Distinct mouse models of Stargardt disease display differences in pharmacological targeting of ceramides and inflammatory responses
We wanted to transcriptomically compare single cells taken from the retinas of C57BL6/J mice, Abca4-/-Rdh8-/- (dKO) mutant mice, and Abca4PV/PVRdh8-/- (PVR) mutant mice before and at an early (24-hour post-bleach) timepoint post-bleaching, to see if the two Abca4 mutants displayed different transcriptomic shifts in response to intense light, which initiates retinal degeneration. Additionally, we wanted to determine if the two mutants had different transcriptional responses to the immunomodulatory drug maraviroc. The wild-type C57BL6/J mice were included as controls for each treatment condition. We profiled the three cohorts of mice under three separate treatment conditions: without light-induced degeneration, 24-hours post-bleaching with intense light and vehicle treatment, and 24-hours post-bleaching with intense light and maraviroc treatment. Single cell isolates were collected, fixed, barcoded, and sequenced using kits from Parse Biosciences. Overall design: In each of the six experimental groups, pairs of retinae from three mice were independly processed; the resulting single-cell suspensions were fixed and frozen using the Parse Biosciences WT V2 chemistry fixation kit instructions. For each of the 27 independent biological samples, the fixed cells were simultaneously thawed and suspensions were loaded into independent wells on a Parse Biosciences WT V2 barcoding plate for library preparation (~2,360 cells/ biological sample), according to the manufacturer's instructons.