a single immune response and suggest that endogenous immunomodulators may contribute to diverse immune responses to distinct MDs. Standard treatment for MDs relies on corticosteroids, with side effects such as muscle weakness and atrophy. Therefore, the modulation of the immune response is a relevant therapeutic option for MDs, but we still need new strategies without or with minor side effects. Cell therapy, as mesenchymal stem cell (MSC) transplantation, could be a strategy for many rare diseases to improve tissue repairing/regenerating. In addition, MSCs mediate favorable effects through paracrine signaling, which stimulate tissue healing by its anti-inflammatory, anti-fibrotic and anti-angiogenic properties. In this project we will use immortalized myoblasts from different MDs, culturing them on a new 3D-bioprinted scaffold, recreating the muscle stem cell niche microenvironment; then we will set up co-cultures with immune cells and MSCs to study inflammation and its modulation by MSCs as a common feature among several MDs. We will focus on the investigation of inflammation markers and related pathways analyzing secretome and transcriptome, as well as the miRNA profile. This step is crucial in understanding both the role of inflammation in muscle disorders and the immunomodulatory action of MSCs to find possible common key actors and druggable targets. The use of a 3D-system will also enable drug repositioning and testing of multiple compounds, as chemical libraries and natural drugs, in a high-throughput manner. The reduction in costs and time expenditure assured by these strategies make them interesting, especially for those diseases for which the canonical drug discovery process would be long and expensive. The project results could be also considered as a proof-of-concept of an innovative tool in drug testing, not only for rare disease, overcoming the needs of animal experimentation.I documenti in SFERA sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.
