Myocilin Regulates Cell Proliferation and Survival

Margetis, Joe; Kwon, Heung Sun; Cojocaru, Radu; Tomarev, Stanislav I.

doi:10.1074/jbc.m113.547091

Cited by 24 publications

(19 citation statements)

References 72 publications

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“…Initially, to assess for potential differences in how eGLuc2-tagged MYOC is secreted and handled within the cell, we compared the secretion and intracellular profile of untagged WT MYOC and WT MYOC FLAG (FT, used extensively in past experiments 2,37,38 ) to WT MYOC eGLuc2. We found that indeed, all three variants are readily secreted from transfected cells and detectable in the media 48 hours after initial transfection (Fig.…”

Section: Resultsmentioning

confidence: 99%

A Novel Luciferase Assay For Sensitively Monitoring Myocilin Variants in Cell Culture

Zadoo

Nguyen

Zode

et al. 2016

Invest. Ophthalmol. Vis. Sci.

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PurposePrimary open angle glaucoma–associated mutations in myocilin (MYOC) cause protein “nonsecretion,” rendering secreted MYOC difficult to detect using conventional techniques. This study focused on developing and using an assay that can quickly and easily detect mutant MYOC secretion.MethodsWe fused Gaussia luciferase (eGLuc2) to MYOC variants and expressed the constructs in HEK-293T and NTM-5 cells. Secreted and intracellular levels of MYOC eGLuc2 variants were evaluated by Western blotting and compared to untagged and FLAG-tagged MYOC constructs. Secreted and soluble intracellular MYOC eGLuc2 were measured by a GLuc assay. The secretion of nine additional MYOC mutants was assayed in conditioned media from transfected cells to test the applicability of the assay for monitoring other MYOC variants.ResultsMyocilin eGLuc2 behaved similarly to untagged and FLAG-tagged MYOC with respect to secretion, soluble intracellular levels, and in response to drug treatment. The GLuc assay could sensitively detect Y437H MYOC secretion 30 minutes after media change. Gaussia luciferase fused variants followed anticipated trends; nonpathogenic variants (D208E, G244V) were secreted at wild-type (WT) levels, whereas predicted disease-causing variants (C245Y, G246R, E300K, Y437H, I477N) demonstrated substantial secretion defects. Secretion defects caused by the C245Y, G246R, and Y437H mutations were partially rescued by permissive growth temperature. Interestingly, however, this increase in secretion was independent of newly synthesized protein.ConclusionsFusion of eGLuc2 to MYOC does not significantly change the behavior of MYOC. This newly developed MYOC reporter system can be used to study engineered MYOC variants and potentially to identify modulators of MYOC secretion and function.

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Section: Resultsmentioning

confidence: 99%

A Novel Luciferase Assay For Sensitively Monitoring Myocilin Variants in Cell Culture

Zadoo

Nguyen

Zode

et al. 2016

Invest. Ophthalmol. Vis. Sci.

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“…In this network, MYOC is shown to interact with PXDN, which was included in our input gene list, and NEFM (neurofilament, medium polypeptide) and CD44 , which were not. All the three genes have been shown to be upregulated in response to MYOC overexpression in HEK293 cells (Joe et al, 2014). NEFM is thought to be important in neural processes and the radial outgrowth of axons during development (Yuan et al, 2012).…”

Section: Pathway Analysis and Functional Annotationmentioning

confidence: 99%

“…MYOC overexpression induces upregulated expression of PGR (Joe et al, 2014). Like estrogen, progesterone has also been shown to play an important role in neural development, with high levels of PGR expression in the neurons and radial glial cells of developing zebrafish (Diotel et al, 2011).…”

Section: Pathway Analysis and Functional Annotationmentioning

confidence: 99%

Characterizing the “POAGome”: A bioinformatics-driven approach to primary open-angle glaucoma

Danford

Verkuil

Choi

et al. 2017

Progress in Retinal and Eye Research

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Primary open-angle glaucoma (POAG) is a genetically, physiologically, and phenotypically complex neurodegenerative disorder. This study addressed the expanding collection of genes associated with POAG, referred to as the “POAGome.” We used bioinformatics tools to perform an extensive, systematic literature search and compiled 542 genes with confirmed associations with POAG and its related phenotypes (normal tension glaucoma, ocular hypertension, juvenile open-angle glaucoma, and primary congenital glaucoma). The genes were classified according to their associated ocular tissues and phenotypes, and functional annotation and pathway analyses were subsequently performed. Our study reveals that no single molecular pathway can encompass the pathophysiology of POAG. The analyses suggested that inflammation and senescence may play pivotal roles in both the development and perpetuation of the retinal ganglion cell degeneration seen in POAG. The TGF-β signaling pathway was repeatedly implicated in our analyses, suggesting that it may be an important contributor to the manifestation of POAG in the anterior and posterior segments of the globe. We propose a molecular model of POAG revolving around TGF-β signaling, which incorporates the roles of inflammation and senescence in this disease. Finally, we highlight emerging molecular therapies that show promise for treating POAG.

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“…230, 231 Myocilin is expressed in anterior segment tissues including the TM, 232, 233 . Recent studies have implicated myocilin in cell proliferation and survival through the ERK1/2 pathway, which involves the aforementioned MMPs, 234 as well as in programmed RGC cell death. 235 While interacting partner(s) of myocilin and precise biological function remains elusive, it is an attractive de novo therapeutic target for glaucoma patients with inherited mutations.…”

Section: Selected Promising New Directions and Emerging Targetsmentioning

confidence: 99%

Discovery of Molecular Therapeutics for Glaucoma: Challenges, Successes, and Promising Directions

Donegan

Lieberman

2015

J. Med. Chem.

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Glaucoma, a heterogeneous ocular disorder affecting ~60 million people worldwide, is characterized by painless neurodegeneration of retinal ganglion cells (RGCs), resulting in irreversible vision loss. Available therapies, which decrease the common causal risk factor of elevated intraocular pressure, delay, but cannot prevent, RGC death and blindness. Notably, it is changes in the anterior segment of the eye, particularly in the drainage of aqueous humor fluid, which are believed to bring about changes in pressure. Thus, it is primarily this region whose properties are manipulated in current and emerging therapies for glaucoma. Here, we focus on the challenges associated with developing treatments, review the available experimental methods to evaluate the therapeutic potential of new drugs, describe the development and evaluation of emerging Rho-kinase inhibitors and adenosine receptor ligands that offer the potential to improve aqueous humor outflow and protect RGCs simultaneously, and present new targets and approaches on the horizon.

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Myocilin Regulates Cell Proliferation and Survival

Cited by 24 publications

References 72 publications

A Novel Luciferase Assay For Sensitively Monitoring Myocilin Variants in Cell Culture

A Novel Luciferase Assay For Sensitively Monitoring Myocilin Variants in Cell Culture

Characterizing the “POAGome”: A bioinformatics-driven approach to primary open-angle glaucoma

Discovery of Molecular Therapeutics for Glaucoma: Challenges, Successes, and Promising Directions

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