Profiling muscle stem cell adhesion using extracellular matrix peptide arrays

Aging is associated with a higher incidence of disease, which severely affects the quality of life and increases pressure on healthcare systems. Declining skeletal muscle (SkM) mass and function coupled with a reduced tissue healing capacity is one of the main drivers of loss of independence in the elderly. Our previous work revealed that aged SkM contains lower levels of certain extracellular matrix (ECM) components that are critical for proper adhesion of muscle stem cells (MuSCs), which are responsible for muscle regeneration. These observations support the notion that peptides derived from ECM proteins containing the consensus sequence for activation of adhesion receptors could have therapeutic value for the treatment of age associated MuSC dysfunction. Here we performed a medium-throughput screen using CelluSpots peptide arrays generated with the Intavis Peptide Synthesizer and a slide spotting robot to access ECM molecule derived motif’s capacity to promote adhesion of MuSC-derived primary myoblasts. Using this experimental paradigm, we identified 5 peptide sequences derived from 4 different ECM components that significantly increase adhesion of primary myoblasts. Using biotin modifications of these candidate peptides in conjunction with fluorescent streptavidin, we were able to demonstrate that they can be used as adhesion receptor probes that strongly bind the plasma membrane of primary myoblasts. In addition, we developed 2D cell culture systems to further assess effects on cell adhesion and successfully integrated an adhesion peptide to the basal lamina after in vivo delivery. Altogether, this project will lay the foundation for the development of much needed therapeutic interventions for degenerative age-associated skeletal muscle conditions.

Name thesis:degree_name

M. Sc.

Level thesis:degree_level

Maîtrise

Discipline thesis:degree_discipline

Pharmacologie

Grantor dc:publisher

Université de Sherbrooke

Year dc:date.issued

2023

Author dc:creator

• Leblanc, Élizabeth

Advisor dc:contributor.advisor

• Bentzinger, Conrad Florian

• Cellules souches musculaires
• Matrice extracellulaire
• Niche de cellules souches
• Adhésion
• Séquences peptidiques
• Synthèse peptidique
• Muscle stem cells
• Myoblasts
• Extracellular matrix
• Adhesion motifs
• Stem cell niche
• Peptide sequences
• Peptide synthesis

Language dc:language.iso

en

DOI

https://doi.org/10.71892/11143/1408

OAI identifier oai:identifier

oai:usherbrooke.scholaris.ca:11143/23990