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Agathe Hribek, Thomas Clahsen, Sebastian E Siebelmann, Ludwig M Heindl, Björn Bachmann, Claus Cursiefen, Mario Matthaei; A model of Fuchs Endothelial Corneal Dystrophy – related extracellular matrix expression by miR-29 knockdown in vitro. Invest. Ophthalmol. Vis. Sci. 2019;60(9):2181. doi: https://doi.org/.
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The miR-29 family is known to be significantly downregulated in the endothelium of Fuchs endothelial corneal dystrophy (FECD)-patients and functionally involved in the regulation of ECM homeostasis. Thus, in this study the impact of reduced miR-29 expression on extracellular matrix (ECM) deposition in corneal endothelial cells (CECs) was further investigated.
To uncover the impact of miR-29 on ECM genes related to FECD, a siRNA based miR-29 knockdown was performed in HCEC-12, a human corneal endothelium derived cell line. Success of the knockdown experiment as well as the expression of well-known ECM associated miR-29 targets were examined by qPCR. A corresponding immunofluorescence staining of FECD and control endothelium was used to analyze ECM components on protein level and to identify their staining pattern.
Knockdown of miR-29a (n= 3) in HCEC-12 cell culture model was highly significant (p ≤ 0.0001) and temporally constant. Downregulation of miR-29 led to a significant upregulation of COL3A1 (p ≤ 0.001) 96h after cell transfection. Furthermore, a trend for upregulation of COL1A1 and COL4A1 was detected in knockdown samples. Increased ECM expression of individual miR-29 target proteins was localized in the center of FECD-endothelium by immunofluorescence staining.
Knockdown of miR-29a in corneal endothelial cells induces a FECD-like expression of ECM-proteins in vitro. Further studies will deepen our knowledge about the impact of miR-29 on CECs and help to establish miR-29 as a therapeutic target in FECD.
This abstract was presented at the 2019 ARVO Annual Meeting, held in Vancouver, Canada, April 28 - May 2, 2019.
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