Melatonin induces autophagy via an mTOR-dependent pathway and enhances clearance of mutant-TGFBIp.
The hallmark of granular corneal dystrophy type 2 (GCD2) is the deposit of mutant transforming growth factor-β (TGF-β)-induced protein (TGFBIp) in the cornea. We have recently shown that there is a delay in autophagic degradation of mutant-TGFBIp via impaired autophagic flux in GCD2 corneal fibroblasts. We hypothesized that melatonin can specifically induce autophagy and consequently eliminate mutant-TGFBIp in GCD corneal fibroblasts. Our results show that melatonin activates autophagy in both wild-type (WT) and GCD2-homozygous (HO) corneal fibroblast cell lines via the mammalian target of rapamycin (mTOR)-dependent pathway. Melatonin treatment also led to increased levels of beclin 1, which is involved in autophagosome formation and maturation. Furthermore, melatonin significantly reduced the amounts of mutant- and WT-TGFBIp. Treatment with melatonin counteracted the autophagy-inhibitory effects of bafilomycin A1, a potent inhibitor of autophagic flux, demonstrating that melatonin enhances activation of autophagy and increases degradation of TGFBIp. Cotreatment with melatonin and rapamycin, an autophagy inducer, had an additive effect on mutant-TGFBIp clearance compared to treatment with either drug alone. Treatment with the selective melatonin receptor antagonist luzindole did not block melatonin-induced autophagy. Given its ability to activate autophagy, melatonin is a potential therapeutic agent for GCD2.
Corneal Dystrophy Research Institute; Department of Ophthalmology, Yonsei University College of Medicine, Seoul, South Korea.
Seung-Il
SI
Choi
Kyu Seo
KS
Kim
Jun-Young
JY
Oh
Jun-Yup
JY
Jin
Ga-Hyun
GH
Lee
Eung Kweon
EK
Kim
D000818:Animals; D001343:Autophagy / Q000187:drug effects*; D001483:Base Sequence; D015153:Blotting, Western; D002478:Cells, Cultured; D017931:DNA Primers; D006801:Humans; D008550:Melatonin / Q000494:pharmacology*; D046529:Microscopy, Electron, Transmission; D058570:TOR Serine-Threonine Kinases / Q000378:metabolism*; D053773:Transforming Growth Factor beta1 / Q000378:metabolism*
D017931:DNA Primers; D053773:Transforming Growth Factor beta1; C546842:MTOR protein, human; D058570:TOR Serine-Threonine Kinases; D008550:Melatonin
(mTOR); LC3; TGFBIp; autophagy; granular corneal dystrophy type 2; melatonin