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Comparison of Transcript and Protein Abundance in Human Donor Retinas Support Extending Post-Mortem Interval from 12- to 18-hours to Expand the Donor Pool for Biomedical Research

ABSTRACT: Extending the post-mortem interval (PMI) from 12- to 18-hours was done to see if transcript and protein abundance in the human retina changed. Donor eyes were recovered from post-mortem donors (n=7 pairs) and stored at 2–8°C. Only donor eyes with ocular cooling within 8 hours of death were examined. The OD retina was preserved at 12 hours and the OS retina at 18 hours post-mortem. Retinas were flash frozen with liquid nitrogen. The macular region superior to the fovea was used for RNA sequencing (RNA-seq). RNA library preparations were performed simultaneously, and the sequencing of all samples were conducted on the same chip. The inferior region was processed for isobaric labeling (TMT) quantitative mass spectrometry. Proteins were isolated from samples and labeled with 16-plex TMTPro isobaric tags. RNA-seq data were processed using DESeq2 (Bioconductor) and proteomic data were analyzed using the OHSU Comet/PAW pipeline. RNA-seq identified 22,446 transcripts, with only 5 transcripts showing significant differential expression (adjusted-P<0.1 and a Log2FoldChange>1) between 12 and 18 hours. Proteomic analysis identified 6,109 proteins, with no significant differences in abundance (all adj-P > 0.05, 1% FDR). Retinal cell-type specific markers and markers relevant for disease research, such as age-related macular degeneration and glaucoma, remained stable across both collection time points. Extending the PMI cutoff from 12 to 18 hours did not significantly impact transcriptomic or proteomic integrity in human donor retinas. This collection period extension greatly increases the availability of donor eyes for biomedical research.

INSTRUMENT(S):

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Retina

SUBMITTER: Phillip Wilmarth

LAB HEAD: Khoa D. Tran

PROVIDER: PXD068030 | Pride | 2026-04-07

REPOSITORIES: Pride

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Dataset's files

Source:

			Action	DRS
	2024.04_UP000005640_9606_Homo_sapiens_canonical_both.fasta	Fasta
	File_list.xlsx	Xlsx
	MSConvert_GUI_1732471350.0370173_log.txt	Txt
	PAW_protein_grouper.log	Other
	PAW_results.log	Other

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Comparison of Transcript and Protein Abundance in Human Donor Retinas Support Extending Post-Mortem Interval from 12- to 18-hours to Expand the Donor Pool for Biomedical Research

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