Serafina Pacilio
Engineering 3D Models to Investigate LGMDD2 Transportin 3 Related: Insights into Myogenic Processes and Contractile Dysfunction
Autori
- SERAFINA PACILIO (DEPARTMENT OF BIOMEDICAL AND NEUROMOTOR SCIENCES DIBINEM, ALMA MATER STUDIORUM-UNIVERSITÀ DI BOLOGNA, ITALY – PHD STUDENT )
- ROBERTA COSTA (DEPARTMENT OF BIOMEDICAL AND NEUROMOTOR SCIENCES DIBINEM, ALMA MATER STUDIORUM-UNIVERSITÀ DI BOLOGNA, ITALY – RESEARCHER)
- MARIA TERESA RODIA (DEPARTMENT OF BIOMEDICAL AND NEUROMOTOR SCIENCES DIBINEM, ALMA MATER STUDIORUM-UNIVERSITÀ DI BOLOGNA, ITALY – TECHNICIAN )
- SARA LOMBARDI (DEPARTMENT OF BIOMEDICAL AND NEUROMOTOR SCIENCES DIBINEM, ALMA MATER STUDIORUM-UNIVERSITÀ DI BOLOGNA, ITALY – RESEARCHER )
- LUANA DI LISA (DEPARTMENT OF CHEMISTRY “”GIACOMO CIAMICIAN”” ALMA MATER STUDIORUM-UNIVERSITÀ DI BOLOGNA, ITALY – PHD STUDENT )
- GHISLAIN BANOS (UNIVERSITÉ PARIS EST CRÉTEIL, INSERM, EFS, IMRB, F-94010 CRÉTEIL, FRANCE – PHD STUDENT )
- NATHALIE DIDER (UNIVERSITÉ PARIS EST CRÉTEIL, INSERM, EFS, IMRB, F-94010 CRÉTEIL, FRANCE – RESEARCHER)
- EDOARDO MALFATTI (UNIVERSITÉ PARIS EST CRÉTEIL, INSERM, U955, IMRB, F-94010 CRÉTEIL, FRANCE; REFERENCE CENTER FOR NEUROMUSCULAR DISORDERS, APHP HENRI MONDOR UNIVERSITY HOSPITAL, FRANCE – PROFESSOR )
- GIOVANNA CENACCHI (DEPARTMENT OF BIOMEDICAL AND NEUROMOTOR SCIENCES DIBINEM, ALMA MATER STUDIORUM-UNIVERSITÀ DI BOLOGNA, ITALY – PROFESSOR )
- MARIA LETIZIA FOCARETE (DEPARTMENT OF CHEMISTRY “”GIACOMO CIAMICIAN”” ALMA MATER STUDIORUM-UNIVERSITÀ DI BOLOGNA, ITALY – PROFESSOR)
Presentatore
SERAFINA PACILIO
Modalità
Oral Communication
Abstract
Limb girdle muscular dystrophies (LGMD) constitute a group of inherited myopathies characterized by muscle fibers degeneration and progressive muscle weakness. Our study focuses on an autosomal dominant form of LGMDD2 linked to transportin 3 (TNPO3) deficiency. We applied tissue engineering to create 3D tissue-like structure by combining myoblast, biomaterials and biophysical elements. We employed immortalized human myoblasts derived from LGMDD2 patients cultured in two 3D scaffolds. The 3D collagen bio-printed hydrogel, recapitulating the native tissue, proved valuable for investigating myogenic processes and exploring TNPO3 in disease development. The 3D micropillar system, resembling the muscle structure, facilitates real-time monitoring of cell contractility. Analysis of genes and proteins related to myogenesis, and muscle-specific proteins, TNPO3, was performed. Several dysregulations were found during the myogenic process in LGMDD2 compared to control, with early differentiation and increased expression of Murf-1 and p62, indicators of atrophy and autophagy. Dysregulation in LGMDD2 was confirmed by morphological studies which revealed reduced maturation, few striations, and α-actinin aggregates. To evaluate cell contractility, electrical stimulation was applied on the micropillars 3D model using 5V with 0.5 Hz to mimic twitch contraction, and 5V with 10 Hz to mimic tetanic contraction under live imaging to monitor micropillars deflections. LGMDD2 cells exhibited greater contractile force compared to control. However, their contractile force was irregular in frequency and amplitude suggesting functional abnormalities, possibly due to abnormal calcium release. In conclusion our 3D models are able to faithfully modelize muscular dystrophies and could be used for calcium imaging and drug repurposing approaches.