RNA-Seq-based transcriptome analysis of Murine Rechallenged Dry Eye Model

Abstract

Purpose : To investigate the gene expression changes of rechallenged dry eye disease (DED) in a
murine model.

Methods : Female C57BL/6 mice aged 6-8 weeks were used for this study. Acute DED was induced
for 14 consecutive days in a controlled desiccative environment for our acute DED group.
Rechallenged DED was developed by transferring the aforementioned acute DED to a standard
vivarium for 3 weeks, followed by an additional 14 days in a controlled desiccative environment.
Samples of cornea, conjunctiva, and cervical lymph nodes from each group were collected for
comparative analysis of differentially expressed gene (DEG) analysis and Gene Ontology analysis
by RNA sequencing.

Results : DEG analysis by RNA sequencing identified characteristic DEGs of the rechallenged DED
group when compared to acute DED group: cornea, 360 genes (up-regulated DEG; 201 genes, down-
regulated DEG; 159 genes); conjunctiva, 4,746 genes (up-regulated DEG; 2082 genes, down-
regulated DEG; 2664 genes); cervical lymph nodes, 956 genes (up-regulated DEG; 497 genes,
down-regulated DEG; 459 genes). Gene Ontology analysis identified Go Terms related to
chemokine activity, inflammatory response, positive regulation of angiogenesis, and innate immune
response in the rechallenged DED group compared to the acute DED group.

Conclusions : The results of this study suggest that immune interactions and inflammation-related Go
Terms may be major contributors to DED, particularly in the setting of repetitive exacerbation of
DED. These findings may imply that genes involved in DED pathogenesis may be dynamic and that
chronic DED may carry unique pathologic underpinnings and features when juxtaposed against the
initial acute phase.

This abstract was presented at the 2024 ARVO Annual Meeting, held in Seattle, WA, May 5-9, 2024.