Novel methods for treating glaucoma

Abstract

The present disclosure relates generally to the treatment of glaucoma and / or the reduction of elevated intraocular pressure (IOP). In particular, the present disclosure relates to compositions and methods for preventing or treating glaucoma, including primary glaucoma, by inhibiting factors associated with ocular lymphangiogenesis.

Description

[Technical Field]

[0001] [CROSS-REFERENCE TO RELATED APPLICATIONS] This application claims the benefit of U.S. Provisional Patent Application No. 63 / 486,665, filed February 23, 2023, the entirety of which is incorporated herein by reference for all purposes.

[0002] [Statement of Government Funding] This invention was made with funding from the United States Federal Government under Award Nos. EY028995 and EY017392 awarded by the National Institutes of Health. The United States Federal Government reserves certain rights in this invention.

[0003] The present disclosure relates generally to the treatment or prevention of glaucoma and / or the reduction of elevated intraocular pressure (IOP). In particular, the present disclosure relates to compositions and methods for preventing or treating glaucoma, particularly primary glaucoma, by inhibiting factors associated with intraocular lymphangiogenesis and Schlemm's canal.

[0004] [Incorporated by reference] The contents of the XML file are named "23091-006WO1_ST26_2024-02-19", were created on February 19, 2024, are 43.7KB in size, and are incorporated herein by reference in their entirety. [Background technology]

[0005] Glaucoma is a group of eye diseases that cause damage to the optic nerve. If left untreated, glaucoma can lead to vision loss and even blindness. Approximately 80 million people worldwide suffer from glaucoma, with approximately 3 million people in the United States suffering from glaucoma.

[0006] Glaucoma is classified as primary or secondary. Primary glaucoma is characterized by increased resistance to aqueous humor outflow, resulting in elevated intraocular pressure (IOP), and ultimately optic nerve damage, in the absence of any other underlying underlying disease. Primary open-angle glaucoma (POAG) is a subset of glaucoma defined by an open, normal-appearing anterior chamber angle and elevated intraocular pressure (IOP) in the absence of any other underlying disease. In contrast, secondary glaucoma is caused by other medical conditions. A particular type of secondary glaucoma is neovascular glaucoma (NVG), characterized by the proliferation of fibrovascular tissue in the anterior chamber angle (see Non-Patent Document 1). A common predisposing factor for NVG is usually retinal ischemia, but in some cases it may be associated with other ocular or extraocular diseases.

[0007] Intraocular pressure (IOP) is measured in millimeters of mercury (mmHg). Normal intraocular pressure in humans is between 12 and 21 mmHg, and IOP above 21 mmHg is considered higher than normal or elevated. Elevated IOP in primary glaucoma is primarily caused by dysregulated drainage of aqueous humor (the clear fluid that fills the anterior surface of the eye). Schlemm's canal is a key structure that maintains aqueous humor drainage and IOP. Schlemm's canal is a core component of the conventional aqueous humor outflow pathway, accounting for 70% to 90% of the total aqueous humor outflow from the human eye. Furthermore, the endothelial cell layer of Schlemm's canal is one of the major sites of resistance to aqueous humor outflow and a major determinant of IOP (Non-Patent Document 2). Schlemm's canal in glaucomatous human eyes is significantly constricted compared to healthy eyes (Non-Patent Document 3). Resistance due to Schlemm's canal constriction and other morphological changes increases with age or under pathological conditions, leading to elevated IOP. Abnormally elevated IOP causes optic nerve damage and vision loss. This is exemplified by the fact that Schlemm's canal contraction alone accounts for approximately 50% of the total aqueous humor outflow loss observed in eyes with primary open-angle glaucoma (POAG) (Non-Patent Document 4). Furthermore, the density of Schlemm's canal wall holes is reduced by one-fifth in glaucomatous eyes compared to normal eyes (Non-Patent Document 5). Therefore, Schlemm's canal is an important ocular structure involved in the pathogenesis of glaucoma. However, the specific mechanism by which Schlemm's canal regulates aqueous humor outflow in both normal and glaucomatous eyes remains unknown.

[0008] Several studies have suggested that the lymphangiogenesis pathway may be a mechanism that plays a role in the pathogenesis of glaucoma. Lymphangiogenesis is the process by which new lymphatic vessels are formed from pre-existing blood or lymphatic endothelial progenitor cells. For example, researchers have found that genetic disruption of lymphatic vessels may contribute to elevated IOP. Therefore, it has previously been proposed that inducing or activating lymphangiogenesis may be a potential treatment for glaucoma (see, for example, Non-Patent Document 6, Non-Patent Document 7, and Non-Patent Document 8).

[0009] Early treatment of glaucoma can slow or halt the progression of the disease. The goal of this treatment is to reduce elevated IOP and protect the optic nerve from damage. The most common treatments for glaucoma include eye drops that reduce the amount of fluid produced in the eye or improve fluid drainage from the eye via the uveoscleral outflow pathway, oral medications that lower IOP, laser therapy (e.g., laser trabeculoplasty and laser iridotomy), and surgery (e.g., implants, trabeculectomy, canaloplasty). One such surgical procedure, canaloplasty, aims to lower IOP by catheterizing and dilating Schlemm's canal using a microcatheter and further introducing a suture. The suture has been shown to apply circumferential tension to the inner wall of Schlemm's canal, resulting in canal dilation and effectively increasing aqueous humor outflow and IOP reduction (Non-Patent Document 9). However, canaloplasty is associated with several complications, including microhyphema (12.1%), hypotony (0.6%), and bleeding in the anterior chamber, which is common postoperatively due to the rapid reduction in IOP (Id.). Therefore, current glaucoma treatment options have limited effectiveness and are associated with side effects or complications.

[0010] Therefore, there is an urgent need to develop new therapies for the treatment of glaucoma, including primary glaucoma. [Prior art documents] [Non-patent literature]

[0011] [Non-Patent Document 1] Wand M., Neovascular glaucoma. In: Ritch R, Shields MB, Krupin T, eds. The Glaucomas - Clinical Science. 2nd. Ed. St. Louis: Mosby; 1996: 1073-1129. Ch 51 [Non-patent document 2] Maeepea, O. & Bill, A. Pressures in the juxtacanalicular tissue and Schlemm's canal in monkeys. Exp Eye Res. 54(6):879-883(1992 Jun) [Non-patent document 3] Gabelt, BT & Kaufman, PL Changes in aqueous humor dynamics with age and glaucoma. Prog Retin Eye Res. 24:612-637(2005) [Non-patent document 4] Allingham, RR et al. Schlemm's canal and primary open angle glaucoma: correlation between Schlemm's canal dimensions and outflow facility. Exp. Eye Res. 62:101-109(1996) [Non-Patent Document 5] Johnson, M. et al. The pore density in the inner wall endothelium of Schlemm's canal of glaucomatous eyes. IOVS. 43:2950-2955(2002) [Non-patent document 6] Aspelund, A. et al. The Schlemm's canal is a VEGF-C / VEGFR-3-responsive lymphatic-like vessel. J Clin Invest. 124(9):3975-3986(2014 Sep) [Non-Patent Document 7] Thomson, B.R. et al. A lymphatic defect causes ocular hypertension and glaucoma in mice. J Clin Invest. 124(10):4320-4324(2014 Oct; Epub 2014 Sep 9)

Non-Patent Document 8

Non-Patent Document 9

Summary of the Invention

[0012] Surprisingly, it has been discovered that inhibiting lymphangiogenic factors ("pro-lymphangiogenic factors"), which mediate the formation or maintenance of lymphatic vessels and are associated with Schlemm's canal in the eye, effectively reduces intraocular pressure (IOP) and provides an effective treatment for glaucoma. The lymphatic system regulates fluid balance and immune function, and lymphatic vessels transport lymph containing immune cells and other components. The eye is composed of alternating lymphatic vessel-rich and lymphatic vessel-depleted tissues (Chen, L. Ocular lymphatics: state-of-the-art review. Lymphology. 42(2):66-76(2009 Jun)). As reported herein, targeting pro-lymphangiogenic factors associated with Schlemm's canal in the eye (e.g., but not limited to, key pro-lymphangiogenic factors such as VEGFR-3, VLA-1, Ang-2, or ITGA5) significantly reduced IOP in a mouse model of glaucoma (see, e.g., Figures 1A, 2A, 3A, 4A, and 5A). This finding directly contradicts previous suggestions in the art that reduction of IOP elevation, and therefore treatment of glaucoma, including primary glaucoma, could be achieved by stimulating lymphangiogenesis.

[0013] It is highly unexpected that inhibiting lymphangiogenesis-promoting factors associated with Schlemm's canal not only successfully reduces IOP but also alleviates several characteristic symptoms associated with glaucoma. The extent to which lymphangiogenesis regulates the progression or severity of complex ocular diseases, such as primary glaucoma, was previously unknown. Unlike most organs in the body that normally have lymphatic vessels, the eye, under normal physiological conditions, contains a heterogeneous collection of tissues that are either rich in lymphatic vessels (e.g., the conjunctiva) or completely lack lymphatic vessels (e.g., the cornea) (Chen, L. Ocular lymphatics: state-of-the-art review. Lymphology. 42(2):66-76 (2009 Jun)). Furthermore, while some ocular tissues (e.g., the cornea) can be induced to form lymphatic vessels, other ocular tissues (e.g., limbal lymphatic vessels) possess classic lymphatic vessel-specific markers, such as LYVE-1, that are absent in Schlemm's canal (Id.). Therefore, a prevalent molecular or mechanistic strategy in the art has been to generate new lymphatic vessels by activating lymphangiogenesis in response to or to reduce elevated IOP (see, e.g., WO 2015110701; Clahsen, T. et al. The novel role of lymphatic vessels in the pathogenesis of ocular diseases. Prog Retin Eye Res. 96:101157 (2023 Sep; Epub 2023 Feb 8); Non-patent literature 6; Non-patent literature 7; Non-patent literature 8). Conversely, however, as described herein, administration of inhibitors of lymphangiogenic factors that induce or maintain lymphangiogenesis has been found to lower IOP and attenuate several features of glaucomatous damage, including corneal edema (see, e.g., Figures 1B and 2B), thinning of the retinal nerve fiber layer (see, e.g., Figure 2C), and retinal ganglion cell (RGC) death (see, e.g., Figure 2D).

[0014] As shown herein, inhibition of lymphangiogenic factors acts somewhat similarly to surgical procedures such as canaloplasty to target and enhance the drainage function of existing lymphatic and lymphatic-like structures (e.g., Schlemm's canal). Administration of lymphangiogenic factor inhibitors to the eye increases the permeability of the structures, thereby allowing for increased aqueous humor outflow and reduced IOP elevation (see, e.g., Figures 1C, 3B-3C, 4B-4C, and 5B). As a key component of the conventional outflow pathway, Schlemm's canal is a critical structure in regulating aqueous humor drainage and IOP. While Schlemm's canal has been suggested to play a role in glaucoma, it has not previously been understood whether activating or inhibiting specific lymphatic mechanisms in Schlemm's canal can prevent or treat glaucoma, particularly primary glaucoma. It has been surprisingly and unexpectedly discovered that molecular inhibition of lymphangiogenic factors can treat glaucoma, including primary glaucoma, by inhibiting Schlemm's canal cell function. For example, inhibition of lymphangiogenic factors VEGFR-3, VLA-1, Ang-2, or ITGA5 reduces Schlemm's canal endothelial cell adhesion (see, e.g., Figures 1C, 3B, 4B, and 5B) and reduces Schlemm's canal tube formation (see, e.g., Figures 3C and 4C).

[0015] Without wishing to be bound by any theory, it has been proposed that inhibiting lymphangiogenic factors targets a specific function of Schlemm's canal to increase permeability, which contributes to enhanced aqueous humor drainage through Schlemm's canal, resulting in a measurable reduction in IOP and a reduction in glaucomatous damage. Furthermore, without wishing to be bound by any theory, a reduction in Schlemm's canalicular cell adhesion and tube formation contributes to increased Schlemm's canal permeability in vivo, allowing for increased outflow function and providing a reduction in IOP and a reduction in some parameters of glaucomatous damage.

[0016] The discoveries described herein provide an advancement in glaucoma treatment by providing methods of preventing or treating glaucoma, including primary glaucoma such as primary open-angle glaucoma (POAG), primary angle-closure glaucoma (PACG), primary congenital glaucoma, and primary normal-tension glaucoma, as well as other IOP-mediated ocular diseases through the inhibition of key lymphangiogenic factors.

[0017] In one aspect, the present disclosure provides a method of preventing or treating glaucoma in an eye of a subject in need thereof, comprising administering to the eye a therapeutically effective amount of an inhibitor of lymphangiogenesis promoting factor, hi some embodiments, the glaucoma is primary glaucoma selected from POAG, PACG, primary congenital glaucoma, and primary normal-tension glaucoma.

[0018] In one aspect, the present disclosure provides a method of inhibiting lymphangiogenesis in the eye of a subject experiencing elevated IOP, comprising administering to the eye a therapeutically effective amount of an inhibitor of lymphangiogenesis-promoting factors. In some embodiments, the subject has elevated IOP greater than about 21 mmHg. In some embodiments, the subject has elevated IOP greater than about 25 mmHg. In some embodiments, the subject has elevated IOP greater than about 30 mmHg. In some embodiments, after administration of the inhibitor, the subject's IOP in the treated eye is reduced by at least about 1 mmHg, at least about 2 mmHg, at least about 5 mmHg, at least about 10 mmHg, or at least about 15 mmHg.

[0019] In another aspect, the disclosure provides a method for inhibiting lymphangiogenesis in the eye of a subject having or suspected of having normal tension glaucoma, the method comprising administering to the eye a therapeutically effective amount of an inhibitor of a lymphangiogenesis promoting factor.

[0020] In one aspect, the present disclosure provides a method of inhibiting lymphangiogenesis in the eye of a subject at risk of developing glaucoma, comprising administering to the eye a therapeutically effective amount of an inhibitor of a lymphangiogenesis-promoting factor. In some embodiments, the glaucoma is primary glaucoma selected from POAG, PACG, primary congenital glaucoma, and primary normal-tension glaucoma.

[0021] In one aspect, the present disclosure provides a method for increasing Schlemm's canal permeability in a subject's eye having elevated IOP, comprising administering a therapeutically effective amount of an inhibitor of lymphangiogenesis promoting factor to the eye to increase Schlemm's canal permeability, increase aqueous humor outflow, and lower IOP in the eye. In some embodiments, the subject has elevated IOP greater than about 21 mmHg. In some embodiments, the subject has elevated IOP greater than about 25 mmHg. In some embodiments, the subject has elevated IOP greater than about 30 mmHg. In some embodiments, after administration of the inhibitor, the subject's IOP in the treated eye is reduced by at least about 1 mmHg, at least about 2 mmHg, at least about 5 mmHg, at least about 10 mmHg, or at least about 15 mmHg.

[0022] In another aspect, the present disclosure provides a method for increasing Schlemm's canal permeability in the eye of a subject having or suspected of having normal-tension glaucoma, the method comprising administering to the eye a therapeutically effective amount of an inhibitor of lymphangiogenesis promoting factor, thereby increasing Schlemm's canal permeability, increasing aqueous humor outflow, and reducing IOP of the eye.

[0023] In one aspect, the present disclosure provides a method for modulating one or more physiological functions of Schlemm's canal in an eye, comprising administering to the eye a therapeutically effective amount of an inhibitor of lymphangiogenesis promoting factor, thereby modulating one or more physiological functions of Schlemm's canal. In some embodiments, the modulated physiological function comprises inhibiting cell adhesion, proliferation, migration, or tube formation in Schlemm's canal.

[0024] In one aspect, the present disclosure provides a method for increasing aqueous humor outflow from the anterior chamber to Schlemm's canal in an eye of a subject in need thereof, the method comprising administering to the eye a therapeutically effective amount of an inhibitor of lymphangiogenesis promoting factor, thereby increasing aqueous humor outflow from Schlemm's canal.

[0025] In one aspect, the present disclosure provides a method for reducing corneal edema in an eye of a subject in need thereof, the method comprising administering to the eye a therapeutically effective amount of an inhibitor of lymphangiogenesis promoting factor, thereby reducing corneal edema.

[0026] In one aspect, the present disclosure provides a method for reducing retinal nerve fiber layer thinning in an eye of a subject in need thereof, the method comprising administering to the eye a therapeutically effective amount of an inhibitor of lymphangiogenesis promoting factor, thereby reducing retinal nerve fiber layer thinning.

[0027] In one aspect, the present disclosure provides a method for reducing retinal ganglion cell (RGC) death in the eye of a subject in need thereof, the method comprising administering to the eye a therapeutically effective amount of an inhibitor of lymphangiogenesis promoting factor, thereby reducing RGC death.

[0028] As provided herein, the method provides for administering an inhibitor of a lymphangiogenesis factor to the eye of a subject in need thereof. In some embodiments, the lymphangiogenesis factor is associated with Schlemm's canal. In some embodiments, the lymphangiogenesis factor is a factor that induces lymphangiogenesis. In some embodiments, the lymphangiogenesis factor is a factor that maintains or patterns lymphangiogenesis. In alternative embodiments, the lymphangiogenesis inhibitor may be an agent that activates or stimulates an anti-lymphangiogenic factor, as further described below.

[0029] In some embodiments, the lymphangiogenic factor targeted for inhibition is a vascular endothelial growth factor / vascular endothelial growth factor receptor (VEGF / VEGFR) family member. In some embodiments, the VEGF / VEGFR family member is selected from the group consisting of VEGF-A, VEGF-B, VEGF-C, VEGF-D, VEGF-E, PIGF, VEGFR-1, VEGFR-2, and VEGFR-3. In some embodiments, the VEGF / VEGFR family member is VEGF-A. In some embodiments, the VEGF / VEGFR family member is VEGF-C. In some embodiments, the VEGF / VEGFR family member is VEGF-D. In some embodiments, the VEGF / VEGFR family member is VEGFR-3.

[0030] In some embodiments, the lymphangiogenesis promoting factor targeted for inhibition is an integrin family member. In some embodiments, the integrin family member is selected from the group consisting of very late antigen-1 (VLA-1), integrin alpha 5 (ITGA5), and integrin alpha 9 (ITGA9). In some embodiments, the integrin family member is VLA-1. In some embodiments, the integrin family member is ITGA5. In some embodiments, the integrin family member is ITGA9.

[0031] In some embodiments, the lymphangiogenesis promoting factor targeted for inhibition is an angiopoietin family member. In some embodiments, the angiopoietin family member is selected from the group consisting of angiopoietin-1 (ANGPT1), angiopoietin-2 (ANGPT2 or Ang-2), and Tie2 / TEK. In some embodiments, the angiopoietin family member is Ang-2.

[0032] In some embodiments, two or more inhibitors targeting two or more lymphangiogenesis-promoting factors are administered to an eye in need thereof. In some embodiments, the two or more inhibitors target two or more of VEGFR-3, VLA-1, ITGA5, ITGA9, and Ang-2. In some embodiments, the targeted two or more lymphangiogenesis-promoting factors include VEGFR-3 and VLA-1. In some embodiments, the targeted two or more lymphangiogenesis-promoting factors include VEGFR-3 and ITGA-5. In some embodiments, the targeted two or more lymphangiogenesis-promoting factors include VEGFR-3 and ITGA-9. In some embodiments, the targeted two or more lymphangiogenesis-promoting factors include VEGFR-3 and Ang-2. In some embodiments, the targeted two or more lymphangiogenesis-promoting factors include VLA-1 and ITGA-5. In some embodiments, the targeted two or more lymphangiogenesis-promoting factors include VLA-1 and ITGA-9. In some embodiments, the two or more targeted lymphangiogenesis promoting factors comprise VLA-1 and Ang-2. In some embodiments, the two or more targeted lymphangiogenesis promoting factors comprise ITGA-5 and ITGA-9. In some embodiments, the two or more targeted lymphangiogenesis promoting factors comprise ITGA-5 and Ang-2. In some embodiments, the two or more targeted lymphangiogenesis promoting factors comprise ITGA-9 and Ang-2.

[0033] Alternatively, in addition to the above lymphangiogenesis-promoting factors, other factors may be used, but are not limited to, cytokines / chemokines, such as interleukin 8, interferon gamma, members of the CCR7 / SLC axis, such as CCR7 and SLC; extracellular matrix proteins, such as CCBE1; transcription factors, such as Sox18 and Hhex; induction molecules, such as neuropilin 2 and SEMA7A; FGFs, such as FGF-2; protein tyrosine phosphatases (PTPs), such as PTPN14; members of platelet factors, such as platelet-derived growth factor (PDGF- Lymphangiogenic factors, or combinations of lymphangiogenic factors, including platelet factor 4 (BB) or platelet factor 4; lymphatic endothelial cell (LEC) polarity factors (e.g., Celsr1, Vangl2, Pdk2, and Fat4); members of the Notch family; or ARAF, SOS1, Apelin, KIF11, REELIN, CALCRL, GJC2 (gap junction protein gamma-2), Rasip1, or FBXL7 (F-box and leucine-rich repeat protein 7), can be targeted for inhibition to inhibit or prevent lymphangiogenesis for the above-described methods.

[0034] Examples of lymphangiogenesis factor inhibitors suitable for administration to an eye in need thereof include, but are not limited to, (1) drugs capable of disrupting the gene encoding a lymphangiogenesis factor, including eliminating or reducing expression of the lymphangiogenesis factor through genome manipulation or knockout; (2) siRNAs capable of degrading the mRNA encoding a lymphangiogenesis factor, thereby reducing expression of the lymphangiogenesis factor; (3) antagonistic antibodies against lymphangiogenesis factors that inhibit or prevent the function of the lymphangiogenesis factor; or (4) small molecules capable of inhibiting or preventing the function of the lymphangiogenesis factor. In some embodiments, the lymphangiogenesis factor inhibitor is an siRNA. In some embodiments, the lymphangiogenesis factor inhibitor is an antagonistic antibody. Other strategies known in the art can also be used to disrupt the function or expression of a lymphangiogenesis factor or a combination of lymphangiogenesis factors.

[0035] The inhibitor of lymphangiogenesis factor can be administered systemically or locally to an eye in need thereof via any suitable route, hi some embodiments, the inhibitor of lymphangiogenesis factor is administered locally to an eye in need thereof via intracameral injection, subconjunctival injection, intravitreal injection, suprachoroidal injection, peribulbar injection, or retrobulbar injection.

[0036] In some embodiments, the inhibitor of lymphangiogenesis promoting factor is formulated as eye drops, a depot, a bolus, an inhibitor-loaded contact lens, a suspension, a solution, an ophthalmic gel, or an ointment.

[0037] In an alternative aspect, the lymphangiogenesis inhibitor may be an agent that activates or stimulates an anti-lymphangiogenic factor that inhibits lymphangiogenesis, as described further below. In some embodiments, the lymphangiogenesis inhibitor is selected from the group consisting of: (1) an agent that activates the expression of an anti-lymphangiogenic factor, (2) an agonist antibody that activates an anti-lymphangiogenic factor, (3) a microRNA or mimic that inhibits lymphangiogenesis, and (4) a small molecule that activates the function of an anti-lymphangiogenic factor. In some embodiments, the eye in need of treatment is treated with a second therapy. In some embodiments, the second therapy is selected from the group consisting of eye drops, oral medications, laser therapy, and surgery. In some embodiments, the second therapy is selected from eye drops that reduce the amount of fluid produced in the eye or improve fluid drainage from the eye, oral medications that lower IOP, laser therapy (e.g., laser trabeculoplasty and laser iridotomy), and surgery (e.g., implants, trabeculectomy). In some embodiments, the second treatment is surgery. In some embodiments, the surgery is canaloplasty.

[0038] It is to be understood that both the foregoing general description and the following detailed description are exemplary and explanatory only and are not restrictive of the invention. Moreover, the accompanying drawings, which are incorporated herein by reference and serve to explain embodiments of the invention and, together with the description, serve to explain the principles of the invention.

[0039] The drawings referenced herein form part of this specification. The features shown in the drawings illustrate only some embodiments of the application and do not illustrate all embodiments of the application unless the detailed description expressly indicates otherwise, and no interpretation to the contrary should be implied by the reader of the specification. [Brief explanation of the drawings]

[0040] [Figure 1] Figures 1A-1C show that siRNA-mediated anti-VEGFR-3 treatment significantly reduced IOP and protected the eye from glaucomatous damage, as demonstrated in a mouse model of glaucoma. Intraocular hypertension was induced in normal eyes by laser photocoagulation of the episcleral vein. Anti-VEGFR-3 siRNA or control scrambled siRNA was administered topically via subconjunctival injection one day after laser. Figure 1A shows that IOP in VEGFR-3-specific siRNA-treated eyes was significantly reduced after treatment compared to the control condition (P<0.05). IOP, measured in millimeters of mercury (mmHg), is represented on the y-axis for the control (gray) and treatment (black) conditions, relative to the number of days after laser photocoagulation on the x-axis. Figure 1B shows that VEGFR-3-specific siRNA treatment significantly reduced corneal edema (*P<0.05) as measured in vivo by optical coagulation (OCT). Central corneal thickness is expressed in millimeters (mm) on the y-axis with control (black) and treatment (gray) conditions on the x-axis. Figure 1C shows summary data for human Schlemm's canal cells showing that anti-VEGFR-3 siRNA treatment significantly inhibits Schlemm's canal cell functions, such as adhesion. Fluorescence intensity, measured in relative units, is expressed on the y-axis with control (black) and treatment (gray) conditions on the x-axis (*P<0.05). [Figure 2]Figures 2A-2D show that anti-VLA-1 antibody treatment significantly reduces IOP and protects ocular tissue from glaucomatous damage, as demonstrated in a mouse model of glaucoma. Laser-induced intraocular hypertension was induced in the right eye of a normal eye. Anti-VLA-1 antibody or control was administered topically via subconjunctival injection, starting 1 day after laser. Figure 2A shows that IOP was significantly reduced in anti-VLA-1 antibody-treated eyes compared to the control condition (p<0.05). IOP, measured in millimeters of mercury (mmHg), is represented on the y-axis for the control (gray) and treatment (black) conditions against the number of days after laser photocoagulation on the x-axis. Figures 2B-2D show summary data demonstrating that anti-VLA-1 antibody treatment protected the cornea from edema (Figure 2B) and also reduced RNFL (retinal nerve fiber layer) thinning (Figure 2C) and retinal ganglion cell (RGC) death (Figure 2D). Central corneal thickness and RNFL thickness were measured in vivo by optical coherence tomography (OCT). *P<0.05; ns: not significant. Figure 2B shows that anti-VLA-1 antibody treatment significantly reduced corneal edema (*P<0.05) as measured in vivo by OCT. Central corneal thickness, measured in micrometers (μm), is represented on the y-axis with control (gray) and treatment (black) conditions on the x-axis. Figure 2C shows that anti-VLA-1 antibody treatment significantly reduced RNFL thinning (*P<0.05) as measured in vivo by OCT. RNFL thickness, measured in micrometers (μm), is represented on the y-axis with control (gray) and treatment (black) conditions on the x-axis. Figure 2D shows that anti-VLA-1 antibody treatment significantly reduced RGC death compared to control, untreated eyes (*P<0.05). Relative RGC numbers, measured in percent (%), are represented on the y-axis for control eyes (gray) and treated eyes (black) on the x-axis. In eyes that did not undergo laser photocoagulation, there was no significant difference in the relative number of RGCs between control and VLA-1 antibody-treated mice (ns, not significant). In eyes in which hypertension was induced by laser application, anti-VLA-1 antibody-treated mice had significantly more RGCs (*P<0.05). [Figure 3]Figure 3A shows that anti-VLA-1 treatment via siRNAs significantly reduced IOP (*P<0.05), as demonstrated in a mouse model of glaucoma. Laser-induced intraocular pressure was induced in normal eyes. One day after laser, anti-VLA-1 siRNA or control scrambled siRNA was administered topically via subconjunctival injection. IOP measured in millimeters of mercury (mmHg) three days after laser is represented on the y-axis with control (white) and treatment (gray) conditions on the x-axis. IOP in anti-VLA-1 siRNA-treated eyes was significantly reduced compared to the control condition (*P<0.05). Figures 3B and 3C show summary data from human Schlemm's canal cells, demonstrating that anti-VLA-1 siRNA treatment inhibits Schlemm's canal cell functions, such as adhesion (Figure 3B) and tube formation (Figure 3C) (*P<0.05). Figure 3B shows summary data for human Schlemm's canal cells demonstrating that anti-VLA-1 siRNA treatment significantly inhibits Schlemm's canal cell adhesion (*P<0.05). Fluorescence intensity is represented in relative units on the y-axis with control (white) and treatment (gray) conditions on the x-axis. Figure 3C shows that anti-VLA-1 siRNA treatment significantly inhibited Schlemm's canal cell tube formation (*P<0.05). The number of meshes is represented in relative percentage (%) on the y-axis with control (white) and treatment (gray) conditions on the x-axis. Significantly fewer (*P<0.05) meshes were observed in the anti-VLA-1 siRNA-treated condition compared to the control condition. [Figure 4]Figures 4A-4C show that siRNA-mediated anti-Ang-2 treatment significantly reduced IOP, as demonstrated in a mouse model of glaucoma. Laser-induced intraocular hypertension was induced in normal eyes. One day after laser irradiation, anti-Ang-2 siRNA or control scrambled siRNA was administered topically via subconjunctival injection. Figure 4A shows that IOP was significantly reduced in anti-Ang-2 siRNA-treated eyes compared to the control condition (P<0.05). IOP, measured in millimeters of mercury (mmHg), is represented on the y-axis for the control (gray) and treatment (black) conditions, relative to the number of days after laser photocoagulation on the x-axis. (Figures 4B and 4C) Summary data from human Schlemm's canal cells show that anti-Ang-2 siRNA treatment inhibits Schlemm's canal cell functions, such as adhesion (Figure 4B) and tube formation (Figure 4C) (*P<0.05). Figure 4B shows summary data for human Schlemm's canal cells demonstrating that anti-Ang-2 siRNA treatment significantly inhibits Schlemm's canal cell adhesion (*P<0.05). Fluorescence intensity is represented in relative units on the y-axis with control (white) and treatment (gray) conditions on the x-axis. Figure 4C shows that anti-Ang-2 siRNA treatment significantly inhibited Schlemm's canal cell tube formation (*P<0.05). The number of meshes is represented as relative percentage (%) on the y-axis with control (white) and treatment (gray) conditions on the x-axis. Significantly fewer (*P<0.05) meshes were observed in the anti-Ang-2 siRNA treatment condition compared to the control condition. [Figure 5]Figure 5A shows that siRNA-mediated anti-ITGA5 treatment significantly reduced IOP (*P<0.05), as demonstrated in a mouse model of glaucoma. Laser-induced intraocular hypertension was induced in normal eyes. One day after laser, anti-ITGA5 siRNA or control scrambled siRNA was administered topically via subconjunctival injection. IOP measured in millimeters of mercury (mmHg) on ​​day 3 after laser is represented on the y-axis for control (white) and treatment (gray) conditions, relative to the number of days after laser photocoagulation on the x-axis. IOP in anti-ITGA5 siRNA-treated eyes was significantly reduced compared to the control condition (*P<0.05). Figure 5B shows summary data from human Schlemm's canal cells, demonstrating that anti-ITGA5 siRNA treatment inhibits Schlemm's canal cell functions, such as adhesion (*P<0.05). Fluorescence intensity is represented in relative units on the y-axis for control (white) and treatment (gray) conditions on the x-axis. DETAILED DESCRIPTION OF THE INVENTION

[0041] The following detailed description of exemplary embodiments of the present application refers to the accompanying drawings, which form a part of the specification. The drawings illustrate specific exemplary embodiments in which the present application may be practiced. The detailed description, including the drawings, describes these embodiments in sufficient detail to enable those skilled in the art to practice the application. Those skilled in the art may further utilize other embodiments of the present application and may make logical, mechanical, and other changes without departing from the spirit or scope of the present application. Therefore, readers of the following detailed description should not interpret the description in a limiting sense, and only the appended claims should define the scope of the embodiments of the present application.

[0042] In this application, the use of the singular includes the plural unless specifically stated otherwise. In this application, the use of "or" means "and / or" unless specifically stated otherwise. Furthermore, the use of the term "including," as well as other forms such as "includes" and "included," is not limiting. Furthermore, the section headings used herein are for organizational purposes only and should not be construed as limiting the subject matter described.

[0043] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this disclosure belongs. Although any methods and materials similar or equivalent to those described herein can also be used in the practice or testing of this disclosure, some illustrative methods and materials are described herein.

[0044] All publications and patents cited herein are incorporated by reference as if each individual publication or patent was specifically and individually indicated to be incorporated by reference to disclose and describe the methods and / or materials in connection with which the publication is cited. The citation of any publication is for its disclosure prior to the filing date and does not constitute an admission that the present disclosure is not entitled to antedate such publication by virtue of prior disclosure. Further, the dates of publication provided may be different from the actual publication dates, which may need to be independently confirmed.

[0045] As will be apparent to those skilled in the art upon reading this disclosure, each individual embodiment described and illustrated herein has distinct components and features that are readily distinguishable or combinable with the features of any of the other several embodiments without departing from the scope or spirit of the disclosure. Any recited method can be carried out in the order of events recited or in any other order that is logically possible.

[0046] definition The following definitions are provided to assist the reader. Unless otherwise defined, all terms, notations, and other scientific or medical terms or nomenclature of the art used herein are intended to have the meanings commonly understood by one of ordinary skill in the chemical and pharmaceutical arts. In some cases, terms with commonly understood meanings are defined herein for clarity and / or ease of reference, and the inclusion of such definitions herein should not necessarily be deemed to indicate a substantial departure from the definition of the term as commonly understood in the art.

[0047] As used herein, the singular forms "a," "an," and "the" include plural references unless the context clearly dictates otherwise.

[0048] As used herein, in the context of a numerical value or range described herein, the term "about" means approximately or near, and means ±10% of the stated or claimed numerical value or range.

[0049] As used herein, the term "administering" means providing a pharmaceutical agent or composition to a subject, and includes, but is not limited to, administration by a medical professional and self-administration.

[0050] As used herein, "antibody" encompasses natural and non-natural immunoglobulins, including, for example, single-chain antibodies, chimeric antibodies (e.g., humanized mouse antibodies), and heteroconjugate antibodies (e.g., bispecific antibodies). Antibody fragments include those that bind to antigens (e.g., Fab', F(ab')2, Fab, Fv, and rIgG). See, for example, Pierce Catalog and Handbook, 1994-1995 (Pierce Chemical Co., Rockford, Ill.), Kuby, J., Immunology, 3 rdEd., W. H. Freeman & Co., New York (1998). The term antibody also includes bivalent or bispecific molecules, diabodies, triabodies, and tetrabodies. The term "antibody" further includes both polyclonal and monoclonal antibodies.

[0051] In this disclosure, terms such as "comprises," "comprised," "comprising," "contains," "containing," and the like have the meaning ascribed to them in U.S. Patent law, which are inclusive, i.e., open-ended, and do not exclude additional, unrecited elements or method steps. Terms such as "consisting essentially of" and "consists essentially of" have the meaning ascribed to them in U.S. Patent law, which permit the inclusion of additional components or steps that do not materially affect the basic and novel characteristics of the claimed invention. The terms "consists of" and "consisting of" have the meaning ascribed to them in U.S. Patent law, i.e., these terms are closed-ended.

[0052] As used herein, the term "inhibitor" refers to a biological or chemical substance that interferes with or otherwise reduces the physiological and / or biochemical action of another biological or chemical molecule. In some embodiments, the inhibitor specifically binds to the other molecule. For example, expression of a lymphatic vessel promoting factor is inhibited by at least about 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, or 50% by administration of an inhibitor described in the present invention. In some embodiments, the lymphatic vessel promoting factor is inhibited by at least about 60%, 70%, or 80% by administration of an inhibitor described in the present invention. In some embodiments, the lymphatic vessel promoting factor is inhibited by at least about 85%, 90%, or 95% by administration of an inhibitor described in the present invention.

[0053] As used herein, the term "small interfering RNA (siRNA)" (also known as short interfering RNA or silencing RNA) refers to a class of double-stranded RNA and / or non-coding RNA molecules. The siRNAs described herein can be used to silence protein-coding genes. The siRNAs described herein can be used to target the mRNA of a specific gene to produce a gene silencing effect. For example, siRNAs can interfere with the expression of a specific gene having a complementary nucleotide sequence by degrading the mRNA after transcription and preventing translation. An siRNA comprises two RNA strands that are sufficiently complementary to hybridize to form a double-stranded structure under appropriate conditions, where one siRNA strand (the antisense strand) comprises a region of complementarity that is substantially complementary to the target sequence, and the other siRNA strand (the sense strand) comprises a region complementary to the antisense strand. In some embodiments, the double-stranded siRNA structure is 15 to 30, or 25 to 30, or 18 to 25, or 19 to 24, or 19 to 21, or 19, 20, or 21 base pairs in length. In some embodiments, the siRNA duplex is 19 base pairs in length. In some embodiments, the siRNA duplex is 21 base pairs in length. Each strand of the siRNA duplex can be the same length or different lengths. The siRNA described herein can include one or more single-stranded overhangs of one or more nucleotides. In some embodiments, at least one end of the dsRNA has a single-stranded nucleotide overhang of 1 to 4, typically 1 or 2, nucleotides. In some embodiments, the siRNA is chemically modified to enhance stability. The nucleic acid molecules described in the present invention can be synthesized and / or modified by methods established in the art, such as those described in "Current protocols in nucleic acid chemistry," Beaucage, SL et al. (Eds.), John Wiley & Sons, Inc., New York, NY, USA, which is incorporated herein by reference.

[0054] The siRNA can be encoded by a nucleic acid sequence, and the nucleic acid sequence can also include a promoter. The nucleic acid sequence can also include a polyadenylation signal. In some embodiments, the polyadenylation signal is a synthetic minimal polyadenylation signal.

[0055] As used herein, the term "antisense strand" refers to the strand of an siRNA that includes a sequence region that is substantially complementary to a target sequence.

[0056] As used herein, the term "sense strand" refers to the strand of an siRNA that includes a region that is substantially complementary to a region of the antisense strand.

[0057] As used herein, the terms "subject," "host," and "patient" are used interchangeably. As used herein, a "patient," "host," or "subject" to be treated is typically a human patient, although it should be understood that the methods described herein are effective with respect to other animals, such as mammals. In certain embodiments, the patient, host, or subject is human.

[0058] As used herein, the term "therapeutically effective amount" means an amount of an agent sufficient to prevent, treat, alleviate and / or ameliorate the symptoms and / or underlying causes of any disorder or disease, or an amount of an agent sufficient to produce a desired effect on a cell.

[0059] The terms "treatment," "treat," or "treating" refer to a method of reducing the damage caused by a disease or a symptom of a disease or the severity of a disease or a symptom of a disease. Thus, in the disclosed methods, treatment can refer to a 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, or 100% reduction in the damage caused by a disease or a symptom of a disease or the severity of a disease or a symptom of a disease. For example, a method of treating a disease is considered therapeutic if one or more symptoms of the disease are reduced by 10% compared to a control. Thus, reduction can be a 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, or any percentage reduction between 10% and 100% compared to native or control levels. It is understood that treatment does not necessarily refer to a cure or complete elimination of the disease, condition, or symptoms of the disease or condition.

[0060] Throughout this disclosure, various aspects of the present invention may be presented in a range format. It should be understood that the description in range format is merely for convenience and does not limit the scope of the invention. The description of a range should be considered to have specifically disclosed all possible subranges in addition to individual numerical values ​​within that range. For example, the description of a range such as 1 to 6 should be considered to have specifically disclosed subranges such as 1 to 3, 1 to 4, 1 to 5, 2 to 4, 2 to 6, 3 to 6, etc., as well as individual numerical values ​​within that range, e.g., 1, 2, 2.7, 3, 4, 5, 5.3, and 6. This applies regardless of the breadth of the range.

[0061] Lymphangiogenesis and glaucoma As used herein, the term "lymphangiogenesis" refers to the process of forming new lymphatic vessels from pre-existing blood or lymphatic endothelial progenitor cells. The lymphatic system plays an important role in maintaining fluid balance and immune function in the body. Lymphatic vessels are responsible for transporting lymph, which contains immune cells and other important components, throughout the body.

[0062] Lymphangiogenesis occurs during normal development, wound healing, and certain pathological conditions such as cancer and inflammation. This process involves multiple facets, including lymphatic endothelial cell activation, vascular patterning and maintenance, and expression of growth factors such as vascular endothelial growth factors and receptors (e.g., VEGF-C, VEGF-D, VEGFR-3), and angiopoietin factors that promote lymphatic vessel formation, patterning, and maintenance. The lymphatic network permeates many tissues in the body, and its dysfunction has been found in a wide range of disorders, including cancer metastasis, inflammatory and immune disorders, tissue and organ (heart and kidney) transplant rejection, obesity, hypertension, and lymphedema (Chen L. Ocular lymphatics: state-of-the-art review. Lymphology. 42:66-76 (2009)).

[0063] Some studies suggest that lymphangiogenesis may be involved in the pathogenesis of glaucoma. For example, studies suggest that lymphatic vessels may be involved in the drainage of aqueous humor, which is important for maintaining normal intraocular pressure. Researchers have found that destruction of lymphatic vessels may contribute to elevated intraocular pressure (IOP) (Non-Patent Documents 6, 7, and 8), and is an important risk factor for glaucoma. It has been proposed that inducing or activating lymphangiogenesis could potentially treat glaucoma (see, for example, International Publication No. 2015110701, which is incorporated herein by reference).

[0064] The present disclosure is based on the surprising discovery that inhibiting lymphangiogenic factors reduces IOP and protects the eye from glaucomatous damage. Accordingly, in one aspect, the present disclosure provides a method for preventing or treating glaucoma. In one embodiment, the method comprises administering a therapeutically effective amount of an inhibitor of lymphangiogenic factors to an eye in need thereof, thereby reducing IOP in the eye. In some embodiments, the present disclosure provides a method for preventing or treating primary glaucoma. In some embodiments, the primary glaucoma is primary open-angle glaucoma (POAG). In some embodiments, the primary glaucoma is angle-closure glaucoma. In some embodiments, the primary glaucoma is congenital glaucoma. In some embodiments, the primary glaucoma is normal-tension glaucoma.

[0065] In some embodiments, the present disclosure provides a method of preventing or treating an IOP-mediated ocular disorder, in some embodiments, the IOP-mediated ocular disorder is selected from the group consisting of secondary glaucoma, glaucomatous optic neuropathy (GON), ocular hypertension, open-angle glaucoma, angle-closure glaucoma, and congenital glaucoma.

[0066] Lymphangiogenesis and Schlemm's Canal As a key component of the conventional outflow pathway, Schlemm's canal is a critical structure in the regulation of aqueous humor drainage and IOP. The canal itself accounts for 70% to 90% of total aqueous humor outflow in humans. The endothelial cells lining the canal are the primary site of resistance to aqueous humor outflow and a major determinant of overall IOP. Increased canal resistance in response to several pathological insults results in elevated IOP. Therefore, Schlemm's canal may play an important role in the pathogenesis of glaucoma.

[0067] Previously, several lymphatic vessel markers, such as PROX-1 and VEGF-C, have been demonstrated to be involved in the development of Schlemm's canal (Non-Patent Document 6), suggesting that Schlemm's canal may exhibit a lymphatic vessel-like phenotype. Although glaucoma is associated with dysregulation of the lymphatic system, other studies have concluded that there is no similarity between lymphatic vessels and aqueous humor drainage vessels (Chen, L. Ocular lymphatics: state-of-the-art review. Lymphology. 42(2):66-76(2009 June)).

[0068] Lymphangiogenesis promoting factor In some aspects of the present invention, lymphangiogenic factors are targeted to reduce, inhibit, or prevent ocular lymphangiogenesis. As used herein, lymphangiogenic factors refer to genes, RNAs, or proteins involved in the formation or maintenance of lymphatic vessels. Lymphangiogenic factors can induce (pro-lymphatic) or inhibit (anti-lymphatic) lymphangiogenesis. Thus, in some embodiments, inhibitors of lymphangiogenic factors inhibit pro-lymphangiogenic factors, and pro-lymphangiogenic factors induce or maintain ocular lymphangiogenesis. In some embodiments, inhibitors of ocular lymphangiogenesis activate anti-lymphangiogenic factors, and anti-lymphangiogenic factors inhibit ocular lymphangiogenesis.

[0069] In some embodiments, the lymphangiogenesis promoting factors disclosed herein are members of the VEGF / VEGFR family. Examples of VEGF / VEGFR family members that are lymphangiogenesis promoting factors include VEGF-A, VEGF-B, VEGF-C, VEGF-D, VEGF-E, VEGFR-1, VEGFR-2, and VEGFR-3. In some embodiments, the lymphangiogenesis promoting factor is selected from the group consisting of VEGF-C, VEGF-D, and VEGFR-3. In some embodiments, the lymphangiogenesis promoting factor is VEGFR-3.

[0070] In some embodiments, the lymphangiogenesis promoter disclosed herein is an integrin. Examples of integrins that are lymphangiogenesis promoters include VLA-1, integrin alpha 5 (ITGA5), and integrin alpha 9 (ITGA9). In some embodiments, the lymphangiogenesis promoter is VLA-1. In some embodiments, the lymphangiogenesis promoter is ITGA5. In some embodiments, the lymphangiogenesis promoter is ITGA9.

[0071] In some embodiments, the lymphangiogenesis promoter disclosed herein is an angiopoietin. Examples of angiopoietins that are lymphangiogenesis promoters include angiopoietin 1 (ANGPT1), angiopoietin-2 (ANGPT2 or Ang-2), and Tie2 / TEK. In some embodiments, the lymphangiogenesis promoter is Ang-2.

[0072] In some embodiments, the lymphangiogenesis promoting factors disclosed herein are cytokines / chemokines. Examples of cytokines / chemokines that are lymphangiogenesis promoting factors include interleukin-8, interferon-γ, CCR7, and SLC.

[0073] In some embodiments, the lymphangiogenesis promoting factors disclosed herein are extracellular matrix proteins. An example of an extracellular matrix protein that is a lymphangiogenesis promoting factor is CCBE1.

[0074] In some embodiments, the lymphangiogenesis promoting factors disclosed herein are transcription factors. Examples of transcription factors that are lymphangiogenesis promoting factors include Sox18 and Hhex.

[0075] In some embodiments, the lymphangiogenesis promoting factors disclosed herein are inductive molecules. Examples of inductive molecules that are lymphangiogenesis promoting factors include neuropilin 2 and SEMA7A.

[0076] In some embodiments, the lymphangiogenesis promoting factor disclosed herein is an FGF. An example of an FGF that is a lymphangiogenesis promoting factor is FGF-2.

[0077] In some embodiments, the lymphangiogenesis promoting factors disclosed herein are protein tyrosine phosphatases (PTPs). Examples of PTPs that are lymphangiogenesis promoting factors include PTPN14.

[0078] In some embodiments, the lymphangiogenesis-promoting factors disclosed herein are platelet factors. Examples of platelet factors that are lymphangiogenesis-promoting factors include platelet factor 4 or platelet-derived growth factor (e.g., PDGF-BB).

[0079] In some embodiments, the lymphangiogenesis-promoting factors disclosed herein are LEC polarization factors (e.g., Celsr1, Vangl2, Pdk2, and Fat4). Examples of LEC polarization factors that are lymphangiogenesis-promoting factors include Celsr1, Vangl2, Pdk2, and Fat4.

[0080] In some embodiments, the lymphangiogenesis promoting factors disclosed herein are members of the Notch family.

[0081] In some embodiments, the lymphangiogenesis promoting factor disclosed herein is ARAF, SOS1, Apelin, KIF11, REELIN, CALCRL, GJC2 (gap junction protein gamma-2), Rasip1, or FBXL7 (F-box and leucine-rich repeat protein 7).

[0082] In some embodiments, the lymphangiogenic factor is a major lymphangiogenic factor, hi some embodiments, the major lymphangiogenic factor is selected from the group consisting of VEGFR-3, VEGF-C, VEGF-D, VEGF-A, VLA-1, angiopoietin-2, Tie-2, and integrin alpha 5.

[0083] In some embodiments, the lymphangiogenesis promoting factor is a secondary lymphangiogenesis promoting factor selected from the group consisting of VEGF-B, VEGF-E, PIGF, VEGFR-1, VEGFR-2, ANGPT1, ITGA9, TEK, interleukin-8, interferon-gamma, CCR7, SLC, CCBE1, Sox18, Hhex, neuropilin 2, and SEMA7, FGF-2, PTPN14, PDGF-BB, platelet factor 4, Celsr1, Vangl2, Pdk2, Fat4, Notch family members, ARAF, SOS1, Apelin, KIF11, REELIN, CALCRL, GJC2, Rasip1, and FBXL7.

[0084] VEGF / VEGFR family The vascular endothelial growth factor (VEGF) family regulates blood and lymphatic processes. In some embodiments, the lymphangiogenesis promoter inhibited as disclosed herein is a member of the VEGF / VEGFR family. Examples of VEGF / VEGFR family members that are lymphangiogenesis promoters include VEGF-A, VEGF-B, VEGF-C, VEGF-D, VEGF-E, VEGFR-1, VEGFR-2, and VEGFR-3. In some embodiments, the lymphangiogenesis promoter is selected from the group consisting of VEGF-C, VEGF-D, and VEGFR-3.

[0085] Vascular endothelial growth factor receptor 3 (VEGFR-3) In some embodiments, the lymphangiogenesis-promoting factor inhibited as disclosed herein is VEGFR-3. VEGFR-3 belongs to the VEGF family. Previous studies have shown that VEGFR-3 mediates lymphangiogenesis in the cornea and other tissues, and its inhibition suppresses transplant rejection, tumor growth, and metastasis (Yuen, D. et al. Combined blockade of VEGFR-2 and VEGFR-3 inhibits inflammatory lymphangiogenesis in early and middle stages. Invest Ophthalmol Vis Sci. 52(5):2593-2597(2011 Apr 20)). However, the potential role and specificity of VEGFR-3 in primary glaucoma have not yet been elucidated.

[0086] Integrin family In some embodiments, the lymphangiogenesis-promoting factor inhibited as disclosed herein is an integrin, such as β1 integrin. Integrins are heterodimeric transmembrane receptors that link the actin cytoskeleton to the ECM and influence gene expression (Vigneault, F. et al. Control of integrin genes expression in the eye. Progress in Retinal and Eye Res. 26:99-161(2007)). Examples of β1 integrins that are lymphangiogenesis-promoting factors include VLA-1, integrin alpha 5 (ITGA5), and integrin alpha 9 (ITGA9). In some embodiments, the lymphangiogenesis-promoting factor is VLA-1. In some embodiments, the lymphangiogenesis-promoting factor is ITGA5. In some embodiments, the lymphangiogenesis-promoting factor is ITGA9.

[0087] Integrin α5 (ITGA5) In some embodiments, the lymphangiogenesis-promoting factor inhibited as disclosed herein is ITGA5. Integrin alpha 5 (ITGA5) belongs to the integrin alpha chain family, which mediates cell surface adhesion and signal transduction. ITGA5 is a preprotein that is proteolytically cleaved to generate the light and heavy chains that make up the alpha 5 subunit. It has previously been reported that ITGA-5 mediates corneal inflammatory lymphangiogenesis, which is suppressed by ITGA-5 blockade (Dietrich T, et al. Inhibition of Inflammatory Lymphangiogenesis by Integrin 5 Blockade. Am J Pathol. 2007 Jul;171(1):361-72).

[0088] Very late antigen 1 (VLA-1) In some embodiments, the lymphangiogenesis-promoting factor inhibited as disclosed herein is VLA-1. VLA-1 (very late antigen-1), also known as integrin α1β1, is a receptor for collagen and laminin. VLA-1 is expressed on lymphatic endothelial cells (LECs). Previously, VLA-1 has been reported to mediate corneal inflammatory lymphangiogenesis, which is suppressed by VLA-1 blockade (Grimaldo, S. et al. Very late antigen-1 mediates corneal lymphangiogenesis. Invest Ophthalmol Vis Sci. 52(7):4808-4812(2011 Jul 1)). It has also been reported that lymphangiogenesis associated with corneal transplantation is suppressed in VLA-1 knockout mice (Chen, L. et al. Very late antigen 1 blockade markedly promotes survival of corneal allografts. Arch Ophthalmol. 125:783-788(2007)). However, the molecular and cellular mechanisms of VLA-1 involvement in other ocular diseases, such as glaucoma, have not yet been elucidated.

[0089] Angiopoietin family In some embodiments, the lymphangiogenesis promoting factor inhibited as disclosed herein is an angiopoietin. The angiopoietin (ANGPT) family is a signaling pathway that includes ligands that activate Tie2 / TEK (e.g., angiopoietin 1 (ANGPT1), angiopoietin 2 (ANGPT2 or Ang-2)). Examples of angiopoietins that are lymphangiogenesis factors include angiopoietin 1 (ANGPT1), angiopoietin-2 (ANGPT2 or Ang-2), and Tie2 / TEK. In some embodiments, the lymphangiogenesis promoting factor is Ang-2.

[0090] Angiopoietin-2 (Ang-2) (ANGPT2) In some embodiments, the lymphangiogenic factor inhibited as disclosed herein is Ang-2. Angiopoietin-2 (Ang-2 or ANGPT2) belongs to the angiopoietin-Tie family. The specific function of Ang-2 in the lymphatic system is not yet fully understood. Previous studies have reported that Ang-2 deficiency leads to lymphatic vessel defects during development and inflammation (Dellinger, M. et al. Defective remodeling and maturation of the lymphatic vasculature in angiopoietin-2-deficient mice. Dev Biol. 319:309-320(2008); Yuen, D. et al. Role of angiopoietin-2 in corneal lymphangiogenesis. Invest Ophthalmol Vis Sci. 55:3320-3327(2014); Zheng, W. et al. Angiopoietin-2 regulates the transformation and integrity of lymphatic endothelial cell junctions. Genes Dev. 28:1592-1603(2014)). However, the role of Ang-2 in glaucoma remains unclear.

[0091] Cytokines / chemokines In some embodiments, the lymphangiogenic factors inhibited as disclosed herein are cytokines / chemokines. Cytokines or chemokines are involved in lymphangiogenesis (Sainz-Jaspeado, M. & Claesson-Welsh, L. Cytokines regulating lymphangiogenesis. Curr Opin in Immunol. 53:58-63(2018)). Chemokines are small protein cytokines that act as chemotactic agents and have also been shown to be involved in the lymphatic system (Farnsworth, RH et al. The Interplay Between Lymphatic Vessels and Chemokines. Front Immunol. 10:518(2019 Apr 12)). Examples of cytokines / chemokines that are lymphangiogenic factors include interleukin-8, interferon-γ, CCR7, and SLC.

[0092] Extracellular matrix proteins In some embodiments, the lymphangiogenic factor inhibited as disclosed herein is an extracellular matrix protein. An example of an extracellular matrix protein that is a lymphangiogenic factor is CCBE1. CCBE1 binds to the ECM and contributes to VEGF-C-mediated VEGFR-3 activation (Brouillard, P. et al. Genetics of lymphatic anomalies. J Clin Invest. 124(3):898-904 (2014 Mar; Epub 2014 Mar 3)).

[0093] transcription factors In some embodiments, the lymphangiogenesis-promoting factor inhibited as disclosed herein is a transcription factor. Examples of transcription factors that are lymphangiogenesis factors include Sox18 and Hhex. Some transcription factors act upstream or downstream of VEGFR-3 (Brouillard, P. et al. Genetics of lymphatic anomalies. J Clin Invest. 124(3):898-904 (2014 Mar; Epub 2014 Mar 3)).

[0094] guiding molecule In some embodiments, the lymphangiogenic factors inhibited as disclosed herein are inductive molecules. Examples of inductive molecules that are lymphangiogenic factors include neuropilin 2 and SEMA7A.

[0095] Fibroblast growth factor (FGF) family In some embodiments, the lymphangiogenic factor inhibited as disclosed herein is a member of the fibroblast growth factor (FGF) family. FGFs are mitogens that regulate various biological processes, including cell proliferation, differentiation, and survival, and are involved in several diseases (Xie, Y. et al. FGF / FGFR signaling in health and disease. Signal Transduct Target Ther. 5(1):181(2020 Sep 2)). An example of an FGF that is a lymphangiogenic factor is FGF-2.

[0096] Protein tyrosine phosphatase (PTP) In some embodiments, the lymphangiogenesis-promoting factor inhibited as disclosed herein is a protein tyrosine phosphatase (PTP). In some embodiments, the lymphangiogenesis factor is PTPN14. PTPN14 is a phosphatase that interacts with VEGFR3, which is activated by VEGF-C (Oliver, G. et al. The Lymphatic Vasculature in the 21st Century: Novel Functional Roles in Homeostasis and Disease. Cell. 182(2):270-296(2020 Jul 23)).

[0097] platelet factor In some embodiments, the lymphangiogenic factor inhibited as disclosed herein is a platelet factor. Examples of platelet factors that are lymphangiogenic factors include platelet factor 4 or platelet-derived growth factor (e.g., PDGF-BB). Platelet factors are associated with the lymphatic vasculature (Ma, W. et al. Platelet factor 4 is a biomarker for lymphatic-promoted disorders. JCI Insight. 5(13):e135109(2020 Jul 9)). In some embodiments, the lymphangiogenic factor is platelet factor 4 (PF4). In some embodiments, the lymphangiogenic factor is platelet-derived growth factor (PDGF-BB).

[0098] Lymphatic endothelial cell (LEC) polarity factors In some embodiments, the lymphangiogenesis-promoting factor inhibited as disclosed herein is an LEC polarity factor. Lymphatic endothelial cell (LEC) polarity factors control planar cell polarity and lymphatic vessel organization. For example, Fat4 acts as a cell polarity regulator required for vascular morphogenesis of developing lymphatic vessels (Betterman, KL et al. Atypical cadherin FAT4 orchestrates lymphatic endothelial cell polarity in response to flow. J Clin Invest. 130(6):3315-3328(2020 Jun 1)). Examples of LEC polarity factors that are lymphangiogenesis factors include Celsr1, Vangl2, Pdk2, and Fat4.

[0099] Notch Family In some embodiments, the lymphangiogenesis-promoting factors disclosed herein are members of the Notch family. The Notch pathway, consisting of receptors (Notch1-4) and ligands (Dll1 / 3 / 4 and Jagged1 / 2), induces intercellular signaling and regulates cell development, proliferation, and differentiation. The Notch pathway is also involved in lymphangiogenesis and lymphatic differentiation (Niessen, K. et al. The Notch1-Dll4 signaling pathway regulates mouse postnatal lymphatic development. Blood. 118(7):1989-1997 (2011 Aug 18)).

[0100] Lymphangiogenesis promoting factor inhibitors In some embodiments, the inhibitor of lymphatic vessel promoting factor is selected from the group consisting of a polynucleotide, a protein, a polypeptide, and a gene editing composition. The inhibitor of lymphatic vessel promoting factor can be a nucleotide (e.g., siRNA), a protein (e.g., antibody, recombinant protein), a peptide, a small molecule, or a gene editing composition (e.g., CRISPR / Cas, gRNA). In some embodiments, the inhibitor of lymphatic vessel promoting factor is selected from the group consisting of a gene disrupting agent, siRNA, antagonist antibody, recombinant protein, and small molecule.

[0101] In some embodiments, the inhibitor of a lymphangiogenesis factor is an agent that disrupts the gene encoding the lymphangiogenesis factor. In some embodiments, the inhibitor of a lymphangiogenesis factor is an siRNA directed against a lymphangiogenesis factor. In some embodiments, the inhibitor of a lymphangiogenesis factor is an antagonist antibody or recombinant protein directed against a lymphangiogenesis factor. In some embodiments, the inhibitor of a lymphangiogenesis factor is a small molecule that inhibits the function of a lymphangiogenesis factor.

[0102] In some embodiments, the inhibitor of a lymphangiogenesis promoting factor is an agent that disrupts a gene for a lymphangiogenesis promoting factor. In some embodiments, the gene disrupting agent comprises a gene editing composition. In some embodiments, the gene editing composition comprises CRISPR / Cas9 and a gRNA. The gene editing compositions described in the present invention can be produced and used according to methods established in the art, for example, the method described in "Genome engineering using the CRISPR-Cas9 system," Ran, FA et al. Nature Protocols 11(8)(2013), which is incorporated herein by reference.

[0103] In some embodiments, the lymphangiogenesis inhibitor is an agent that activates expression of an anti-lymphangiogenic factor. In some embodiments, the lymphangiogenesis inhibitor is an agonist antibody to an anti-lymphangiogenic factor. In some embodiments, the lymphangiogenesis inhibitor is a small molecule that activates the function of an anti-lymphangiogenic factor.

[0104] Exemplary siRNA molecules for inhibiting specific lymphangiogenic factors are shown in Table 1.

[0105] [Table 1] TIFF2026507009000003.tif162170

[0106] siRNAs are commercially available from several suppliers, including Thermo Fisher Scientific, Qiagen, Origene, etc. The example above is from Thermo Fisher Scientific. Additional examples include siRNAs from other companies, such as Qiagen, such as VLA-1 (5'-TCACAGAAGTAAAGGAGAAA-3') (SEQ ID NO: 27) and ITGA-9 (5'-AAGAAGAAAGTC GTACTATAG-3') (SEQ ID NO: 28). Other exemplary siRNAs include VEGFR-3 (Origene, catalog number SR301631), Ang-2 (Santa Cruz Biotechnology Inc., catalog number sc-39305; Origene, catalog number SR300199), ITGA5 (Santa Cruz Biotechnology, Inc., catalog number sc-29372; Biorbyt Ltd, catalog number orb1865790; Origene, catalog number SR320702), ITGA9 (Biorbyt Ltd, catalog number orb1865788; Origene, catalog number TR312090; Abbexa, catalog number abx920905), VEGF-A (MyBioSource, catalog number MBS8229845; Biorbyt Ltd, catalog number orb1863180), VEGF-C (Biorbyt Ltd., catalog number orb1863178; SignalChem, catalog number V812-911; Santa Cruz Biotechnology, Inc., catalog number sc-39842), and those specific for VEGF-D (Santa Cruz Biotechnology, Inc., catalog number sc-39844).

[0107] Antagonist antibodies against target lymphangiogenic factors can also be used to practice the methods described herein. Exemplary antagonist antibodies for inhibiting specific lymphangiogenic factors are listed in Table 2.

[0108] [Table 2]

[0109] Antibodies are commercially available from several suppliers, including Thermo Fisher Scientific, Abcam, Santa Cruz Biotechnology, Inc., etc. Examples of the above are from ImClone Systems Inc., BD Pharmingen / Biosciences, Thermo Fisher Scientific, Chemicon, BioCell, and Santa Cruz Biotechnology, Inc. Additional exemplary antibody inhibitors of lymphangiogenesis include anti-ITGA-9 antibodies (clone Y9A2, clone ASP5094) (see, e.g., Emori et al. Constitutive Activation of Integrin α9 Augments Self-Directed Hyperplastic and Proinflammatory Properties of Fibroblast-like Synoviocytes of Rheumatoid Arthritis. J Immunol. 199(10):3427-3436 (2017 Nov 15)).

[0110] In some embodiments, the inhibitor of lymphangiogenesis is selected from small non-coding RNAs, such as miR-184, miR-126, miR-31, miR-181a, miR-132, miR-194, miR-186, miR-99a, miR-92a, and miR-466; MEK inhibitors (e.g., trametinib), integrin inhibitors (e.g., JSM6427), angiopoietin inhibitors (e.g., L1-10), statins, VEGF family inhibitors (e.g., soluble VEGFR-2, soluble VEGFR-3, VEGF-C / VEGF-D ligand trap, aflibercept, bevacizumab, brolucizumab, ranibizumab, pegaptanib sodium), and / or faricimab-svoa.

[0111] In some embodiments, the inhibitor of ocular lymphangiogenesis is formulated as eye drops, a depot, a bolus, an inhibitor-loaded contact lens, a suspension, a solution, an ophthalmic gel, or an ointment.

[0112] Anti-lymphangiogenic factors In an alternative aspect, the inhibitor of lymphangiogenesis can be an agent that activates or stimulates an anti-lymphangiogenic factor that inhibits lymphangiogenesis, as further described below. In some embodiments, the inhibitor of lymphangiogenesis is selected from the group consisting of: (1) an agent that activates expression of an anti-lymphangiogenic factor, (2) an agonist antibody that activates an anti-lymphangiogenic factor, (3) a microRNA or mimic that inhibits lymphangiogenesis, and (4) a small molecule that activates the function of an anti-lymphangiogenic factor.

[0113] In some embodiments, the inhibitor of lymphangiogenesis is selected from small non-coding RNAs, such as miR-184, miR-126, miR-31, miR-181a, miR-132, miR-194, miR-186, miR-99a, miR-92a, and miR-466; MEK inhibitors (e.g., trametinib), integrin inhibitors (e.g., JSM6427), angiopoietin inhibitors (e.g., L1-10), statins, VEGF family inhibitors (e.g., soluble VEGFR-2, soluble VEGFR-3, VEGF-C / VEGF-D ligand trap, aflibercept, bevacizumab, brolucizumab, ranibizumab, pegaptanib sodium), and / or faricimab-svoa.

[0114] In some embodiments, the anti-lymphangiogenic factor is selected from the group consisting of microRNA-184, microRNA-126, miR-31, miR-181a, microRNA-132, miR-194, miR-186, miR-99a, miR-92a, and miR-466, endostatin, and a VEGF family inhibitor (e.g., soluble VEGFR-2, soluble VEGFR-3).

[0115] Treatment method In some embodiments, the methods disclosed herein treat glaucoma, including primary glaucoma, by modulating the function of Schlemm's canal (SC) in the eye. Schlemm's canal is a ring-shaped canal located in the angle of the anterior chamber of the eye. Schlemm's canal is responsible for the drainage of aqueous humor, a clear fluid that circulates within the eye and maintains intraocular pressure. In some embodiments, the glaucoma is primary glaucoma. In some embodiments, the primary glaucoma is primary open-angle glaucoma (POAG). In some embodiments, the primary glaucoma is angle-closure glaucoma. In some embodiments, the primary glaucoma is congenital glaucoma. In some embodiments, the primary glaucoma is normal-tension glaucoma.

[0116] In some embodiments, the present disclosure provides a method of preventing or treating an IOP-mediated ocular disorder, in some embodiments, the IOP-mediated ocular disorder is selected from the group consisting of secondary glaucoma, glaucomatous optic neuropathy (GON), ocular hypertension, open-angle glaucoma, angle-closure glaucoma, and congenital glaucoma.

[0117] In some embodiments, the methods disclosed herein inhibit SC cell adhesion in the eye. In some embodiments, the methods disclosed herein inhibit SC cell function (e.g., adhesion, proliferation, migration, tube formation) in the eye. In some embodiments, the methods disclosed herein increase aqueous humor outflow from the anterior chamber of the eye.

[0118] In the methods disclosed herein, the inhibitor of lymphatic vessel promoting factor can be administered to an eye in need thereof via any suitable route, either systemically or locally. In some embodiments, the inhibitor of lymphatic vessel promoting factor is administered locally via intracameral injection, subconjunctival injection, intravitreal injection, suprachoroidal injection, peribulbar injection, or retrobulbar injection.

[0119] In some embodiments, the methods disclosed herein protect the eye from glaucomatous damage, such as corneal edema, retinal ganglion cell loss, and retinal nerve fiber layer thinning. In some embodiments, the glaucomatous damage is corneal edema. In some embodiments, the glaucomatous damage is retinal ganglion cell loss. In some embodiments, the glaucomatous damage is retinal nerve fiber layer thinning.

[0120] In some embodiments, the eye in need of treatment is treated with a second therapy. In some embodiments, the second therapy is selected from the group consisting of eye drops, oral medications, laser therapy, and surgery. In some embodiments, the second therapy is selected from eye drops that reduce the amount of fluid produced in the eye or improve fluid drainage from the eye, oral medications that lower IOP, laser therapy (e.g., laser trabeculoplasty and laser iridotomy), and surgery (e.g., implants, trabeculectomy). In some embodiments, the second therapy is surgery. In some embodiments, the surgery is canaloplasty.

[0121] In one aspect, the present disclosure provides a method of treating glaucoma in an eye of a subject in need thereof, comprising administering to the eye a therapeutically effective amount of an inhibitor of a lymphangiogenic factor associated with Schlemm's canal, wherein the lymphangiogenic factor induces or maintains lymphangiogenesis in the eye (a "pro-lymphangiogenic factor"). In some embodiments, the subject has primary glaucoma. In some embodiments, the subject has primary open-angle glaucoma. In some embodiments, the subject has primary angle-closure glaucoma. In some embodiments, the subject has primary congenital glaucoma. In some embodiments, the subject has primary normal-tension glaucoma.

[0122] In another aspect, the present disclosure provides a method of preventing glaucoma in an eye of a subject in need thereof, comprising administering to the eye a therapeutically effective amount of an inhibitor of lymphangiogenesis promoting factor. In some embodiments, the subject has primary glaucoma. In some embodiments, the subject has primary open-angle glaucoma. In some embodiments, the subject has primary angle-closure glaucoma. In some embodiments, the subject has primary congenital glaucoma. In some embodiments, the subject has primary normal-tension glaucoma.

[0123] In another aspect, the present disclosure provides a method for inhibiting lymphangiogenesis in the eye of a subject experiencing elevated intraocular pressure (IOP), the method comprising administering to the eye a therapeutically effective amount of an inhibitor of a lymphangiogenesis promoting factor, wherein administration of the inhibitor results in a decrease in IOP in the eye.

[0124] In another aspect, the present disclosure provides a method for inhibiting lymphangiogenesis in the eye of a subject at risk of developing glaucoma, comprising administering a therapeutically effective amount of an inhibitor of lymphangiogenesis promoting factor to the eye, wherein administration of the inhibitor results in a decrease in IOP of the eye. In some embodiments, the glaucoma is primary glaucoma.

[0125] In another aspect, the present disclosure provides a method for increasing Schlemm's canal permeability in the eye of a subject experiencing elevated IOP, comprising administering to the eye a therapeutically effective amount of an inhibitor of lymphangiogenesis promoting factor, wherein administration of the inhibitor reduces IOP in the eye.

[0126] In another aspect, the disclosure provides a method for inhibiting Schlemm's canal cell activity in the eye of a subject experiencing elevated IOP, comprising administering to the eye a therapeutically effective amount of an inhibitor of lymphangiogenesis promoting factor, wherein the cell activity is selected from the group consisting of inhibition of cell adhesion, inhibition of cell proliferation, inhibition of cell migration, or inhibition of tube formation.

[0127] In another aspect, the present disclosure provides a method for increasing aqueous humor outflow in Schlemm's canal of an eye of a subject experiencing elevated IOP, comprising administering to the eye a therapeutically effective amount of an inhibitor of lymphangiogenesis promoting factor, wherein administration results in increased aqueous humor outflow in Schlemm's canal of the eye and reduced IOP.

[0128] In another aspect, the present disclosure provides a method for reducing corneal edema in an eye of a subject in need thereof, the method comprising administering to the eye a therapeutically effective amount of an inhibitor of lymphangiogenesis promoting factor, wherein administration of the inhibitor results in a reduction of corneal edema.

[0129] In another aspect, the present disclosure provides a method for reducing retinal nerve fiber layer thinning in an eye of a subject in need thereof, the method comprising administering to the eye a therapeutically effective amount of an inhibitor of lymphangiogenesis promoting factor, wherein administration of the inhibitor results in a reduction in nerve fiber thinning.

[0130] In another aspect, the present disclosure provides a method for reducing retinal ganglion cell (RGC) death in the eye of a subject in need thereof, the method comprising administering to the eye a therapeutically effective amount of an inhibitor of lymphangiogenesis promoting factor, wherein administration of the inhibitor results in a reduction in retinal ganglion cell (RGC) death.

[0131] In some embodiments, the subject has primary glaucoma selected from the group consisting of primary open-angle glaucoma, primary angle-closure glaucoma, primary congenital glaucoma, and primary normal-tension glaucoma. In some embodiments, the subject has primary open-angle glaucoma. In some embodiments, the subject has primary angle-closure glaucoma. In some embodiments, the subject has primary congenital glaucoma. In some embodiments, the subject has primary normal-tension glaucoma. In some embodiments, the subject is human.

[0132] In some embodiments, the lymphangiogenesis promoting factor is a VEGF / VEGFR family member. In some embodiments, the VEGF / VEGFR family member is selected from the group consisting of VEGF-A, VEGF-B, VEGF-C, VEGF-D, VEGF-E, PIGF, VEGFR-1, VEGFR-2, and VEGFR-3. In some embodiments, the VEGF / VEGFR family member is VEGF-A. In some embodiments, the VEGF / VEGFR family member is VEGF-C. In some embodiments, the VEGF / VEGFR family member is VEGF-D. In some embodiments, the VEGF / VEGFR family member is VEGFR-3.

[0133] In some embodiments, the lymphangiogenesis promoting factor is an integrin family member. In some embodiments, the integrin family member is selected from the group consisting of very late antigen-1 (VLA-1), integrin alpha 5 (ITGA5), and integrin alpha 9 (ITGA9). In some embodiments, the integrin family member is VLA-1. In some embodiments, the integrin family member is ITGA5. In some embodiments, the integrin family member is ITGA9.

[0134] In some embodiments, the lymphangiogenesis promoting factor is an angiopoietin family member. In some embodiments, the angiopoietin family member is selected from the group consisting of angiopoietin-1 (ANGPT1), angiopoietin-2 (ANGPT2 or Ang-2), and Tie2 / TEK. In some embodiments, the angiopoietin family member is Ang-2.

[0135] In some embodiments, the lymphangiogenesis promoting factor is selected from the group consisting of: (i) a cytokine or chemokine; (ii) an extracellular matrix protein; (iii) a transcription factor; (iv) an inductive molecule; (v) a fibroblast growth factor (FGF); (vi) a protein tyrosine phosphatase (PTP); (vii) a platelet factor member, or (viii) a platelet-derived growth factor; (ix) an LEC polarity factor; (x) a Notch family member; and (xi) a combination of any of (i)-(x).

[0136] In some embodiments, the lymphangiogenesis promoting factor is selected from the group consisting of: (i) a cytokine / chemokine selected from the group consisting of interleukin-8, interferon-γ, CCR7, and SLC; (ii) an extracellular matrix protein, which is CCBE1; (iii) a transcription factor, which is selected from the group consisting of Sox18 and Hhex; (iv) an inducer molecule, which is selected from the group consisting of neuropilin 2 and SEMA7A; (v) an FGF, which is FGF-2; (vi) a protein tyrosine phosphatase (PTP), which is PTPN14; (vi (i) a member of the platelet factors that is platelet factor 4, or (viii) a platelet-derived growth factor including PDGF-BB; (ix) an LEC polarity factor selected from the group consisting of Celsr1, Vangl2, Pdk2, and Fat4; (x) a member of the Notch family; (xi) ARAF, SOS1, Apelin, KIF11, REELIN, CALCRL, GJC2 (gap junction protein gamma-2), Rasip1, or FBXL7 (F-box and leucine-rich repeat protein 7); and (xii) any one combination of (i) to (xi).

[0137] In some embodiments, the inhibitor of lymphangiogenesis factor is selected from the group consisting of: (i) an agent capable of disrupting the gene for the lymphangiogenesis factor, thereby eliminating or reducing expression of the lymphangiogenesis factor; (ii) an siRNA capable of degrading the mRNA for the lymphangiogenesis factor, thereby reducing expression of the lymphangiogenesis factor; (iii) an antagonist antibody to the lymphangiogenesis factor that inhibits or prevents the function of the lymphangiogenesis factor; (iv) a small molecule capable of inhibiting or preventing the function of the lymphangiogenesis factor; and (v) any combination of (i)-(iv).

[0138] In some embodiments, the inhibitor of lymphangiogenesis factor is administered locally via intracameral injection, via subconjunctival injection, via intravitreal injection, via suprachoroidal injection, via peribulbar injection, or via retrobulbar injection. In some embodiments, the inhibitor of lymphangiogenesis factor is formulated as eye drops, a depot, a bolus, an inhibitor-loaded contact lens, a suspension, a solution, an eye drop gel, or an ointment.

[0139] In some embodiments, the methods described herein further include treating the eye with a second therapy. In some embodiments, the second therapy is selected from the group consisting of eye drops, oral medications, laser therapy, and surgery. In some embodiments, the second therapy is selected from the group consisting of eye drops that reduce the amount of fluid produced in the eye or improve fluid drainage from the eye, oral medications that lower IOP, laser therapy, including laser trabeculoplasty or laser iridotomy, and surgery, including implants or trabeculectomy.

[0140] In some embodiments, prior to administration of the inhibitor, the subject has an IOP elevation of greater than about 21 mmHg. In some embodiments, prior to administration of the inhibitor, the subject has an IOP elevation of greater than about 25 mmHg. In some embodiments, prior to administration of the inhibitor, the subject has an IOP elevation of greater than about 30 mmHg.

[0141] In some embodiments, after administration of the inhibitor, the subject's IOP in the treated eye is reduced by at least about 2 mmHg. In some embodiments, after administration of the inhibitor, the subject's IOP in the treated eye is reduced by at least about 5 mmHg. In some embodiments, after administration of the inhibitor, the subject's IOP in the treated eye is reduced by at least about 10 mmHg. In some embodiments, after administration of the inhibitor, the subject's IOP in the treated eye is reduced by at least about 15 mmHg.

[0142] In one aspect, the disclosure provides a method of treating glaucoma in an eye of a subject in need thereof, comprising administering to the eye a therapeutically effective amount of an inhibitor of a lymphangiogenesis factor selected from the group consisting of VEGFR-3, VLA-1, ITGA5, and Ang-2. In some embodiments, the lymphangiogenesis factor is VEGFR-3. In some embodiments, the lymphangiogenesis factor is VLA-1. In some embodiments, the lymphangiogenesis factor is ITGA5. In some embodiments, the lymphangiogenesis factor is Ang-2. In some embodiments, the subject has primary glaucoma. In some embodiments, the subject has primary open-angle glaucoma. In some embodiments, the subject has primary angle-closure glaucoma. In some embodiments, the subject has primary congenital glaucoma. In some embodiments, the subject has primary normal-tension glaucoma.

[0143] In another aspect, the disclosure provides a method of preventing glaucoma in an eye of a subject in need thereof, comprising administering to the eye a therapeutically effective amount of an inhibitor of a lymphangiogenesis factor selected from the group consisting of VEGFR-3, VLA-1, ITGA5, and Ang-2. In some embodiments, the lymphangiogenesis factor is VEGFR-3. In some embodiments, the lymphangiogenesis factor is VLA-1. In some embodiments, the lymphangiogenesis factor is ITGA5. In some embodiments, the lymphangiogenesis factor is Ang-2. In some embodiments, the glaucoma is primary glaucoma. In some embodiments, the glaucoma is primary open-angle glaucoma. In some embodiments, the glaucoma is primary angle-closure glaucoma. In some embodiments, the glaucoma is primary congenital glaucoma. In some embodiments, the glaucoma is primary normal-tension glaucoma.

[0144] In another aspect, the present disclosure provides a method of inhibiting lymphangiogenesis in the eye of a subject experiencing elevated intraocular pressure (IOP), comprising administering to the eye a therapeutically effective amount of an inhibitor of a lymphangiogenesis factor selected from the group consisting of VEGFR-3, VLA-1, ITGA5, and Ang-2, wherein administration of the inhibitor results in a decrease in IOP in the eye. In some embodiments, the lymphangiogenesis factor is VEGFR-3. In some embodiments, the lymphangiogenesis factor is VLA-1. In some embodiments, the lymphangiogenesis factor is ITGA5. In some embodiments, the lymphangiogenesis factor is Ang-2.

[0145] In another aspect, the disclosure provides a method of inhibiting lymphangiogenesis in the eye of a subject at risk of developing glaucoma, comprising administering to the eye a therapeutically effective amount of an inhibitor of a lymphangiogenesis factor selected from the group consisting of VEGFR-3, VLA-1, ITGA5, and Ang-2, wherein administration of the inhibitor results in a decrease in IOP of the eye. In some embodiments, the lymphangiogenesis factor is VEGFR-3. In some embodiments, the lymphangiogenesis factor is VLA-1. In some embodiments, the lymphangiogenesis factor is ITGA5. In some embodiments, the lymphangiogenesis factor is Ang-2. In some embodiments, the glaucoma is primary glaucoma. In some embodiments, the glaucoma is primary open-angle glaucoma. In some embodiments, the glaucoma is primary angle-closure glaucoma. In some embodiments, the glaucoma is primary congenital glaucoma. In some embodiments, the glaucoma is primary normal tension glaucoma.

[0146] In another aspect, the present disclosure provides a method for increasing Schlemm's canal permeability in an eye of a subject experiencing elevated IOP, comprising administering to the eye a therapeutically effective amount of an inhibitor of a lymphangiogenesis factor selected from the group consisting of VEGFR-3, VLA-1, ITGA5, and Ang-2, wherein administration of the inhibitor reduces IOP in the eye. In some embodiments, the lymphangiogenesis factor is VEGFR-3. In some embodiments, the lymphangiogenesis factor is VLA-1. In some embodiments, the lymphangiogenesis factor is ITGA5. In some embodiments, the lymphangiogenesis factor is Ang-2.

[0147] In another aspect, the present disclosure provides a method of inhibiting Schlemm's canal cell activity in the eye of a subject experiencing elevated IOP, comprising administering to the eye a therapeutically effective amount of an inhibitor of a lymphangiogenesis factor selected from the group consisting of VEGFR-3, VLA-1, ITGA5, and Ang-2, wherein the cellular activity is selected from the group consisting of inhibiting cell adhesion, inhibiting cell proliferation, inhibiting cell migration, or inhibiting angiogenesis. In some embodiments, the lymphangiogenesis factor is VEGFR-3. In some embodiments, the lymphangiogenesis factor is VLA-1. In some embodiments, the lymphangiogenesis factor is ITGA5. In some embodiments, the lymphangiogenesis factor is Ang-2.

[0148] In another aspect, the present disclosure provides a method for increasing aqueous humor outflow in Schlemm's canal of an eye in a subject experiencing elevated IOP, comprising administering to the eye a therapeutically effective amount of an inhibitor of a lymphangiogenesis factor selected from the group consisting of VEGFR-3, VLA-1, ITGA5, and Ang-2, wherein administration of the inhibitor results in increased aqueous humor outflow in Schlemm's canal of the eye and reduced IOP. In some embodiments, the lymphangiogenesis factor is VEGFR-3. In some embodiments, the lymphangiogenesis factor is VLA-1. In some embodiments, the lymphangiogenesis factor is ITGA5. In some embodiments, the lymphangiogenesis factor is Ang-2.

[0149] In another aspect, the present disclosure provides a method of reducing corneal edema in an eye of a subject in need thereof, comprising administering to the eye a therapeutically effective amount of an inhibitor of a lymphangiogenesis factor selected from the group consisting of VEGFR-3, VLA-1, ITGA5, and Ang-2, wherein administration of the inhibitor results in reduction of corneal edema. In some embodiments, the lymphangiogenesis factor is VEGFR-3. In some embodiments, the lymphangiogenesis factor is VLA-1. In some embodiments, the lymphangiogenesis factor is ITGA5. In some embodiments, the lymphangiogenesis factor is Ang-2.

[0150] In another aspect, the present disclosure provides a method for reducing retinal nerve fiber layer (RNFL) thinning in an eye of a subject in need thereof, the method comprising administering to the eye a therapeutically effective amount of an inhibitor of a lymphangiogenesis factor selected from the group consisting of VEGFR-3, VLA-1, ITGA5, and Ang-2, wherein administration of the inhibitor results in a reduction in RNFL thinning. In some embodiments, the lymphangiogenesis factor is VEGFR-3. In some embodiments, the lymphangiogenesis factor is VLA-1. In some embodiments, the lymphangiogenesis factor is ITGA5. In some embodiments, the lymphangiogenesis factor is Ang-2.

[0151] In another aspect, the disclosure provides a method of reducing retinal ganglion cell (RGC) death in the eye of a subject in need thereof, the method comprising administering to the eye a therapeutically effective amount of an inhibitor of a lymphangiogenesis factor selected from the group consisting of VEGFR-3, VLA-1, ITGA5, and Ang-2, wherein administration of the inhibitor results in reduced RGC death. In some embodiments, the lymphangiogenesis factor is VEGFR-3. In some embodiments, the lymphangiogenesis factor is VLA-1. In some embodiments, the lymphangiogenesis factor is ITGA5. In some embodiments, the lymphangiogenesis factor is Ang-2.

[0152] In some embodiments, the subject has primary glaucoma selected from the group consisting of primary open-angle glaucoma, primary angle-closure glaucoma, primary congenital glaucoma, and primary normal-tension glaucoma. The subject has primary open-angle glaucoma. In some embodiments, the subject has primary angle-closure glaucoma. In some embodiments, the subject has primary congenital glaucoma. In some embodiments, the subject has primary normal-tension glaucoma. In some embodiments, the subject is human.

[0153] In some embodiments, the inhibitor of lymphangiogenesis factor is selected from the group consisting of: (i) an agent capable of disrupting the gene for the lymphangiogenesis factor, thereby eliminating or reducing expression of the lymphangiogenesis factor; (ii) an siRNA capable of degrading the mRNA for the lymphangiogenesis factor, thereby reducing expression of the lymphangiogenesis factor; (iii) an antagonist antibody to the lymphangiogenesis factor that inhibits or prevents the function of the lymphangiogenesis factor; (iv) a small molecule capable of inhibiting or preventing the function of the lymphangiogenesis factor; and (v) any combination of (i)-(iv).

[0154] In some embodiments, the inhibitor of lymphangiogenesis factor is administered locally via intracameral injection, via subconjunctival injection, via intravitreal injection, via suprachoroidal injection, via peribulbar injection, or via retrobulbar injection. In some embodiments, the inhibitor of lymphangiogenesis factor is formulated as eye drops, a depot, a bolus, an inhibitor-loaded contact lens, a suspension, a solution, an eye drop gel, or an ointment.

[0155] In some embodiments, the methods described herein further include treating the eye with a second therapy. In some embodiments, the second therapy is selected from the group consisting of eye drops, oral medications, laser therapy, and surgery. In some embodiments, the second therapy is selected from the group consisting of eye drops that reduce the amount of fluid produced in the eye or improve fluid drainage from the eye, oral medications that lower IOP, laser therapy, including laser trabeculoplasty or laser iridotomy, and surgery, including implants or trabeculectomy.

[0156] In some embodiments, prior to administration of the inhibitor, the subject has an IOP elevation of greater than about 21 mmHg. In some embodiments, prior to administration of the inhibitor, the subject has an IOP elevation of greater than about 25 mmHg. In some embodiments, prior to administration of the inhibitor, the subject has an IOP elevation of greater than about 30 mmHg.

[0157] In some embodiments, after administration of the inhibitor, the subject's IOP in the treated eye is reduced by at least about 1 mmHg. In some embodiments, after administration of the inhibitor, the subject's IOP in the treated eye is reduced by at least about 2 mmHg. In some embodiments, after administration of the inhibitor, the subject's IOP in the treated eye is reduced by at least about 5 mmHg. In some embodiments, after administration of the inhibitor, the subject's IOP in the treated eye is reduced by at least about 10 mmHg. In some embodiments, after administration of the inhibitor, the subject's IOP in the treated eye is reduced by at least about 15 mmHg.

[0158] In some embodiments, the lymphangiogenesis promoter is VEGFR-3. In some embodiments, the lymphangiogenesis promoter is VLA-1. In some embodiments, the lymphangiogenesis promoter is ITAG5. In some embodiments, the lymphangiogenesis promoter is Ang-2.

[0159] In some embodiments, the inhibitor is an siRNA. In some embodiments, the inhibitor is an antagonist antibody. In some embodiments, the inhibitor disrupts expression of a lymphangiogenesis promoter gene. In some embodiments, the inhibitor that disrupts expression of a lymphangiogenesis promoter gene is a gene editing composition. In some embodiments, the gene editing composition comprises CRISPR / Cas9 and a gRNA. In some embodiments, the subject is a human.

[0160] In an alternative aspect, the present disclosure provides a method of treating primary glaucoma in an eye of a subject in need thereof, comprising administering to the eye a therapeutically effective amount of an agent that activates or stimulates lymphatic factors that inhibit or prevent lymphangiogenesis in the eye ("anti-lymphangiogenic factors").

[0161] In some embodiments, the subject has primary glaucoma selected from the group consisting of primary open-angle glaucoma, primary angle-closure glaucoma, primary congenital glaucoma, and primary normal-tension glaucoma. In some embodiments, the subject has primary open-angle glaucoma. The subject has primary angle-closure glaucoma. In some embodiments, the subject has primary congenital glaucoma. In some embodiments, the subject has primary normal-tension glaucoma. In some embodiments, the subject is human.

[0162] In another aspect, the present disclosure provides a method for preventing primary glaucoma in an eye of a subject in need thereof, the method comprising administering to the eye a therapeutically effective amount of an anti-lymphangiogenic factor.

[0163] In another aspect, the present disclosure provides a method for inhibiting lymphangiogenesis in the eye of a subject experiencing elevated intraocular pressure (IOP), comprising administering to the eye a therapeutically effective amount of an agent that activates or stimulates an anti-lymphangiogenic factor, wherein administration of the agent results in a decrease in IOP in the eye.

[0164] In another aspect, the present disclosure provides a method for inhibiting lymphangiogenesis in the eye of a subject at risk of developing primary glaucoma, the method comprising administering to the eye a therapeutically effective amount of an agent that activates or stimulates an anti-lymphangiogenic factor, wherein administration of the agent results in a decrease in IOP of the eye.

[0165] In another aspect, the present disclosure provides a method for increasing Schlemm's canal permeability in the eye of a subject experiencing elevated IOP, comprising administering to the eye a therapeutically effective amount of an agent that activates or stimulates an anti-lymphangiogenic factor, wherein administration of the agent reduces IOP in the eye.

[0166] In another aspect, the disclosure provides a method for inhibiting Schlemm's canal cell activity in the eye of a subject experiencing elevated IOP, comprising administering to the eye a therapeutically effective amount of an agent that activates or stimulates an anti-lymphangiogenic factor, wherein the cellular activity is selected from the group consisting of inhibiting cell adhesion, inhibiting cell proliferation, inhibiting cell migration, or inhibiting tube formation.

[0167] In another aspect, the present disclosure provides a method for increasing aqueous humor outflow in Schlemm's canal of an eye of a subject experiencing elevated IOP, comprising administering to the eye a therapeutically effective amount of an agent that activates or stimulates an anti-lymphangiogenic factor, wherein administration of the agent results in increased aqueous humor outflow in Schlemm's canal of the eye and a decrease in IOP.

[0168] In another aspect, the present disclosure provides a method for reducing corneal edema in the eye of a subject in need thereof, the method comprising administering to the eye a therapeutically effective amount of an inhibitor of an agent that activates or stimulates anti-lymphangiogenic factors, wherein administration of the agent results in a reduction of corneal edema.

[0169] In another aspect, the present disclosure provides a method for reducing retinal nerve fiber layer (RNFL) thinning in the eye of a subject in need thereof, the method comprising administering to the eye a therapeutically effective amount of an agent that activates or stimulates an anti-lymphangiogenic factor, wherein administration of the agent results in a reduction in RNFL thinning.

[0170] In another aspect, the present disclosure provides a method for reducing retinal ganglion cell (RGC) death in the eye of a subject in need thereof, the method comprising administering to the eye a therapeutically effective amount of an agent that activates or stimulates an anti-lymphangiogenic factor, wherein administration of the agent results in a reduction in RGC death.

[0171] In some embodiments of the aspects described herein, the subject has primary glaucoma selected from the group consisting of primary open-angle glaucoma, primary angle-closure glaucoma, primary congenital glaucoma, and primary normal-tension glaucoma. The subject has primary open-angle glaucoma. In some embodiments, the subject has primary angle-closure glaucoma. In some embodiments, the subject has primary congenital glaucoma. In some embodiments, the subject has primary normal-tension glaucoma. In some embodiments, the subject is human.

[0172] In some embodiments, the agent is administered locally via intracameral injection, via subconjunctival injection, via intravitreal injection, via suprachoroidal injection, via peribulbar injection, or via retrobulbar injection. In some embodiments, the agent is formulated as eye drops, a depot, a bolus, an inhibitor-loaded contact lens, a suspension, a solution, an eye drop gel, or an ointment.

[0173] In some embodiments, the methods described herein further include treating the eye with a second therapy. In some embodiments, the second therapy is selected from the group consisting of eye drops, oral medications, laser therapy, and surgery. In some embodiments, the second therapy is selected from the group consisting of eye drops that reduce the amount of fluid produced in the eye or improve fluid drainage from the eye, oral medications that lower IOP, laser therapy, including laser trabeculoplasty or laser iridotomy, and surgery, including implants or trabeculectomy.

[0174] In some embodiments, prior to administration of the agent, the subject has an IOP elevation of greater than about 21 mmHg. In some embodiments, prior to administration of the agent, the subject has an IOP elevation of greater than about 25 mmHg. In some embodiments, prior to administration of the agent, the subject has an IOP elevation of greater than about 30 mmHg.

[0175] In some embodiments, after administration of the agent, the subject's IOP in the treated eye is reduced by at least about 2 mmHg. In some embodiments, after administration of the agent, the subject's IOP in the treated eye is reduced by at least about 5 mmHg. In some embodiments, after administration of the agent, the subject's IOP in the treated eye is reduced by at least about 10 mmHg. In some embodiments, after administration of the agent, the subject's IOP in the treated eye is reduced by at least about 15 mmHg.

[0176] In some embodiments, the administered agent is selected from the group consisting of: (i) an anti-lymphangiogenic factor; (ii) an agent that activates the expression of an anti-lymphangiogenic factor; (iii) an agonist antibody that activates an anti-lymphangiogenic factor; (iv) a microRNA or mimic that inhibits lymphangiogenesis; and (v) a small molecule that activates the function of an anti-lymphangiogenic factor.

[0177] In some embodiments, the anti-lymphangiogenic factor is selected from the group consisting of microRNA-184, microRNA-126, miR-31, miR-181a, microRNA-132, miR-194, miR-186, miR-99a, miR-92a, and miR-466, endostatin, and a VEGF family inhibitor (e.g., soluble VEGFR-2, soluble VEGFR-3).

[0178] Pharmaceutical Composition In one aspect, the present invention provides a pharmaceutical ophthalmological composition comprising an inhibitor of lymphangiogenic factor as described herein and a pharmaceutically acceptable carrier. Pharmaceutical compositions comprising a lymphangiogenic factor inhibitor are useful for treating glaucoma, including primary glaucoma or IOP-mediated disorders. Such pharmaceutical compositions are formulated based on the mode of delivery. One example is a composition formulated for direct delivery to the eye. In some embodiments, delivery is local, and the mode of delivery is selected from the group consisting of eye drops, intracameral injection, subconjunctival injection, intravitreal injection, suprachoroidal injection, peribulbar injection, and retrobulbar injection. In some embodiments, the pharmaceutical ophthalmological composition is a liquid. In an alternative embodiment, a pharmaceutical ophthalmological composition comprises an activator of anti-lymphangiogenic factor as described herein and a pharmaceutically acceptable carrier.

[0179] The dosage administered will, of course, vary depending on known factors such as the pharmacodynamic properties of the particular agent and its mode and route of administration, age, health, nature and extent of symptoms, type of concurrent treatment, frequency of treatment, and the desired effect. The pharmaceutical compositions featured herein are administered in dosages sufficient to inhibit expression of lymphatic vessel promoting factors.

[0180] In some embodiments, the pharmaceutical ophthalmological composition comprises at least one siRNA. In some embodiments, a suitable dose of siRNA will be in the range of 0.000001 milligrams to 100.0 milligrams per recipient eye per single dose, including 0.000001 milligrams, 0.00001 milligrams, 0.0001 milligrams, 0.0005 milligrams, 0.001 milligrams, 0.005 milligrams, 0.01 milligrams, 0.05 milligrams, 0.1 milligrams, 0.5 milligrams, 1 milligram, 5 milligrams, 10 milligrams, 50 milligrams, or 100 milligrams. In some embodiments, the pharmaceutical composition is contained within a volume of about 1 μL to about 500 μL, including 1.0 μL, 3.0 μL, 5.0 μL, 10.0 μL, 15.0 μL, 20.0 μL, 25.0 μL, 50.0 μL, 100.0 μL, 200.0 μL, 250.0 μL, 300.0 μL, or 500.0 μL. For example, siRNA may be administered at a single dose of 0.05 μg / eye / 50 μL, 0.25 μg / eye / 50 μL, 0.5 μg / eye / 50 μL, 5 μg / eye / 50 μL, 50 μg / eye / 50 μL, 100 μg / eye / 50 μL, 250 μg / eye / 50 μL, 500 μg / eye / 50 μL, 750 μg / eye / 50 μL, 950 μg / eye / 50 μL, or 1000 μg / eye / 50 μL. In some embodiments, the volume of the pharmaceutical composition is limited by the mode of delivery (e.g., 100 μL for intravitreal injection).

[0181] In some embodiments, the pharmaceutical ophthalmological composition comprises at least one antibody. In some embodiments, the pharmaceutical composition is a dosage form containing about 0.01 mg to about 2000 mg, about 0.1 mg to about 1000 mg, about 1 mg to about 800 mg, about 10 mg to about 600 mg, or about 100 mg to about 500 mg of active compound per unit dosage form, and, optionally, about 0.01 mg to about 1000 mg, about 0.1 mg to about 500 mg, about 1 mg to about 200 mg, or about 10 mg to about 100 mg of an additional active agent. Examples include dosage forms containing at least 0.01 mg, 0.1 mg, 1 mg, 5 mg, 10 mg, 25 mg, 50 mg, 100 mg, 200 mg, 250 mg, 300 mg, 400 mg, 500 mg, 600 mg, 700 mg, or 750 mg of the active compound or a salt thereof. For example, the antibody can be administered at a single dose of 50 μg / eye / 50 μL, 100 μg / eye / 50 μL, 250 μg / eye / 50 μL, 500 μg / eye / 50 μL, or 1000 μg / eye / 50 μL. In some embodiments, the volume of the pharmaceutical composition is limited by the mode of delivery (e.g., 100 μL for intravitreal injection).

[0182] The pharmaceutical compositions described herein may be administered once daily, weekly, twice weekly, monthly, bimonthly, trimonthly, or six-monthly. Alternatively, the pharmaceutical compositions may be administered as two, three, or more subdoses over a period of time, or at appropriate intervals using continuous infusion or controlled-release delivery. In this case, the dose of lymphatic vessel promoting factor inhibitor contained in each subdose must be correspondingly reduced to achieve the total dose over a given period. Dosage units can also be formulated for delivery over several days using conventional sustained-release formulations that provide sustained release of lymphatic vessel promoting factor inhibitor over a period of several days or months. Sustained-release formulations are well known in the art and are particularly useful for delivering drugs to specific sites, such as those used by the agents of the present invention. In this embodiment, the dosage unit contains a corresponding multiple of a single dose.

[0183] Those skilled in the art will understand that certain factors, including but not limited to, the severity of the disease or disorder, previous treatment, the subject's general health and / or age, and other existing diseases, can affect the dosage and timing required to effectively treat a subject. Furthermore, treatment of a subject with a therapeutically effective amount of a composition can include a single treatment or a series of treatments. The effective dosage and in vivo half-life of each siRNA or antibody or other compound encompassed by the present invention can be estimated using conventional methodologies or based on in vivo tests using appropriate animal models, as described elsewhere herein.

[0184] Carriers include sterile excipients and diluents and must be of sufficiently high purity and sufficiently low toxicity to ocular tissues to be suitable for administration to the patient being treated. Carriers may be inert or may have pharmaceutical benefits of their own. The amount of carrier used with the compound is sufficient to provide a practical amount of material for administration per unit dose of the compound.

[0185] Classes of carriers include, but are not limited to, binders, buffers, coloring agents, diluents, disintegrants, emulsifiers, glidants, lubricants, preservatives, stabilizers, surfactants, and wetting agents.Some carriers may be classified into two or more classes.Optional active agents may be included in the pharmaceutical composition and do not substantially interfere with the activity of the ocular lymphangiogenesis inhibitor of the present invention.

[0186] Embodiment 1. A method of treating glaucoma in the eye of a subject in need thereof, comprising administering to the eye a therapeutically effective amount of an inhibitor of a lymphangiogenic factor associated with Schlemm's canal, wherein the lymphangiogenic factor induces or maintains lymphangiogenesis in the eye (a "pro-lymphangiogenic factor").

[0187] 2. The method of embodiment 1, wherein the subject has primary glaucoma.

[0188] 3. The method of embodiment 2, wherein the subject has primary open-angle glaucoma.

[0189] 4. The method of embodiment 2, wherein the subject has primary angle-closure glaucoma.

[0190] 5. The method of embodiment 2, wherein the subject has primary congenital glaucoma.

[0191] 6. The method of embodiment 2, wherein the subject has primary normal-tension glaucoma.

[0192] 7. A method for preventing glaucoma in the eye of a subject in need thereof, comprising administering to the eye a therapeutically effective amount of an inhibitor of lymphangiogenesis promoting factor.

[0193] 8. A method for inhibiting lymphangiogenesis in the eye of a subject at risk of developing glaucoma, comprising administering to the eye a therapeutically effective amount of an inhibitor of a lymphangiogenesis promoting factor, wherein administration of the inhibitor results in a decrease in IOP of the eye.

[0194] 9. A method for inhibiting lymphangiogenesis in the eye of a subject experiencing elevated intraocular pressure (IOP), comprising administering to the eye a therapeutically effective amount of an inhibitor of a lymphangiogenesis promoting factor, wherein administration of the inhibitor results in a decrease in IOP in the eye.

[0195] 10. A method for increasing Schlemm's canal permeability in the eye of a subject experiencing elevated IOP, comprising administering to the eye a therapeutically effective amount of an inhibitor of lymphangiogenesis promoting factor, wherein administration of the inhibitor reduces IOP in the eye.

[0196] 11. A method for inhibiting Schlemm's canal cell activity in the eye of a subject experiencing elevated IOP, comprising administering to the eye a therapeutically effective amount of an inhibitor of lymphangiogenesis promoting factor, wherein the cell activity is selected from the group consisting of inhibition of cell adhesion, inhibition of cell proliferation, inhibition of cell migration, or inhibition of tube formation.

[0197] 12. A method for increasing aqueous humor outflow in Schlemm's canal of an eye of a subject experiencing elevated IOP, comprising administering to the eye a therapeutically effective amount of an inhibitor of lymphangiogenesis promoting factor, wherein administration results in increased aqueous humor outflow in Schlemm's canal of the eye and a decrease in IOP.

[0198] 13. A method for reducing corneal edema in the eye of a subject in need thereof, comprising administering to the eye a therapeutically effective amount of an inhibitor of lymphangiogenesis promoting factor, wherein administration of the inhibitor results in a reduction of corneal edema.

[0199] 14. A method for reducing retinal nerve fiber layer thinning in the eye of a subject in need thereof, comprising administering to the eye a therapeutically effective amount of an inhibitor of lymphangiogenesis promoting factor, wherein administration of the inhibitor results in a reduction in nerve fiber thinning.

[0200] 15. A method for reducing retinal ganglion cell (RGC) death in the eye of a subject in need thereof, comprising administering to the eye a therapeutically effective amount of an inhibitor of lymphangiogenesis promoting factor, wherein administration of the inhibitor results in a reduction in retinal ganglion cell (RGC) death.

[0201] 16. The method of embodiments 9-15, wherein the subject has primary glaucoma.

[0202] 17. The method of embodiment 16, wherein the subject has primary open-angle glaucoma.

[0203] 18. The method of embodiment 16, wherein the subject has primary angle-closure glaucoma.

[0204] 19. The method of embodiment 16, wherein the subject has primary congenital glaucoma.

[0205] 20. The method of embodiment 16, wherein the subject has primary normal-tension glaucoma.

[0206] 21. The method of embodiments 1-20, wherein the subject is a human.

[0207] 22. The method of embodiments 1-21, wherein the lymphangiogenesis promoting factor is a VEGF / VEGFR family member.

[0208] 23. The method of embodiment 22, wherein the VEGF / VEGFR family member is selected from the group consisting of VEGF-A, VEGF-B, VEGF-C, VEGF-D, VEGF-E, P1GF, VEGFR-1, VEGFR-2, and VEGFR-3.

[0209] 24. The method of embodiment 23, wherein the VEGF / VEGFR family member is VEGF-A.

[0210] 25. The method of embodiment 23, wherein the VEGF / VEGFR family member is VEGF-C.

[0211] 26. The method of embodiment 23, wherein the VEGF / VEGFR family member is VEGF-D.

[0212] 27. The method of embodiment 23, wherein the VEGF / VEGFR family member is VEGFR-3.

[0213] 28. The method of embodiments 1-21, wherein the lymphangiogenesis promoting factor is an integrin family member.

[0214] 29. The method of embodiment 28, wherein the integrin family member is selected from the group consisting of very late antigen-1 (VLA-1), integrin alpha 5 (ITGA5), and integrin alpha 9 (ITGA9).

[0215] 30. The method of embodiment 29, wherein the integrin family member is VLA-1.

[0216] 31. The method of embodiment 29, wherein the integrin family member is ITGA5.

[0217] 32. The method of embodiment 29, wherein the integrin family member is ITGA9.

[0218] 33. The method of embodiments 1-21, wherein the lymphangiogenesis promoting factor is an angiopoietin family member.

[0219] 34. The method of embodiment 33, wherein the angiopoietin family member is selected from the group consisting of angiopoietin-1 (ANGPT1), angiopoietin-2 (ANGPT2 or Ang-2), and Tie2 / TEK.

[0220] 35. The method of embodiment 34, wherein the angiopoietin family member is Ang-2.

[0221] 36. The method of embodiments 1-21, wherein the lymphangiogenesis promoting factor is selected from the group consisting of: (i) cytokines or chemokines; (ii) extracellular matrix proteins; (iii) transcription factors; (iv) guiding molecules; (v) fibroblast growth factor (FGF); (vi) protein tyrosine phosphatases (PTPs); (vii) a platelet factor member, or (viii) a platelet-derived growth factor; (ix) LEC polarity factor; (x) a Notch family member; and (xi) Any combination of (i) to (xi).

[0222] 37. The method of embodiments 1-21, wherein the lymphangiogenesis promoting factor is selected from the group consisting of: (i) a cytokine / chemokine selected from the group consisting of interleukin-8, interferon-γ, CCR7, and SLC; (ii) an extracellular matrix protein, CCBE1; (iii) a transcription factor selected from the group consisting of Sox18 and Hhex; (iv) an inducer molecule selected from the group consisting of neuropilin 2 and SEMA7A; (v) FGF, which is FGF-2; (vi) protein tyrosine phosphatase (PTP), which is PTPN14; (vii) a member of the platelet factors, which is platelet factor 4, or (viii) a platelet-derived growth factor, including PDGF-BB; (ix) an LEC polarity factor selected from the group consisting of Celsr1, Vangl2, Pdk2, and Fat4; (x) Notch family members; (xi) ARAF, SOS1, Apelin, KIF11, REELIN, CALCRL, GJC2 (gap junction protein gamma-2), Rasip1, or FBXL7 (F-box and leucine-rich repeat protein 7); and (xii) Any one combination of (i) to (xi).

[0223] 38. The method of embodiments 1-37, wherein the inhibitor of lymphangiogenesis promoting factor is selected from the group consisting of: (i) an agent capable of disrupting the gene for a lymphangiogenesis promoting factor, thereby eliminating or reducing the expression of the lymphangiogenesis promoting factor; (ii) siRNA that can degrade the mRNA of lymphangiogenesis-promoting factors, thereby reducing the expression of lymphangiogenesis-promoting factors; (iii) antagonist antibodies against lymphangiogenesis promoting factors that inhibit or prevent the function of lymphangiogenesis promoting factors; (iv) a small molecule capable of inhibiting or preventing the function of lymphangiogenesis promoting factors; and (v) Any combination of (i) to (iv).

[0224] 39. The method of embodiments 1-38, wherein the inhibitor of lymphangiogenesis promoting factor is administered locally via intracameral injection, via subconjunctival injection, via intravitreal injection, via suprachoroidal injection, via peribulbar injection, or via retrobulbar injection.

[0225] 40. The method of any one of embodiments 1 to 37, wherein the inhibitor of lymphangiogenesis promoting factor is formulated as eye drops, a depot, a bolus, an inhibitor-loaded contact lens, a suspension, a solution, an eye drop gel, or an ointment.

[0226] 41. The method of embodiments 1-40, further comprising treating the eye with a second therapy.

[0227] 42. The method of embodiment 41, wherein the second treatment is selected from the group consisting of eye drops, oral medications, laser therapy, and surgery.

[0228] 43. The method of embodiment 41 or 42, wherein the second treatment is selected from the group consisting of eye drops that reduce the amount of fluid produced in the eye or improve drainage of fluid from the eye, oral medications that lower IOP, laser therapy, including laser trabeculoplasty or laser iridotomy, and surgery, including implants or trabeculectomy.

[0229] 44. The method of embodiments 1-43, wherein, prior to administration of the inhibitor, the subject has an IOP elevation of greater than about 21 mmHg.

[0230] 45. The method of embodiments 1-43, wherein, prior to administration of the inhibitor, the subject has an IOP elevation of greater than about 25 mmHg.

[0231] 46. ​​The method of embodiments 1-43, wherein, prior to administration of the inhibitor, the subject has an IOP elevation of greater than about 30 mmHg.

[0232] 47. The method of embodiments 1-43, wherein after administration of the inhibitor, the subject's IOP in the treated eye is reduced by at least about 2 mmHg.

[0233] 48. The method of embodiments 1-43, wherein after administration of the inhibitor, the subject's IOP in the treated eye is reduced by at least about 5 mmHg.

[0234] 49. The method of embodiments 1-43, wherein after administration of the inhibitor, the subject's IOP in the treated eye is reduced by at least about 10 mmHg.

[0235] 50. The method of embodiments 1-43, wherein after administration of the inhibitor, the subject's IOP in the treated eye is reduced by at least about 15 mmHg.

[0236] 51. A method for treating glaucoma in the eye of a subject in need thereof, comprising administering to the eye a therapeutically effective amount of an inhibitor of a lymphangiogenic factor selected from the group consisting of VEGFR-3, VLA-1, ITGA5, and Ang-2.

[0237] 52. The method of embodiment 51, wherein the subject has primary glaucoma.

[0238] 53. The method of embodiment 52, wherein the subject has primary open-angle glaucoma.

[0239] 54. The method of embodiment 52, wherein the subject has primary angle-closure glaucoma.

[0240] 55. The method of embodiment 52, wherein the subject has primary congenital glaucoma.

[0241] 56. The method of embodiment 52, wherein the subject has primary normal-tension glaucoma.

[0242] 57. A method for preventing glaucoma in the eye of a subject in need thereof, comprising administering to the eye a therapeutically effective amount of an inhibitor of a lymphangiogenic factor selected from the group consisting of VEGFR-3, VLA-1, ITGA5, and Ang-2.

[0243] 58. A method for inhibiting lymphangiogenesis in the eye of a subject at risk of developing glaucoma, comprising administering to the eye a therapeutically effective amount of an inhibitor of a lymphangiogenesis promoter selected from the group consisting of VEGFR-3, VLA-1, ITGA5, and Ang-2, wherein administration of the inhibitor results in a decrease in IOP of the eye.

[0244] 59. A method for inhibiting lymphangiogenesis in the eye of a subject experiencing elevated intraocular pressure (IOP), comprising administering to the eye a therapeutically effective amount of an inhibitor of a lymphangiogenesis promoter selected from the group consisting of VEGFR-3, VLA-1, ITGA5, and Ang-2, wherein administration of the inhibitor results in a decrease in IOP in the eye.

[0245] 60. A method for increasing Schlemm's canal permeability in the eye of a subject experiencing elevated IOP, comprising administering to the eye a therapeutically effective amount of an inhibitor of a lymphangiogenesis promoter selected from the group consisting of VEGFR-3, VLA-1, ITGA5, and Ang-2, wherein administration of the inhibitor reduces IOP in the eye.

[0246] 61. A method for inhibiting Schlemm's canal cell activity in the eye of a subject experiencing elevated IOP, comprising administering to the eye a therapeutically effective amount of an inhibitor of a lymphangiogenic factor selected from the group consisting of VEGFR-3, VLA-1, ITGA5, and Ang-2, wherein the cellular activity is selected from the group consisting of inhibition of cell adhesion, inhibition of cell proliferation, inhibition of cell migration, or inhibition of tube formation.

[0247] 62. A method for increasing aqueous humor outflow in Schlemm's canal of an eye of a subject experiencing elevated IOP, comprising administering to the eye a therapeutically effective amount of an inhibitor of a lymphangiogenesis promoter selected from the group consisting of VEGFR-3, VLA-1, ITGA5, and Ang-2, wherein administration of the inhibitor results in increased aqueous humor outflow in Schlemm's canal of the eye and a decrease in IOP.

[0248] 63. A method for reducing corneal edema in the eye of a subject in need thereof, comprising administering to the eye a therapeutically effective amount of an inhibitor of a lymphangiogenic factor selected from the group consisting of VEGFR-3, VLA-1, ITGA5, and Ang-2, wherein administration of the inhibitor results in a reduction of corneal edema.

[0249] 64. A method for reducing retinal nerve fiber layer (RNFL) thinning in the eye of a subject in need thereof, comprising administering to the eye a therapeutically effective amount of an inhibitor of a lymphangiogenic factor selected from the group consisting of VEGFR-3, VLA-1, ITGA5, and Ang-2, wherein administration of the inhibitor results in a reduction in RNFL thinning.

[0250] 65. A method for reducing retinal ganglion cell (RGC) death in the eye of a subject in need thereof, comprising administering to the eye a therapeutically effective amount of an inhibitor of a lymphangiogenesis promoter selected from the group consisting of VEGFR-3, VLA-1, ITGA5, and Ang-2, wherein administration of the inhibitor results in a reduction of RGC death.

[0251] 66. The method of embodiments 59-66, wherein the subject has primary glaucoma.

[0252] 67. The method of embodiment 67, wherein the subject has primary open-angle glaucoma.

[0253] 68. The method of embodiment 67, wherein the subject has primary angle-closure glaucoma.

[0254] 69. The method of embodiment 67, wherein the subject has primary congenital glaucoma.

[0255] 70. The method of embodiment 67, wherein the subject has primary normal-tension glaucoma.

[0256] 71. The method of embodiments 57-70, wherein the subject is a human.

[0257] 72. The method of embodiments 51-71, wherein the inhibitor of lymphangiogenesis promoting factors is selected from the group consisting of: (i) an agent capable of disrupting the gene for a lymphangiogenesis promoting factor, thereby eliminating or reducing the expression of the lymphangiogenesis promoting factor; (ii) siRNA that can degrade the mRNA of lymphangiogenesis-promoting factors, thereby reducing the expression of lymphangiogenesis-promoting factors; (iii) antagonist antibodies against lymphangiogenesis promoting factors that inhibit or prevent the function of lymphangiogenesis promoting factors; (iv) a small molecule capable of inhibiting or preventing the function of lymphangiogenesis promoting factors; and (v) Any combination of (i) to (iv).

[0258] 73. The method of embodiments 51-72, wherein the inhibitor of lymphangiogenesis promoting factor is administered locally via intracameral injection, via subconjunctival injection, via intravitreal injection, via suprachoroidal injection, via peribulbar injection, or via retrobulbar injection.

[0259] 74. The method of embodiments 51-73, wherein the inhibitor of lymphangiogenesis promoting factor is formulated as eye drops, a depot, a bolus, an inhibitor-loaded contact lens, a suspension, a solution, an eye drop gel, or an ointment.

[0260] 75. The method of any one of embodiments 51-74, further comprising treating the eye with a second therapy.

[0261] 76. The method of embodiment 75, wherein the second treatment is selected from the group consisting of eye drops, oral medications, laser therapy, and surgery.

[0262] 77. The method of embodiment 75 or 76, wherein the second treatment is selected from the group consisting of eye drops that reduce the amount of fluid produced in the eye or improve drainage of fluid from the eye, oral medications that lower IOP, laser therapy, including laser trabeculoplasty or laser iridotomy, and surgery, including implants or trabeculectomy.

[0263] 78. The method of embodiments 51-77, wherein, prior to administration of the inhibitor, the subject has an IOP elevation of greater than about 21 mmHg.

[0264] 79. The method of embodiments 51-77, wherein, prior to administration of the inhibitor, the subject has an IOP elevation of greater than about 25 mmHg.

[0265] 80. The method of embodiments 51-77, wherein, prior to administration of the inhibitor, the subject has an IOP elevation of greater than about 30 mmHg.

[0266] 81. The method of embodiments 51-80, wherein after administration of the inhibitor, the subject's IOP in the treated eye is reduced by at least about 2 mmHg.

[0267] 82. The method of embodiments 51-80, wherein after administration of the inhibitor, the subject's IOP in the treated eye is reduced by at least about 5 mmHg.

[0268] 83. The method of embodiments 51-80, wherein after administration of the inhibitor, the subject's IOP in the treated eye is reduced by at least about 10 mmHg.

[0269] 84. The method of embodiments 51-80, wherein after administration of the inhibitor, the subject's IOP in the treated eye is reduced by at least about 15 mmHg.

[0270] 85. The method of embodiments 51-84, wherein the lymphangiogenesis promoting factor is VEGFR-3.

[0271] 86. The method of embodiments 51-84, wherein the lymphangiogenesis promoting factor is VLA-1.

[0272] 87. The method of embodiments 51 to 80, wherein the lymphangiogenesis promoting factor is ITAG5.

[0273] 88. The method of embodiments 51-84, wherein the lymphangiogenesis promoting factor is Ang-2.

[0274] 89. The method of embodiments 1-88, wherein the inhibitor is an siRNA.

[0275] 90. The method of embodiments 1-88, wherein the inhibitor is an antagonist antibody.

[0276] 91. The method of embodiments 1-88, wherein the inhibitor interferes with the expression of a gene for a lymphangiogenesis promoting factor.

[0277] 92. The method of embodiments 1-91, wherein the subject is a human.

[0278] 93. A method of treating glaucoma in the eye of a subject in need thereof, comprising administering to the eye a therapeutically effective amount of an agent that activates or stimulates a lymphatic factor that inhibits or prevents lymphangiogenesis in the eye (an "anti-lymphangiogenic factor").

[0279] 94. The method of embodiment 93, wherein the subject has primary glaucoma.

[0280] 95. The method of embodiment 94, wherein the subject has primary open-angle glaucoma.

[0281] 96. The method of embodiment 94, wherein the subject has primary angle-closure glaucoma.

[0282] 97. The method of embodiment 94, wherein the subject has primary congenital glaucoma.

[0283] 98. The method of embodiment 94, wherein the subject has primary normal-tension glaucoma.

[0284] 99. A method for preventing glaucoma in an eye of a subject in need thereof, comprising administering to the eye a therapeutically effective amount of an anti-lymphangiogenic factor.

[0285] 100. A method for inhibiting lymphangiogenesis in the eye of a subject at risk of developing primary glaucoma, comprising administering to the eye a therapeutically effective amount of an agent that activates or stimulates an anti-lymphangiogenic factor, wherein administration of the agent results in a decrease in IOP of the eye.

[0286] 101. A method for inhibiting lymphangiogenesis in the eye of a subject experiencing elevated intraocular pressure (IOP), comprising administering to the eye a therapeutically effective amount of an agent that activates or stimulates an anti-lymphangiogenic factor, wherein administration of the agent results in a decrease in IOP in the eye.

[0287] 102. A method for increasing Schlemm's canal permeability in the eye of a subject experiencing elevated IOP, comprising administering to the eye a therapeutically effective amount of an agent that activates or stimulates an anti-lymphangiogenic factor, wherein administration of the agent reduces IOP in the eye.

[0288] 103. A method for inhibiting Schlemm's canal cell activity in the eye of a subject experiencing elevated IOP, comprising administering to the eye a therapeutically effective amount of an agent that activates or stimulates an anti-lymphangiogenic factor, wherein the cellular activity is selected from the group consisting of inhibition of cell adhesion, inhibition of cell proliferation, inhibition of cell migration, or inhibition of tube formation.

[0289] 104. A method for increasing aqueous humor outflow in Schlemm's canal of an eye of a subject experiencing elevated IOP, comprising administering to the eye a therapeutically effective amount of an agent that activates or stimulates an anti-lymphangiogenic factor, wherein administration of the agent results in increased aqueous humor outflow in Schlemm's canal of the eye and a decrease in IOP.

[0290] 105. A method for reducing corneal edema in the eye of a subject in need thereof, comprising administering to the eye a therapeutically effective amount of an inhibitor of an agent that activates or stimulates anti-lymphangiogenic factor, wherein administration of the agent results in a reduction of corneal edema.

[0291] 106. A method for reducing retinal nerve fiber layer (RNFL) thinning in the eye of a subject in need thereof, comprising administering to the eye a therapeutically effective amount of an agent that activates or stimulates an anti-lymphangiogenic factor, wherein administration of the agent results in a reduction in RNFL thinning.

[0292] 107. A method for reducing retinal ganglion cell (RGC) death in the eye of a subject in need thereof, comprising administering to the eye a therapeutically effective amount of an agent that activates or stimulates an anti-lymphangiogenic factor, wherein administration of the agent results in a reduction in RGC death.

[0293] 108. The method of embodiments 100-107, wherein the subject has primary glaucoma.

[0294] 109. The method of embodiment 108, wherein the subject has primary open-angle glaucoma.

[0295] 110. The method of embodiment 108, wherein the subject has primary angle-closure glaucoma.

[0296] 111. The method of embodiment 108, wherein the subject has primary congenital glaucoma.

[0297] 112. The method of embodiment 108, wherein the subject has primary normal-tension glaucoma.

[0298] 113. The method of embodiments 93-112, wherein the subject is a human.

[0299] 114. The method of embodiments 93 to 113, wherein the agent is administered locally via intracameral injection, via subconjunctival injection, via intravitreal injection, via suprachoroidal injection, via peribulbar injection, or via retrobulbar injection.

[0300] 115. The method of embodiments 93-114, wherein the agent is formulated as eye drops, a depot, a bolus, an inhibitor-loaded contact lens, a suspension, a solution, an eye drop gel, or an ointment.

[0301] 116. The method of embodiments 93-115, further comprising treating the eye with a second therapy.

[0302] 117. The method of embodiment 116, wherein the second treatment is selected from the group consisting of eye drops, oral medications, laser therapy, and surgery.

[0303] 118. The method of embodiment 116 or 117, wherein the second treatment is selected from the group consisting of eye drops that reduce the amount of fluid produced in the eye or improve drainage of fluid from the eye, oral medications that lower IOP, laser therapy, including laser trabeculoplasty or laser iridotomy, and surgery, including implants or trabeculectomy.

[0304] 119. The method of embodiments 93-118, wherein, prior to administration of the agent, the subject has an IOP elevation of greater than about 21 mmHg.

[0305] 120. The method of embodiments 93-118, wherein, prior to administration of the agent, the subject has an IOP elevation of more than about 25 mmHg.

[0306] 121. The method of embodiments 93-118, wherein, prior to administration of the agent, the subject has an IOP elevation of more than about 30 mmHg.

[0307] 122. The method of embodiments 93-121, wherein after administration of the agent, the subject's IOP in the treated eye is reduced by at least about 2 mmHg.

[0308] 123. The method of embodiments 93-121, wherein after administration of the agent, the subject's IOP in the treated eye is reduced by at least about 5 mmHg.

[0309] 124. The method of embodiments 93-121, wherein after administration of the agent, the subject's IOP in the treated eye is reduced by at least about 10 mmHg.

[0310] 125. The method of embodiments 93-121, wherein after administration of the agent, the subject's IOP in the treated eye is reduced by at least about 15 mmHg.

[0311] 126. The method of embodiments 93-125, wherein the administered agent is selected from the group consisting of: (i) anti-lymphangiogenic factors; (ii) drugs that activate the expression of anti-lymphangiogenic factors; (iii) agonist antibodies that activate anti-lymphangiogenic factors; (iv) a microRNA or mimic that inhibits lymphangiogenesis; and (v) small molecules that activate the function of antilymphangiogenic factors;

[0312] 127. The method of embodiments 93-126, wherein the anti-lymphangiogenic factor is selected from the group consisting of microRNA-184, microRNA-126, miR-31, miR-181a, microRNA-132, miR-194, miR-186, miR-99a, miR-92a, and miR-466, endostatin, and VEGF family inhibitors (e.g., soluble VEGFR-2, soluble VEGFR-3). [Example]

[0313] Example 1. VEGFR-3 blockade reduces IOP and protects the eye from glaucomatous damage This example shows that siRNA-mediated anti-VEGFR-3 treatment significantly reduced IOP and protected the eye from glaucomatous damage, as demonstrated in a mouse model of glaucoma.

[0314] method material Intraocular hypertension was induced in normal eyes by laser photocoagulation of episcleral veins. Anti-VEGFR-3 siRNA (sequence (5'->3') sense: CCAGCAUCCUGACCAUCCAtt (SEQ ID NO: 1); sequence (5'->3') antisense: UGGAUGGUCAGGAUGCUGGag (SEQ ID NO: 2)) or control scrambled siRNA was administered subconjunctivally one day after laser photocoagulation.

[0315] Laser photocoagulation In vivo data were obtained using a well-established mouse model of glaucoma involving laser-induced occlusion of episcleral veins in normal eyes (Zhang, L. et al. Establishment and Characterization of an Acute Model of Ocular Hypertension by Laser-Induced Occlusion of Episcleral Veins. Invest Ophthalmol Vis Sci. 58(10):3879-3886(2017 Aug 1)). Briefly, mice were randomized and underwent unilateral laser photocoagulation (532 nm, OcuLight TX; IRIDEX Corporation, Mountain View, CA, USA) in the right eye. The non-lasered left eye with normal IOP served as a control for this procedure.

[0316] Intraocular pressure (IOP) measurement Intraocular pressure (IOP) was measured with a noninvasive TonoLab tonometer (Icare Lab, Helsinki, Finland) under light general anesthesia with 2% isoflurane.

[0317] Optical coherence tomography (OCT) for measuring central corneal thickness Central corneal thickness was measured using optical coherence tomography (OCT) according to a previously reported method (Zhang, L. et al. Establishment and Characterization of an Acute Model of Ocular Hypertension by Laser-Induced Occlusion of Episcleral Veins. Invest Ophthalmol Vis Sci. 58(10):3879-3886 (2017 Aug 1)). Quadrant scans along four axes were performed to scan the central cornea, and data along the 0°–180° axis were used for analysis.

[0318] result As shown in Figure 1A, IOP in VEGFR-3-specific siRNA-treated eyes was significantly reduced after treatment compared to control conditions. This effect is specific, as treatment with scrambled siRNA did not affect IOP. VEGFR-3-specific siRNA treatment significantly reduced corneal edema as measured in vivo by OCT (Figure 1B). As shown in Figure 1C, summary data from human Schlemm's canal cells indicate that anti-VEGFR-3 siRNA treatment inhibits Schlemm's canal cell functions, such as adhesion ( * P<0.05).

[0319] conclusion While not wishing to endorse any one theory, it has been proposed that inhibiting VEGFR-3, a key lymphangiogenic factor, targets specific lymphatic functions of Schlemm's canal and increases its permeability, which contributes to enhanced aqueous humor movement through Schlemm's canal, resulting in measurable lowering of IOP and reduced glaucomatous damage.

[0320] Example 2. VLA-1 blockade reduces IOP and protects the eye from glaucomatous damage This example shows that anti-VLA-1 antibody treatment significantly reduces IOP and protects ocular tissue from glaucomatous damage, as demonstrated in a mouse model of glaucoma.

[0321] method material Laser-induced intraocular hypertension was induced in normal eyes. Anti-VLA-1 antibody (clone Ha31 / 8) or control was administered topically via subconjunctival injection starting 1 day after laser.

[0322] Laser photocoagulation In vivo data were obtained using a well-established mouse model of glaucoma involving laser-induced occlusion of episcleral veins in normal eyes (Zhang, L. et al. Establishment and Characterization of an Acute Model of Ocular Hypertension by Laser-Induced Occlusion of Episcleral Veins. Invest Ophthalmol Vis Sci. 58(10):3879-3886(2017 Aug 1)). Briefly, mice were randomized and underwent unilateral laser photocoagulation (532 nm, OcuLight TX; IRIDEX Corporation, Mountain View, CA, USA) in the right eye. The non-lasered left eye with normal IOP served as a control for this procedure.

[0323] Intraocular pressure (IOP) measurement Intraocular pressure (IOP) was measured with a noninvasive TonoLab tonometer (Icare Lab, Helsinki, Finland) under light general anesthesia with 2% isoflurane.

[0324] Optical coherence tomography (OCT) for measuring central corneal thickness and retinal nerve fiber layer thickness Central corneal thickness was measured using optical coherence tomography (OCT) according to a previously reported method (Zhang, L. et al. Establishment and Characterization of an Acute Model of Ocular Hypertension by Laser-Induced Occlusion of Episcleral Veins. Invest Ophthalmol Vis Sci. 58(10):3879-3886 (2017 Aug 1)). Quadrant scans along four axes were performed to scan the central cornea, and data along the 0°–180° axis were used for analysis.

[0325] For measurement of retinal nerve fiber layer (RNFL) thickness, cross-sectional retinal images were acquired with OCT after pupil dilation with 1% tropicamide eye drops. A rectangular scan sequence was used to generate a single en face image of the retina. RNFL thickness was measured in four quadrants of the en face image.

[0326] Measurement of relative retinal ganglion cell (RGC) density The relative number of retinal ganglion cells (RGCs) was assessed as previously described (Zhang, L. et al. Establishment and Characterization of an Acute Model of Ocular Hypertension by Laser-Induced Occlusion of Episcleral Veins. Invest Ophthalmol Vis Sci. 58(10):3879-3886(2017 Aug 1)). Briefly, whole-mount retinal specimens were harvested 7 days after laser application, fixed in methanol, and immunostained for Brn3a antibody. Samples were imaged, and percentage scores were obtained by normalizing to the normal control condition, which was defined as 100%.

[0327] result Compared to control conditions, IOP was significantly reduced in anti-VLA-1 antibody-treated eyes (Figure 2A). Summary data show that anti-VLA-1 antibody treatment protected the cornea from edema (Figure 2B) and also reduced RNFL (retinal nerve fiber layer) thinning (Figure 2C) and retinal ganglion cell (RGC) death (Figure 2D). Central corneal thickness and RNFL thickness were measured in vivo by optical coherence tomography (OCT). * P < 0.05; ns not significant).

[0328] Example 3. VLA-1 blockade reduces IOP and inhibits Schlemm's canal cell function. This example shows that siRNA-mediated anti-VLA-1 treatment significantly reduced IOP, as demonstrated in a mouse model of glaucoma (FIG. 3A).

[0329] method material Laser-induced intraocular pressure was induced in normal eyes. Two sets of anti-VLA-1 siRNAs were used. To test IOP in vivo, the following anti-VLA-1 siRNA set was used: (sequence (5'->3') sense: GGAUCAACUUUAGUCACCAtt (SEQ ID NO: 9); sequence (5'->3') antisense: UGGUGACUAAAGUUGAUCCaa (SEQ ID NO: 10)). One day after laser, anti-VLA-1 siRNA or control scrambled siRNA was administered topically via subconjunctival injection. To test Schlemm's canal cell function in vitro, the following anti-VLA-1 siRNA set was used: (sequence (5'->3') sense: GGACUUUAAUCUUACCGAUtt (SEQ ID NO: 5); sequence (5'->3') antisense: AUCGGUAAGAUUAAAGUCCaa (SEQ ID NO: 6)).

[0330] Laser photocoagulation In vivo data were obtained using a well-established mouse model of glaucoma involving laser-induced occlusion of episcleral veins in normal eyes (Zhang, L. et al. Establishment and Characterization of an Acute Model of Ocular Hypertension by Laser-Induced Occlusion of Episcleral Veins. Invest Ophthalmol Vis Sci. 58(10):3879-3886(2017 Aug 1)). Briefly, mice were randomized and underwent unilateral laser photocoagulation (532 nm, OcuLight TX; IRIDEX Corporation, Mountain View, CA, USA) in the right eye. The non-lasered left eye with normal IOP served as a control for this procedure.

[0331] Intraocular pressure (IOP) measurement Intraocular pressure (IOP) was measured with a noninvasive TonoLab tonometer (Icare Lab, Helsinki, Finland) under light general anesthesia with 2% isoflurane.

[0332] Schlemm's canal cell adhesion assay Schlemm's canal cell adhesion assays were performed according to a previously reported method (Altiok, E. et al. Integrin Alpha-9 Mediates Lymphatic Valve Formation in Corneal Lymphangiogenesis. Invest Ophthalmol Vis Sci. 56(11):6313-6319 (2015 October)). Briefly, 48 hours after siRNA transfection with either anti-VLA-1 or control scrambled siRNA, Schlemm's canal cells were seeded onto collagen IV-coated plates. The plates were incubated at 37°C for 30 minutes, washed with PBS, and incubated with calcein (1 μg / mL) at room temperature for 30 minutes. The plates were then washed with PBS, and fluorescence intensity was measured using a microplate reader.

[0333] Schlemm's canal cell tube formation assay Schlemm's canal cell tube formation assay was performed according to a previously reported method (Altiok, E. et al. Integrin Alpha-9 Mediates Lymphatic Valve Formation in Corneal Lymphangiogenesis. Invest Ophthalmol Vis Sci. 56(11):6313-6319 (2015 October)). Briefly, 48 hours after siRNA transfection with either anti-VLA-1 or control scrambled siRNA, Schlemm's canal cells were seeded onto plates containing solidified Matrigel. Tube formation was imaged 6 hours after seeding using an inverted microscope.

[0334] result IOP, measured 3 days after laser, was significantly reduced in anti-VLA-1 siRNA-treated eyes compared to control conditions. This effect is specific, as treatment with scrambled siRNA did not affect IOP. Figures 3B and 3C show summary data from human Schlemm's canal cells showing that anti-VLA-1 siRNA treatment inhibits Schlemm's canal cell functions such as adhesion (Figure 3B) and tube formation (Figure 3C). * P<0.05).

[0335] conclusion While not wishing to endorse any one theory, it has been proposed that inhibiting the key lymphangiogenic factor VLA-1 targets specific lymphatic functions of Schlemm's canal and increases its permeability, which contributes to enhanced aqueous humor movement through the canal, resulting in a measurable reduction in IOP.

[0336] Example 4. Ang-2 blockade reduces IOP and inhibits Schlemm's canal cell function This example shows that siRNA-mediated anti-Ang-2 treatment significantly reduced IOP, as demonstrated in a mouse model of glaucoma.

[0337] method material Laser-induced intraocular pressure was induced in normal eyes. Two sets of anti-Ang-2 siRNA were used. To examine IOP in vivo, the following anti-Ang-2 siRNA set was used: (Sequence (5'->3') sense: CCUCAGGAAUGAAUCAGAAtt (SEQ ID NO: 17); Sequence (5'->3') antisense: UUCUGAUUCAUUCCUGAGGtA (SEQ ID NO: 18)). One day after laser, anti-Ang-2 siRNA or control scrambled siRNA was administered locally via subconjunctival injection. To examine Schlemm's canal cell function in vitro, the following anti-Ang-2 siRNA set was used: (Sequence (5'->3') sense: GCAGAGAGAUGAAGAUCUAtt (SEQ ID NO: 15); Sequence (5'->3') antisense: UAGAUCUUCAUCUCUCUGCaa (SEQ ID NO: 16)).

[0338] Laser photocoagulation In vivo data were obtained using a well-established mouse model of glaucoma involving laser-induced occlusion of episcleral veins in normal eyes (Zhang, L. et al. Establishment and Characterization of an Acute Model of Ocular Hypertension by Laser-Induced Occlusion of Episcleral Veins. Invest Ophthalmol Vis Sci. 58(10):3879-3886(2017 Aug 1)). Briefly, mice were randomized and underwent unilateral laser photocoagulation (532 nm, OcuLight TX; IRIDEX Corporation, Mountain View, CA, USA) in the right eye. The non-lasered left eye with normal IOP served as a control for this procedure.

[0339] Intraocular pressure (IOP) measurement Intraocular pressure (IOP) was measured with a noninvasive TonoLab tonometer (Icare Lab, Helsinki, Finland) under light general anesthesia with 2% isoflurane.

[0340] Schlemm's canal cell adhesion assay Schlemm's canal cell adhesion assays were performed according to a previously reported method (Altiok, E. et al. Integrin Alpha-9 Mediates Lymphatic Valve Formation in Corneal Lymphangiogenesis. Invest Ophthalmol Vis Sci. 56(11):6313-6319 (2015 October)). Briefly, 48 hours after siRNA transfection with either anti-Ang-2 or control scrambled siRNA, Schlemm's canal cells were seeded onto collagen IV-coated plates. The plates were incubated at 37°C for 30 minutes, washed with PBS, and incubated with calcein (1 μg / mL) at room temperature for 30 minutes. The plates were then washed with PBS, and fluorescence intensity was measured using a microplate reader.

[0341] Schlemm's canal cell tube formation assay Schlemm's canal cell tube formation assay was performed according to a previously reported method (Altiok, E. et al. Integrin Alpha-9 Mediates Lymphatic Valve Formation in Corneal Lymphangiogenesis. Invest Ophthalmol Vis Sci. 56(11):6313-6319 (2015 October)). Briefly, 48 hours after siRNA transfection with either anti-Ang-2 or control scrambled siRNA, Schlemm's canal cells were seeded onto plates containing solidified Matrigel. Tube formation was imaged 6 hours after seeding using an inverted microscope.

[0342] result Compared to control conditions, IOP was significantly reduced in anti-Ang-2 siRNA-treated eyes (Figure 4A). This effect is specific, as treatment with scrambled siRNA did not affect IOP. Summary data from human Schlemm's canal cells (Figure 4B) show that anti-Ang-2 siRNA treatment inhibits Schlemm's canal cell functions such as adhesion (Figure 4B) and tube formation (Figure 4C). * P<0.05).

[0343] conclusion While not wishing to endorse any one theory, it has been proposed that inhibiting Ang-2, a key lymphangiogenic factor, targets specific lymphatic functions of Schlemm's canal and increases its permeability, which contributes to enhanced aqueous humor movement through the canal, resulting in a measurable reduction in IOP.

[0344] Example 5. ITGA5 blockade reduces IOP and inhibits Schlemm's canal cell function This example shows that siRNA-mediated anti-ITGA5-1 treatment significantly reduced IOP, as demonstrated in a mouse model of glaucoma.

[0345] method material Laser-induced intraocular pressure was induced in normal eyes. Two sets of anti-ITGA5 siRNA were used. To examine IOP in vivo, the following anti-ITGA5 siRNA set was used: (Sequence (5'->3') sense: CCCUUAUGGACGAAAUUUAtt (SEQ ID NO: 13); Sequence (5'->3') antisense: UAAAUUUCGUCCAUAAGGGct (SEQ ID NO: 14)). Anti-ITGA5 siRNA or control scrambled siRNA was administered topically via subconjunctival injection. To examine Schlemm's canal cell function in vitro, the following anti-ITGA5 siRNA set was used: (Sequence (5'->3') sense: CCUUCCAACUUGAACGGAAtt (SEQ ID NO: 11); Sequence (5'->3') antisense: UUCCGUUCAAGUUGGAAGGac (SEQ ID NO: 12)).

[0346] Laser photocoagulation In vivo data were obtained using a well-established mouse model of glaucoma involving laser-induced occlusion of episcleral veins in normal eyes (Zhang, L. et al. Establishment and Characterization of an Acute Model of Ocular Hypertension by Laser-Induced Occlusion of Episcleral Veins. Invest Ophthalmol Vis Sci. 58(10):3879-3886(2017 Aug 1)). Briefly, mice were randomized and underwent unilateral laser photocoagulation (532 nm, OcuLight TX; IRIDEX Corporation, Mountain View, CA, USA) in the right eye. The non-lasered left eye with normal IOP served as a control for this procedure.

[0347] Intraocular pressure (IOP) measurement Intraocular pressure (IOP) was measured with a noninvasive TonoLab tonometer (Icare Lab, Helsinki, Finland) under light general anesthesia with 2% isoflurane.

[0348] Schlemm's canal cell adhesion assay Schlemm's canal cell adhesion assays were performed as previously described (Altiok, E. et al. Integrin Alpha-9 Mediates Lymphatic Valve Formation in Corneal Lymphangiogenesis. Invest Ophthalmol Vis Sci. 56(11):6313-6319(2015 October)). Briefly, 48 hours after siRNA transfection with either anti-lymphangiogenic factors or control scrambled siRNA, Schlemm's canal cells were seeded onto collagen IV-coated plates. The plates were incubated at 37°C for 30 minutes, washed with PBS, and incubated with calcein (1 μg / mL) at room temperature for 30 minutes. The plates were then washed with PBS, and fluorescence intensity was measured using a microplate reader.

[0349] result IOP was measured 3 days after laser and was significantly reduced in anti-ITGA5 siRNA-treated eyes compared to control conditions (Figure 5A). This effect is specific, as treatment with scrambled siRNA did not affect IOP. Summary data from human Schlemm's canal cells show that anti-ITGA5 siRNA treatment inhibits Schlemm's canal cell functions, such as adhesion (Figure 5B). * P<0.05).

[0350] conclusion While not wishing to endorse any one theory, it has been proposed that inhibiting ITGA5, a key lymphangiogenic factor, targets specific lymphatic functions of Schlemm's canal and increases its permeability, which contributes to enhanced aqueous humor movement through the canal, resulting in a measurable reduction in IOP.

[0351] Various embodiments have been described herein with reference to the accompanying drawings. However, it will be apparent that various modifications and changes may be made and further embodiments may be implemented without departing from the broader scope of the invention as set forth in the appended claims. Moreover, other embodiments will be apparent to those skilled in the art from consideration of the specification and practice of one or more embodiments of the invention disclosed herein. Accordingly, it is intended that the application and examples herein be considered exemplary only, with the true scope and spirit of the invention being indicated by the following listing of exemplary claims.

Claims

1. A method for treating glaucoma in the eye of a subject in need thereof, comprising administering to the eye a therapeutically effective amount of an inhibitor of a lymphangiogenic factor associated with Schlemm's canal, wherein the lymphangiogenic factor induces or maintains lymphangiogenesis in the eye (a "lymphangiogenic factor").

2. The method of claim 1 , wherein the subject has primary glaucoma.

3. 3. The method of claim 2, wherein the subject has primary open-angle glaucoma.

4. 3. The method of claim 2, wherein the subject has primary angle-closure glaucoma.

5. The method of claim 2, wherein the subject has primary congenital glaucoma.

6. The method of claim 2, wherein the subject has primary normal-tension glaucoma.

7. A method for preventing glaucoma in an eye of a subject in need thereof, comprising administering to said eye a therapeutically effective amount of an inhibitor of lymphangiogenesis promoting factor.

8. 1. A method for inhibiting lymphangiogenesis in the eye of a subject at risk of developing glaucoma, comprising administering to the eye a therapeutically effective amount of an inhibitor of a lymphangiogenesis promoting factor, wherein administration of the inhibitor results in a decrease in IOP in the eye.

9. 1. A method for inhibiting lymphangiogenesis in the eye of a subject experiencing elevated intraocular pressure (IOP), comprising administering to the eye a therapeutically effective amount of an inhibitor of lymphangiogenesis promoting factors, wherein administration of the inhibitor results in a decrease in IOP in the eye.

10. A method for increasing Schlemm's canal permeability in the eye of a subject experiencing elevated IOP, comprising administering to the eye a therapeutically effective amount of an inhibitor of lymphangiogenesis promoting factor, wherein administration of the inhibitor reduces IOP in the eye.

11. A method for inhibiting Schlemm's canal cell activity in the eye of a subject experiencing elevated IOP, comprising administering to the eye a therapeutically effective amount of an inhibitor of lymphangiogenesis promoting factor, wherein the cell activity is selected from the group consisting of inhibition of cell adhesion, inhibition of cell proliferation, inhibition of cell migration, or inhibition of tube formation.

12. A method for increasing aqueous humor outflow in Schlemm's canal of the eye of a subject experiencing elevated IOP, comprising administering to the eye a therapeutically effective amount of an inhibitor of lymphangiogenesis promoting factor, wherein the administration results in an increase in aqueous humor outflow in Schlemm's canal of the eye and a decrease in IOP.

13. A method for reducing corneal edema in the eye of a subject in need thereof, comprising administering to the eye a therapeutically effective amount of an inhibitor of lymphangiogenesis promoting factor, wherein administration of the inhibitor results in a reduction of corneal edema.

14. A method for reducing thinning of the retinal nerve fiber layer in the eye of a subject in need thereof, comprising administering to the eye a therapeutically effective amount of an inhibitor of lymphangiogenesis promoting factor, wherein administration of the inhibitor results in a reduction in thinning of the nerve fibers.

15. A method for reducing retinal ganglion cell (RGC) death in the eye of a subject in need thereof, comprising administering to the eye a therapeutically effective amount of an inhibitor of lymphangiogenesis promoting factor, wherein administration of the inhibitor results in a reduction in retinal ganglion cell (RGC) death.

16. The method of any one of claims 9 to 15, wherein the subject has primary glaucoma.

17. 17. The method of claim 16, wherein the subject has primary open-angle glaucoma.

18. 17. The method of claim 16, wherein the subject has primary angle-closure glaucoma.

19. 17. The method of claim 16, wherein the subject has primary congenital glaucoma.

20. 17. The method of claim 16, wherein the subject has primary normal-tension glaucoma.

21. The method of any one of claims 1 to 20, wherein the subject is a human.

22. The method according to any one of claims 1 to 21, wherein the lymphangiogenesis promoting factor is a VEGF / VEGFR family member.

23. 23. The method of claim 22, wherein the VEGF / VEGFR family member is selected from the group consisting of VEGF-A, VEGF-B, VEGF-C, VEGF-D, VEGF-E, PIGF, VEGFR-1, VEGFR-2, and VEGFR-3.

24. 24. The method of claim 23, wherein the VEGF / VEGFR family member is VEGF-A.

25. 24. The method of claim 23, wherein the VEGF / VEGFR family member is VEGF-C.

26. 24. The method of claim 23, wherein the VEGF / VEGFR family member is VEGF-D.

27. 24. The method of claim 23, wherein the VEGF / VEGFR family member is VEGFR-3.

28. The method according to any one of claims 1 to 21, wherein the lymphangiogenesis promoting factor is an integrin family member.

29. 29. The method of claim 28, wherein the integrin family member is selected from the group consisting of very late antigen-1 (VLA-1), integrin alpha 5 (ITGA5), and integrin alpha 9 (ITGA9).

30. 30. The method of claim 29, wherein the integrin family member is VLA-1.

31. 30. The method of claim 29, wherein the integrin family member is ITGA5.

32. 30. The method of claim 29, wherein the integrin family member is ITGA9.

33. The method according to any one of claims 1 to 21, wherein the lymphangiogenesis promoting factor is a member of the angiopoietin family.

34. 34. The method of claim 33, wherein the angiopoietin family member is selected from the group consisting of angiopoietin-1 (ANGPT1), angiopoietin-2 (ANGPT2 or Ang-2), and Tie2 / TEK.

35. 35. The method of claim 34, wherein the angiopoietin family member is Ang-2.

36. The method of any one of claims 1 to 21, wherein the lymphangiogenesis promoting factor is selected from the group consisting of: (i) a cytokine or chemokine; (ii) extracellular matrix proteins; (iii) transcription factors; (iv) guiding molecule; (v) fibroblast growth factor (FGF); (vi) protein tyrosine phosphatase (PTP); (vii) a platelet factor member, or (viii) a platelet-derived growth factor; (ix) LEC polarity factor; (x) a Notch family member; and (xi) Any combination of (i) to (xi).

37. The method of any one of claims 1 to 21, wherein the lymphangiogenesis promoting factor is selected from the group consisting of: (i) a cytokine / chemokine selected from the group consisting of interleukin-8, interferon-γ, CCR7, and SLC; (ii) an extracellular matrix protein that is CCBE1; (iii) a transcription factor selected from the group consisting of Sox18 and Hhex; (iv) an inducer molecule selected from the group consisting of neuropilin 2 and SEMA7A; (v) an FGF that is FGF-2; (vi) a protein tyrosine phosphatase (PTP) that is PTPN14; (vii) a member of the platelet factors, which is platelet factor 4, or (viii) a platelet-derived growth factor, including PDGF-BB; (ix) an LEC polarity factor selected from the group consisting of Celsr1, Vangl2, Pdk2, and Fat4; (x) a member of the Notch family; (xi) ARAF, SOS1, Apelin, KIF11, REELIN, CALCRL, GJC2 (gap junction protein gamma-2), Rasip1, or FBXL7 (F-box and leucine-rich repeat protein 7); and (xii) Any one combination of (i) to (xi).

38. The method of any one of claims 1 to 37, wherein the inhibitor of lymphangiogenesis promoting factor is selected from the group consisting of: (i) an agent capable of disrupting the gene encoding the lymphangiogenesis promoting factor, thereby eliminating or reducing the expression of the lymphangiogenesis promoting factor; (ii) siRNA capable of degrading the mRNA of the lymphangiogenesis promoting factor, thereby reducing the expression of the lymphangiogenesis promoting factor; (iii) an antagonist antibody against the lymphangiogenesis promoting factor, which inhibits or prevents the function of the lymphangiogenesis promoting factor; (iv) a small molecule capable of inhibiting or preventing the function of the lymphangiogenesis promoting factor; and (v) Any combination of (i) to (iv).

39. 39. The method of any one of claims 1 to 38, wherein the inhibitor of lymphangiogenesis promoting factor is administered locally via intracameral injection, via subconjunctival injection, via intravitreal injection, via suprachoroidal injection, via peribulbar injection, or via retrobulbar injection.

40. 38. The method of any one of claims 1 to 37, wherein the inhibitor of lymphangiogenesis promoting factor is formulated as an eye drop, a depot, a bolus, an inhibitor-loaded contact lens, a suspension, a solution, an eye drop gel, or an ointment.

41. 41. The method of any one of claims 1 to 40, further comprising treating the eye with a second therapy.

42. 42. The method of claim 41, wherein the second treatment is selected from the group consisting of eye drops, oral medications, laser therapy, and surgery.

43. 43. The method of claim 41 or 42, wherein the second treatment is selected from the group consisting of eye drops that reduce the amount of fluid produced in the eye or improve drainage of fluid from the eye, oral medications that lower IOP, laser therapy including laser trabeculoplasty or laser iridotomy, and surgery including implants or trabeculectomy.

44. 44. The method of any one of claims 1 to 43, wherein prior to administration of the inhibitor, the subject has an IOP elevation of greater than about 21 mmHg.

45. 44. The method of any one of claims 1 to 43, wherein the subject has an IOP elevation of greater than about 25 mmHg prior to administration of the inhibitor.

46. 44. The method of any one of claims 1 to 43, wherein prior to administration of the inhibitor, the subject has an IOP elevation of greater than about 30 mmHg.

47. 44. The method of any one of claims 1 to 43, wherein the subject's IOP in the treated eye is reduced by at least about 2 mmHg following administration of the inhibitor.

48. 44. The method of any one of claims 1 to 43, wherein the subject's IOP in the treated eye is reduced by at least about 5 mmHg following administration of the inhibitor.

49. 44. The method of any one of claims 1 to 43, wherein the subject's IOP in the treated eye is reduced by at least about 10 mmHg after administration of the inhibitor.

50. 44. The method of any one of claims 1 to 43, wherein the subject's IOP in the treated eye is reduced by at least about 15 mmHg following administration of the inhibitor.

51. 1. A method of treating glaucoma in an eye of a subject in need thereof, comprising administering to said eye a therapeutically effective amount of an inhibitor of a lymphangiogenesis promoter selected from the group consisting of VEGFR-3, VLA-1, ITGA5, and Ang-2.

52. 52. The method of claim 51, wherein the subject has primary glaucoma.

53. 53. The method of claim 52, wherein the subject has primary open-angle glaucoma.

54. 53. The method of claim 52, wherein the subject has primary angle-closure glaucoma.

55. 53. The method of claim 52, wherein the subject has primary congenital glaucoma.

56. 53. The method of claim 52, wherein the subject has primary normal tension glaucoma.

57. 1. A method of preventing glaucoma in an eye of a subject in need thereof, comprising administering to said eye a therapeutically effective amount of an inhibitor of a lymphangiogenesis promoter selected from the group consisting of VEGFR-3, VLA-1, ITGA5, and Ang-2.

58. 1. A method of inhibiting lymphangiogenesis in the eye of a subject at risk of developing glaucoma, comprising administering to the eye a therapeutically effective amount of an inhibitor of a lymphangiogenesis promoting factor selected from the group consisting of VEGFR-3, VLA-1, ITGA5, and Ang-2, wherein administration of the inhibitor results in a decrease in IOP in the eye.

59. 1. A method for inhibiting lymphangiogenesis in the eye of a subject experiencing elevated intraocular pressure (IOP), comprising administering to the eye a therapeutically effective amount of an inhibitor of a lymphangiogenesis promoting factor selected from the group consisting of VEGFR-3, VLA-1, ITGA5, and Ang-2, wherein administration of the inhibitor results in a decrease in IOP in the eye.

60. 1. A method for increasing Schlemm's canal permeability in the eye of a subject experiencing elevated IOP, comprising administering to the eye a therapeutically effective amount of an inhibitor of lymphangiogenesis promoters selected from the group consisting of VEGFR-3, VLA-1, ITGA5, and Ang-2, wherein administration of the inhibitor reduces IOP in the eye.

61. 1. A method for inhibiting Schlemm's canal cell activity in the eye of a subject experiencing elevated IOP, comprising administering to the eye a therapeutically effective amount of an inhibitor of a lymphangiogenesis promoter selected from the group consisting of VEGFR-3, VLA-1, ITGA5, and Ang-2, wherein the cell activity is selected from the group consisting of inhibition of cell adhesion, inhibition of cell proliferation, inhibition of cell migration, or inhibition of tube formation.

62. 1. A method for increasing aqueous humor outflow in Schlemm's canal of an eye of a subject experiencing elevated IOP, comprising administering to the eye a therapeutically effective amount of an inhibitor of lymphangiogenesis promoting factors selected from the group consisting of VEGFR-3, VLA-1, ITGA5, and Ang-2, wherein administration of the inhibitor results in an increase in aqueous humor outflow in Schlemm's canal and a decrease in IOP in the eye.

63. 1. A method of reducing corneal edema in an eye of a subject in need thereof, comprising administering to the eye a therapeutically effective amount of an inhibitor of lymphangiogenesis promoters selected from the group consisting of VEGFR-3, VLA-1, ITGA5, and Ang-2, wherein administration of the inhibitor results in a reduction of corneal edema.

64. 1. A method of reducing retinal nerve fiber layer (RNFL) thinning in an eye of a subject in need thereof, comprising administering to the eye a therapeutically effective amount of an inhibitor of lymphangiogenesis promoters selected from the group consisting of VEGFR-3, VLA-1, ITGA5, and Ang-2, wherein administration of the inhibitor results in a reduction in RNFL thinning.

65. 1. A method of reducing retinal ganglion cell (RGC) death in the eye of a subject in need thereof, comprising administering to the eye a therapeutically effective amount of an inhibitor of a lymphangiogenesis promoter selected from the group consisting of VEGFR-3, VLA-1, ITGA5, and Ang-2, wherein administration of the inhibitor results in a reduction of RGC death.

66. 67. The method of any one of claims 59 to 66, wherein the subject has primary glaucoma.

67. 68. The method of claim 67, wherein the subject has primary open-angle glaucoma.

68. 68. The method of claim 67, wherein the subject has primary angle-closure glaucoma.

69. 68. The method of claim 67, wherein the subject has primary congenital glaucoma.

70. 68. The method of claim 67, wherein the subject has primary normal tension glaucoma.

71. The method of any one of claims 57 to 70, wherein the subject is a human.

72. 72. The method of any one of claims 51 to 71, wherein the inhibitor of lymphangiogenesis promoting factor is selected from the group consisting of: (i) an agent capable of disrupting the gene encoding the lymphangiogenesis promoting factor, thereby eliminating or reducing the expression of the lymphangiogenesis promoting factor; (ii) siRNA capable of degrading the mRNA of the lymphangiogenesis promoting factor, thereby reducing the expression of the lymphangiogenesis promoting factor; (iii) an antagonist antibody against the lymphangiogenesis promoting factor, which inhibits or prevents the function of the lymphangiogenesis promoting factor; (iv) a small molecule capable of inhibiting or preventing the function of the lymphangiogenesis promoting factor; and (v) Any combination of (i) to (iv).

73. 73. The method of any one of claims 51 to 72, wherein the inhibitor of lymphangiogenesis promoting factor is administered locally via intracameral injection, via subconjunctival injection, via intravitreal injection, via suprachoroidal injection, via peribulbar injection, or via retrobulbar injection.

74. 74. The method of any one of claims 51 to 73, wherein the inhibitor of lymphangiogenesis promoting factor is formulated as an eye drop, a depot, a bolus, an inhibitor-loaded contact lens, a suspension, a solution, an eye drop gel, or an ointment.

75. 75. The method of any one of claims 51 to 74, further comprising treating the eye with a second therapy.

76. 76. The method of claim 75, wherein the second treatment is selected from the group consisting of eye drops, oral medications, laser therapy, and surgery.

77. 77. The method of claim 75 or 76, wherein the second treatment is selected from the group consisting of eye drops that reduce the amount of fluid produced in the eye or improve drainage of fluid from the eye, oral medications that lower IOP, laser therapy including laser trabeculoplasty or laser iridotomy, and surgery including implants or trabeculectomy.

78. 78. The method of any one of claims 51-77, wherein prior to administration of the inhibitor, the subject has an IOP elevation of greater than about 21 mmHg.

79. 78. The method of any one of claims 51-77, wherein the subject has an IOP elevation of greater than about 25 mmHg prior to administration of the inhibitor.

80. 78. The method of any one of claims 51-77, wherein the subject has an IOP elevation of greater than about 30 mmHg prior to administration of the inhibitor.

81. 81. The method of any one of claims 51-80, wherein the subject's IOP in the treated eye is reduced by at least about 2 mmHg after administration of the inhibitor.

82. 81. The method of any one of claims 51-80, wherein the subject's IOP in the treated eye is reduced by at least about 5 mmHg after administration of the inhibitor.

83. 81. The method of any one of claims 51-80, wherein the subject's IOP in the treated eye is reduced by at least about 10 mmHg after administration of the inhibitor.

84. 81. The method of any one of claims 51-80, wherein the subject's IOP in the treated eye is reduced by at least about 15 mmHg following administration of the inhibitor.

85. The method of any one of claims 51 to 84, wherein the lymphangiogenesis promoting factor is VEGFR-3.

86. The method of any one of claims 51 to 84, wherein the lymphangiogenesis promoting factor is VLA-1.

87. The method of any one of claims 51 to 80, wherein the lymphangiogenesis promoting factor is ITAG5.

88. The method of any one of claims 51 to 84, wherein the lymphangiogenesis promoting factor is Ang-2.

89. The method of any one of claims 1 to 88, wherein the inhibitor is an siRNA.

90. The method of any one of claims 1 to 88, wherein the inhibitor is an antagonist antibody.

91. 89. The method of any one of claims 1 to 88, wherein the inhibitor interferes with the expression of the gene for the lymphangiogenesis promoting factor.

92. The method of any one of claims 1 to 91, wherein the subject is a human.

93. A method of treating glaucoma in an eye of a subject in need thereof, comprising administering to the eye a therapeutically effective amount of an agent that activates or stimulates lymphatic factors that inhibit or prevent lymphangiogenesis in the eye ("anti-lymphangiogenic factors").

94. 94. The method of claim 93, wherein the subject has primary glaucoma.

95. 95. The method of claim 94, wherein the subject has primary open-angle glaucoma.

96. 95. The method of claim 94, wherein the subject has primary angle-closure glaucoma.

97. 95. The method of claim 94, wherein the subject has primary congenital glaucoma.

98. 95. The method of claim 94, wherein the subject has primary normal tension glaucoma.

99. 1. A method of preventing glaucoma in an eye of a subject in need thereof, comprising administering to the eye a therapeutically effective amount of an anti-lymphangiogenic factor.

100. 1. A method for inhibiting lymphangiogenesis in the eye of a subject at risk of developing glaucoma, comprising administering to the eye a therapeutically effective amount of an agent that activates or stimulates an anti-lymphangiogenic factor, wherein administration of the agent results in a decrease in IOP in the eye.

101. 1. A method for inhibiting lymphangiogenesis in the eye of a subject experiencing elevated intraocular pressure (IOP), comprising administering to the eye a therapeutically effective amount of an agent that activates or stimulates an anti-lymphangiogenic factor, wherein administration of the agent results in a decrease in IOP in the eye.

102. 1. A method for increasing Schlemm's canal permeability in the eye of a subject experiencing elevated IOP, comprising administering to the eye a therapeutically effective amount of an agent that activates or stimulates an anti-lymphangiogenic factor, wherein administration of the agent reduces IOP in the eye.

103. A method for inhibiting Schlemm's canal cell activity in the eye of a subject experiencing elevated IOP, comprising administering to the eye a therapeutically effective amount of an agent that activates or stimulates an anti-lymphangiogenic factor, wherein the cell activity is selected from the group consisting of inhibition of cell adhesion, inhibition of cell proliferation, inhibition of cell migration, or inhibition of tube formation.

104. A method for increasing aqueous humor outflow in Schlemm's canal of the eye of a subject experiencing elevated IOP, comprising administering to the eye a therapeutically effective amount of an agent that activates or stimulates an anti-lymphangiogenic factor, wherein administration of the agent results in an increase in aqueous humor outflow in Schlemm's canal of the eye and a decrease in IOP.

105. A method for reducing corneal edema in the eye of a subject in need thereof, comprising administering to the eye a therapeutically effective amount of an inhibitor of an agent that activates or stimulates an anti-lymphangiogenic factor, wherein administration of the agent results in a reduction of corneal edema.

106. 1. A method for reducing retinal nerve fiber layer (RNFL) thinning in the eye of a subject in need thereof, comprising administering to the eye a therapeutically effective amount of an agent that activates or stimulates an anti-lymphangiogenic factor, wherein administration of the agent results in a reduction in RNFL thinning.

107. A method for reducing retinal ganglion cell (RGC) death in the eye of a subject in need thereof, comprising administering to the eye a therapeutically effective amount of an agent that activates or stimulates an anti-lymphangiogenic factor, wherein administration of the agent results in a reduction in RGC death.

108. The method of any one of claims 101 to 107, wherein the subject has primary glaucoma.

109. 109. The method of claim 108, wherein the subject has primary open-angle glaucoma.

110. 109. The method of claim 108, wherein the subject has primary angle-closure glaucoma.

111. 109. The method of claim 108, wherein the subject has primary congenital glaucoma.

112. 109. The method of claim 108, wherein the subject has primary normal tension glaucoma.

113. The method of any one of claims 93 to 112, wherein the subject is a human.

114. 114. The method of any one of claims 93 to 113, wherein the agent is administered locally via intracameral injection, via subconjunctival injection, via intravitreal injection, via suprachoroidal injection, via peribulbar injection, or via retrobulbar injection.

115. 115. The method of any one of claims 93 to 114, wherein the medicament is formulated as eye drops, a depot, a bolus, an inhibitor-loaded contact lens, a suspension, a solution, an eye drop gel, or an ointment.

116. 116. The method of any one of claims 93 to 115, further comprising treating the eye with a second therapy.

117. 117. The method of claim 116, wherein the second treatment is selected from the group consisting of eye drops, oral medications, laser therapy, and surgery.

118. 118. The method of claim 116 or 117, wherein the second treatment is selected from the group consisting of eye drops that reduce the amount of fluid produced in the eye or improve drainage of fluid from the eye, oral medications that lower IOP, laser therapy including laser trabeculoplasty or laser iridotomy, and surgery including implants or trabeculectomy.

119. 119. The method of any one of claims 93 to 118, wherein prior to administration of the agent, the subject has an IOP elevation of greater than about 21 mmHg.

120. 119. The method of any one of claims 93 to 118, wherein the subject has an IOP elevation of greater than about 25 mmHg prior to administration of the agent.

121. 119. The method of any one of claims 93 to 118, wherein the subject has an IOP elevation of greater than about 30 mmHg prior to administration of the agent.

122. 122. The method of any one of claims 93 to 121, wherein the subject's IOP in the treated eye is reduced by at least about 2 mmHg after administration of the agent.

123. 122. The method of any one of claims 93 to 121, wherein the subject's IOP in the treated eye is reduced by at least about 5 mmHg after administration of the agent.

124. 122. The method of any one of claims 93 to 121, wherein the subject's IOP in the treated eye is reduced by at least about 10 mmHg after administration of the agent.

125. 122. The method of any one of claims 93 to 121, wherein the subject's IOP in the treated eye is reduced by at least about 15 mmHg after administration of the agent.

126. 126. The method of any one of claims 93 to 125, wherein the agent administered is selected from the group consisting of: (i) anti-lymphangiogenic factors; (ii) an agent that activates the expression of said anti-lymphangiogenic factor; (iii) an agonist antibody that activates the anti-lymphangiogenic factor; (iv) a microRNA or mimic that inhibits lymphangiogenesis; and (v) a small molecule that activates the function of said anti-lymphangiogenic factor.

127. 127. The method of any one of claims 93 to 126, wherein the anti-lymphangiogenic factor is selected from the group consisting of microRNA-184, microRNA-126, miR-31, miR-181a, microRNA-132, miR-194, miR-186, miR-99a, miR-92a, and miR-466, endostatin, and a VEGF family inhibitor (e.g., soluble VEGFR-2, soluble VEGFR-3).

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