Matrix Metallopeptidase 19 (MMP19) and Optic Nerve Disease

描述（由申请人提供）：视神经疾病是一个主要的公共卫生问题。包括青光眼和空洞性视盘异常（CODA）在内的几种视神经疾病的定义特征是视神经头部的凹陷或凹陷。然而，在这些疾病中导致视盘凹陷的生物学途径还不完全清楚。因此，迫切需要阐明导致视神经组织损失（挖掘）的生物学机制，以提高我们对疾病过程的理解，并开发新的治疗视神经疾病（包括CODA和青光眼）的方法。我们最近发现基质金属肽酶19（MMP 19）基因突变与CODA有关。具体来说，我们发现了MMP 19基因上游的一个6 kb的DNA序列，该序列在CODA患者中是三重的。此外，我们已经证明，该序列是一个积极的增强子和上调MMP 19的表达。我们还发现MMP 19在视神经乳头中特异性表达，在那里发生CODA异常。此外，基因表达数据显示，在患有青光眼的人类患者和动物的视神经中，MMP 19的表达大大增加。总之，这些发现支持我们的假设，MMP 19的异常调节促进先天性凹陷的CODA视盘。我们将通过以下研究目的来验证这一假设：目的1：检测青光眼患者的MMP 19突变。我们将用下一代测序技术检测一组青光眼患者的MMP 19拷贝数变异、调控序列突变和编码序列突变。目的2：鉴定调控MMP 19表达的特异性增强子元件和转录因子。我们将1）通过诱变研究鉴定MMP 19增强子元件的特异性DNA序列，2）使用DNA下拉试验和质谱法分离与MMP 19增强子结合的转录因子。这一建议将阐明CODA的发病机制，并为青光眼的视神经杯状形成过程提供见解。这些研究可能为未来研究开发新的视力挽救疗法提供基础。

DESCRIPTION (provided by applicant): Optic nerve diseases are a major public health problem. The defining feature of several optic nerve diseases, including glaucoma and cavitary optic disc anomaly (CODA), is excavation or cupping of the head of the optic nerve. The biological pathways that lead to excavation of the optic disc in these diseases, however, are incompletely understood. Consequently, there is a critical need to clarify the biological mechanisms that lead to loss of tissue (excavation) of the optic nerve to improve our understanding of disease processes and to develop new treatments for optic nerve diseases including CODA and glaucoma. We recently discovered that mutation of the matrix metallopeptidase 19 (MMP19) gene is associated with CODA. Specifically, we discovered a 6 kb DNA sequence upstream of the MMP19 gene that is triplicated in patients with CODA. Moreover, we have proven that this sequence is an active enhancer and upregulates expression of MMP19. We have also shown that MMP19 is specifically expressed in the optic nerve head, where the abnormalities of CODA occur. Moreover, gene expression data shows that MMP19 expression is greatly increased in the optic nerves of both human patients and animals with glaucoma. Together these findings support our hypothesis that abnormal regulation of MMP19 promotes congenital excavation of the optic disc in CODA. We will test this hypothesis with the following research aims: Aim 1: Test glaucoma patients for MMP19 mutations. We will test a cohort of glaucoma patients for copy number variations, regulatory sequence mutations, and coding sequence mutations in MMP19 with next generation sequencing. Aim 2: Identify the specific enhancer element and transcription factors that regulate MMP19 expression. We will 1) identify the specific DNA sequence of the MMP19 enhancer element via mutagenesis studies and 2) isolate transcription factor(s) that bind to the MMP19 enhancer using a DNA pull down assay and mass spectrometry. This proposal will clarify mechanisms of disease in CODA and provide insights in the process of optic nerve cupping in glaucoma. These investigations may provide the foundation for future studies to develop new sight-saving therapies.