组蛋白去乙酰化酶抑制延缓色素性视网膜炎突变小鼠的光感受器丧失：单细胞 RNA 测序和 CUT&Tag 的见解。

HDAC inhibition delays photoreceptor loss in mutant mice of retinitis pigmentosa: insights from scRNA-seq and CUT&Tag.

机构信息

Kunming Medical University, Kunming, Yunnan, China.

Key Laboratory of Yunnan Province, Yunnan Eye Institute, Affiliated Hospital of Yunnan University, Yunnan University, Kunming, Yunnan, China.

出版信息

PeerJ. 2023 Jul 12;11:e15659. doi: 10.7717/peerj.15659. eCollection 2023.

DOI:10.7717/peerj.15659

PMID:37456870

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC10349563/

Abstract

PURPOSE

This research aimed to ascertain the neuroprotective effect of histone deacetylase (HDAC) inhibition on retinal photoreceptors in mice, a model of retinitis pigmentosa (RP)

METHODS

Single-cell RNA-sequencing (scRNA-seq) explored HDAC and poly (ADP-ribose) polymerase (PARP)-related gene expression in both -mutant and wild-type (WT) mice. The CUT&Tag method was employed to examine the functions of HDAC in mice. Organotypic retinal explant cultures from WT and mice were exposed to the HDAC inhibitor SAHA (suberoylanilide hydroxamic acid) postnatally, from day 5 to day 11. The terminal deoxynucleotidyl transferase-mediated nick-end labeling (TUNEL) assay was applied to quantify the percentage of photoreceptor loss in the outer nuclear layer (ONL). HDAC activity was confirmed to be inhibited by SAHA through an HDAC activity assay. Moreover, the study evaluated PARP activity, a key driver of the initial response to DNA damage during photoreceptor degeneration, following HDAC inhibition.

RESULTS

The scRNA-seq revealed that diverse roles of HDAC and PARP isoforms in photoreceptor cell death. HDAC-related genes appeared to regulate cell death and primary immunodeficiency. Alterations in HDAC activity were consistent with the TUNEL-positive cells in the ONL at different time points. Notably, SAHA significantly postponed photoreceptor loss and decreased HDAC and PARP activity, thereby implicating both in the same degenerative pathway.

CONCLUSIONS

This study highlights that the interaction between HDAC inhibition and PARP can delay photoreceptor cell death, proposing a promising therapeutic approach for RP.

摘要

目的

本研究旨在确定组蛋白去乙酰化酶（HDAC）抑制对色素性视网膜炎（RP）模型小鼠视网膜光感受器的神经保护作用。

方法

单细胞 RNA 测序（scRNA-seq）探索了 HDAC 和多聚（ADP-核糖）聚合酶（PARP）相关基因在 -突变和野生型（WT）小鼠中的表达。采用 CUT&Tag 方法研究 HDAC 在小鼠中的功能。从第 5 天到第 11 天，对 WT 和小鼠的器官型视网膜外植体培养物进行 HDAC 抑制剂 SAHA（丁酰羟肟酸）处理。采用末端脱氧核苷酸转移酶介导的缺口末端标记（TUNEL）测定法来量化外核层（ONL）中光感受器损失的百分比。通过 HDAC 活性测定证实 SAHA 抑制了 HDAC 活性。此外，该研究还评估了 HDAC 抑制后对光感受器变性过程中 DNA 损伤初始反应的关键驱动因素 PARP 活性。

结果

scRNA-seq 显示 HDAC 和 PARP 同工型在光感受器细胞死亡中的多种作用。HDAC 相关基因似乎调节细胞死亡和原发性免疫缺陷。HDAC 活性的改变与不同时间点 ONL 中的 TUNEL 阳性细胞一致。值得注意的是，SAHA 显著延迟了光感受器的损失，并降低了 HDAC 和 PARP 活性，这表明两者在相同的退行性途径中。