Quantitative proteomic analysis of aqueous humor after rabbit lensectomy reveals differences in coagulation and immunomodulatory proteins

• Published in: Molecular omics 2020-04, Vol.16 (2), p.126-137
• Main Authors: Young, Jonathon B, Keppel, Theodore R, Waas, Matthew, Salmon, Alexander E, Buchberger, Amanda Rae, Skumatz, Christine M.B, Gundry, Rebekah L, Kassem, Iris S
• Format: Article
• Language: English
• Subjects: Animals; Aqueous Humor - metabolism; Disease Models, Animal; Eye Proteins - metabolism; Fibrinogen - metabolism; Inflammation - etiology; Inflammation - metabolism; Lens Implantation, Intraocular - adverse effects; Lens, Crystalline - surgery; Male; Proteomics - methods; Rabbits; Retinol-Binding Proteins - metabolism; Sutures - adverse effects; Transforming Growth Factor beta2 - metabolism; Up-Regulation
• ISSN: 2515-4184; 2515-4184
• DOI: 10.1039/c9mo00169g

Compared to adults, children experience increased postoperative scarring and inflammation following intraocular surgery. While the underlying causes of the exaggerated immune response in children are not understood, proteins play key roles in postoperative scarring and wound healing processes. To identify and quantify proteins associated with the robust postoperative immune response, this study applied quantitative proteomics approaches to a juvenile rabbit model of lensectomy with intraocular lens (IOL) insertion. Twenty-six 6-7 week-old New Zealand white rabbits underwent unilateral portions of lensectomy with IOL insertion including: anterior chamber paracentesis, corneal incision with wound suture, lensectomy only, and lensectomy with IOL insertion. Aqueous humor was collected immediately prior and three days after each procedure. Semi-quantitative protein discovery was achieved by label-free quantitation using data dependent and data independent acquisition modes. Based on the discovery results, targeted quantitation by parallel reaction monitoring of 3 proteins of interest, fibrinogen-beta chain, transforming growth factor beta-2, and retinol binding protein 3, was used to confirm the observed quantitative trends. Total protein concentration levels increased with each progressive surgical step of lensectomy with IOL insertion. Proteins related to the complement and coagulation cascades were found to increase in relative abundance, while proteins related to ocular immunosuppression decreased in abundance following surgery. These data provide insights into the postoperative response by providing the first surgical step-wise views of the AH proteome before and after surgery. Overall, this work provides the foundation for future investigations targeting specific proteins for therapeutic interventions aimed at minimizing postoperative complications after pediatric intraocular surgery. Proteomics of juvenile rabbit aqueous humor was evaluated before and after surgery to determine proteins responsible for postoperative outcomes.