Hypoxia-Induced Scleral HIF-2α Upregulation Contributes to Rises in MMP-2 Expression and Myopia Development in Mice

Purpose: Scleral hypoxia is a key factor that induces hypoxia-inducible factor-1α (HIF-1α) upregulation, and this response contributes to myopia progression. Currently, we aim to determine if the different HIF subtypes, including HIF-1α and HIF-2α, mediate hypoxia-induced myopia development through promoting scleral MMP-2 expression and collagen degradation. Methods: Our study included: (1) time-course of scleral HIF-2α, MMP-2, and COL1α1 expression during form-deprivation myopia (FDM) development was determined in C57BL/6J mice. (2) The effect of silencing either HIF-1Α or HIF-2A on hypoxia-induced alterations in MMP-2 expression was analyzed in cultured human scleral fibroblasts (HSFs) under a hypoxic condition (i.e. 1% oxygen). (3) To knock-down either HIF-1α or HIF-2α expression in the sclera, we performed Sub-Tenon's capsule injection of an adeno-associated virus (AAV)8-packaged Cre overexpression vector (AAV8-Cre) in HIF-1αfl/fl or HIF-2αfl/fl mice. HIF-1α, HIF-2α, MMP-2, and COL1α1 expression were analyzed by Western blot or quantitative real-time PCR (qRT-PCR). In addition, the effects of scleral HIF-2α knock-down on normal refractive development and FDM development were evaluated. Results: The time-dependent increases in scleral HIF-2α mimicked the HIF-1α expression profiles as we previously described. Hypoxia significantly promoted MMP-2 expression in HSFs, and this upregulation was solely alleviated by HIF-2A rather than HIF-1A silencing. Scleral HIF-2α knockdown significantly inhibited form-deprivation (FD)-induced MMP-2 upregulation and declines in COL1α1 accumulation and myopia development. Although scleral HIF-1α knockdown also significantly suppressed FD-induced declines in COL1α1 accumulation, it did not abrogate scleral MMP-2 upregulation. Conclusions: HIF-2α rather than HIF-1α induces myopia development through upregulating MMP-2 and promoting collagen degradation in the sclera.