Abstract
Purpose :
Peptide therapy developments have gained prominence in recent decades. The capacity to modulate the fibrosis process following trabeculectomy surgery is critical for patient care. We synthesized a novel cell-penetrating peptide (VRF2019), conjugated it to mitomycin C, and then used multi-omics approaches to assess its influence on biological and cellular processes in primary human tenon fibroblast cells
Methods :
Human primary tenon fibroblast cells (control) were treated with the conjugate and peptide alone. We performed RNA sequencing on the treated fibroblast cells using the Illumina platform to acquire a global transcriptome signature. We also investigated the kinome of conjugate-treated cells using the Protein Profiler. We investigated the differential expression of coding and non-coding (mRNA and lncRNA) genes, as well as the pathways altered with conjugate treatment.
Results :
Global transcriptome profile of peptide and conjugate-treated fibroblast cells showed changes in both canonical and non-canonical fibrotic pathways. The TGF- β and MAP kinase pathways were downregulated in cells treated with peptide or conjugate. Long Non-coding RNAs that regulate canonical fibrotic pathways are also downregulated such as LcSNHG5, LINC01504 ncRNA, PEG3, LUCAT1, and MAGI1-IT1. Similar to the transcriptomic effect, the targeted phospho-kinase array data demonstrated dysregulated phospho-signaling in non-canonical fibrotic pathways. Phosphorylation of Proteins like ERK1/2, MSK1/2, and AKT1/2/3, were downregulated in peptide treated than conjugate treated. After peptide and drug conjugate treatment, the most significantly differentially regulated pathways include the apoptosis signalling pathway, TGF-β signalling pathway, MAPK signalling pathway, Toll-like receptor signalling pathway, FoxO signalling, and p53 signalling pathway.
Conclusions :
The transcriptomic and targeted kinome data following peptide and drug conjugate therapy revealed an effect on profibrotic gene downregulation of TGF-β and MAP kinase, which may aid in delaying the fibrosis progression
This abstract was presented at the 2024 ARVO Annual Meeting, held in Seattle, WA, May 5-9, 2024.