Introduction: Age-related macular degeneration (AMD) is the leading cause of irreversible vision loss among people over 55 years of age globally, being neovascular AMD (nAMD) its most aggressive form. Its treatment consists of the use of drugs that block vascular endothelial growth factor (anti-VEGF). Proteomics may allow the identification of differentially expressed proteins between responders and non-responders to each anti-VEGF drug. Thus, the objective of Pharmacoproteomics in the development of personalised medicine in Age-related Macular Degeneration (PHARPRO-AMD) is to find new proteomic biomarkers, predictive of response to antiangiogenic treatment in patients with nAMD. Methods and analysis: PHARPRO-AMD is a nationwide, multicentre, prospective, observational study. Treatment-naïve patients with nAMD starting anti-VEGF therapy will be enrolled and followed up for 2 years. During this period, clinical variables will be gathered to classify treatment response. In addition, blood, tear and vitreous and aqueous humour samples will be collected and will undergo a ZenoSWATH proteomic analysis. Relevant biomarkers identified and response classification will be used to perform a multivariate logistic regression and construct receiver operating characteristic curves. Results: The study is expected to identify a panel of proteomic biomarkers predictive of anti-VEGF treatment response. Integrating data from invasive and non-invasive biological samples may enhance clinical applicability. Once validated, these biomarkers could support the design of future clinical trials on biomarker-guided therapies, helping to optimise treatment regimens and improve visual outcomes. Conclusions: The PHARPRO-AMD study aims to provide proof-of-concept for biomarker-guided anti-VEGF therapy in nAMD, potentially improving vision outcomes. A notable limitation is the exclusion of patients with visual acuity above 73 Early Treatment of Diabetic Retinopathy Study letters, a criterion chosen to reduce potential ceiling effects and improve response assessment accuracy. Ethics and dissemination: Approved by the Galician Network of Ethics Committees, with nationwide validity. Anonymised data will be deposited in open-access repositories and published in peer-reviewed journals. Trial registration number: Spanish Clinical Studies Registry (REec) (0033-2024-OBS).

Objectives: To characterize the one-year functional, anatomical, and molecular responses to intravitreal Ranibizumab in treatment-naïve patients with neovascular age-related macular degeneration (nAMD), and to identify systemic immune and miRNA signatures associated with treatment response. Methods: This prospective longitudinal observational study included 44 treatment-naïve patients with nAMD. Patients received up to four monthly intravitreal Ranibizumab injections, followed by a treat-and-extend regimen. Best-corrected visual acuity using ETDRS letters, central retinal thickness by optical coherence tomography, fluorescein angiography, and OCT angiography were assessed at baseline and 12 months. Peripheral blood samples were collected at both time points to quantify seven circulating cytokines using an IMMULITE chemiluminescent immunoassay and to profile 37 candidate miRNAs by TaqMan OpenArray RT-qPCR from leukocyte-derived RNA. Treatment response was classified using composite anatomical and functional criteria, including intraretinal/subretinal fluid resolution, ≥25% central retinal thickness reduction, and a ≥5 ETDRS letter gain. Results: At one year, patients showed significant central retinal thickness reduction and overall visual stabilization, although good and poor responders differed according to composite response criteria. Statin use was numerically more frequent among poor responders, although this difference was not statistically significant. Soluble IL-2R increased significantly over time in the overall cohort, mainly driven by good responders who showed higher median levels at both visits. IL-8 also increased globally, without significant between-group differences. Among differentially expressed miRNAs, miR-3121 was the only candidate reaching statistical significance and was downregulated in good responders. ROC analysis showed moderate discriminative performance for miR-3121, with an AUC of 0.76. Conclusions: One-year response to Ranibizumab in nAMD may involve systemic immune activation and miRNA regulation. miR-3121 emerges as a candidate biomarker of treatment response, supporting further validation in larger independent cohorts.

Background: Linezolid commonly causes hematologic toxicity, especially thrombocytopenia, during extended treatment. This effect is linked to the inhibition of mitochondrial protein synthesis, which impairs oxidative phosphorylation and cellular energy production. Although reduced complex IV activity has been observed in long-term therapy, the connection between treatment duration and mitochondrial dysfunction in humans remains insufficiently defined. Methods: Forty patients were included and stratified by treatment duration: 2–7 days (group 1), 8–14 days (group 2) and more than 14 days (group 3). To evaluate mitochondrial function in peripheral blood mononuclear cells (PBMCs) from patients treated with linezolid for different durations, Seahorse XF was used. To assess protein expression changes associated with mitochondrial toxicity, LC-MS/MS was used. Differentially expressed proteins were identified in R based on fold-change thresholds (|log₂FC|≥0.58) and p < 0.05, and results were visualized with volcano plots and STRING interaction networks. Results: All the groups were similar in terms of variables and characteristics. Mitochondrial respiration declined progressively with treatment duration and platelet counts showed a parallel reduction. Proteomic analysis identified one cluster of downregulated protein in group 2 of patients and two in group 3; they were involved in mitochondrial ATP synthesis—mainly subunits of complexes I and IV of the respiratory chain—supporting a concordant functional and molecular pattern of mitochondrial impairment. Conclusions: Prolonged linezolid treatment was associated with progressive mitochondrial dysfunction, as reflected by both functional assays and proteomic profiles. Concordant changes in respiration and protein expression support mitochondrial involvement in linezolid-related toxicity and suggest that, in addition to previously reported alterations in complex IV, complex I proteins may also be affected in treated patients.