Work

Proteomic profiling of adipose-derived mesenchymal stromal cell response to novel compounds targeting the Laminin Receptor – PEDF interaction

Year: 2025

Type: article

Abstract: This study characterizes the proteomic responses of adipose-derived mesenchymal stromal cells (ASC) to novel compounds that target the interaction between pigment epithelium-derived factor (PEDF) and the 37 KDa Laminin Receptor (LR). Building on previous research, we introduced the second-generation analog, 02–09, for comparative analysis with C3, the original compound. C3 was identified through in silico screening and experimental validation, showing anti-inflammatory activity. Analog 02–09 was synthesized by modifying C3's aromatic ring, resulting in similar binding affinities to LR. The distinct molecular effects of C3 and 02–09, along with their potential as anti-inflammatory therapies from prior work, suggested their multi-functional utility. Proteomic profiling of ASC, under both undifferentiated and chondrogenic differentiation conditions, was performed using LC-MS/MS, with controls including the PEDF-derived peptide p18 and the cartilage-regenerative compound Kartogenin (Krt). Both C3 and 02–09 induced significant proteomic modulation compared to controls, with enrichment in pathways related to integrin β1 signaling, extracellular matrix (ECM) organization , and TGF-β signaling . C3 uniquely activated the nuclear receptors meta-pathway, whereas 02–09 was associated with laminin interactions . Protein-protein interaction and transcription factor enrichment analyses revealed distinct and overlapping regulatory networks for LR-targeting compounds. During chondrogenic differentiation, C3 primarily enhanced proteins linked to a chondrogenic phenotype with minimal fibrogenic activity, while 02–09 showed an influence on both chondrogenic and fibrogenic pathways. Comparative analysis of the proteomic response to C3 or 02–09 with a single-cell RNA-sequencing dataset confirmed that C3's modulated proteins primarily correlated with a chondrogenic differentiation phenotype, with minimal fibrogenic influence. The proteins modulated by 02–09 correlated with a dual profile (both chondrogenic and fibrogenic potential). Our results suggest that C3 has potential as a targeted chondrogenic agent, while 02–09 may serve as a multifaceted modulator of both cartilage repair and ECM protection. These findings indicate the utility of these compounds for regenerative medicine and the development of disease-modifying therapies for inflammatory and immune-related conditions. • Proteomics showed ASC responses to LR–PEDF-targeting molecules. • Analog 02-09 and C3 displayed distinct differentiation profiles. • C3 promoted chondrogenic phenotype with minimal fibrogenic activity. • 02–09 induced chondrogenic and fibrogenic signatures. • LR-targeting modulated ECM, integrin, and TGF-β signaling pathways.

Source: Biochemistry and Biophysics Reports

DOI: 10.1016/j.bbrep.2025.102377