Abstract
Purpose :
Epigenetic factors play dynamic roles in neuronal development, progenitor cell proliferation, differentiation, and neural regeneration. Histone modification by histone deacetylases (HDACs) represents a key mechanism by which neurons regulate transcriptional profile during proliferation and differentiation. Accumulative studies have highlighted the therapeutic potential of HDACs inhibitors in neurological disorder. Therefore, we aimed to find HDAC that correlates with retinal ganglion cells (RGCs) development, which may provide a target of HDAC inhibitor in retinal degenerative diseases.
Methods :
HDACs (members 1-6, 8-11) mRNA levels in retinal ganglion cells (RGCs) isolated from mouse at embryonic (E16) and postnatal (P0, P10) stages were analyzed by quantitative polymerase chain reaction (qPCR). PC12 cells were transduced with lentivirus carrying shRNA against HDAC11. Over 25 different transductants were generated with HDAC11 mRNA level knockdown ranged 20% to 70%. The transductants were subjected to proliferation assay, evaluated by 5-ethynyl-2’-deoxyuridine (EdU) incorporation before and after treatment of nerve growth factor (NGF; 30 ng/ml). Neurite outgrowth was quantitatively assessed 3 days after NGF-induction of neural differentiation.
Results :
Initial screening of HDACs mRNA revealed a unique temporal increase of HDAC11 from E16 to P10, indicating an inhibitory contribution of HDAC11 to RGC growth capacity. To test this hypothesis, we established HDAC11 knockdown cell lines using neuron-like PC12 cells and examined their performance in proliferation and differentiation. In the absence of NGF, transduced cell line with about 50% knockdown in HDAC11 showed significantly increased number of dividing cells compared to naïve control PC12 cells while it also showed significantly more neurite outgrowth following NGF treatment, suggesting HDAC11 deficiency can enhance both proliferation and differentiation.
Conclusions :
Our data show that HDAC11 may play a role in mediating neural progenitor cell proliferation and neuronal differentiation. These findings may suggest a novel target for promoting neuronal regeneration.
This abstract was presented at the 2022 ARVO Annual Meeting, held in Denver, CO, May 1-4, 2022, and virtually.