Systematic Interrogation of the Stiffness-sensitive Transcriptome in Mesenchymal Stromal Cells Reveals an Immunomodulatory LncRNA CYTOR

Previous studies have demonstrated that the physical properties of biomaterials can be adjusted to program therapeutically relevant functions in encapsulated mesenchymal stromal cells (MSCs). However, the intracellular pathways affected by mechanical cues in this cell type have not been thoroughly investigated. An understanding of the pertinent factors involved would not only aid in the rational design of substrates but also uncover targetable mechanisms that can facilitate the engineering of MSCs for cell therapies. Here, a bone marrow-mimetic hydrogel is employed to systematically explore the stiffness-responsive transcriptome of MSCs. High matrix rigidity impedes integrin-collagen adhesions which yields changes in cell morphology that are characterized by a contractile network of actin proximal to the cell membrane. This results in a suppression of extracellular matrix (ECM)-regulatory genes involved in the remodeling of collagen fibrils and an upregulation of secreted immunomodulatory factors. Moreover, matrix stiffness induces the expression of nuclear factor kappa B (NF-κB) and activator protein 1 (AP-1) regulons, suggesting that these transcription factors serve as mechanotransducers. In addition, an investigation of long non-coding RNAs reveals that CYTOR contributes to these stiffness-driven changes in gene abundance. Knockdown of CYTOR using antisense oligonucleotides enhances the expression of numerous mechanoresponsive cytokines and chemokines to levels exceeding what is achievable by modulating matrix stiffness alone. Taken together, these findings reveal previously unexplored mechanisms of mechanotransduction that inform novel strategies for enhancing the efficacy of MSC-based therapies.

Department dc:contributor.department

Molecular Genetics

Year dc:date.issued

2024

Author dc:creator

• Lim, Justin Jisu

Advisor dc:contributor.advisor

• Blencowe, Benjamin J

• biomaterials
• lncRNA
• mechanotransduction
• mesenchymal stromal cells
• rna-seq

Handle dc:identifier.uri

http://hdl.handle.net/1807/140730

OAI identifier oai:identifier

oai:utoronto.scholaris.ca:1807/140730