HSP90β Regulates the Inflammatory Response of the Ocular Lens through the NF-kB Pathway

Abstract

Purpose : HSP90β is an important molecular chaperone and plays an fundamental role in many cellular processes. Whether it plays a role in lens remains to be studied. Here we show that HSP90β can regulate the inflammatory response in the ocular lens through direct interaction with p65 of the NF-kB pathway.

Methods : CRISPR/Cas9 technology and shRNA-mediated silence were used to knockdown HSP90β in mouse lens epithelial cell line, aTN4-1. QRT-PCR was used to analyze the mRNA expression of the inflammatory cytokines. Western blot was used to analyze expression levels of NF-kB pathway components. Immunofluorescence and Co-IP assays were used to verify the interactions between HSP90β and p65 in human lens epithelial cell line FHL124.

Results : Knockdown of HSP90β in aTN4-1 leads to increased expression levels of inflammatory cytokines. Immunofluorescence and Co-IP assays reveal that HSP90β regulates the inflammatory response of lens epithelial cells through interaction with p65 of NF-kB. In HSP90β knockdown cells, we also observed that the expression levels of NF-kB pathway related proteins were increased.

Conclusions : Our results demonstrated that HSP90β plays a fundamental role in regulating lens inflammatory response. Through interactions with p65, HSP90β regulates the microenvironment of the ocular lens through NF-kB. [Supported by National Natural Science Foundation of China (Grants 82271071, 81970787, 81770910) and the Joint Key Project from the Natural Science Foundation of Guangdong Province and Guangzhou City (2019B1515120014)，and the Fundamental Funds of the State Key Laboratory of Ophthalmology of Zhongshan Ophthalmic Center (3030901010111)].

This abstract was presented at the 2023 ARVO Annual Meeting, held in New Orleans, LA, April 23-27, 2023.