TY - JOUR
T1 - Mitochondrial Profile and Responses to TGF-β Ligands in Keratoconus
AU - Sarker-Nag, Akhee
AU - Hutcheon, Audrey E.K.
AU - Karamichos, Dimitrios
N1 - Funding Information:
This work was supported by the National Institutes of Health/National Eye Institute grants EY023568 and EY020886 (DK), and, in part, by an unrestricted grant (DMEI) from Research to Prevent Blindness (New York, NY USA).
Publisher Copyright:
© 2016 Taylor & Francis.
PY - 2016/7/2
Y1 - 2016/7/2
N2 - Purpose: Keratoconus (KC) is a complex corneal dystrophy with multifactorial etiology. Previous studies have shown evidence of mitochondrial abnormalities in KC; however, the exact cause of these abnormalities remains unknown. The aim of this study was to identify if transforming growth factor-β (TGF-β) isoforms play a role in the regulation of mitochondrial proteins in human KC cells (HKC). Materials and Methods: Human corneal fibroblasts (HCF) and HKC were isolated and cultured for 4 weeks in three different conditions: (a) Control: MEM + 10%FBS, (b) MEM + 10%FBS + TGF-β1 and (c) MEM + 10%FBS + TGF-β3. All samples were processed for mitochondrial damage analysis using real-time PCR. Results: We quantified and analyzed 84 mitochondrial and five housekeeping genes in HCFs and HKCs. Our data showed that when TGF-β1 and/or TGF-β3 were compared with control in HCFs, nine genes were significantly different; however, no genes were significantly regulated by the TGF-β isoforms in HKCs. Significant differences were also seen in seven genes when HFCs were compared with HKCs, in all three conditions. Conclusions: Overall, our data support the growing consensus that mitochondrial dysfunction is a key player in KC disease. These in vitro data show clear links between mitochondrial function and TGF-β isoforms, with TGF-β1 severely disrupting KC-mitochondrial function, while TGF-β3 maintained it, thus suggesting that TGF-β may play a role in KC-disease treatment.
AB - Purpose: Keratoconus (KC) is a complex corneal dystrophy with multifactorial etiology. Previous studies have shown evidence of mitochondrial abnormalities in KC; however, the exact cause of these abnormalities remains unknown. The aim of this study was to identify if transforming growth factor-β (TGF-β) isoforms play a role in the regulation of mitochondrial proteins in human KC cells (HKC). Materials and Methods: Human corneal fibroblasts (HCF) and HKC were isolated and cultured for 4 weeks in three different conditions: (a) Control: MEM + 10%FBS, (b) MEM + 10%FBS + TGF-β1 and (c) MEM + 10%FBS + TGF-β3. All samples were processed for mitochondrial damage analysis using real-time PCR. Results: We quantified and analyzed 84 mitochondrial and five housekeeping genes in HCFs and HKCs. Our data showed that when TGF-β1 and/or TGF-β3 were compared with control in HCFs, nine genes were significantly different; however, no genes were significantly regulated by the TGF-β isoforms in HKCs. Significant differences were also seen in seven genes when HFCs were compared with HKCs, in all three conditions. Conclusions: Overall, our data support the growing consensus that mitochondrial dysfunction is a key player in KC disease. These in vitro data show clear links between mitochondrial function and TGF-β isoforms, with TGF-β1 severely disrupting KC-mitochondrial function, while TGF-β3 maintained it, thus suggesting that TGF-β may play a role in KC-disease treatment.
KW - Cornea
KW - keratoconus
KW - mitochondria
KW - mitochondrial dysfunction
KW - transforming growth factor-β
UR - http://www.scopus.com/inward/record.url?scp=84945206600&partnerID=8YFLogxK
U2 - 10.3109/02713683.2015.1078361
DO - 10.3109/02713683.2015.1078361
M3 - Article
C2 - 26430764
AN - SCOPUS:84945206600
SN - 0271-3683
VL - 41
SP - 900
EP - 907
JO - Current Eye Research
JF - Current Eye Research
IS - 7
ER -