Testing a Prespecified Matrix Transcriptional Programme in Myeloid Cells from Proliferative Vitreoretinopathy Membranes: A Donor-Aware Computational Single-Cell Study

Abstract

Whether myeloid cells in proliferative vitreoretinopathy (PVR) membranes directly transcribe a coherent extracellular matrix (ECM) programme remains uncertain. We tested a prespecified seven-gene programme in a discovery cohort (GSE294329; five PVR donors) and an independent epiretinal PVR cohort (SCP2582; five donors), with GSE165784 (five public proliferative diabetic retinopathy [PDR] sample accessions of uncertain patient independence) providing separate disease context. Frozen cell states were evaluated using within-donor or within-accession summaries of rank scores, pseudobulk burden, detection, and ambient-corrected expression. In GSE294329, cluster 1 met the locked UCell discovery rule, but total ECM7 burden and detection were lower than in other myeloid cells in all five donors. Exact seven-gene replication was unevaluable because CCN2/CTGF was absent from the processed SCP2582 features; no eligible state passed the amended six-gene gate. In PDR, no myeloid state passed the direct-ECM gate, whereas stromal cells had higher ECM7 burden and detection in all five accessions. Secondary SAM-like states occurred in PDR clusters 1 and 6 only and did not rescue the direct-ECM result. An exploratory fixed-cell SCP2582 sensitivity analysis changed cluster membership, but no alternative donor-eligible cluster met the necessary raw gene-coherence criterion. An independently defined collagen-panel sensitivity yielded different patterns in the two PVR cohorts. These data do not demonstrate a reproducible ECM7-expressing myeloid state under the specified rules; small biological cohorts, a restricted gene panel, and indirect use of outcome genes during ambient correction limit broader exclusion.