Compositions and methods for ocular diseases

Abstract

The present disclosure relates, in some aspects, to methods and compositions for determining an anatomical response of an eye of a subject to treatment with a VEGF antagonist comprising determining a VEGF level in aqueous humor of the subject or in combination with determining a bl-CMT level in the subject wherein the determined VEGF level is above a VEGF threshold / range or in combination with the determined bl-CMT level is above a bl-CMT threshold / range, the eye is shown to have a good anatomical response to treatment via an intravitreal, suprachoroidal and / or subretinal VEGF antagonist wherein the determined VEGF level is below a VEGF critical value / range or the determined bl-CMT level is below a bl-CMT critical value / range, the eye is indicated to have an undesirable anatomical response to treatment via an intravitreal, suprachoroidal and / or subretinal VEGF antagonist. VEGF levels and / or bl-CMT levels in aqueous humor can be used as companion diagnostic indicators for predicting the response of a subject to an anti-VEGF treatment.

Description

Technical Field

[0001] The present disclosure relates in some aspects to methods and compositions for retinal vein occlusion treatment and companion diagnostics. Background Art

[0002] Retinal vein occlusion (RVO) is one of the most common retinal vascular diseases [1, 2]. It is caused by partial or complete blockage of venous blood flow, resulting in increased venous pressure and subsequent retinal microvascular leakage behind the occlusion site [3]. Improved methods for treating RVO and its symptoms are needed. Methods and compositions that meet these needs and other needs are provided herein.

[0003] RVO can include occlusion of the main retinal vein, called central retinal vein occlusion (CRVO), and / or occlusion of small veins, called branch retinal vein occlusion (BRVO). The pathological mechanism of RVO is a series of changes that occur after retinal vein occlusion, including hypoxia, increased vascular permeability, inflammation, and neovascularization, and involves a complex interaction between multiple vascular and inflammatory mediators.

[0004] Vascular endothelial growth factor (VEGF), a potent mediator of angiogenesis and vascular permeability, is a key molecule involved in the pathophysiology of RVO [4, 5], and intravitreal anti-VEGF agents are currently considered the first-line treatment for macular edema (ME) secondary to RVO (RVO-ME) [6]. However, on the other hand, a considerable proportion of eyes with RVO-ME are inadequately responsive to anti-VEGF agents, notably, they usually respond well to intravitreal glucocorticoids, and visual outcomes are negatively correlated with disease duration [7]. These findings suggest that RVO-ME can be divided into two endotypes, one in which VEGF plays a central role and therefore responds well to intravitreal anti-VEGF agents (tentatively referred to as the “anti-VEGF-responsive endotype”), and the other in which VEGF is less relevant and responds poorly to anti-VEGF agents (tentatively referred to as the “anti-VEGF-resistant endotype”). Prediction of treatment response to anti-VEGF agents at baseline could avoid unnecessary anti-VEGF treatment in the latter endotype and improve the overall visual outcome of RVO-ME.

[0005] Companion diagnostics are a set of diagnostic tests that predict the safety and / or efficacy of a specific treatment and are increasingly being considered as a means to improve the precision of cancer treatment [8]. Pathophysiological therapeutic targets themselves are often ideal companion biomarkers for treatments directed against that target. Modi, A. et al. supportively described a customized treatment approach in which the type of intravitreal drug was personalized based on aqueous humor cytokine levels (VEGF, interleukin (IL) 6, and IL-8) and was shown to be beneficial in cases of refractory RVO-ME [9]. Several early studies in the literature have explored the association between intraocular VEGF levels and response to anti-VEGF agents. Campochiaro et al. reported that baseline aqueous humor VEGF levels were inversely correlated with improvement in visual acuity (VA) after three monthly IVR injections

[10] . Similarly, Park, SP et al. detected higher baseline aqueous humor VEGF levels in patients who did not respond to a single IVB injection

[11] . However, these results contradict the general rule that upregulation of pathogenic molecules is generally associated with a favorable response to treatments targeting that molecule.

[0006] A comprehensive review of intraocular biomarkers of RVO found that a group of cytokines including VEGF, IL-6, IL-8, IL-10, monocyte chemoattractant protein-1 (MCP-1), interferon-γ-induced protein 10 (IP-10), etc. can be used as potential concomitant biomarkers for anti-VEGF therapy

[12] . The invention disclosed herein addresses whether aqueous humor biomarkers and imaging biomarkers alone or in combination have predictive value for the response to intravitreal anti-VEGF agents. Summary of the invention

[0007] In order to evaluate whether aqueous humor biomarkers and imaging biomarkers, alone or in combination, have predictive value for the response to intravitreal anti-VEGF preparations, the present disclosure provides, in certain aspects, a method for predicting the anatomical response of a treatment target eye to an anti-VEGF preparation. In some embodiments, provided herein is the use of an anti-VEGF therapeutic agent in the preparation of a medicament for treating retinal vein occlusion (RVO) in a subject, wherein the VEGF level in the subject's aqueous humor is above a critical value, and the critical value is between 10 pg / mL and 100 pg / mL. In some embodiments, provided herein is the use of an anti-VEGF therapeutic agent in the preparation of a medicament for treating retinal vein occlusion (RVO) in a subject, wherein the VEGF level in the subject's aqueous humor is above a VEGF critical value, and the VEGF critical value is between 10 pg / mL and 100 pg / mL; and wherein the bl-CMT level in the subject is above the bl-CMT critical value, and the bl-CMT critical value is between about 250 μm and about 600 μm. In some embodiments, the anti-VEGF therapeutic is a VEGF antagonist which can be any therapeutic modality, such as an antibody, peptide or protein therapy, gene therapy, nucleic acid, small molecule, or any combination thereof.

[0008] In some aspects, the present invention provides a method of treating retinal vein occlusion (RVO) in a subject, comprising administering to the subject an effective amount of an anti-VEGF therapeutic agent, wherein the subject's VEGF level in the aqueous humor is above a VEGF cutoff value, and the VEGF cutoff value is between 10 pg / mL and 100 pg / mL.

[0009] In one aspect, the invention provides a method of treating RVO in a subject as disclosed herein, wherein the anti-VEGF therapeutic comprises:

[0010] An anti-VEGF antibody or an antigen-binding fragment thereof, optionally the antigen-binding fragment is Fab, Fab', F(ab')2 or Fv;

[0011] A VEGF receptor (VEGFR) fusion protein, optionally the VEGFR fusion protein is a VEGFR-Fc immunoadhesin; or

[0012] An anti-VEGF gene therapy agent, optionally the anti-VEGF gene therapy agent comprises a nucleic acid encoding a VEGF antagonist, a nucleic acid encoding a protein that downregulates VEGF / VEGFR expression, and / or a nucleic acid targeting a nucleic acid and / or protein involved in VEGF / VEGFR expression, optionally the anti-VEGF gene therapy agent comprises a viral or non-viral vector, optionally the viral vector is an adeno-associated virus (AAV) vector.

[0013] In another aspect, the invention provides a method of treating RVO in a subject as disclosed herein, wherein the anti-VEGF therapeutic is administered or is intended to be administered intravitreally, suprachoroidally and / or subretinally.

[0014] In another aspect, the invention provides a method of treating RVO in a subject as disclosed herein, wherein the subject has macular edema secondary to retinal vein occlusion (ME-RVO), optionally the retinal vein occlusion is a central retinal vein occlusion (CRVO) and / or a branch retinal vein occlusion (BRVO).

[0015] In another aspect, the invention provides a method of treating RVO in a subject as disclosed herein, wherein the subject has macular edema secondary to retinal vein occlusion (ME-RVO), optionally the retinal vein occlusion is a central retinal vein occlusion (CRVO) and / or a branch retinal vein occlusion (BRVO).

[0016] In another aspect, the invention provides a method of treating RVO in a subject as disclosed herein, wherein the subject has been treated with the anti-VEGF therapeutic or another anti-VEGF therapeutic.

[0017] In another aspect, the invention provides a method of treating RVO in a subject as disclosed herein, wherein the subject has not been treated with the anti-VEGF therapeutic or another anti-VEGF therapeutic.

[0018] In another aspect, the present invention provides a method of treating RVO in a subject as disclosed herein, wherein the subject has been treated with an anti-inflammatory agent or an immunosuppressant or a combination thereof, wherein the VEGF level in the subject's aqueous humor is above a threshold value, optionally the anti-inflammatory agent or immunosuppressant is selected from glucocorticoids, methotrexate, cyclosporin A, tacrolimus, azathioprine, or a biologic targeting a pro-inflammatory molecule such as interleukin-6, interleukin-8, and tumor necrosis factor alpha.

[0019] In another aspect, the present invention provides a method of treating RVO in a subject as disclosed herein, wherein the subject has not been treated with an anti-inflammatory agent or an immunosuppressant, or a combination thereof, wherein the subject's VEGF level in the aqueous humor is above a threshold value, optionally the anti-inflammatory agent or immunosuppressant is selected from glucocorticoids, methotrexate, cyclosporin A, tacrolimus, azathioprine, or a biologic targeting a pro-inflammatory molecule such as interleukin-6, interleukin-8, and tumor necrosis factor alpha.

[0020] In another aspect, the present invention provides a method of treating RVO in a subject as disclosed herein, wherein the subject has been treated with a steroid, which may be selected as a corticosteroid, further selected as a glucocorticoid.

[0021] In another aspect, the present invention provides a method of treating RVO in a subject as disclosed herein, wherein the subject has not been treated with a steroid, the steroid may be selected as a corticosteroid, further selected as a glucocorticoid.

[0022] In another aspect, the invention provides a method of treating RVO in a subject as disclosed herein, wherein the VEGF cutoff is between 30 pg / mL and 90 pg / mL.

[0023] In another aspect, the invention provides a method of treating RVO in a subject as disclosed herein, wherein the VEGF cutoff is about 35 pg / mL, 40 pg / mL, 45 pg / mL, 50 pg / mL, 55 pg / mL, 60 pg / mL, 65 pg / mL, 70 pg / mL, 75 pg / mL, 80 pg / mL or 85 pg / mL.

[0024] In another aspect, the invention provides a method of treating RVO in a subject as disclosed herein, wherein the subject has an anatomical response to an anti-VEGF therapeutic as measured by a change in central macular thickness (CMT) or foveal thickness (CFT), wherein CMT or CFT is defined as the perpendicular distance between the internal limiting membrane and the retinal pigment epithelium at the center of the fovea, also described in some cases as foveal thickness (CFT).

[0025] In another aspect, the invention provides a method of treating RVO in a subject as disclosed herein, wherein the subject has an anatomical response to an anti-VEGF therapeutic as measured by a relative change in central macular thickness (CMT), wherein the relative change in CMT is defined by the change in CMT divided by the baseline central macular thickness (bl-CMT).

[0026] In another aspect, the invention provides a method of treating RVO in a subject as disclosed herein, wherein the subject has a baseline central macular thickness (bl-CMT) level above a bl-CMT cutoff value, wherein the bl-CMT cutoff value is between about 250 μm and about 600 μm.

[0027] In another aspect, the present invention provides a method of treating retinal vein occlusion (RVO) in a subject as disclosed herein, comprising:

[0028] (a) determining a level of VEGF in aqueous humor collected from the subject and a level of bl-CMT in the subject, wherein the determined level of VEGF is above a VEGF cutoff value and the determined level of bl-CMT is above a bl-CMT cutoff value,

[0029] (b) administering to the subject an effective amount of an anti-VEGF therapeutic agent.

[0030] In another aspect, the invention provides a method of treating RVO in a subject as disclosed herein, wherein the VEGF cutoff is between 10 pg / mL and 100 pg / mL.

[0031] In another aspect, the invention provides a method of treating RVO in a subject as disclosed herein, wherein the bl-CMT cutoff is between about 250 μm and about 600 μm.

[0032] In another aspect, the invention provides a method of treating RVO in a subject as disclosed herein,

[0033] In another aspect, the invention provides a method of treating RVO in a subject as disclosed herein, wherein the anatomical response to the anti-VEGF therapeutic is measured by changes in CMT, which is defined as the perpendicular distance between the internal limiting membrane and the retinal pigment epithelium at the center of the fovea.

[0034] In another aspect, the invention provides a method of treating RVO in a subject as disclosed herein, wherein the anatomical response to the anti-VEGF therapeutic is measured by a relative change in CMT, wherein the relative change in CMT is defined by dividing the change in CMT by bl-CMT.

[0035] In another aspect, the invention provides a method of treating RVO in a subject as disclosed herein, wherein the determined VEGF level is above the VEGF cutoff value of between 10 pg / mL and 100 pg / mL and the determined bl-CMT level is above the bl-CMT cutoff value of between about 250 μm and about 600 μm, indicating that the eye has a good anatomical response to treatment with an anti-VEGF therapeutic agent.

[0036] In another aspect, the invention provides a method of treating RVO in a subject as disclosed herein, wherein the determined VEGF level is above a VEGF cutoff value and the determined bl-CMT level is above a bl-CMT cutoff value, predicting a change in CFT of <-200 μm.

[0037] In another aspect, the invention provides a method of treating RVO in a subject as disclosed herein, wherein the VEGF cutoff is between 30 pg / mL and 90 pg / mL.

[0038] In another aspect, the invention provides a method of treating RVO in a subject as disclosed herein, wherein the VEGF cutoff is about 35 pg / mL, 40 pg / mL, 45 pg / mL, 50 pg / mL, 55 pg / mL, 60 pg / mL, 65 pg / mL, 70 pg / mL, 75 pg / mL, 80 pg / mL or 85 pg / mL.

[0039] In another aspect, the invention provides a method of treating RVO in a subject as disclosed herein, wherein the bl-CMT cutoff is between 250 μm and 600 μm.

[0040] In another aspect, the invention provides a method of treating RVO in a subject as disclosed herein, wherein the bl-CMT cutoff is about 250 μm, 300 μm, 350 μm, 400 μm, 450 μm, 500 μm, 550 μm or 600 μm.

[0041] In another aspect, the invention provides a method of treating RVO in a subject as disclosed herein, wherein the level of VEGF is determined by an immunoassay, optionally an enzyme immunoassay.

[0042] In another aspect, the invention provides a method of treating RVO in a subject as disclosed herein, wherein the subject is in need of treatment for retinal vein occlusion (RVO).

[0043] In another aspect, the invention provides a method of treating RVO in a subject as disclosed herein, wherein the subject is in need of treatment for macular edema secondary to retinal vein occlusion (ME-RVO), optionally the retinal vein occlusion is a central retinal vein occlusion (CRVO) and / or a branch retinal vein occlusion (BRVO).

[0044] In another aspect, the invention provides a method of treating RVO in a subject as disclosed herein, further comprising administering the anti-VEGF therapeutic or another anti-VEGF therapeutic to the subject intravitreally, suprachoroidally and / or subretinally when the determined VEGF level is above the VEGF cutoff value and when the determined bl-CMT level is above the bl-CMT cutoff value.

[0045] In another aspect, the invention provides a method of treating RVO in a subject as disclosed herein, wherein the determined VEGF level is above 90 pg / mL, and the anti-VEGF therapeutic or another anti-VEGF therapeutic is administered to the subject intravitreally, suprachoroidally and / or subretinally.

[0046] In another aspect, the invention provides a method of treating RVO in a subject as disclosed herein, wherein prior to the determining step, the subject has not been treated with the anti-VEGF therapeutic or another anti-VEGF therapeutic.

[0047] In another aspect, the invention provides a method of treating RVO in a subject as disclosed herein, wherein prior to the determining step, the subject has been treated with the anti-VEGF therapeutic agent or another anti-VEGF therapeutic agent.

[0048] In another aspect, the invention provides a method of treating RVO in a subject as disclosed herein, wherein prior to the determining step, the subject has not received steroid therapy, the steroid can be selected as a corticosteroid, further selected as a glucocorticoid.

[0049] In another aspect, the invention provides a method of treating RVO in a subject as disclosed herein, wherein prior to the determining step, the subject has been receiving steroid therapy, the steroid may be selected as a corticosteroid, further selected as a glucocorticoid.

[0050] In another aspect, the present invention provides a method of treating RVO in a subject as disclosed herein, wherein the subject has not been treated with an anti-inflammatory agent or an immunosuppressant, or a combination thereof, optionally wherein the anti-inflammatory agent or immunosuppressant is a glucocorticoid, methotrexate, cyclosporin A, tacrolimus, azathioprine, or a biologic targeting a pro-inflammatory molecule such as interleukin-6, interleukin-8, tumor necrosis factor alpha, or the like.

[0051] In another aspect, the present invention provides a method of treating RVO in a subject as disclosed herein, wherein the subject has been treated with an anti-inflammatory agent or an immunosuppressant or a combination thereof, optionally the anti-inflammatory agent or immunosuppressant is a glucocorticoid, methotrexate, cyclosporin A, tacrolimus, azathioprine, or a biologic targeting a pro-inflammatory molecule such as interleukin-6, interleukin-8, tumor necrosis factor alpha, etc.

[0052] In another aspect, the invention provides a method of treating RVO in a subject as disclosed herein, wherein the subject is selected to initiate, continue or resume use of an anti-VEGF drug or other anti-VEGF drug, and / or to discontinue treatment with a steroid, which may be a corticosteroid, further selected as a glucocorticoid.

[0053] In another aspect, the invention provides a method of treating RVO in a subject as disclosed herein, wherein the subject chooses to discontinue use of an anti-VEGF drug or other anti-VEGF drug, and / or chooses to initiate, continue or resume treatment with a steroid, which may be a corticosteroid, further selected as a glucocorticoid.

[0054] In another aspect, the invention provides a method of treating RVO in a subject as disclosed herein.

[0055] In another aspect, the invention provides a method of treating RVO in a subject as disclosed herein, wherein the subject is a human.

[0056] In another aspect, the invention provides a method of treating RVO in a subject as disclosed herein, wherein the anti-VEGF therapeutic is formulated for intravitreal, suprachoroidal and / or subretinal administration, optionally further comprising mixing the anti-VEGF therapeutic with a pharmaceutically acceptable excipient or carrier.

[0057] In another aspect, the present invention provides a use of an anti-VEGF therapeutic agent in the preparation of a medicament for treating retinal vein occlusion (RVO) in a subject, wherein the VEGF level in the subject's aqueous humor is above a cutoff value, and the cutoff value is between 10 pg / mL and 100 pg / mL.

[0058] In another aspect, the present invention provides use of an anti-VEGF therapeutic as disclosed herein in the preparation of a medicament for treating RVO in a subject, wherein the anti-VEGF therapeutic comprises:

[0059] An anti-VEGF antibody or an antigen-binding fragment thereof, optionally the antigen-binding fragment is Fab, Fab', F(ab')2 or Fv;

[0060] A VEGF receptor (VEGFR) fusion protein, optionally the VEGFR fusion protein is a VEGFR-Fc immunoadhesin; or

[0061] An anti-VEGF gene therapy agent, optionally the anti-VEGF gene therapy agent comprises a nucleic acid encoding a VEGF antagonist, a nucleic acid encoding a protein that downregulates VEGF / VEGFR expression, and / or a nucleic acid targeting a nucleic acid and / or protein involved in VEGF / VEGFR expression, optionally the anti-VEGF gene therapy agent comprises a viral vector or a non-viral vector, optionally the viral vector is an adeno-associated virus (AAV) vector.

[0062] In another aspect, the present invention provides use of an anti-VEGF therapeutic as disclosed herein in the preparation of a medicament for treating RVO in a subject, wherein the anti-VEGF therapeutic is administered or is to be administered intravitreally, suprachoroidally and / or subretinally.

[0063] In another aspect, the invention provides use of an anti-VEGF therapeutic as disclosed herein in the preparation of a medicament for treating RVO in a subject, wherein the subject suffers from macular edema secondary to retinal vein occlusion (ME-RVO), optionally wherein the retinal vein occlusion is a central retinal vein occlusion (CRVO) and / or a branch retinal vein occlusion (BRVO).

[0064] In another aspect, the invention provides use of an anti-VEGF therapeutic as disclosed herein in the preparation of a medicament for treating RVO in a subject, wherein the subject has or has not been treated with the anti-VEGF therapeutic or another anti-VEGF therapeutic.

[0065] In another aspect, the present invention provides the use of an anti-VEGF therapeutic as disclosed herein in the preparation of a medicament for treating RVO in a subject, wherein the subject has or has not been treated with an anti-inflammatory agent or an immunosuppressant or a combination thereof, wherein the subject's VEGF level in the aqueous humor is above a threshold value, optionally the anti-inflammatory agent or immunosuppressant is selected from glucocorticoids, methotrexate, cyclosporin A, tacrolimus, azathioprine, or a biological agent targeting a pro-inflammatory molecule such as interleukin-6, interleukin-8 and tumor necrosis factor alpha.

[0066] In another aspect, the present invention provides the use of an anti-VEGF therapeutic agent as disclosed herein in the preparation of a medicament for treating RVO in a subject, wherein the subject has or has not received steroid therapy, and the steroid can be selected as a corticosteroid, further selected as a glucocorticoid.

[0067] In another aspect, the present invention provides use of an anti-VEGF therapeutic as disclosed herein in the preparation of a medicament for treating RVO in a subject, wherein the cutoff value is between 30 pg / mL and 90 pg / mL.

[0068] In another aspect, the invention provides use of an anti-VEGF therapeutic as disclosed herein in the preparation of a medicament for treating RVO in a subject, wherein the cutoff value is about 35 pg / mL, 40 pg / mL, 45 pg / mL, 50 pg / mL, 55 pg / mL, 60 pg / mL, 65 pg / mL, 70 pg / mL, 75 pg / mL, 80 pg / mL or 85 pg / mL.

[0069] In another aspect, the invention provides use of an anti-VEGF therapeutic as disclosed herein in the preparation of a medicament for treating RVO in a subject, wherein the subject has an anatomical response to the anti-VEGF therapeutic as measured by a change in central macular thickness (CMT) or foveal thickness (CFT), or as measured by a relative change in central macular thickness (CMT), wherein CMT or CFT is defined as the perpendicular distance between the internal limiting membrane and the retinal pigment epithelium at the center of the fovea, wherein the relative change in CMT is defined by the change in CMT divided by the baseline central macular thickness (bl-CMT).

[0070] In another aspect, the present invention provides use of an anti-VEGF therapeutic as disclosed herein in the preparation of a medicament for treating RVO in a subject, wherein the subject has a baseline central macular thickness (bl-CMT) level above a bl-CMT cutoff value, wherein the bl-CMT cutoff value is between about 250 μm and about 600 μm.

[0071] In another aspect, the invention provides use of an anti-VEGF therapeutic in the preparation of a kit for treating retinal vein occlusion (RVO), wherein the kit comprises: i) the anti-VEGF therapeutic, and ii) instructions for administering the anti-VEGF therapeutic to a subject having a VEGF level in aqueous humor above a cutoff value, and wherein the cutoff value is between 10 pg / mL and 100 pg / mL.

[0072] In another aspect, the present invention provides the use of an anti-VEGF therapeutic as disclosed herein in the preparation of a kit for treating RVO, wherein the kit further comprises instructions for not administering the anti-VEGF therapeutic to a subject whose VEGF level in the aqueous humor is not above a critical value, and / or instructions for administering an anti-inflammatory agent or an immunosuppressant or a combination thereof to a subject whose VEGF level in the aqueous humor is above a critical value, optionally the anti-inflammatory agent or immunosuppressant is selected from glucocorticoids, methotrexate, cyclosporin A, tacrolimus, azathioprine, or a biological agent targeting a pro-inflammatory molecule such as interleukin-6, interleukin-8, and tumor necrosis factor-α.

[0073] In another aspect, the present invention provides use of an anti-VEGF therapeutic as disclosed herein in the preparation of a kit for treating RVO, wherein the subject whose VEGF level in aqueous humor is above a cut-off value has a bl-CMT level above a bl-CMT cut-off value, and wherein the bl-CMT cut-off value is between about 250 μm and about 600 μm.

[0074] In another aspect, the invention provides a kit comprising i) an anti-VEGF therapeutic and ii) instructions for administering the anti-VEGF therapeutic to a subject having VEGF levels in aqueous humor above a cutoff value, wherein the cutoff value is between 10 pg / mL and 100 pg / mL.

[0075] In another aspect, the present invention provides a kit as disclosed herein, further comprising instructions for not administering an anti-VEGF therapeutic agent to a subject whose VEGF level in the aqueous humor is not above a threshold value, and / or instructions for administering an anti-inflammatory agent or an immunosuppressant or a combination thereof to a subject whose VEGF level in the aqueous humor is above a threshold value, optionally the anti-inflammatory agent or immunosuppressant is selected from glucocorticoids, methotrexate, cyclosporin A, tacrolimus, azathioprine, or a biologic targeting a pro-inflammatory molecule such as interleukin-6, interleukin-8, and tumor necrosis factor-α.

[0076] In another aspect, the invention provides a kit as disclosed herein, wherein the subject having VEGF levels in aqueous humor above a cut-off value has bl-CMT levels above a bl-CMT cut-off value, and wherein the bl-CMT cut-off value is between about 250 μm and about 600 μm.

[0077] In another aspect, the invention provides a kit comprising: i) reagents for determining VEGF levels in a subject's aqueous humor, and ii) instructions comprising a cutoff value for VEGF levels, and optionally instructions for comparing the determined VEGF levels to the cutoff value, wherein the cutoff value is between 10 pg / mL and 100 pg / mL.

[0078] In another aspect, the present invention provides a kit as disclosed herein, further comprising a first standard sample having a VEGF level substantially equal to a cut-off value.

[0079] In another aspect, the present invention provides a kit as disclosed herein, further comprising a second standard sample having a VEGF level above a critical value.

[0080] In another aspect, the present invention provides a kit as disclosed herein, further comprising a third standard sample having a VEGF level below a critical value.

[0081] In another aspect, the invention provides a kit as disclosed herein, wherein the agent is an anti-VEGF antibody or an antigen-binding fragment thereof, optionally the antigen-binding fragment is Fab, Fab', F(ab')2 or Fv.

[0082] In another aspect, the present invention provides a kit as disclosed herein, comprising an enzyme reactant, a magnetic separation reagent, a stabilizer, a standard, one or more concentrated wash buffers and / or a substrate buffer.

[0083] In another aspect, the present invention provides a kit as disclosed herein, wherein the enzyme reactant comprises a VEGF-specific antibody I labeled with alkaline phosphatase, and wherein the labeling ratio of the VEGF antibody I to the alkaline phosphatase is 1:0.75.

[0084] In another aspect, the present invention provides a kit as disclosed herein, wherein the magnetic separation reagent contains 0.05 mass % magnetic particles, wherein the magnetic particles are coated with VEGF-specific antibody II, wherein the magnetic reagent further comprises a blocking agent, wherein the blocking agent is a mutant alkaline phosphatase.

[0085] In another aspect, the present invention provides a kit as disclosed herein, wherein the VEGF-specific antibody I and / or the VEGF-specific antibody II is an anti-VEGF165 / VEGFA antibody.

[0086] In another aspect, the present invention provides a kit as disclosed herein, wherein the stabilizer comprises one or more agents selected from the group consisting of Tris, NaCl, Tween 20, bovine serum albumin, mutant alkaline phosphatase and ProClin 300.

[0087] In another aspect, the invention provides a kit as disclosed herein, wherein the wash buffer comprises Tris, NaCl, Tween-20, Triton-100 and ProClin 300.

[0088] In another aspect, the present invention provides a composition comprising i) aqueous humor isolated from a subject, and ii) an agent that interacts directly or indirectly with VEGF in the isolated aqueous humor, wherein the isolated aqueous humor comprises a VEGF level above a cutoff value, and the cutoff value is between 10 pg / mL and 100 pg / mL.

[0089] In another aspect, the invention provides a composition as disclosed herein, wherein the agent is an anti-VEGF antibody or an antigen-binding fragment thereof, optionally the antigen-binding fragment is Fab, Fab', F(ab')2 or Fv.

[0090] In another aspect, the present invention provides a composition as disclosed herein, wherein the reagent comprises a VEGF-specific antibody I labeled with alkaline phosphatase, and wherein the labeling ratio of the VEGF antibody I to the alkaline phosphatase is 1:0.75.

[0091] In another aspect, the present invention provides a composition as disclosed herein, wherein the reagent comprises 0.05 mass % of magnetic particles, wherein the magnetic particles are coated with VEGF-specific antibody II, wherein the reagent further comprises a blocking agent, and wherein the blocking agent is a mutant alkaline phosphatase.

[0092] In another aspect, the present invention provides a composition as disclosed herein, wherein the VEGF-specific antibody I and / or the VEGF-specific antibody II is an anti-VEGF165 / VEGFA antibody.

[0093] In another aspect, the present invention provides an anti-VEGF therapeutic for treating retinal vein occlusion (RVO) in a subject, wherein the level of VEGF in the subject's aqueous humor is above a cutoff value, and the cutoff value is between 10 pg / mL and 100 pg / mL.

[0094] In another aspect, the present invention provides an anti-VEGF therapeutic as disclosed herein for use in treating RVO in a subject, the anti-VEGF therapeutic comprising:

[0095] An anti-VEGF antibody or an antigen-binding fragment thereof, optionally the antigen-binding fragment is Fab, Fab', F(ab')2 or Fv;

[0096] A VEGF receptor (VEGFR) fusion protein, optionally wherein the VEGFR fusion protein is a VEGFR-Fc immunoadhesin; or

[0097] An anti-VEGF gene therapy agent comprising a nucleic acid encoding a VEGF antagonist, a nucleic acid encoding a protein that downregulates VEGF / VEGFR expression, and / or a nucleic acid targeting a nucleic acid and / or protein involved in VEGF / VEGFR expression, optionally comprising a viral vector or a non-viral vector, optionally the viral vector is an adeno-associated virus (AAV) vector.

[0098] In another aspect, the present invention provides an anti-VEGF therapeutic as disclosed herein for use in treating RVO in a subject, wherein the anti-VEGF therapeutic is administered or is intended to be administered intravitreally, suprachoroidally and / or subretinally.

[0099] In another aspect, the invention provides an anti-VEGF therapeutic as disclosed herein for use in treating RVO in a subject, wherein the subject suffers from macular edema secondary to retinal vein occlusion (ME-RVO), optionally the retinal vein occlusion is a central retinal vein occlusion (CRVO) and / or a branch retinal vein occlusion (BRVO).

[0100] In another aspect, the invention provides an anti-VEGF therapeutic as disclosed herein for use in treating RVO in a subject, wherein the subject has or has not been treated with the anti-VEGF therapeutic or another anti-VEGF therapeutic.

[0101] In another aspect, the present invention provides an anti-VEGF therapeutic as disclosed herein for use in treating RVO in a subject, wherein the subject has been treated with an anti-inflammatory agent or an immunosuppressant or a combination thereof, wherein the subject's VEGF level in the aqueous humor is above a threshold value, optionally the anti-inflammatory agent or immunosuppressant is selected from glucocorticoids, methotrexate, cyclosporin A, tacrolimus, azathioprine, or a biologic targeting a pro-inflammatory molecule such as interleukin-6, interleukin-8, and tumor necrosis factor-α.

[0102] In another aspect, the present invention provides an anti-VEGF therapeutic as disclosed herein for use in treating RVO in a subject, wherein the subject has not been treated with an anti-inflammatory agent or an immunosuppressant, or a combination thereof, wherein the subject's VEGF level in the aqueous humor is above a threshold value, optionally the anti-inflammatory agent or immunosuppressant is selected from glucocorticoids, methotrexate, cyclosporin A, tacrolimus, azathioprine, or a biologic targeting a pro-inflammatory molecule such as interleukin-6, interleukin-8, and tumor necrosis factor-α.

[0103] In another aspect, the invention provides an anti-VEGF therapeutic as disclosed herein for use in treating RVO in a subject, wherein the cutoff value is between 30 pg / mL and 90 pg / mL.

[0104] In another aspect, the invention provides an anti-VEGF therapeutic as disclosed herein for use in treating RVO in a subject, wherein the cutoff value is about 35 pg / mL, 40 pg / mL, 45 pg / mL, 50 pg / mL, 55 pg / mL, 60 pg / mL, 65 pg / mL, 70 pg / mL, 75 pg / mL, 80 pg / mL or 85 pg / mL.

[0105] In another aspect, the invention provides an anti-VEGF therapeutic as disclosed herein for use in treating RVO in a subject, wherein the subject has an anatomical response to the anti-VEGF therapeutic as measured by a change in central macular thickness (CMT) or foveal thickness (CFT), or as measured by a relative change in central macular thickness (CMT), wherein the relative change in CMT is defined by dividing the change in CMT by the baseline central macular thickness (bl-CMT).

[0106] In another aspect, the present invention provides an anti-VEGF therapeutic as disclosed herein for use in treating RVO in a subject, wherein the subject's baseline central macular thickness (bl-CMT) level is above the bl-CMT cutoff value, and wherein the bl-CMT cutoff value is between about 250 μm and about 600 μm.

[0107] In another aspect, the invention provides a method for predicting an anatomical response of an eye of a subject to treatment with an anti-VEGF therapeutic agent, comprising determining a VEGF level in aqueous humor collected from the subject, wherein a determined VEGF level above a cutoff value indicates that the eye has a good anatomical response to treatment with the anti-VEGF therapeutic agent, wherein a determined VEGF level not above the cutoff value indicates that the eye has a poor anatomical response to treatment with the anti-VEGF therapeutic agent, and wherein the cutoff value is between 10 pg / mL and 100 pg / mL.

[0108] In another aspect, the present invention provides a method for predicting the anatomical response of a subject's eye to treatment with an anti-VEGF therapeutic as disclosed herein, wherein the anatomical response to the anti-VEGF therapeutic is measured by a change in central macular thickness (CMT) or foveal thickness (CFT), wherein CMT or CFT is defined as the perpendicular distance between the inner limiting membrane and the retinal pigment epithelium at the center of the fovea; or wherein the anatomical response to the anti-VEGF therapeutic is measured by a relative change in central macular thickness (CMT), which is defined by dividing the change in CMT by the baseline central macular thickness (bl-CMT).

[0109] In another aspect, the present invention provides a method for predicting the anatomical response of an eye of a subject to treatment with an anti-VEGF therapeutic agent as disclosed herein, wherein a determined VEGF level above a cut-off value predicts a change in CFT of <-200 μm.

[0110] In another aspect, the present invention provides a method for predicting the anatomical response of a subject's eye to treatment with an anti-VEGF therapeutic agent as disclosed herein, wherein a determined VEGF level below a cut-off value predicts a change in CFT > -200 μm.

[0111] In another aspect, the invention provides a method for predicting the anatomical response of an eye of a subject to treatment with an anti-VEGF therapeutic agent as disclosed herein, wherein the cut-off value is between 30 pg / mL and 90 pg / mL.

[0112] In another aspect, the invention provides a method for predicting the anatomical response of a subject's eye to treatment with an anti-VEGF therapeutic agent as disclosed herein, wherein the cutoff value is about 35 pg / mL, 40 pg / mL, 45 pg / mL, 50 pg / mL, 55 pg / mL, 60 pg / mL, 65 pg / mL, 70 pg / mL, 75 pg / mL, 80 pg / mL or 85 pg / mL.

[0113] In another aspect, the invention provides a method for predicting the anatomical response of a subject's eye to treatment with an anti-VEGF therapeutic agent as disclosed herein, wherein a determined VEGF level above 90 pg / mL predicts that the eye has a good anatomical response to treatment with the anti-VEGF therapeutic agent.

[0114] In another aspect, the invention provides a method for predicting an anatomical response of an eye of a subject to treatment with an anti-VEGF therapeutic agent as disclosed herein, wherein a determined VEGF level below 30 pg / mL predicts that the eye will have a poor anatomical response to treatment with the anti-VEGF therapeutic agent.

[0115] In another aspect, the invention provides a method as disclosed herein for predicting the anatomical response of a subject's eye to treatment with an anti-VEGF therapeutic agent, wherein the VEGF level is determined by an immunoassay, optionally an enzyme immunoassay.

[0116] In another aspect, the invention provides a method as disclosed herein for predicting the anatomical response of an eye of a subject to treatment with an anti-VEGF therapeutic agent, wherein the subject is in need of treatment for retinal vein occlusion (RVO).

[0117] In another aspect, the invention provides a method as disclosed herein for predicting the anatomical response of an eye of a subject to treatment with an anti-VEGF therapeutic agent, wherein the subject is in need of treatment for macular edema secondary to retinal vein occlusion (ME-RVO), optionally the retinal vein occlusion is a central retinal vein occlusion (CRVO) and / or a branch retinal vein occlusion (BRVO).

[0118] In another aspect, the present invention provides a method for predicting the anatomical response of a subject's eye to treatment with an anti-VEGF therapeutic agent as disclosed herein, the method further comprising administering the anti-VEGF therapeutic agent or another anti-VEGF therapeutic agent to the subject intravitreally, suprachoroidally and / or subretinally when the determined VEGF level is above a threshold value.

[0119] In another aspect, the invention provides a method for predicting the anatomical response of a subject's eye to treatment with an anti-VEGF therapeutic as disclosed herein, wherein the VEGF level determined is above 90 pg / mL and the anti-VEGF therapeutic or another anti-VEGF therapeutic is administered to the subject intravitreally, suprachoroidally and / or subretinally.

[0120] In another aspect, the present invention provides a method for predicting the anatomical response of a subject's eye to treatment with an anti-VEGF therapeutic agent as disclosed herein, the method further comprising administering to the subject a treatment other than the anti-VEGF therapeutic agent when the determined VEGF level is below a threshold value.

[0121] In another aspect, the invention provides a method for predicting the anatomical response of a subject's eye to treatment with an anti-VEGF therapeutic as disclosed herein, wherein the VEGF level determined is less than 30 pg / mL and the anti-VEGF therapeutic or another anti-VEGF therapeutic is not administered intravitreally, suprachoroidally and / or subretinally to the subject.

[0122] In another aspect, the invention provides a method as disclosed herein for predicting the anatomical response of a subject's eye to treatment with an anti-VEGF therapeutic agent, wherein prior to the determining step, the subject has not been treated with the anti-VEGF therapeutic agent or another anti-VEGF therapeutic agent.

[0123] In another aspect, the invention provides a method as disclosed herein for predicting the anatomical response of a subject's eye to treatment with an anti-VEGF therapeutic agent, wherein prior to the determining step, the subject has been treated with the anti-VEGF therapeutic agent or another anti-VEGF therapeutic agent.

[0124] In another aspect, the present invention provides a method for predicting the anatomical response of a subject's eye to treatment with an anti-VEGF therapeutic agent as disclosed herein, wherein prior to the determining step, the subject has not received steroid treatment, the steroid may be selected as a corticosteroid, further selected as a glucocorticoid.

[0125] In another aspect, the present invention provides a method for predicting the anatomical response of a subject's eye to treatment with an anti-VEGF therapeutic agent as disclosed herein, wherein prior to the determining step, the subject has been receiving steroid therapy, which steroid may be selected as a corticosteroid, further selected as a glucocorticoid.

[0126] In another aspect, the present invention provides a method as disclosed herein for predicting the anatomical response of a subject's eye to treatment with an anti-VEGF therapeutic agent, wherein the subject is selected to initiate, continue or resume use of an anti-VEGF drug or other anti-VEGF drug, and / or to discontinue treatment with a steroid, which may be a corticosteroid, further selected as a glucocorticoid.

[0127] In another aspect, the invention provides a method as disclosed herein for predicting the anatomical response of a subject's eye to treatment with an anti-VEGF therapeutic agent, wherein the subject has chosen to discontinue use of the anti-VEGF drug or other anti-VEGF drug, and / or has chosen to initiate, continue or resume treatment with a steroid, which may be a corticosteroid, further selected as a glucocorticoid.

[0128] In another aspect, the present invention provides a method as disclosed herein for predicting the anatomical response of an eye of a subject to treatment with an anti-VEGF therapeutic agent, wherein the subject is a human.

[0129] In another aspect, the present invention provides a method for predicting the anatomical response of a subject's eye to treatment with an anti-VEGF therapeutic as disclosed herein, wherein the anti-VEGF therapeutic is formulated for intravitreal, suprachoroidal and / or subretinal administration, optionally further comprising mixing the anti-VEGF therapeutic with a pharmaceutically acceptable excipient or carrier.

[0130] In another aspect, the present invention provides a method for predicting an anatomical response of an eye of a subject to treatment with an anti-VEGF therapeutic agent as disclosed herein, the method further comprising determining a bl-CMT level in the subject, wherein a determined bl-CMT level above a bl-CMT cut-off value indicates that the eye has a good anatomical response to treatment with the anti-VEGF therapeutic agent, wherein a determined bl-CMT level not above a bl-CMT cut-off value indicates that the eye has a poor anatomical response to treatment with the anti-VEGF therapeutic agent, and wherein the bl-CMT cut-off value is between about 250 μm and about 600 μm.

[0131] In another aspect, the invention provides a method of predicting an anatomical response of an eye of a subject to treatment with an anti-VEGF therapeutic agent, the method comprising determining a level of VEGF in aqueous humor collected from the subject and a level of bl-CMT in the subject, wherein the determined level of VEGF is above a VEGF cutoff value and the determined level of bl-CMT is above a bl-CMT cutoff value, indicating that the eye has a favorable anatomical response to treatment with the anti-VEGF therapeutic agent; and

[0132] Wherein the determined VEGF level is not above the VEGF cut-off value or the determined bl-CMT level is not above the bl-CMT cut-off value, it indicates that the eye has a poor anatomical response to treatment with the anti-VEGF therapeutic agent.

[0133] In another aspect, the invention provides a method for predicting the anatomical response of an eye of a subject to treatment with an anti-VEGF therapeutic agent as disclosed herein, wherein the VEGF cut-off value is between 10 pg / mL and 100 pg / mL.

[0134] In another aspect, the present invention provides a method for predicting the anatomical response of an eye of a subject to treatment with an anti-VEGF therapeutic agent as disclosed herein, wherein the bl-CMT cutoff value is between about 250 μm and about 600 μm.

[0135] In another aspect, the present invention provides a method for predicting the anatomical response of a subject's eye to treatment with an anti-VEGF therapeutic agent as disclosed herein, wherein the anatomical response to the anti-VEGF therapeutic agent is measured by changes in CMT, which is defined as the perpendicular distance between the internal limiting membrane and the retinal pigment epithelium at the center of the fovea.

[0136] In another aspect, the present invention provides a method for predicting the anatomical response of a subject's eye to treatment with an anti-VEGF therapeutic agent as disclosed herein, wherein the anatomical response to the anti-VEGF therapeutic agent is measured by a relative change in CMT, wherein the relative change in CMT is defined by dividing the change in CMT by bl-CMT.

[0137] In another aspect, the present invention provides a method for predicting the anatomical response of a subject's eye to treatment with an anti-VEGF therapeutic agent as disclosed herein, wherein the determined VEGF level is above a VEGF cutoff value of 10 pg / mL to 100 pg / mL and the determined bl-CMT level is above a bl-CMT cutoff value of about 250 μm to about 600 μm, indicating that the eye has a good anatomical response to treatment with the anti-VEGF therapeutic agent.

[0138] In another aspect, the present invention provides a method for predicting the anatomical response of a subject's eye to treatment with an anti-VEGF therapeutic agent as disclosed herein, wherein the determined VEGF level is above a VEGF cutoff value and the determined bl-CMT level is above a bl-CMT cutoff value, predicting a change in CFT of <-200 μm.

[0139] In another aspect, the present invention provides a method for predicting the anatomical response of a subject's eye to treatment with an anti-VEGF therapeutic agent as disclosed herein, wherein the determined VEGF level is below a VEGF cutoff value or the determined bl-CMT level is below a bl-CMT cutoff value, predicting a change in CFT of >-200 μm.

[0140] In another aspect, the invention provides a method for predicting the anatomical response of an eye of a subject to treatment with an anti-VEGF therapeutic agent as disclosed herein, wherein the VEGF cut-off value is between 30 pg / mL and 90 pg / mL.

[0141] In another aspect, the invention provides a method for predicting the anatomical response of a subject's eye to treatment with an anti-VEGF therapeutic agent as disclosed herein, wherein the VEGF cutoff value is about 35 pg / mL, 40 pg / mL, 45 pg / mL, 50 pg / mL, 55 pg / mL, 60 pg / mL, 65 pg / mL, 70 pg / mL, 75 pg / mL, 80 pg / mL or 85 pg / mL.

[0142] In another aspect, the present invention provides a method for predicting the anatomical response of an eye of a subject to treatment with an anti-VEGF therapeutic agent as disclosed herein, wherein the bl-CMT cut-off value is between 250 μm and 600 μm.

[0143] In another aspect, the invention provides a method for predicting the anatomical response of a subject's eye to treatment with an anti-VEGF therapeutic agent as disclosed herein, wherein the bl-CMT cutoff is about 250 μm, 300 μm, 350 μm, 400 μm, 450 μm, 500 μm, 550 μm or 600 μm.

[0144] In another aspect, the invention provides a method as disclosed herein for predicting the anatomical response of a subject's eye to treatment with an anti-VEGF therapeutic agent, wherein the level of VEGF is determined by an immunoassay, optionally an enzyme immunoassay.

[0145] In another aspect, the invention provides a method as disclosed herein for predicting the anatomical response of an eye of a subject to treatment with an anti-VEGF therapeutic agent, wherein the subject is in need of treatment for retinal vein occlusion (RVO).

[0146] In another aspect, the invention provides a method as disclosed herein for predicting the anatomical response of an eye of a subject to treatment with an anti-VEGF therapeutic agent, wherein the subject is in need of treatment for macular edema secondary to retinal vein occlusion (ME-RVO), optionally the retinal vein occlusion is a central retinal vein occlusion (CRVO) and / or a branch retinal vein occlusion (BRVO).

[0147] In another aspect, the invention provides a method for predicting an anatomical response of an eye of a subject to treatment with an anti-VEGF therapeutic as disclosed herein, the method further comprising administering the anti-VEGF therapeutic or another anti-VEGF therapeutic to the subject intravitreally, suprachoroidally and / or subretinally when the determined VEGF level is above a VEGF cutoff value and when the determined bl-CMT level is above a bl-CMT cutoff value.

[0148] In another aspect, the invention provides a method for predicting the anatomical response of a subject's eye to treatment with an anti-VEGF therapeutic as disclosed herein, wherein the VEGF level determined is above 90 pg / mL and the anti-VEGF therapeutic or another anti-VEGF therapeutic is administered to the subject intravitreally, suprachoroidally and / or subretinally.

[0149] In another aspect, the present invention provides a method for predicting the anatomical response of a subject's eye to treatment with an anti-VEGF therapeutic agent as disclosed herein, the method further comprising administering to the subject a treatment other than the anti-VEGF therapeutic agent when the determined VEGF level is below the VEGF threshold.

[0150] In another aspect, the invention provides a method for predicting the anatomical response of a subject's eye to treatment with an anti-VEGF therapeutic as disclosed herein, wherein the VEGF level determined is less than 30 pg / mL and the anti-VEGF therapeutic or another anti-VEGF therapeutic is not administered to the subject intravitreally, suprachoroidally and / or subretinally.

[0151] In another aspect, the invention provides a method as disclosed herein for predicting the anatomical response of a subject's eye to treatment with an anti-VEGF therapeutic agent, wherein the treatment other than the anti-VEGF therapeutic agent is a steroid, optionally a corticosteroid, further optionally a glucocorticoid.

[0152] In another aspect, the invention provides a method as disclosed herein for predicting the anatomical response of a subject's eye to treatment with an anti-VEGF therapeutic agent, wherein prior to the determining step, the subject has or has not been treated with the anti-VEGF therapeutic agent or another anti-VEGF therapeutic agent.

[0153] In another aspect, the present invention provides a method for predicting the anatomical response of a subject's eye to treatment with an anti-VEGF therapeutic agent as disclosed herein, wherein prior to the determining step, the subject has or has not received steroid treatment, the steroid may be selected as a corticosteroid, further selected as a glucocorticoid.

[0154] In another aspect, the present invention provides a method as disclosed herein for predicting the anatomical response of a subject's eye to treatment with an anti-VEGF therapeutic agent, wherein the subject has or has not been treated with an anti-inflammatory agent or an immunosuppressant or a combination thereof, optionally the anti-inflammatory agent or immunosuppressant is selected from glucocorticoids, methotrexate, cyclosporin A, tacrolimus, azathioprine or a biologic targeting a pro-inflammatory molecule such as interleukin-6, interleukin-8 and tumor necrosis factor-α.

[0155] In another aspect, the present invention provides a method as disclosed herein for predicting the anatomical response of a subject's eye to treatment with an anti-VEGF therapeutic agent, wherein the subject is selected to initiate, continue or resume use of an anti-VEGF drug or other anti-VEGF drug, and / or to discontinue treatment with a steroid, which may be a corticosteroid, further selected as a glucocorticoid.

[0156] In another aspect, the invention provides a method as disclosed herein for predicting the anatomical response of a subject's eye to treatment with an anti-VEGF therapeutic agent, wherein the subject has chosen to discontinue use of the anti-VEGF drug or other anti-VEGF drug, and / or has chosen to initiate, continue or resume treatment with a steroid, which may be a corticosteroid, further selected as a glucocorticoid.

[0157] In another aspect, the present invention provides a method as disclosed herein for predicting the anatomical response of an eye of a subject to treatment with an anti-VEGF therapeutic agent, wherein the subject is a human.

[0158] In another aspect, the present invention provides a method as disclosed herein for predicting the anatomical response of a subject's eye to treatment with an anti-VEGF therapeutic agent,

[0159] In another aspect, the present invention provides a method for predicting the anatomical response of a subject's eye to treatment with an anti-VEGF therapeutic as disclosed herein, wherein the anti-VEGF therapeutic is formulated for intravitreal, suprachoroidal and / or subretinal administration, optionally further comprising mixing the anti-VEGF therapeutic with a pharmaceutically acceptable excipient or carrier. BRIEF DESCRIPTION OF THE DRAWINGS

[0160] The accompanying drawings illustrate certain features and advantages of the invention. These embodiments are not intended to limit the scope of the appended claims in any way.

[0161] Figure 1 Changes in central foveal thickness (CFT, defined as the perpendicular distance between the internal limiting membrane and the retinal pigment epithelium at the center of the fovea) or maximum parafoveal retinal thickness (MRT) 4 weeks after intravitreal injection without foveal involvement compared with baseline plotted against baseline VEGF levels in aqueous humor are described.

[0162] Figure 2 Comparison of baseline aqueous humor cytokine concentrations and clinical parameters between the two subgroups. Figures AH: Comparison of aqueous humor interleukin (IL)-6, IL-8, interferon gamma-induced protein-10 (IP-10), IL-10, monocyte chemoattractant protein-1 (MCP-1), vascular endothelial growth factor (VEGF) detected by magnetic particle-based chemiluminescent enzyme immunoassay kit (MECLEIA), baseline central macular thickness (bl-CMT) and best corrected visual acuity (bl-BCVA) (logarithm of the minimum angle of resolution (logMAR) scale) between the untreated and treated groups.

[0163] Figure 3 Receiver operating characteristic (ROC) curves were used to identify critical values ​​and evaluate regression models. Figure 3 A and Figure 3 B: Cut-off values ​​of aqueous humor interleukin (IL)-8 and vascular endothelial growth factor (VEGF) detected by a magnetic particle-based chemiluminescent enzyme immunoassay kit (VEGF-MECLEIA) for predicting ATR as measured by change in central macular thickness (ΔCMT) / baseline central macular thickness (bl-CMT). Figure 3C: Area under the curve (AUC) of the multivariate logistic regression model for VEGF+bl-CMT. Figure 3 D and Figure 3 E: AUC of the multivariate logistic regression model for VEGF and bl-CMT treatment response.

[0164] Figure 4 These are representative cases of the good response (GR) group and the adverse response (PR) group. Figure 4 A and Figure 4 B is the baseline fundus photographs and optical coherence tomography (OCT) scans of representative patients in the GR group and the untreated group. Before intravitreal injection of ranibizumab (IVR), the baseline central macular thickness (bl-CMT) and baseline central retinal thickness (bl-CRT) were 404 μm and 357 μm, respectively; VEGF in aqueous humor detected by magnetic particle-based chemiluminescent enzyme immunoassay kit (VEGF-MPCLEIA) was 129.8 pg / ml. Figure 4 C and Figure 4 D is the fundus photograph and OCT scan of this case 5 weeks after IVR. The CMT and CRT decreased to 148 μm and 195 μm, respectively, and the retinal edema completely subsided. Figure 4 E and Figure 4 F is the baseline fundus photographs and OCT scans of representative cases in the PR and treatment groups. The bl-CMT and bl-CRT were 714 μm and 405 μm, respectively. Figure 4 G and Figure 4 H is a fundus photograph and OCT scan of the same patient 5 weeks after anti-VEGF injection, with CMT and CRT of 730 μm and 402 μm, respectively. DETAILED DESCRIPTION

[0165] All publications, including patent documents, scientific articles, and databases, mentioned in this application are incorporated by reference in their entirety for all purposes to the same extent as if each individual publication was individually incorporated by reference. If definitions set forth herein are contrary to or inconsistent with definitions set forth in patents, applications, published applications, and other publications incorporated herein by reference, definitions set forth herein take precedence over definitions incorporated herein by reference.

[0166] The section headings used herein are for organizational purposes only and are not to be construed as limiting the subject matter described.

[0167] In some embodiments, provided herein is the use of VEGF levels in aqueous humor as a companion biomarker for anti-VEGF treatment in RVO, and methods and compositions related to the use. In some embodiments, the level of VEGF in the aqueous humor of a subject can be used to predict the subject's responsiveness to one or more anti-VEGF therapeutic agents. In some embodiments, VEGF levels in aqueous humor that are above the VEGF cutoff or in a range between about 10 pg / mL and about 100 pg / mL can be used as a companion diagnostic to predict a patient's anatomical treatment response to anti-VEGF. In some embodiments, the anti-VEGF therapeutic agent herein is an agent that targets a signaling pathway mediated by VEGF, such as by VEGFR. In some embodiments, the anti-VEGF therapeutic agent targets VEGF. In some embodiments, the anti-VEGF therapeutic agent targets VEGFR.

[0168] In some embodiments, provided herein is the use of aqueous humor VEGF levels in combination with bl-CMT levels as a companion biomarker for anti-VEGF treatment in RVO, and methods and compositions related to the use. In some embodiments, the combination of VEGF levels in the subject's aqueous humor and bl-CMT levels in the subject can be used to predict the subject's responsiveness to one or more anti-VEGF therapeutics. In some embodiments, the VEGF level in the aqueous humor is above the VEGF cutoff or in the range between about 10 pg / mL and about 100 pg / mL, and the bl-CMT level in the subject is above the bl-CMT cutoff or in the range between about 250 μm and about 600 μm, which can be used as a companion diagnostic to predict the patient's anatomical treatment response to anti-VEGF. In some embodiments, the anti-VEGF therapeutic agent herein is an agent that targets a signaling pathway mediated by VEGF, such as by VEGFR. In some embodiments, the anti-VEGF therapeutic agent targets VEGF. In some embodiments, the anti-VEGF therapeutic agent targets VEGFR.

[0169] In any of the embodiments herein, the anti-VEGF therapeutic may comprise an anti-VEGF antibody or an antigen-binding fragment thereof, optionally the antigen-binding fragment is a Fab, Fab', F(ab')2, or Fv.

[0170] In any of the embodiments herein, the anti-VEGF therapeutic may comprise a VEGF receptor (VEGFR) fusion protein, optionally the VEGFR fusion protein is a VEGFR-Fc immunoadhesin.

[0171] In any embodiment herein, the anti-VEGF therapeutic agent may comprise an anti-VEGF gene therapy agent. In any embodiment herein, the anti-VEGF therapeutic agent may comprise a nucleic acid encoding a VEGF antagonist. In any embodiment herein, the anti-VEGF therapeutic agent may comprise a nucleic acid encoding a VEGFR antagonist. In any embodiment herein, the anti-VEGF therapeutic agent may comprise a nucleic acid encoding a protein or nucleic acid that downregulates VEGF and / or VEGFR expression. In any embodiment herein, the anti-VEGF therapeutic agent may comprise a nucleic acid that downregulates VEGF and / or VEGFR expression.

[0172] In any of the embodiments herein, the anti-VEGF therapeutic may comprise a viral vector or a non-viral vector, optionally the viral vector is an adeno-associated virus (AAV) vector.

[0173] In any of the embodiments herein, the anti-VEGF therapeutic agent may comprise a nucleic acid (DNA or RNA) that targets a nucleic acid or protein involved in VEGF and / or VEGFR expression. In any of the embodiments herein, the anti-VEGF therapeutic agent may comprise a VEGF antisense oligodeoxynucleotide and / or a VEGFR antisense oligodeoxynucleotide.

[0174] In any embodiment herein, the anti-VEGF therapeutic agent may be administered or prepared for administration intravitreally, suprachoroidally and / or subretinally. In any embodiment herein, the subject may have macular edema secondary to retinal vein occlusion (ME-RVO), optionally central retinal vein occlusion (CRVO) and / or branch retinal vein occlusion (BRVO).

[0175] In any embodiment herein, the subject may have been treated with the anti-VEGF therapeutic agent or another anti-VEGF therapeutic agent. In any embodiment herein, the subject may not have been treated with the anti-VEGF therapeutic agent or another anti-VEGF therapeutic agent. In any embodiment herein, the subject may have been treated with steroids, which may be selected as corticosteroids, further selected as glucocorticoids.

[0176] In any of the embodiments herein, the subject may not have been receiving steroid therapy, and the steroid may be selected as a corticosteroid, further selected as a glucocorticoid.

[0177] In any embodiment herein, the subject may have been treated with an anti-inflammatory agent or an immunosuppressant, or a combination thereof, optionally selected from glucocorticoids, methotrexate, cyclosporin A, tacrolimus, azathioprine, or a biological agent targeting a pro-inflammatory molecule such as interleukin-6, interleukin-8, and tumor necrosis factor-α. In any embodiment herein, the subject may not have been treated with an anti-inflammatory agent or an immunosuppressant, or a combination thereof, optionally selected from glucocorticoids,

[0178] In any embodiment herein, VEGF threshold value can be between 30pg / mL and 90pg / mL. In any embodiment herein, VEGF threshold value can be about 35pg / mL, 40pg / mL, 45pg / mL, 50pg / mL, 55pg / mL, 60pg / mL, 65pg / mL, 70pg / mL, 75pg / mL, 80pg / mL or 85pg / mL.

[0179] In any of the embodiments herein, a subject may have an anatomical response to an anti-VEGF therapeutic agent, or what is referred to as an "anatomical therapeutic response (ATR)", such as measured by a change in foveal thickness (CFT, defined as the perpendicular distance between the inner limiting membrane and the retinal pigment epithelium at the center of the fovea), or a change in central macular thickness (CMT, defined as the perpendicular distance between the inner limiting membrane and the retinal pigment epithelium at the center of the fovea), a relative change in CMT (defined as the change in CMT divided by the baseline central macular thickness (bl-CMT), i.e., ΔCMT / bl-CMT), a change in mean macular thickness (MMT, defined as the mean retinal thickness of all 9 areas of the ETDRS map), or a change in maximum parafoveal retinal thickness (MRT), or other measurements derived from any of the above measurements. In any of the embodiments herein, a subject may have an anatomical response to an anti-VEGF therapeutic agent as measured by ΔCFT / CFT. In some embodiments, the anatomical response or ATR to an anti-VEGF therapeutic agent is measured by a change in CFT or CMT. In some embodiments, the anatomical response or ATR to an anti-VEGF therapeutic agent is measured by a relative change in CMT. A relative CMT change (ΔCMT / bl-CMT) ≤ -25% after intravitreal injection was classified as a good anatomic response or a good ATR. A relative CMT change (ΔCMT / bl-CMT) > -25% after intravitreal injection was classified as a poor anatomic response or a poor ATR.

[0180] In some embodiments, provided herein is the use of an anti-VEGF therapeutic agent in the preparation of a kit for treating retinal vein occlusion (RVO), wherein the kit comprises: i) an anti-VEGF therapeutic agent and ii) instructions for administering the anti-VEGF therapeutic agent to a subject whose VEGF level in aqueous humor is above a VEGF cutoff value, and wherein the VEGF cutoff value is between 10 pg / mL and 100 pg / mL. In some embodiments, the kit further comprises instructions for not administering the anti-VEGF therapeutic agent to a subject whose VEGF level in aqueous humor is not above a VEGF cutoff value. In any embodiment herein, the kit may comprise instructions for administering a steroid to a subject whose VEGF level in aqueous humor is not above a VEGF cutoff value, and the steroid may be selected as a corticosteroid, further selected as a glucocorticoid. In any of the embodiments herein, the kit may include instructions for administering an anti-inflammatory agent or an immunosuppressant, or a combination thereof, to a subject whose VEGF level in aqueous humor is above a threshold value, optionally the anti-inflammatory agent or immunosuppressant is selected from glucocorticoids, methotrexate, cyclosporin A, tacrolimus, azathioprine, or a biologic targeting a pro-inflammatory molecule such as interleukin-6, interleukin-8, and tumor necrosis factor-α.

[0181] In some embodiments, provided herein is a use of an anti-VEGF therapeutic agent in the preparation of a kit for treating retinal vein occlusion (RVO), wherein the kit comprises: i) the anti-VEGF therapeutic agent and ii) instructions for administering the anti-VEGF therapeutic agent to a subject having a VEGF level in aqueous humor above a VEGF cutoff value between 10 pg / mL and 100 pg / mL and a bl-CMT level above a bl-CMT cutoff value between about 250 μm and about 600 μm. In some embodiments, the kit further comprises instructions for not administering the anti-VEGF therapeutic agent to a subject having a VEGF level in aqueous humor not above a VEGF cutoff value or a bl-CMT level not above a bl-CMT cutoff value. In any embodiment herein, the kit may comprise instructions for administering a steroid to a subject having a VEGF level in aqueous humor not above a VEGF cutoff value or a bl-CMT level not above a bl-CMT cutoff value, the steroid being selected as a corticosteroid, further selected as a glucocorticoid. In any of the embodiments herein, the kit may include instructions for administering an anti-inflammatory agent or an immunosuppressant, or a combination thereof, to a subject whose VEGF level in the aqueous humor is above a cutoff value or whose bl-CMT level is not above a cutoff value for bl-CMT, optionally the anti-inflammatory agent or immunosuppressant is selected from glucocorticoids, methotrexate, cyclosporin A, tacrolimus, azathioprine, or a biologic targeting a pro-inflammatory molecule such as interleukin-6, interleukin-8, and tumor necrosis factor-α.

[0182] In some embodiments, provided herein is a kit comprising i) an anti-VEGF therapeutic and ii) instructions for administering the anti-VEGF therapeutic to a subject whose VEGF level in aqueous humor is above a VEGF cutoff value, wherein the VEGF cutoff value is between 10 pg / mL and 100 pg / mL. In any embodiments herein, the kit may include instructions for not administering the anti-VEGF therapeutic to a subject whose VEGF level in aqueous humor is not above the VEGF cutoff value. In any embodiments herein, the kit may include instructions for administering a steroid to a subject whose VEGF level in aqueous humor is not above the VEGF cutoff value, and the steroid may be selected as a corticosteroid, further selected as a glucocorticoid.

[0183] In some embodiments, provided herein is a kit comprising: i) reagents for determining VEGF levels in a subject's aqueous humor, and ii) instructions comprising a VEGF cutoff value for the VEGF level, and optionally instructions for comparing the determined VEGF level to the VEGF cutoff value, wherein the VEGF cutoff value is between 10 pg / mL and 100 pg / mL.

[0184] In any embodiment herein, the kit may include a first standard sample having a VEGF level substantially equal to a VEGF critical value. In any embodiment herein, the kit may include a second standard sample having a VEGF level higher than a VEGF critical value. In any embodiment herein, the kit may include a third standard sample having a VEGF level lower than a VEGF critical value.

[0185] In any of the embodiments herein, the agent can be an anti-VEGF antibody or an antigen binding fragment thereof, optionally the antigen binding fragment is a Fab, Fab', F(ab')2 or Fv.

[0186] In any embodiment herein, the kit may include an enzyme reactant, a magnetic separation reagent, a stabilizer, a standard, one or more concentrated washing buffers and / or a substrate buffer. In any embodiment herein, the enzyme reactant may include a VEGF-specific antibody I labeled with alkaline phosphatase, wherein the labeling ratio of the VEGF antibody I to the alkaline phosphatase is 1:0.75. In any embodiment herein, the magnetic separation reagent may contain 0.05% by mass of magnetic particles, wherein these magnetic particles are coated with VEGF-specific antibody II, wherein the magnetic reagent also includes a blocking agent, and wherein the blocking agent is a mutant alkaline phosphatase. In any embodiment herein, the VEGF-specific antibody I and / or the VEGF-specific antibody II may be an anti-VEGF165 / VEGFA antibody. In any embodiment herein, the stabilizer may include one or more of Tris, NaCl, Tween20, bovine serum albumin, mutant alkaline phosphatase, and ProClin 300. In any of the embodiments herein, the wash buffer may comprise Tris, NaCl, Tween-20, Triton-100 and ProClin300.

[0187] In some embodiments, provided herein is a composition comprising i) aqueous humor isolated from a subject, and ii) an agent that interacts directly or indirectly with VEGF in the isolated aqueous humor, wherein the isolated aqueous humor comprises a VEGF level above a VEGF cutoff value, and the VEGF cutoff value is between 10 pg / mL and 100 pg / mL.

[0188] In some embodiments, provided herein is a composition comprising i) aqueous humor isolated from a subject, and ii) an agent that interacts directly or indirectly with VEGF in the isolated aqueous humor, wherein the isolated aqueous humor comprises a VEGF level that is no greater than a VEGF cutoff value, and the VEGF cutoff value is between 10 pg / mL and 100 pg / mL.

[0189] In any embodiment herein, the reagent may be an anti-VEGF antibody or an antigen-binding fragment thereof, optionally the antigen-binding fragment is Fab, Fab', F(ab')2 or Fv. In any embodiment herein, the reagent may comprise a VEGF-specific antibody I labeled with alkaline phosphatase, and wherein the labeling ratio of the VEGF antibody I to the alkaline phosphatase is 1:0.75. In any embodiment herein, the reagent may comprise 0.05% by mass of magnetic particles, wherein the magnetic particles are coated with a VEGF-specific antibody II, wherein the reagent further comprises a blocker, and wherein the blocker is a mutant alkaline phosphatase. In any embodiment herein, the VEGF-specific antibody I and / or the VEGF-specific antibody II may be an anti-VEGF165 / VEGFA antibody.

[0190] In some embodiments, disclosed herein is an anti-VEGF therapeutic for treating retinal vein occlusion (RVO) in a subject, wherein the subject has a VEGF level in aqueous humor that is above a VEGF cutoff value, and the VEGF cutoff value is between 10 pg / mL and 100 pg / mL.

[0191] In any embodiment herein, the anti-VEGF therapeutic agent may include an anti-VEGF antibody or an antigen binding fragment thereof, optionally the antigen binding fragment is Fab, Fab', F(ab')2 or Fv. In any embodiment herein, the anti-VEGF therapeutic agent may include a VEGF receptor (VEGFR) fusion protein, optionally the VEGFR fusion protein is a VEGFR-Fc immunoadhesin.

[0192] In any embodiment herein, the anti-VEGF therapeutic agent may include an anti-VEGF gene therapy agent comprising a nucleic acid encoding a VEGF antagonist, optionally the anti-VEGF gene therapy agent comprises a viral vector or a non-viral vector, optionally the viral vector is an adeno-associated virus (AAV) vector. In any embodiment herein, the anti-VEGF therapeutic agent may include a nucleic acid encoding a VEGFR antagonist. In any embodiment herein, the anti-VEGF therapeutic agent may include a nucleic acid encoding a protein or nucleic acid that downregulates VEGF and / or VEGFR expression. In any embodiment herein, the anti-VEGF therapeutic agent may include a nucleic acid that downregulates VEGF and / or VEGFR expression. In any embodiment herein, the anti-VEGF therapeutic agent may include a nucleic acid (DNA or RNA) targeting a nucleic acid or protein involved in VEGF and / or VEGFR expression. In any embodiment herein, the anti-VEGF therapeutic agent may include a VEGF antisense oligodeoxynucleotide and / or a VEGFR antisense oligodeoxynucleotide. In any embodiment herein, the anti-VEGF therapeutic agent may be administered or prepared for administration intravitreally, suprachoroidally and / or subretinaly.

[0193] In any of the embodiments herein, the subject may have macular edema secondary to retinal vein occlusion (ME-RVO), optionally the retinal vein occlusion is a central retinal vein occlusion (CRVO) and / or a branch retinal vein occlusion (BRVO).

[0194] In any of the embodiments herein, the subject may have been treated with the anti-VEGF therapeutic or another anti-VEGF therapeutic.

[0195] In any of the embodiments herein, the subject may not have been treated with the anti-VEGF therapeutic or another anti-VEGF therapeutic.

[0196] In any of the embodiments herein, the subject may have been receiving steroid therapy, which steroid may be selected as a corticosteroid, further selected as a glucocorticoid.

[0197] In any of the embodiments herein, the subject may not have been receiving steroid therapy, and the steroid may be selected as a corticosteroid, further selected as a glucocorticoid.

[0198] In any embodiment herein, VEGF critical value can include certain scope, and i.e. VEGF critical value can be VEGF critical range. In any embodiment herein, VEGF critical value can be any value or scope between 10pg / mL and 100pg / mL. In any embodiment herein, VEGF critical value can be any value or scope between 30pg / mL and 90pg / mL. In any embodiment herein, VEGF critical value can be about 35pg / mL, 40pg / mL, 45pg / mL, 50pg / mL, 55pg / mL, 60pg / mL, 65pg / mL, 70pg / mL, 75pg / mL, 80pg / mL or 85pg / mL.

[0199] In any of the embodiments herein, the subject may have an anatomical response to an anti-VEGF therapeutic as measured by a change in CFT.

[0200] In some embodiments, disclosed herein is a method of determining an anatomical response of an eye of a subject to treatment with an anti-VEGF therapeutic agent, comprising determining a VEGF level in aqueous humor collected from the subject, wherein a determined VEGF level above a VEGF cutoff value indicates that the eye has a good anatomical response to treatment with the anti-VEGF therapeutic agent, wherein a determined VEGF level not above a VEGF cutoff value indicates that the eye has a poor anatomical response to treatment with the anti-VEGF therapeutic agent, and wherein the VEGF cutoff value is between 10 pg / mL and 100 pg / mL.

[0201] In any embodiment herein, the anatomical response to an anti-VEGF therapeutic agent can be measured by a change in CFT. In any embodiment herein, a measured VEGF level above a VEGF cutoff value can be associated with a change in CFT <-200 μm and / or used to predict a change in CFT <-200 μm. In any embodiment herein, a measured VEGF level below a VEGF cutoff value can be associated with a change in CFT >-200 μm and / or used to predict a change in CFT >-200 μm.

[0202] In any embodiment herein, VEGF threshold value can be between 30pg / mL and 90pg / mL. In any embodiment herein, VEGF threshold value can be about 35pg / mL, 40pg / mL, 45pg / mL, 50pg / mL, 55pg / mL, 60pg / mL, 65pg / mL, 70pg / mL, 75pg / mL, 80pg / mL or 85pg / mL.

[0203] In any of the embodiments herein, a VEGF level determined to be above 90 pg / mL can be associated with and / or used to predict a good anatomical response of the eye to treatment with an anti-VEGF therapeutic agent.

[0204] In any of the embodiments herein, a VEGF level determined to be less than 30 pg / mL can be associated with and / or used to predict a poor anatomical response of the eye to treatment with an anti-VEGF therapeutic agent.

[0205] In any of the embodiments herein, the level of VEGF may be determined by an immunoassay, optionally an enzyme immunoassay.

[0206] In any embodiment herein, the subject may need treatment for retinal vein occlusion (RVO). In any embodiment herein, the subject may need treatment for macular edema secondary to retinal vein occlusion (ME-RVO), optionally the retinal vein occlusion is a central retinal vein occlusion (CRVO) and / or a branch retinal vein occlusion (BRVO).

[0207] In any of the embodiments herein, when the determined VEGF level is above the VEGF cutoff value, the method may further comprise administering to the subject intravitreally, suprachoroidally, or subretinally an anti-VEGF therapeutic or other anti-VEGF therapeutic.

[0208] In any embodiment herein, the VEGF level determined may be above 90 pg / mL, and the subject is administered an intravitreal, suprachoroidal or subretinal anti-VEGF therapeutic or other anti-VEGF therapeutic. In some embodiments, the subject is not treated with steroids, which may be selected as corticosteroids, further selected as glucocorticoids.

[0209] In any of the embodiments herein, when the determined VEGF level is below the VEGF cutoff value, the method may further comprise administering to the subject a treatment other than the anti-VEGF therapeutic agent.

[0210] In any of the embodiments herein, the VEGF level determined may be less than 30 pg / mL and the subject is not administered intravitreal, suprachoroidal, or subretinal anti-VEGF therapeutics or other anti-VEGF therapeutics.

[0211] In any of the embodiments herein, the treatment other than the anti-VEGF therapeutic may be a steroid, optionally a corticosteroid, further optionally a glucocorticoid.

[0212] In some embodiments, the present invention provides a method of treating retinal vein occlusion (RVO) in a subject, comprising administering to the subject an effective amount of an anti-VEGF therapeutic, wherein the subject has a VEGF level in aqueous humor that is above a VEGF cutoff value, and the VEGF cutoff value is between 10 pg / mL and 100 pg / mL.

[0213] In some embodiments, the invention provides methods of treating RVO in a subject as disclosed herein, wherein the anti-VEGF therapeutic comprises:

[0214] An anti-VEGF antibody or an antigen-binding fragment thereof, optionally the antigen-binding fragment is Fab, Fab', F(ab')2 or Fv;

[0215] A VEGF receptor (VEGFR) fusion protein, optionally the VEGFR fusion protein is a VEGFR-Fc immunoadhesin; or

[0216] An anti-VEGF gene therapy agent, optionally the anti-VEGF gene therapy agent comprises a nucleic acid encoding a VEGF antagonist, a nucleic acid encoding a protein that downregulates VEGF / VEGFR expression, and / or a nucleic acid targeting a nucleic acid and / or protein involved in VEGF / VEGFR expression, optionally the anti-VEGF gene therapy agent comprises a viral vector or a non-viral vector, optionally the viral vector is an adeno-associated virus (AAV) vector.

[0217] In some embodiments, the invention provides methods of treating RVO in a subject as disclosed herein, wherein the anti-VEGF therapeutic is administered or is intended to be administered intravitreally, suprachoroidally, and / or subretinally.

[0218] In some embodiments, the invention provides a method of treating RVO in a subject as disclosed herein, wherein the subject has macular edema secondary to retinal vein occlusion (ME-RVO), optionally wherein the retinal vein occlusion is a central retinal vein occlusion (CRVO) and / or a branch retinal vein occlusion (BRVO).

[0219] In some embodiments, the invention provides a method of treating RVO in a subject as disclosed herein, wherein the subject has macular edema secondary to retinal vein occlusion (ME-RVO), optionally wherein the retinal vein occlusion is a central retinal vein occlusion (CRVO) and / or a branch retinal vein occlusion (BRVO).

[0220] In some embodiments, the invention provides methods of treating RVO in a subject as disclosed herein, wherein the subject has been treated with the anti-VEGF therapeutic or another anti-VEGF therapeutic.

[0221] In some embodiments, the invention provides methods of treating RVO in a subject as disclosed herein, wherein the subject has not been treated with the anti-VEGF therapeutic or another anti-VEGF therapeutic.

[0222] In some embodiments, the present invention provides a method of treating RVO in a subject as disclosed herein, wherein the subject has been treated with an anti-inflammatory agent or an immunosuppressant or a combination thereof, wherein the subject's VEGF level in the aqueous humor is above a threshold value, optionally the anti-inflammatory agent or immunosuppressant is selected from glucocorticoids, methotrexate, cyclosporin A, tacrolimus, azathioprine, or a biologic targeting a pro-inflammatory molecule such as interleukin-6, interleukin-8, and tumor necrosis factor-α.

[0223] In some embodiments, the present invention provides a method of treating RVO in a subject as disclosed herein, wherein the subject has not been treated with an anti-inflammatory agent or an immunosuppressant, or a combination thereof, wherein the subject's VEGF level in the aqueous humor is above a threshold value, optionally the anti-inflammatory agent or immunosuppressant is selected from glucocorticoids, methotrexate, cyclosporin A, tacrolimus, azathioprine, or a biologic targeting a pro-inflammatory molecule such as interleukin-6, interleukin-8, and tumor necrosis factor-α.

[0224] In some embodiments, the present invention provides a method of treating RVO in a subject as disclosed herein, wherein the subject has been treated with a steroid, which may be selected as a corticosteroid, further selected as a glucocorticoid.

[0225] In some embodiments, the present invention provides a method of treating RVO in a subject as disclosed herein, wherein the subject has not been treated with a steroid, the steroid may be selected as a corticosteroid, further selected as a glucocorticoid.

[0226] In some embodiments, the invention provides a method of treating RVO in a subject as disclosed herein, wherein the VEGF cutoff is between 30 pg / mL and 90 pg / mL.

[0227] In some embodiments, the invention provides a method of treating RVO in a subject as disclosed herein, wherein the VEGF cutoff is about 35 pg / mL, 40 pg / mL, 45 pg / mL, 50 pg / mL, 55 pg / mL, 60 pg / mL, 65 pg / mL, 70 pg / mL, 75 pg / mL, 80 pg / mL, or 85 pg / mL.

[0228] In some embodiments, the invention provides a method of treating RVO in a subject as disclosed herein, wherein the subject has an anatomical response to the anti-VEGF therapeutic as measured by a change in central macular thickness (CMT) or foveal thickness (CFT), wherein CMT or CFT is defined as the perpendicular distance between the internal limiting membrane and the retinal pigment epithelium at the center of the fovea, and in some cases is also described as central foveal thickness (CFT).

[0229] In some embodiments, the invention provides a method of treating RVO in a subject as disclosed herein, wherein the subject has an anatomical response to the anti-VEGF therapeutic as measured by a relative change in central macular thickness (CMT), wherein the relative change in CMT is defined by the change in CMT divided by the baseline central macular thickness (bl-CMT).

[0230] In some embodiments, the present invention provides a method of treating RVO in a subject as disclosed herein, wherein the subject has a baseline central macular thickness (bl-CMT) level above a bl-CMT cutoff value, wherein the bl-CMT cutoff value is between about 250 μm and about 600 μm.

[0231] In some embodiments, the present invention provides a method of treating retinal vein occlusion (RVO) in a subject as disclosed herein, comprising:

[0232] (a) determining the level of VEGF in aqueous humor collected from the subject and the level of bl-CMT in the subject, wherein the determined level of VEGF is above a VEGF cutoff value and the determined level of bl-CMT is above a bl-CMT cutoff value,

[0233] (b) administering to the subject an effective amount of an anti-VEGF therapeutic agent.

[0234] In some embodiments, the invention provides a method of treating RVO in a subject as disclosed herein, wherein the VEGF cutoff is between 10 pg / mL and 100 pg / mL.

[0235] In some embodiments, the invention provides a method of treating RVO in a subject as disclosed herein, wherein the bl-CMT cutoff is between about 250 μm and about 600 μm.

[0236] In some embodiments, the present invention provides methods of treating RVO in a subject as disclosed herein,

[0237] In some embodiments, the invention provides a method of treating RVO in a subject as disclosed herein, wherein the anatomical response to the anti-VEGF therapeutic is measured by a change in CMT, which is defined as the perpendicular distance between the internal limiting membrane and the retinal pigment epithelium at the center of the fovea.

[0238] In some embodiments, the invention provides a method of treating RVO in a subject as disclosed herein, wherein the anatomical response to the anti-VEGF therapeutic is measured by a relative change in CMT, wherein the relative change in CMT is defined by the change in CMT divided by bl-CMT.

[0239] In some embodiments, the present invention provides a method of treating RVO in a subject as disclosed herein, wherein the determined VEGF level is above a VEGF cutoff value of 10 pg / mL to 100 pg / mL and the determined bl-CMT level is above a bl-CMT cutoff value of about 250 μm to about 600 μm, indicating that the eye has a good anatomical response to treatment with the anti-VEGF therapeutic agent.

[0240] In some embodiments, the invention provides a method of treating RVO in a subject as disclosed herein, wherein the determined VEGF level is above a VEGF cutoff value and the determined bl-CMT level is above a bl-CMT cutoff value, predicting a change in CFT of <-200 μm.

[0241] In some embodiments, the invention provides a method of treating RVO in a subject as disclosed herein, wherein the VEGF cutoff is between 30 pg / mL and 90 pg / mL.

[0242] In some embodiments, the invention provides a method of treating RVO in a subject as disclosed herein, wherein the VEGF cutoff is about 35 pg / mL, 40 pg / mL, 45 pg / mL, 50 pg / mL, 55 pg / mL, 60 pg / mL, 65 pg / mL, 70 pg / mL, 75 pg / mL, 80 pg / mL, or 85 pg / mL.

[0243] In some embodiments, the invention provides a method of treating RVO in a subject as disclosed herein, wherein the bl-CMT cutoff is between 250 μm and 600 μm.

[0244] In some embodiments, the invention provides a method of treating RVO in a subject as disclosed herein, wherein the bl-CMT cutoff is about 250 μm, 300 μm, 350 μm, 400 μm, 450 μm, 500 μm, 550 μm, or 600 μm.

[0245] In some embodiments, the invention provides methods of treating RVO in a subject as disclosed herein, wherein the VEGF level is determined by an immunoassay, optionally an enzyme immunoassay.

[0246] In some embodiments, the invention provides methods of treating RVO in a subject as disclosed herein, wherein the subject is in need of treatment for retinal vein occlusion (RVO).

[0247] In some embodiments, the invention provides a method of treating RVO in a subject as disclosed herein, wherein the subject is in need of treatment for macular edema secondary to retinal vein occlusion (ME-RVO), optionally the retinal vein occlusion is a central retinal vein occlusion (CRVO) and / or a branch retinal vein occlusion (BRVO).

[0248] In some embodiments, the invention provides a method of treating RVO in a subject as disclosed herein, further comprising administering the anti-VEGF therapeutic or another anti-VEGF therapeutic to the subject intravitreally, suprachoroidally and / or subretinally when the determined VEGF level is above a VEGF cutoff value and when the determined bl-CMT level is above a bl-CMT cutoff value.

[0249] In some embodiments, the invention provides a method of treating RVO in a subject as disclosed herein, wherein the determined VEGF level is above 90 pg / mL, and the anti-VEGF therapeutic or another anti-VEGF therapeutic is administered to the subject intravitreally, suprachoroidally, and / or subretinally.

[0250] In some embodiments, the invention provides methods of treating RVO in a subject as disclosed herein, wherein prior to the determining step, the subject has not been treated with the anti-VEGF therapeutic or another anti-VEGF therapeutic.

[0251] In some embodiments, the invention provides methods of treating RVO in a subject as disclosed herein, wherein prior to the determining step, the subject has been treated with the anti-VEGF therapeutic or another anti-VEGF therapeutic.

[0252] In some embodiments, the present invention provides a method of treating RVO in a subject as disclosed herein, wherein prior to the determining step, the subject has not been treated with a steroid, the steroid can be selected as a corticosteroid, further selected as a glucocorticoid.

[0253] In some embodiments, the present invention provides a method of treating RVO in a subject as disclosed herein, wherein prior to the determining step, the subject has been receiving steroid therapy, the steroid can be selected as a corticosteroid, further selected as a glucocorticoid.

[0254] In some embodiments, the present invention provides a method of treating RVO in a subject as disclosed herein, wherein the subject has not been treated with an anti-inflammatory agent or an immunosuppressant, or a combination thereof, optionally the anti-inflammatory agent or immunosuppressant is a glucocorticoid, methotrexate, cyclosporin A, tacrolimus, azathioprine, or a biologic targeting a pro-inflammatory molecule such as interleukin-6, interleukin-8, and tumor necrosis factor-α.

[0255] In some embodiments, the present invention provides a method of treating RVO in a subject as disclosed herein, wherein the subject has been treated with an anti-inflammatory agent or an immunosuppressant, or a combination thereof, optionally the anti-inflammatory agent or immunosuppressant is a glucocorticoid, methotrexate, cyclosporin A, tacrolimus, azathioprine, or a biologic targeting a pro-inflammatory molecule such as interleukin-6, interleukin-8, and tumor necrosis factor-α.

[0256] In some embodiments, the invention provides a method of treating RVO in a subject as disclosed herein, wherein the subject chooses to initiate, continue or resume treatment with the anti-VEGF therapeutic or another anti-VEGF therapeutic, and / or chooses to discontinue treatment with a steroid, which may be a corticosteroid, further selected as a glucocorticoid.

[0257] In some embodiments, the invention provides methods of treating RVO in a subject as disclosed herein, wherein the subject chooses to discontinue treatment with the anti-VEGF therapeutic or another anti-VEGF therapeutic, and / or chooses to initiate, continue or resume treatment with a steroid, which may be a corticosteroid, further selected as a glucocorticoid.

[0258] In some embodiments, the present invention provides methods of treating RVO in a subject as disclosed herein,

[0259] In some embodiments, the invention provides methods of treating RVO in a subject as disclosed herein, wherein the subject is a human.

[0260] In some embodiments, the present invention provides a method of treating RVO in a subject as disclosed herein, wherein the anti-VEGF therapeutic is formulated for intravitreal, suprachoroidal and / or subretinal administration, optionally further comprising admixing the anti-VEGF therapeutic with a pharmaceutically acceptable excipient or carrier.

[0261] In some embodiments, the present invention provides the use of an anti-VEGF therapeutic agent in the preparation of a medicament for treating retinal vein occlusion (RVO) in a subject, wherein the level of VEGF in the subject's aqueous humor is above a cutoff value, and the cutoff value is between 10 pg / mL and 100 pg / mL.

[0262] In some embodiments, the present invention provides a use of an anti-VEGF therapeutic as disclosed herein in the preparation of a medicament for treating RVO in a subject, wherein the anti-VEGF therapeutic comprises:

[0263] An anti-VEGF antibody or an antigen-binding fragment thereof, optionally the antigen-binding fragment is Fab, Fab', F(ab')2 or Fv;

[0264] A VEGF receptor (VEGFR) fusion protein, optionally the VEGFR fusion protein is a VEGFR-Fc immunoadhesin; or

[0265] An anti-VEGF gene therapy agent, optionally the anti-VEGF gene therapy agent comprises a nucleic acid encoding a VEGF antagonist, a nucleic acid encoding a protein that downregulates VEGF / VEGFR expression, and / or a nucleic acid targeting a nucleic acid and / or protein involved in VEGF / VEGFR expression, optionally the anti-VEGF gene therapy agent comprises a viral vector or a non-viral vector, optionally the viral vector is an adeno-associated virus (AAV) vector.

[0266] In some embodiments, the present invention provides use of an anti-VEGF therapeutic as disclosed herein in the preparation of a medicament for treating RVO in a subject, wherein the anti-VEGF therapeutic is administered or is to be administered intravitreally, suprachoroidally, and / or subretinally.

[0267] In some embodiments, the invention provides use of an anti-VEGF therapeutic as disclosed herein in the preparation of a medicament for treating RVO in a subject, wherein the subject suffers from macular edema secondary to retinal vein occlusion (ME-RVO), optionally the retinal vein occlusion is a central retinal vein occlusion (CRVO) and / or a branch retinal vein occlusion (BRVO).

[0268] In some embodiments, the present invention provides the use of an anti-VEGF therapeutic as disclosed herein in the preparation of a medicament for treating RVO in a subject, wherein the subject has or has not been treated with the anti-VEGF therapeutic or another anti-VEGF therapeutic.

[0269] In some embodiments, the present invention provides the use of an anti-VEGF therapeutic as disclosed herein in the preparation of a medicament for treating RVO in a subject, wherein the subject has or has not been treated with an anti-inflammatory agent or an immunosuppressant or a combination thereof, wherein the VEGF level in the subject's aqueous humor is above a threshold value, optionally the anti-inflammatory agent or immunosuppressant is selected from glucocorticoids, methotrexate, cyclosporin A, tacrolimus, azathioprine, or a biological agent targeting a pro-inflammatory molecule such as interleukin-6, interleukin-8, and tumor necrosis factor-α.

[0270] In some embodiments, the invention provides for the use of an anti-VEGF therapeutic as disclosed herein in the preparation of a medicament for treating RVO in a subject, wherein the subject has or has not been receiving steroid therapy, optionally the steroid is a corticosteroid, and optionally the corticosteroid is a glucocorticoid.

[0271] In some embodiments, the present invention provides use of an anti-VEGF therapeutic as disclosed herein in the preparation of a medicament for treating RVO in a subject, wherein the cutoff value is between 30 pg / mL and 90 pg / mL.

[0272] In some embodiments, the present invention provides use of an anti-VEGF therapeutic as disclosed herein in the preparation of a medicament for treating RVO in a subject, wherein the cutoff value is about 35 pg / mL, 40 pg / mL, 45 pg / mL, 50 pg / mL, 55 pg / mL, 60 pg / mL, 65 pg / mL, 70 pg / mL, 75 pg / mL, 80 pg / mL or 85 pg / mL.

[0273] In some embodiments, the present invention provides use of an anti-VEGF therapeutic as disclosed herein in the preparation of a medicament for treating RVO in a subject, wherein the subject has an anatomical response to the anti-VEGF therapeutic as measured by a change in central macular thickness (CMT) or foveal thickness (CFT), or as measured by a relative change in central macular thickness (CMT), wherein CMT or CFT is defined as the perpendicular distance between the internal limiting membrane and the retinal pigment epithelium at the center of the fovea, and the relative change in CMT is defined as the change in CMT divided by the baseline central macular thickness (bl-CMT).

[0274] In some embodiments, the present invention provides a use of an anti-VEGF therapeutic as disclosed herein in the preparation of a medicament for treating RVO in a subject, wherein the subject has a baseline central macular thickness (bl-CMT) level above a bl-CMT cutoff value, wherein the bl-CMT cutoff value is between about 250 μm and about 600 μm.

[0275] In some embodiments, the present invention provides use of an anti-VEGF therapeutic in the preparation of a kit for treating retinal vein occlusion (RVO), wherein the kit comprises: i) the anti-VEGF therapeutic, and ii) instructions for administering the anti-VEGF therapeutic to a subject whose VEGF level in aqueous humor is above a cutoff value, and wherein the cutoff value is between 10 pg / mL and 100 pg / mL.

[0276] In some embodiments, the present invention provides the use of an anti-VEGF therapeutic as disclosed herein in the preparation of a kit for treating RVO, wherein the kit further comprises instructions for not administering the anti-VEGF therapeutic to a subject whose VEGF level in the aqueous humor is not above a threshold value, and / or instructions for administering an anti-inflammatory agent or an immunosuppressant or a combination thereof to a subject whose VEGF level in the aqueous humor is above a threshold value, optionally the anti-inflammatory agent or immunosuppressant is selected from glucocorticoids, methotrexate, cyclosporin A, tacrolimus, azathioprine, or a biological agent targeting a pro-inflammatory molecule such as interleukin-6, interleukin-8, and tumor necrosis factor-α.

[0277] In some embodiments, the present invention provides use of an anti-VEGF therapeutic as disclosed herein in the preparation of a kit for treating RVO, wherein a subject having VEGF levels in aqueous humor above a cutoff value has a bl-CMT level above a bl-CMT cutoff value, and wherein the bl-CMT cutoff value is between about 250 μm and about 600 μm.

[0278] In some embodiments, the invention provides a kit comprising i) an anti-VEGF therapeutic, and ii) instructions for administering the anti-VEGF therapeutic to a subject having VEGF levels in aqueous humor above a cutoff value, wherein the cutoff value is between 10 pg / mL and 100 pg / mL.

[0279] In some embodiments, the present invention provides a kit as disclosed herein, further comprising instructions for not administering the anti-VEGF therapeutic agent to a subject whose VEGF level in the aqueous humor is not above a threshold value, and / or instructions for administering an anti-inflammatory agent or an immunosuppressant or a combination thereof to a subject whose VEGF level in the aqueous humor is above a threshold value, optionally the anti-inflammatory agent or immunosuppressant is selected from glucocorticoids, methotrexate, cyclosporin A, tacrolimus, azathioprine, or a biologic targeting a pro-inflammatory molecule such as interleukin-6, interleukin-8, and tumor necrosis factor-α.

[0280] In some embodiments, the invention provides a kit as disclosed herein, wherein the subject having VEGF levels in aqueous humor above a cutoff value has bl-CMT levels above a bl-CMT cutoff value, and wherein the bl-CMT cutoff value is between about 250 μm and about 600 μm.

[0281] In some embodiments, the present invention provides a kit comprising: i) reagents for determining VEGF levels in a subject's aqueous humor, and ii) instructions comprising a cutoff value for VEGF levels, and optionally instructions for comparing the determined VEGF levels to the cutoff value, wherein the cutoff value is between 10 pg / mL and 100 pg / mL.

[0282] In some embodiments, the invention provides a kit as disclosed herein, further comprising a first standard sample having a VEGF level substantially equal to a cut-off value.

[0283] In some embodiments, the present invention provides a kit as disclosed herein, further comprising a second standard sample having a VEGF level above a cut-off value.

[0284] In some embodiments, the present invention provides a kit as disclosed herein, further comprising a third standard sample having a VEGF level below a critical value.

[0285] In some embodiments, the invention provides a kit as disclosed herein, wherein the agent is an anti-VEGF antibody or an antigen-binding fragment thereof, optionally the antigen-binding fragment is Fab, Fab', F(ab')2 or Fv.

[0286] In some embodiments, the present invention provides a kit as disclosed herein, comprising an enzyme reactant, a magnetic separation reagent, a stabilizer, a standard, one or more concentrated wash buffers and / or a substrate buffer.

[0287] In some embodiments, the present invention provides a kit as disclosed herein, wherein the enzyme reactant comprises a VEGF-specific antibody I labeled with alkaline phosphatase, wherein the labeling ratio of the VEGF antibody I to the alkaline phosphatase is 1:0.75.

[0288] In some embodiments, the present invention provides a kit as disclosed herein, wherein the magnetic separation reagent contains 0.05 mass % magnetic particles, wherein the magnetic particles are coated with VEGF-specific antibody II, wherein the magnetic reagent further comprises a blocking agent, wherein the blocking agent is a mutant alkaline phosphatase.

[0289] In some embodiments, the present invention provides a kit as disclosed herein, wherein the VEGF-specific antibody I and / or the VEGF-specific antibody II is an anti-VEGF165 / VEGFA antibody.

[0290] In some embodiments, the present invention provides a kit as disclosed herein, wherein the stabilizer comprises one or more agents selected from the group consisting of Tris, NaCl, Tween 20, bovine serum albumin, mutant alkaline phosphatase and ProClin 300.

[0291] In some embodiments, the invention provides a kit as disclosed herein, wherein the wash buffer comprises Tris, NaCl, Tween-20, Triton-100 and ProClin 300.

[0292] In some embodiments, the present invention provides a composition comprising i) aqueous humor isolated from a subject, and ii) an agent that directly or indirectly interacts with VEGF in the isolated aqueous humor, wherein the isolated aqueous humor comprises a VEGF level above a cutoff value, and the cutoff value is between 10 pg / mL and 100 pg / mL.

[0293] In some embodiments, the invention provides a composition as disclosed herein, wherein the agent is an anti-VEGF antibody or an antigen-binding fragment thereof, optionally the antigen-binding fragment is Fab, Fab', F(ab')2 or Fv.

[0294] In some embodiments, the present invention provides a composition as disclosed herein, wherein the reagent comprises a VEGF-specific antibody I labeled with alkaline phosphatase, and wherein the labeling ratio of the VEGF antibody I to the alkaline phosphatase is 1:0.75.

[0295] In some embodiments, the present invention provides a composition as disclosed herein, wherein the reagent comprises 0.05 mass % magnetic particles, wherein the magnetic particles are coated with VEGF-specific antibody II, wherein the reagent further comprises a blocking agent, wherein the blocking agent is a mutant alkaline phosphatase.

[0296] In some embodiments, the present invention provides a composition as disclosed herein, wherein the VEGF-specific antibody I and / or the VEGF-specific antibody II is an anti-VEGF165 / VEGFA antibody.

[0297] In some embodiments, the present invention provides an anti-VEGF therapeutic for treating retinal vein occlusion (RVO) in a subject, wherein the level of VEGF in the subject's aqueous humor is above a cutoff value, and the cutoff value is between 10 pg / mL and 100 pg / mL.

[0298] In some embodiments, the present invention provides an anti-VEGF therapeutic as disclosed herein for treating RVO in a subject, the anti-VEGF therapeutic comprising:

[0299] An anti-VEGF antibody or an antigen-binding fragment thereof, optionally the antigen-binding fragment is Fab, Fab', F(ab')2 or Fv;

[0300] A VEGF receptor (VEGFR) fusion protein, optionally the VEGFR fusion protein is a VEGFR-Fc immunoadhesin; or

[0301] An anti-VEGF gene therapy agent comprising a nucleic acid encoding a VEGF antagonist, a nucleic acid encoding a protein that downregulates VEGF / VEGFR expression, and / or a nucleic acid targeting a nucleic acid and / or protein involved in VEGF / VEGFR expression, optionally comprising a viral vector or a non-viral vector, optionally the viral vector is an adeno-associated virus (AAV) vector.

[0302] In some embodiments, the present invention provides an anti-VEGF therapeutic as disclosed herein for use in treating RVO in a subject, wherein the anti-VEGF therapeutic is administered or is intended to be administered intravitreally, suprachoroidally, and / or subretinally.

[0303] In some embodiments, the invention provides an anti-VEGF therapeutic as disclosed herein for use in treating RVO in a subject, wherein the subject has macular edema secondary to retinal vein occlusion (ME-RVO), optionally the retinal vein occlusion is a central retinal vein occlusion (CRVO) and / or a branch retinal vein occlusion (BRVO).

[0304] In some embodiments, the invention provides an anti-VEGF therapeutic as disclosed herein for use in treating RVO in a subject, wherein the subject has or has not been treated with the anti-VEGF therapeutic or another anti-VEGF therapeutic.

[0305] In some embodiments, the present invention provides an anti-VEGF therapeutic as disclosed herein for use in treating RVO in a subject, wherein the subject has been treated with an anti-inflammatory agent or an immunosuppressant or a combination thereof, wherein the subject's VEGF level in the aqueous humor is above a threshold value, optionally the anti-inflammatory agent or immunosuppressant is selected from glucocorticoids, methotrexate, cyclosporin A, tacrolimus, azathioprine, or a biologic targeting a pro-inflammatory molecule such as interleukin-6, interleukin-8, and tumor necrosis factor-α.

[0306] In some embodiments, the present invention provides an anti-VEGF therapeutic as disclosed herein for use in treating RVO in a subject, wherein the subject has not been treated with an anti-inflammatory agent or an immunosuppressant, or a combination thereof, wherein the subject's VEGF level in the aqueous humor is above a threshold value, optionally the anti-inflammatory agent or immunosuppressant is selected from glucocorticoids, methotrexate, cyclosporin A, tacrolimus, azathioprine, or a biologic targeting a pro-inflammatory molecule such as interleukin-6, interleukin-8, and tumor necrosis factor-α.

[0307] In some embodiments, the present invention provides an anti-VEGF therapeutic as disclosed herein for use in treating RVO in a subject, wherein the cutoff value is between 30 pg / mL and 90 pg / mL.

[0308] In some embodiments, the present invention provides an anti-VEGF therapeutic as disclosed herein for use in treating RVO in a subject, wherein the cutoff value is about 35 pg / mL, 40 pg / mL, 45 pg / mL, 50 pg / mL, 55 pg / mL, 60 pg / mL, 65 pg / mL, 70 pg / mL, 75 pg / mL, 80 pg / mL or 85 pg / mL.

[0309] In some embodiments, the present invention provides an anti-VEGF therapeutic as disclosed herein for use in treating RVO in a subject, wherein the subject has an anatomical response to the anti-VEGF therapeutic as measured by a change in central macular thickness (CMT) or foveal thickness (CFT), or as measured by a relative change in central macular thickness (CMT), wherein the relative change in CMT is defined by the change in CMT divided by the baseline central macular thickness (bl-CMT).

[0310] In some embodiments, the present invention provides an anti-VEGF therapeutic as disclosed herein for use in treating RVO in a subject, wherein the subject's baseline central macular thickness (bl-CMT) level is above the bl-CMT cutoff value, and wherein the bl-CMT cutoff value is between about 250 μm and about 600 μm.

[0311] In some embodiments, the invention provides a method of predicting an anatomical response of an eye of a subject to treatment with an anti-VEGF therapeutic agent, the method comprising assessing or determining a VEGF level in aqueous humor collected from the subject, wherein a determined VEGF level above a cutoff value indicates that the eye has a good anatomical response to treatment with the anti-VEGF therapeutic agent, wherein a determined VEGF level not above the cutoff value indicates that the eye has a poor anatomical response to treatment with the anti-VEGF therapeutic agent, and wherein the cutoff value is between 10 pg / mL and 100 pg / mL.

[0312] In some embodiments, the present invention provides a method of predicting an anatomical response of a subject's eye to treatment with an anti-VEGF therapeutic as disclosed herein, wherein the anatomical response to the anti-VEGF therapeutic is measured by a change in central macular thickness (CMT) or foveal thickness (CFT), wherein CMT or CFT is defined as the perpendicular distance between the internal limiting membrane and the retinal pigment epithelium at the center of the fovea; or wherein the anatomical response to the anti-VEGF therapeutic is measured by a relative change in central macular thickness (CMT), which is defined as the change in CMT divided by the baseline central macular thickness (bl-CMT).

[0313] In some embodiments, the invention provides a method of predicting anatomic response of a subject's eye to treatment with an anti-VEGF therapeutic agent as disclosed herein, wherein a determined VEGF level above a cutoff value predicts a change in CFT of <-200 μm.

[0314] In some embodiments, the invention provides a method of predicting anatomic response of a subject's eye to treatment with an anti-VEGF therapeutic agent as disclosed herein, wherein a determined VEGF level below a cutoff value predicts a change in CFT > -200 μm.

[0315] In some embodiments, the invention provides a method of predicting the anatomical response of an eye of a subject to treatment with an anti-VEGF therapeutic agent as disclosed herein, wherein the cutoff value is between 30 pg / mL and 90 pg / mL.

[0316] In some embodiments, the present invention provides a method of predicting an anatomical response of a subject's eye to treatment with an anti-VEGF therapeutic agent as disclosed herein, wherein the cutoff value is about 35 pg / mL, 40 pg / mL, 45 pg / mL, 50 pg / mL, 55 pg / mL, 60 pg / mL, 65 pg / mL, 70 pg / mL, 75 pg / mL, 80 pg / mL or 85 pg / mL.

[0317] In some embodiments, the invention provides a method of predicting an anatomical response of a subject's eye to treatment with an anti-VEGF therapeutic agent as disclosed herein, wherein a VEGF level determined to be above 90 pg / mL predicts that the eye has a good anatomical response to treatment with the anti-VEGF therapeutic agent.

[0318] In some embodiments, the invention provides a method of predicting an anatomical response of a subject's eye to treatment with an anti-VEGF therapeutic agent as disclosed herein, wherein a VEGF level determined to be less than 30 pg / mL predicts that the eye will have a poor anatomical response to treatment with the anti-VEGF therapeutic agent.

[0319] In some embodiments, the invention provides a method of predicting the anatomical response of a subject's eye to treatment with an anti-VEGF therapeutic agent as disclosed herein, wherein the level of VEGF is determined by an immunoassay, optionally an enzyme immunoassay.

[0320] In some embodiments, the invention provides methods of predicting the anatomical response of an eye of a subject to treatment with an anti-VEGF therapeutic agent as disclosed herein, wherein the subject is in need of treatment for retinal vein occlusion (RVO).

[0321] In some embodiments, the invention provides a method of predicting the anatomical response of an eye of a subject to treatment with an anti-VEGF therapeutic agent as disclosed herein, wherein the subject is in need of treatment for macular edema secondary to retinal vein occlusion (ME-RVO), optionally the retinal vein occlusion is a central retinal vein occlusion (CRVO) and / or a branch retinal vein occlusion (BRVO).

[0322] In some embodiments, the present invention provides a method of predicting an anatomical response of a subject's eye to treatment with an anti-VEGF therapeutic as disclosed herein, the method further comprising administering the anti-VEGF therapeutic or another anti-VEGF therapeutic to the subject intravitreally, suprachoroidally and / or subretinally when the determined VEGF level is above a threshold value.

[0323] In some embodiments, the invention provides a method of predicting an anatomical response of a subject's eye to treatment with an anti-VEGF therapeutic as disclosed herein, wherein the VEGF level determined is above 90 pg / mL and the anti-VEGF therapeutic or another anti-VEGF therapeutic is administered to the subject intravitreally, suprachoroidally and / or subretinally.

[0324] In some embodiments, the present invention provides a method of predicting an anatomical response of a subject's eye to treatment with an anti-VEGF therapeutic agent as disclosed herein, the method further comprising administering to the subject a treatment other than the anti-VEGF therapeutic agent when the determined VEGF level is below a threshold value.

[0325] In some embodiments, the invention provides a method of predicting an anatomical response of an eye of a subject to treatment with an anti-VEGF therapeutic as disclosed herein, wherein the VEGF level determined is less than 30 pg / mL and the anti-VEGF therapeutic or another anti-VEGF therapeutic is not administered intravitreally, suprachoroidally and / or subretinally to the subject.

[0326] In some embodiments, the invention provides a method of predicting an anatomical response of a subject's eye to treatment with an anti-VEGF therapeutic agent as disclosed herein, wherein prior to the determining step, the subject has not been treated with the anti-VEGF therapeutic agent or another anti-VEGF therapeutic agent.

[0327] In some embodiments, the invention provides a method of predicting an anatomical response of a subject's eye to treatment with an anti-VEGF therapeutic agent as disclosed herein, wherein prior to the determining step, the subject has been treated with the anti-VEGF therapeutic agent or another anti-VEGF therapeutic agent.

[0328] In some embodiments, the present invention provides a method for predicting the anatomical response of a subject's eye to treatment with an anti-VEGF therapeutic agent as disclosed herein, wherein prior to the determining step, the subject has not received steroid treatment, the steroid may be selected as a corticosteroid, further selected as a glucocorticoid.

[0329] In some embodiments, the present invention provides a method for predicting the anatomical response of a subject's eye to treatment with an anti-VEGF therapeutic agent as disclosed herein, wherein prior to the determining step, the subject has been receiving steroid therapy, which steroid may be selected as a corticosteroid, further selected as a glucocorticoid.

[0330] In some embodiments, the present invention provides a method of predicting the anatomical response of a subject's eye to treatment with an anti-VEGF therapeutic agent as disclosed herein, wherein the subject is selected to initiate, continue or resume use of an anti-VEGF drug or other anti-VEGF drug, and / or to discontinue treatment with a steroid, which may be a corticosteroid, further selected as a glucocorticoid.

[0331] In some embodiments, the present invention provides a method of predicting the anatomical response of a subject's eye to treatment with an anti-VEGF therapeutic agent as disclosed herein, wherein the subject has chosen to discontinue use of the anti-VEGF drug or other anti-VEGF drug, and / or has chosen to initiate, continue or resume treatment with a steroid, which may be a corticosteroid, further selected as a glucocorticoid.

[0332] In some embodiments, the invention provides methods of predicting the anatomical response of an eye of a subject to treatment with an anti-VEGF therapeutic agent as disclosed herein, wherein the subject is a human.

[0333] In some embodiments, the present invention provides a method of predicting the anatomical response of a subject's eye to treatment with an anti-VEGF therapeutic as disclosed herein, wherein the anti-VEGF therapeutic is formulated for intravitreal, suprachoroidal and / or subretinal administration, optionally further comprising admixing the anti-VEGF therapeutic with a pharmaceutically acceptable excipient or carrier.

[0334] In some embodiments, the present invention provides a method of predicting an anatomical response of an eye of a subject to treatment with an anti-VEGF therapeutic agent as disclosed herein, the method further comprising assessing or determining the level of bl-CMT in the subject, wherein a determined level of bl-CMT above a bl-CMT cutoff value indicates that the eye has a good anatomical response to treatment with the anti-VEGF therapeutic agent, wherein a determined level of bl-CMT not above the bl-CMT cutoff value indicates that the eye has a poor anatomical response to treatment with the anti-VEGF therapeutic agent, and wherein the bl-CMT cutoff value is between about 250 μm and about 600 μm.

[0335] In some embodiments, the invention provides a method of predicting anatomic response of an eye of a subject to treatment with an anti-VEGF therapeutic agent, the method comprising assessing or determining the level of VEGF in aqueous humor collected from the subject and the level of bl-CMT in the subject,

[0336] wherein the determined VEGF level is above a VEGF cutoff value, and the determined bl-CMT level is above a bl-CMT cutoff value, indicating that the eye has a favorable anatomical response to treatment with the anti-VEGF therapeutic agent; and

[0337] Wherein the determined VEGF level is not above the VEGF cut-off value or the determined bl-CMT level is not above the bl-CMT cut-off value, indicating that the eye has a poor anatomical response to treatment with the anti-VEGF therapeutic agent.

[0338] In some embodiments, the invention provides a method of predicting the anatomical response of an eye of a subject to treatment with an anti-VEGF therapeutic agent as disclosed herein, wherein the VEGF cutoff value is between 10 pg / mL and 100 pg / mL.

[0339] In some embodiments, the invention provides a method of predicting anatomic response of an eye of a subject to treatment with an anti-VEGF therapeutic agent as disclosed herein, wherein the bl-CMT cutoff is between about 250 μm and about 600 μm.

[0340] In some embodiments, the present invention provides a method of predicting the anatomical response of a subject's eye to treatment with an anti-VEGF therapeutic agent as disclosed herein, wherein the anatomical response to the anti-VEGF therapeutic agent is measured by a change in CMT, which is defined as the perpendicular distance between the internal limiting membrane and the retinal pigment epithelium at the center of the fovea.

[0341] In some embodiments, the present invention provides a method of predicting the anatomical response of a subject's eye to treatment with an anti-VEGF therapeutic agent as disclosed herein, wherein the anatomical response to the anti-VEGF therapeutic agent is measured by a relative change in CMT, wherein the relative change in CMT is defined by the change in CMT divided by bl-CMT.

[0342] In some embodiments, the present invention provides a method of predicting an anatomical response of an eye of a subject to treatment with an anti-VEGF therapeutic agent as disclosed herein, wherein the determined VEGF level is above a VEGF cutoff value of 10 pg / mL to 100 pg / mL, and the determined bl-CMT level is above a bl-CMT cutoff value of about 250 μm to about 600 μm, indicating that the eye has a good anatomical response to treatment with the anti-VEGF therapeutic agent.

[0343] In some embodiments, the present invention provides a method of predicting an anatomical response of a subject's eye to treatment with an anti-VEGF therapeutic agent as disclosed herein, wherein the determined VEGF level is above a VEGF cutoff value and the determined bl-CMT level is above a bl-CMT cutoff value, predicting a change in CFT of <-200 μm.

[0344] In some embodiments, the present invention provides a method of predicting an anatomical response of a subject's eye to treatment with an anti-VEGF therapeutic agent as disclosed herein, wherein the determined VEGF level is below a VEGF cutoff value or the determined bl-CMT level is below a bl-CMT cutoff value, predicting a change in CFT of >-200 μm.

[0345] In some embodiments, the invention provides a method of predicting an anatomical response of an eye of a subject to treatment with an anti-VEGF therapeutic agent as disclosed herein, wherein the VEGF cutoff is between 30 pg / mL and 90 pg / mL.

[0346] In some embodiments, the invention provides a method of predicting an anatomical response of an eye of a subject to treatment with an anti-VEGF therapeutic agent as disclosed herein, wherein the VEGF cutoff is about 35 pg / mL, 40 pg / mL, 45 pg / mL, 50 pg / mL, 55 pg / mL, 60 pg / mL, 65 pg / mL, 70 pg / mL, 75 pg / mL, 80 pg / mL or 85 pg / mL.

[0347] In some embodiments, the invention provides a method of predicting the anatomical response of an eye of a subject to treatment with an anti-VEGF therapeutic agent as disclosed herein, wherein the bl-CMT cutoff is between 250 μm and 600 μm.

[0348] In some embodiments, the invention provides a method of predicting an anatomical response of a subject's eye to treatment with an anti-VEGF therapeutic agent as disclosed herein, wherein the bl-CMT cutoff is about 250 μm, 300 μm, 350 μm, 400 μm, 450 μm, 500 μm, 550 μm or 600 μm.

[0349] In some embodiments, the invention provides a method of predicting the anatomical response of a subject's eye to treatment with an anti-VEGF therapeutic agent as disclosed herein, wherein the level of VEGF is determined by an immunoassay, optionally an enzyme immunoassay.

[0350] In some embodiments, the invention provides methods of predicting the anatomical response of an eye of a subject to treatment with an anti-VEGF therapeutic agent as disclosed herein, wherein the subject is in need of treatment for retinal vein occlusion (RVO).

[0351] In some embodiments, the invention provides a method of predicting the anatomical response of an eye of a subject to treatment with an anti-VEGF therapeutic agent as disclosed herein, wherein the subject is in need of treatment for macular edema secondary to retinal vein occlusion (ME-RVO), optionally the retinal vein occlusion is a central retinal vein occlusion (CRVO) and / or a branch retinal vein occlusion (BRVO).

[0352] In some embodiments, the invention provides a method of predicting an anatomical response of an eye of a subject to treatment with an anti-VEGF therapeutic as disclosed herein, the method comprising administering the anti-VEGF therapeutic or another anti-VEGF therapeutic to the subject intravitreally, suprachoroidally and / or subretinally when the determined VEGF level is above a VEGF cutoff value and when the determined bl-CMT level is above a bl-CMT cutoff value.

[0353] In some embodiments, the invention provides a method of predicting an anatomical response of a subject's eye to treatment with an anti-VEGF therapeutic as disclosed herein, wherein the VEGF level determined is above 90 pg / mL and the anti-VEGF therapeutic or another anti-VEGF therapeutic is administered to the subject intravitreally, suprachoroidally and / or subretinally.

[0354] In some embodiments, the present invention provides a method of predicting an anatomical response of a subject's eye to treatment with an anti-VEGF therapeutic agent as disclosed herein, the method further comprising administering to the subject a treatment other than the anti-VEGF therapeutic agent when the determined VEGF level is below the VEGF threshold.

[0355] In some embodiments, the invention provides a method of predicting an anatomical response of an eye of a subject to treatment with an anti-VEGF therapeutic as disclosed herein, wherein the VEGF level determined is less than 30 pg / mL and the anti-VEGF therapeutic or another anti-VEGF therapeutic is not administered intravitreally, suprachoroidally and / or subretinally to the subject.

[0356] In some embodiments, the invention provides a method of predicting the anatomical response of a subject's eye to treatment with an anti-VEGF therapeutic as disclosed herein, wherein the treatment other than the anti-VEGF therapeutic is a steroid, optionally a corticosteroid, further optionally a glucocorticoid.

[0357] In some embodiments, the invention provides a method of predicting the anatomical response of a subject's eye to treatment with an anti-VEGF therapeutic agent as disclosed herein, wherein prior to the determining step, the subject has or has not been treated with the anti-VEGF therapeutic agent or another anti-VEGF therapeutic agent.

[0358] In some embodiments, the present invention provides a method for predicting the anatomical response of a subject's eye to treatment with an anti-VEGF therapeutic agent as disclosed herein, wherein prior to the determining step, the subject has or has not received steroid treatment, the steroid may be selected as a corticosteroid, further selected as a glucocorticoid.

[0359] In some embodiments, the present invention provides a method of predicting the anatomical response of a subject's eye to treatment with an anti-VEGF therapeutic agent as disclosed herein, wherein the subject has or has not been treated with an anti-inflammatory agent or an immunosuppressant or a combination thereof, optionally the anti-inflammatory agent or immunosuppressant is selected from glucocorticoids, methotrexate, cyclosporin A, tacrolimus, azathioprine, or a biologic targeting a pro-inflammatory molecule such as interleukin-6, interleukin-8, and tumor necrosis factor-α.

[0360] In some embodiments, the present invention provides a method of predicting the anatomical response of a subject's eye to treatment with an anti-VEGF therapeutic agent as disclosed herein, wherein the subject is selected to initiate, continue or resume use of an anti-VEGF drug or other anti-VEGF drug, and / or to discontinue treatment with a steroid, which may be a corticosteroid, further selected as a glucocorticoid.

[0361] In some embodiments, the present invention provides a method of predicting the anatomical response of a subject's eye to treatment with an anti-VEGF therapeutic agent as disclosed herein, wherein the subject has chosen to discontinue use of the anti-VEGF drug or other anti-VEGF drug, and / or has chosen to initiate, continue or resume treatment with a steroid, which may be a corticosteroid, further selected as a glucocorticoid.

[0362] In some embodiments, the invention provides methods of predicting the anatomical response of an eye of a subject to treatment with an anti-VEGF therapeutic agent as disclosed herein, wherein the subject is a human.

[0363] In some embodiments, the present invention provides a method of predicting the anatomical response of a subject's eye to treatment with an anti-VEGF therapeutic agent as disclosed herein,

[0364] In some embodiments, the present invention provides a method of predicting the anatomical response of a subject's eye to treatment with an anti-VEGF therapeutic as disclosed herein, wherein the anti-VEGF therapeutic is formulated for intravitreal, suprachoroidal and / or subretinal administration, optionally further comprising admixing the anti-VEGF therapeutic with a pharmaceutically acceptable excipient or carrier.

[0365] In any of the embodiments herein, the subject may not have been treated with the anti-VEGF therapeutic or another anti-VEGF therapeutic prior to the determining step.

[0366] In any of the embodiments herein, prior to the determining step, the subject may have been treated with the anti-VEGF therapeutic or another anti-VEGF therapeutic.

[0367] In any of the embodiments herein, prior to the determining step, the subject may not have been receiving steroid therapy, and the steroid may be selected as a corticosteroid, further selected as a glucocorticoid.

[0368] In any of the embodiments herein, prior to the determining step, the subject may have been receiving steroid therapy, which steroid may be selected as a corticosteroid, further selected as a glucocorticoid.

[0369] In any of the embodiments herein, the subject may be selected to initiate, continue or resume use of an anti-VEGF drug or other anti-VEGF drug, and / or to discontinue treatment with a steroid, which may be a corticosteroid, further selected as a glucocorticoid.

[0370] In any of the embodiments herein, the subject may choose to discontinue use of the anti-VEGF drug or other anti-VEGF drug, and / or choose to start, continue or resume treatment with a steroid, which may be a corticosteroid, further selected as a glucocorticoid.

[0371] In any of the embodiments herein, the subject may be a human.In any of the embodiments herein, the anti-VEGF therapeutic may be formulated for intravitreal, suprachoroidal and / or subretinal administration.

[0372] In any of the embodiments herein, the method may further comprise admixing the anti-VEGF therapeutic agent and a pharmaceutically acceptable excipient or carrier.

[0373] The term "about" as used herein refers to the typical error range for each value that is readily known to those skilled in the art. Reference herein to "about" with respect to a value or parameter includes (and describes) embodiments for that value or parameter per se.

[0374] As used herein, an referent without a quantifier before it includes a plural referent unless the context clearly dictates otherwise. For example, an referent without a quantifier before it means "at least one" or "one or more."

[0375] Throughout this disclosure, various aspects are presented in the form of ranges. It should be understood that the description in the form of ranges is only for convenience and brevity and should not be interpreted as an unchangeable limitation on the scope of the present disclosure. Therefore, the description of the range should be considered to have specifically disclosed all possible sub-ranges and each numerical value within the range. For example, if a numerical range is provided, it is understood that each intermediate value between the upper and lower limits of the range and any other specified value or intermediate value within the specified range are included in the present disclosure. The upper and lower limits of these smaller ranges can be independently included in the smaller range, and are also included in the present disclosure, subject to any explicitly excluded restrictions in the described range. If the specified range includes one or both of the upper and lower limits, the range that does not include one or both of the included upper and lower limits is also included in the present disclosure. This applies no matter how large the range is.

[0376] The use of ordinal numbers such as "first", "second", "third", etc. in the claims to modify claim elements does not by itself imply any priority, precedence or order of one claim element relative to another claim element, or a temporal order of performing method acts, but is merely used as a label to distinguish one claim element with a particular name from another claim element with the same name (but with an ordinal number) to distinguish the claim elements. Similarly, the use of a), b), etc. or i), ii), etc. does not by itself imply any priority, precedence or order of steps in the claims. Similarly, the use of these terms in the specification does not by itself imply any required priority, precedence or order.

[0377] References

[0378] 1.Branch Vein Occlusion Study Group.Argon laser scatterphotocoagulation for prevention of neovascularization and vitreous hemorrhagein branch vein occlusion.A randomized clinical trial.Archives ofophthalmology(Chicago,Ill.:1960),1986.104(1):p.34-41.

[0379] 2.The Central Vein Occlusion Study.Baseline and early natural historyreport.Archives of Ophthalmology,1993.111(8):p.1087-1095.

[0380] 3.Christoffersen,N.L.B.and M.Larsen,Pathophysiology and hemodynamicsof branch retinal vein occlusion.Ophthalmology,1999.106(11):p.2054-2062.

[0381] 4.Noma,H.,K.Yasuda,and M.Shimura,Cytokines and Pathogenesis ofCentral Retinal Vein Occlusion.J Clin Med,2020.9(11).

[0382] 5.Noma,H.,K.Yasuda,and M.Shimura,Cytokines and the Pathogenesis ofMacular Edema in Branch Retinal Vein Occlusion.J Ophthalmol,2019.2019:p.5185128.

[0383] 6.Schmidt-Erfurth,U.,et al.,Guidelines for the Management of RetinalVein Occlusion by the European Society of Retina Specialists(EURETINA).Ophthalmologica,2019.242(3):p.123-162.

[0384] 7.Wallsh,J.O.and R.P.Gallemore,Anti-VEGF-Resistant Retinal Diseases:AReview of the Latest Treatment Options.Cells,2021.10(5).

[0385] 8.Rosenbaum,J.N.and P.Weisman,The Evolving Role of CompanionDiagnostics for Breast Cancer in an Era of Next-Generation Omics.Am J Pathol,2017.187(10):p.2185-2198.

[0386] 9.Modi,A.,et al.,Aqueous Humor Cytokines and TherapeuticCustomization in Nonresponding Macular Edema Secondary To Retinal VeinOcclusion.Retin Cases Brief Rep,2021.15(2):p.127-130.

[0387] 10.Campochiaro,P.A.,et al.,Ranibizumab for macular edema due toretinal vein occlusions:implication of VEGF as a critical stimulator.MolTher,2008.16(4):p.791-9.

[0388] 11.Park,S.P.and J.K.Ahn,Changes of aqueous vascular endothelialgrowth factor and pigment epithelium-derived factor following intravitrealbevacizumab for macular oedema secondary to branch retinal veinocclusion.Clinical&experimental ophthalmology,2009.37(5):p.490-5.

[0389] 12.Wang B, Zhang

[0390] Example

[0391] The following examples are for illustrative purposes only and are not intended to limit the scope of the present disclosure.

[0392] Example 1: VEGF in aqueous humor as a companion diagnostic marker

[0393] A magnetic particle-based chemiluminescent enzyme immunoassay (MP CLEIA) kit with high sensitivity, specificity and reproducibility was developed for the detection of VEGF in human aqueous humor. Please refer to CN112557669A, which is incorporated herein by reference in its entirety for all purposes.

[0394] To test the use of VEGF in aqueous humor as a concomitant biomarker for anti-VEGF therapy in RVO, aqueous humor samples were collected in 24 RVO-ME cases before intravitreal injection of anti-VEGF agents. Changes in central macular thickness (CMT) or foveal thickness (CFT) without foveal involvement (CMT or CFT was defined as the perpendicular distance between the internal limiting membrane and the retinal pigment epithelium at the center of the fovea) or maximum parafoveal retinal thickness (MRT) were plotted at 2 to 4 weeks after intravitreal injection compared with baseline plotted against baseline aqueous humor VEGF levels. Figure 1 In the study, we found that the change in CFT / MRT was generally negatively correlated with baseline aqueous humor VEGF levels (Spearman rank correlation coefficient test, p < 0.01). In general, higher baseline aqueous humor VEGF was associated with a better treatment response.

[0395] Eyes with aqueous humor VEGF levels above a cutoff of about 10 pg / mL to 100 pg / mL had significantly better anatomic treatment response as measured by changes in CFT than eyes with lower aqueous humor VEGF levels. Figure 1It was shown that the aqueous VEGF level in the eyes with a change in CFT / MRT of ≥-200 μm was less than 30 pg / mL, and the aqueous VEGF level in the eyes with a change in CFT / MRT of ≤-200 μm was greater than 90 pg / mL. For example, a VEGF level of about 100 pg / mL determined can be associated with a change in CFT between about -250 μm and about 0 μm and / or used to predict a change in CFT between about -250 μm and about 0 μm, and a VEGF level of about 1 pg / mL and about 10 pg / mL determined can be associated with a change in CFT between about -200 μm and about 0 μm and / or used to predict a change in CFT between about -200 μm and about 0 μm. Therefore, the aqueous VEGF cutoff value between about 10 pg / mL and about 100 pg / mL can be used to divide eyes into two groups with different anatomical responses. In other words, measuring VEGF levels in basal aqueous humor (e.g., using the MP-CLEIA kit described in CN112557669A or any other suitable kit) can be used to predict anatomical treatment response to anti-VEGF in patients with RVO-ME as a companion diagnostic.

[0396] Example 2: Baseline Aqueous VEGF Levels and Central Macular Thickness Predict Anatomical Response to Anti-VEGF Treatment in RVO-ME

[0397] To investigate whether aqueous humor biomarkers and anatomical imaging biomarkers have predictive value for treatment response to anti-VEGF therapy in patients with retinal vein occlusion-related macular edema (RVO-ME). We performed a retrospective observational study at 2 tertiary referral centers.

[0398] Participants

[0399] This was a two-center observational study. The medical records of patients with RVO-ME who underwent intravitreal injection of anti-VEGF agents with aqueous humor extraction at Peking Union Medical College Hospital and Xi'an People's Hospital from March 2020 to January 2022 were reviewed. Patients were excluded if: (1) baseline (bl) (≤2 weeks before intravitreal injection) OCT scan centered on the fovea was not available or the quality of the bl-OCT scan was unsatisfactory; (2) ME did not involve the fovea or the central macular thickness (CMT) was ≤250 μm on bl-OCT; (3) there was a history of or concurrent diabetes, retinal vasculitis, uveitis, age-related macular degeneration, high myopia, epiretinal membrane, or other fundus diseases that could cause ME; (4) they received intravitreal or periocular corticosteroid injections.

[0400] Clinical evaluation

[0401] All patients underwent a comprehensive ophthalmologic examination within 2 weeks before intravitreal injection, including best-corrected visual acuity (BCVA), intraocular pressure (IOP), slit-lamp examination, funduscopy, and OCT. Raster or radial OCT scans centered on the fovea were obtained using one of the following OCT systems: Triton , TOPCON Inc, Japan; VanGogh , SVision Imaging Inc, China; RTVue XR Avanti , Optovue , Inc, USA. CMT (foveal retinal thickness) was measured using built-in software.

[0402] Intravitreal injection and aqueous humor extraction

[0403] Before treatment, antibiotic (0.5% levofloxacin) eye drops were applied 4 times a day for at least 3 days. After topical anesthesia and disinfection of the conjunctival sac with 5% povidone-iodine for 30 seconds, about 50ul to 100ul of aqueous humor was collected with a 25-gauge needle syringe, and then 50ul of ranibizumab (0.5mg) (Novartis, Switzerland), Conbercept (0.5mg) (Chengdu Kanghong, China) or Aflibercept (2mg) (Bayer, Germany) were injected intravitreally. After injection, antibiotic ointment was applied and the eyes were repaired for at least 3 hours.

[0404] Aqueous humor test

[0405] Follow the manufacturer's instructions for using a magnetic Performance Assay kit (R&D Systems Inc., USA), through the Luminex xMAPTM platform ( 100 system, Luminexcorporation, USA) was used to detect the concentrations of VEGF, IL-6, IL-8, IL-10, MCP-1 and IP-10 in aqueous humor.

[0406] The VEGF level in aqueous humor was measured using the MPCLEIA kit. The kit only requires 20ul of aqueous humor sample, with a limit of blank (LOB) of approximately 2pg / ml, a limit of detection (LOD) of approximately 5pg / ml, and a linear range of approximately 5pg / ml to 5000pg / ml. The intra- and inter-assay coefficients of variation (CV) were approximately 10% and 15%, respectively.

[0407] Subgroup Assessment of Treatment Response

[0408] According to the treatment history, eyes with RVO were divided into 2 groups. Eyes that had not received anti-VEGF treatment and eyes that had previously received anti-VEGF treatment for more than 3 months were designated as the untreated group; eyes that had previously received anti-VEGF treatment within 3 months were classified as the treated group.

[0409] Definition of treatment response

[0410] Eyes with complete clinical evaluation 2-6 weeks after injection were divided into 2 groups according to the anatomic therapeutic response (ATR) to intravitreal treatment. The ATR was expressed as the relative CMT change (ΔCMT / bl-CMT) after intravitreal injection and was divided into good response (GR) (≤-25%) and poor response (PR) (>-25%).

[0411] Data analysis

[0412] Statistical analysis was performed using IBM SPSS statistics 26.0 (SPSS Inc, Chicago, IL), RStudio version 1.4.1717 (RStudio Team 2021), or GraphPad Prism 8 (GraphPad Prism Software Inc., San Diego, CA). Measurements below the limit of detection were replaced by the 30% LOD value of the detection platform for statistical analysis without changing the rank of the included data. Unmeasured samples were treated as missing values ​​and were not included in the data analysis.

[0413] Normally and non-normally distributed continuous variables were expressed as mean ± standard deviation (SD) and median (interquartile range (IQR)), respectively; discrete variables were expressed as counts (%). Unpaired data were compared using the unpaired Student's t test for normally distributed data, the nonparametric Mann-Whitney U test for non-normally distributed data, and the chi-square test, Fisher test, or Wilcoxon test for discrete variables.

[0414] Receiver operating characteristic (ROC) curves were generated to evaluate the predictive efficacy of different variables and regression models for ATR. The optimal cutoff value was determined as the point where the Youden index (sensitivity + specificity - 1) was maximum. Univariate and multivariate logistic regression models were used to study prognostic factors.

[0415] Univariate and multivariate logistic regression analyses were performed to assess factors associated with ATR using the Wald test. Automated stepwise regression developed a series of logistic regression models to handle variable selection, with covariates added or removed at each step. The stepwise regression procedure examined the deviance value and Akaike information criterion (AIC) for each variable at each step. The preferred model was determined by removing the variable with the highest AIC value at each step. The likelihood ratio test (LRT) was performed to compare the goodness of fit of the two nested models. A p value < 0.05 was considered statistically significant.

[0416] result

[0417] A total of 94 patients (44 females and 50 males) (94 eyes) with a mean age of 53.12 ± 15.67 years were included in this study. Among the 94 eyes with RVO-ME, 41 eyes were BRVO and 53 eyes were CRVO; 80 eyes, 7 eyes, and 7 eyes were treated with ranibizumab (IVR), aflibercept (IVA), and conbercept (IVC), respectively. As described previously, the patients were divided into the untreated group and the treated group, which included 58 and 36 patients, respectively. Table 1 lists the general demographic and clinical characteristics of the untreated group (previous anti-VEGF treatment had minimal or no effect) and the treated group.

[0418] Table 1 General demographic and clinical characteristics of the included patients

[0419]

[0420]

[0421] Abbreviations: BRVO, branch retinal vein occlusion; IVR / IVC / IVA, intravitreal ranibizumab / conbercept / aflibercept; bl, baseline; BCVA, best-corrected visual acuity; LogMAR, logarithm of the minimum angle of resolution; CMT, central macular thickness. *P < .05

[0422] Analysis of cytokines in aqueous humor

[0423] The concentrations of IL-6, IL-8, IL-10, IP-10, MCP-1, and VEGF in aqueous humor of all 94 eyes included were 13.8 (5.1-31.5), 52.5 (22.8-107.8), 2.3 (0.5-5.2), 65.5 (22.9-106.6), 1345.9 (779.2-1745.4), and 75.6 (5.8-204.1), respectively. Correlation analysis revealed extensive associations between cytokines / measurements. In particular, VEGF levels were significantly correlated with all other cytokines / measurements; among all cytokines, IL-10 had the least correlation with the other cytokines.

[0424] Subgroup analysis showed that the levels of VEGF (5.23 (1.56-14.49) vs. 151.44 (62.17-297.86) pg / ml), IL-6 (8.35 (1.57-13.84) vs. 22.00 (6.00-51.84) pg / ml), and IL-8 (34.39 (18.84-70.97) vs. 69.31 (26.26-156.78) pg / ml) in the aqueous humor of the treatment group were significantly higher than those in the control group. pg / ml) and MCP-1 (1150.73 (731.81-1442.02) vs. 1540.65 (936.36-2594.05) pg / ml) were significantly lower than those in the untreated group (Z = -4.83, P < .001; Z = -5.43, P < .001; Z = -4.07, P < .001; Z = -2.57, P = .010; Z = -2.62, P = .009, respectively). The bl-CMT level was significantly higher in the untreated group compared with the treated group (593 (390-824.5) vs. 284 (197-376) μm), but the bl-BCVA (logarithm of the minimum angle of resolution (logMAR) scale) was not significantly different between the two groups (Z = -4.97, P < .001; Z = -0.79, P = .432, respectively) ( Figure 2 ).

[0425] Identification of aqueous humor cytokines associated with treatment response

[0426] Among the 94 included patients, follow-up data from 2 to 6 weeks after injection were obtained for 40 IVR patients. Among these 40 IVR patients, 31 and 9 patients were classified into the GR and PR groups, respectively, based on ATR (ΔCMT / bl-CMT).

[0427] Correlation analysis was performed to identify aqueous humor cytokines that may have predictive value for treatment response and revealed that ΔCMT / bl-CMT was significantly correlated with IL-8 (r = -0.330, P = .040) and VEGF (r = -0.506, P = .001) in aqueous humor, but not with IL-6 (r = -0.312, P = .053), IP-10 (r = -0.259, P = .298), IL-10 (r = -0.081, P = .698), and MCP-1 (r = -0.270, P = .101).

[0428] ROC analysis using Youden index determined that the optimal cut-off values ​​for IL-8 and VEGF were 19.7 pg / ml and 26.0 pg / ml, respectively. Figure 3). Thus, 28 / 3, 25 / 6, and 28 / 3 of 31 patients in the GR group and 4 / 4 (one missing value for IL-8 data), 4 / 5, and 4 / 5 of 9 patients in the PR group had high (≥cutoff value) / low (<cutoff value) levels of IL-8 (n=39) and VEGF (n=40), respectively. Chi-square tests showed significant correlations between ATR category (GR or PR) and levels of IL-8 and VEGF (high or low) in aqueous humor (χ2=7.020, P=.008; χ2=11.176, P<.001, respectively).

[0429] The Wilcoxon-Mann-Whitney U test also showed significant differences in the distribution of ATR (ΔCMT / bl-CMT) data between eyes with high and low IL-8 levels (W = 48, P = .017) and VEGF levels (W = 55, P = .012). Both the chi-square test and the Wilcoxon-Mann-Whitney U test confirmed that the classification of patients based on the cutoff values ​​derived from the ROC was reasonable.

[0430] Univariate analysis further confirmed that ATR (ΔCMT / bl-CMT) was significantly correlated with IL-8 (z=2.396, P=.017) and VEGF (z=2.715, P<.001) in aqueous humor (Table 2).

[0431] Table 2 Univariate and multivariate logistic regression analysis

[0432]

[0433] Abbreviations: IL, interleukin; bl-CMT, baseline central macular thickness. *P < .05; **P < .01; ***P < .001. a Low levels of VEGF were defined as the reference.

[0434] Development and validation of the best prediction model for treatment response

[0435] To establish the best prediction model for treatment response, multivariable logistic stepwise regression analysis was performed with the variables IL-8 (high or low), VEGF (high or low), bl-CMT, disease duration, treatment history (naive or IVR-treated, with or without laser photocoagulation), and disease phenotype (CRVO or BRVO). After stepwise screening and LRT comparison, the variables VEGF and bl-CMT were retained in the model with the lower AIC. In addition, bl-CMT and VEGF were significantly associated with ATR category in multivariable logistic regression analysis (Table 2).

[0436] The predictive efficacy of the above univariate and multivariate logistic regression models was further evaluated by ROC curve analysis and LRT. The AUC of the multivariate logistic regression model for VEGF and bl-CMT was 0.952, while the AUC of the univariate logistic regression model for VEGF and bl-CMT was 0.702 and 0.891, respectively. Figure 3 ). Based on the LRT results, the logistic regression model combining VEGF with bl-CMT showed a higher goodness of fit than any model using VEGF (deviation (D) = -16.18, P < .001) or bl-CMT (D = -7.596, P = .006) alone. These results suggest that aqueous humor biomarkers (e.g., VEGF) and imaging markers (e.g., bl-CMT) can be integrated to improve the accuracy of predicting the anatomical response of a subject's eye to treatment with an anti-VEGF therapeutic agent.

[0437] It is not intended that the present disclosure be limited to the scope of the specific embodiments disclosed, which are provided, for example, to illustrate various aspects of the present disclosure. From the description and teachings herein, various modifications to the described compositions and methods will become apparent. These changes may be implemented without departing from the true scope and spirit of the present disclosure, and these changes are intended to fall within the scope of the present disclosure.

Claims

1. Use of an anti-VEGF therapeutic agent in the preparation of a medicament for treating retinal vein occlusion (RVO) in a subject, wherein the level of VEGF in the subject's aqueous humor is above a cutoff value, and the cutoff value is between 10 pg / mL and 100 pg / mL.

2. The use according to claim 1, wherein the anti-VEGF therapeutic agent comprises: An anti-VEGF antibody or an antigen-binding fragment thereof, optionally the antigen-binding fragment is Fab, Fab', F(ab')2 or Fv; A VEGF receptor (VEGFR) fusion protein, optionally the VEGFR fusion protein is a VEGFR-Fc immunoadhesin; or An anti-VEGF gene therapy agent, optionally the anti-VEGF gene therapy agent comprises a nucleic acid encoding a VEGF antagonist, a nucleic acid encoding a protein that downregulates VEGF / VEGFR expression, and / or a nucleic acid targeting a nucleic acid and / or protein involved in VEGF / VEGFR expression, optionally the anti-VEGF gene therapy agent comprises a viral vector or a non-viral vector, optionally the viral vector is an adeno-associated virus (AAV) vector.

3. The use according to claim 1 or 2, wherein the anti-VEGF therapeutic agent is administered or is to be administered intravitreally, suprachoroidally and / or subretinally.

4. The use according to any one of claims 1 to 3, wherein the subject suffers from macular edema secondary to retinal vein occlusion (ME-RVO), optionally the retinal vein occlusion is central retinal vein occlusion (CRVO) and / or branch retinal vein occlusion (BRVO).

5. The use according to any one of claims 1 to 4, wherein the subject has or has not been treated with the anti-VEGF therapeutic agent or another anti-VEGF therapeutic agent.

6. The use according to any one of claims 1 to 4, wherein the subject has or has not been treated with an anti-inflammatory agent or an immunosuppressant or a combination thereof, wherein the VEGF level in the subject's aqueous humor is above a critical value, optionally the anti-inflammatory agent or immunosuppressant is selected from glucocorticoids, methotrexate, cyclosporin A, tacrolimus, azathioprine, or a biological agent targeting a pro-inflammatory molecule such as interleukin-6, interleukin-8, and tumor necrosis factor-α.

7. The use according to any one of claims 1 to 6, wherein the subject has or has not been receiving steroid treatment, optionally the steroid is a corticosteroid, and optionally the corticosteroid is a glucocorticoid.

8. The use according to any one of claims 1 to 7, wherein the cut-off value is between 30 pg / mL and 90 pg / mL.

9. The method of claim 1 , wherein the cutoff value is about 35 pg / mL, 40 pg / mL, 45 pg / mL, 50 pg / mL, 55 pg / mL, 60 pg / mL, 65 pg / mL, 70 pg / mL, 75 pg / mL, 80 pg / mL or 85 pg / mL.

10. The use of any one of claims 1-9, wherein the subject has an anatomical response to the anti-VEGF therapeutic as measured by a change in central macular thickness (CMT) or foveal thickness (CFT), or as measured by a relative change in central macular thickness (CMT), wherein CMT or CFT is defined as the perpendicular distance between the inner limiting membrane and the retinal pigment epithelium at the center of the fovea, wherein the relative change in CMT is defined by the change in CMT divided by the baseline central macular thickness (bl-CMT).

11. The use according to any one of claims 1 to 10, wherein the subject has a baseline central macular thickness (bl-CMT) level above a bl-CMT cut-off value, wherein the bl-CMT cut-off value is between about 250 μm and about 600 μm.

12. Use of an anti-VEGF therapeutic agent in the preparation of a kit for treating retinal vein occlusion (RVO), wherein the kit comprises: i) the anti-VEGF therapeutic, and ii) instructions for administering the anti-VEGF therapeutic to a subject having a VEGF level in aqueous humor above a cutoff value, and wherein the cutoff value is between 10 pg / mL and 100 pg / mL.

13. The use according to claim 12, wherein the kit further comprises instructions for not administering the anti-VEGF therapeutic agent to a subject whose VEGF level in aqueous humor is not above a critical value, and / or instructions for administering an anti-inflammatory agent or an immunosuppressant or a combination thereof to a subject whose VEGF level in aqueous humor is above a critical value, optionally the anti-inflammatory agent or immunosuppressant is selected from glucocorticoids, methotrexate, cyclosporin A, tacrolimus, azathioprine, or a biological agent targeting a pro-inflammatory molecule such as interleukin-6, interleukin-8, and tumor necrosis factor-α.

14. The use according to claim 12 or 13, wherein the subject having an aqueous humor VEGF level above a cut-off value has a bl-CMT level above a bl-CMT cut-off value, and wherein the bl-CMT cut-off value is between about 250 μm and about 600 μm.

15. A kit comprising i) an anti-VEGF therapeutic, and ii) instructions for administering the anti-VEGF therapeutic to a subject having VEGF levels in aqueous humor above a cutoff value, wherein the cutoff value is between 10 pg / mL and 100 pg / mL.

16. The kit of claim 15, further comprising instructions for not administering the anti-VEGF therapeutic agent to a subject whose VEGF level in aqueous humor is not above a threshold value, and / or instructions for administering an anti-inflammatory agent or an immunosuppressant or a combination thereof to a subject whose VEGF level in aqueous humor is above a threshold value, optionally the anti-inflammatory agent or immunosuppressant is selected from glucocorticoids, methotrexate, cyclosporin A, tacrolimus, azathioprine, or a biologic targeting a pro-inflammatory molecule such as interleukin-6, interleukin-8, and tumor necrosis factor-α.

17. The kit of claim 15 or 16, wherein the subject having an aqueous humor VEGF level above a cut-off value has a bl-CMT level above a bl-CMT cut-off value, and wherein the bl-CMT cut-off value is between about 250 μm and about 600 μm.

18. A kit, comprising: i) reagents for determining the level of VEGF in the aqueous humor of a subject, and ii) instructions comprising a cut-off value for the VEGF level and optionally instructions for comparing the determined VEGF level to the cut-off value, wherein the cut-off value is between 10 pg / mL and 100 pg / mL.

19. The kit according to claim 18, further comprising a first standard sample having a VEGF level substantially equal to a critical value.

20. The kit according to claim 18 or 19, further comprising a second standard sample having a VEGF level higher than a critical value.

21. The kit according to any one of claims 18 to 20, further comprising a third standard sample having a VEGF level below a critical value.

22. The kit of any one of claims 18-21, wherein the agent is an anti-VEGF antibody or an antigen-binding fragment thereof, optionally the antigen-binding fragment is Fab, Fab', F(ab')2 or Fv.

23. The kit according to any one of claims 18 to 22, comprising an enzyme reactant, a magnetic separation reagent, a stabilizer, a standard, one or more concentrated wash buffers and / or a substrate buffer.

24. The kit according to claim 23, wherein the enzyme reactant comprises VEGF-specific antibody I labeled with alkaline phosphatase, and wherein the labeling ratio of VEGF antibody I to alkaline phosphatase is 1:0.

75.

25. The kit according to claim 23 or 24, wherein the magnetic separation reagent comprises 0.05 mass % of magnetic particles, wherein the magnetic particles are coated with VEGF-specific antibody II, wherein the magnetic separation reagent further comprises a blocking agent, wherein the blocking agent is a mutant alkaline phosphatase.

26. The kit according to claim 24 or 25, wherein VEGF-specific antibody I and / or VEGF-specific antibody II is an anti-VEGF165 / VEGFA antibody.

27. The kit according to any one of claims 23 to 26, wherein the stabilizer comprises one or more agents selected from the group consisting of Tris, NaCl, Tween 20, bovine serum albumin, mutant alkaline phosphatase and ProClin 300.

28. The kit according to any one of claims 23 to 27, wherein the wash buffer comprises Tris, NaCl, Tween-20, Triton-100 and ProClin 300.

29. A composition comprising i) aqueous humor isolated from a subject, and ii) an agent that interacts directly or indirectly with VEGF in the isolated aqueous humor, wherein the isolated aqueous humor comprises a VEGF level above a cutoff value, and the cutoff value is between 10 pg / mL and 100 pg / mL.

30. A composition comprising i) aqueous humor isolated from a subject, and ii) an agent that interacts directly or indirectly with VEGF in the isolated aqueous humor, wherein the isolated aqueous humor comprises a VEGF level no greater than a cutoff value, and the cutoff value is between 10 pg / mL and 100 pg / mL.

31. The composition of claim 29 or 30, wherein the agent is an anti-VEGF antibody or an antigen-binding fragment thereof, optionally the antigen-binding fragment is Fab, Fab', F(ab')2 or Fv.

32. The composition according to any one of claims 29-31, wherein the reagent comprises VEGF-specific antibody I labeled with alkaline phosphatase, and wherein the labeling ratio of VEGF antibody I to alkaline phosphatase is 1:0.

75.

33. The composition according to any one of claims 29-32, wherein the reagent comprises 0.05 mass % of magnetic particles, wherein the magnetic particles are coated with VEGF-specific antibody II, wherein the reagent further comprises a blocking agent, wherein the blocking agent is a mutant alkaline phosphatase.

34. The composition according to claim 32 or 33, wherein VEGF-specific antibody I and / or VEGF-specific antibody II is an anti-VEGF165 / VEGFA antibody.

35. An anti-VEGF therapeutic for treating retinal vein occlusion (RVO) in a subject, wherein the level of VEGF in the subject's aqueous humor is above a cutoff value, and the cutoff value is between 10 pg / mL and 100 pg / mL.

36. The anti-VEGF therapeutic agent for use according to claim 35, wherein the anti-VEGF therapeutic agent comprises: An anti-VEGF antibody or an antigen-binding fragment thereof, optionally the antigen-binding fragment is Fab, Fab', F(ab')2 or Fv; A VEGF receptor (VEGFR) fusion protein, optionally the VEGFR fusion protein is a VEGFR-Fc immunoadhesin; or An anti-VEGF gene therapy agent, comprising a nucleic acid encoding a VEGF antagonist, a nucleic acid encoding a protein that downregulates VEGF / VEGFR expression, and / or a nucleic acid targeting a nucleic acid and / or protein involved in VEGF / VEGFR expression, optionally the anti-VEGF gene therapy agent comprises a viral vector or a non-viral vector, optionally the viral vector is an adeno-associated virus (AAV) vector.

37. An anti-VEGF therapeutic for use according to claim 35 or 36, wherein the anti-VEGF therapeutic is administered or is to be administered intravitreally, suprachoroidally and / or subretinally.

38. An anti-VEGF therapeutic for use according to any one of claims 35 to 37, wherein the subject suffers from macular edema secondary to retinal vein occlusion (ME-RVO), optionally the retinal vein occlusion is central retinal vein occlusion (CRVO) and / or branch retinal vein occlusion (BRVO).

39. An anti-VEGF therapeutic for use according to any one of claims 35 to 38, wherein the subject has or has not been treated with the anti-VEGF therapeutic or another anti-VEGF therapeutic.

40. An anti-VEGF therapeutic for use according to any one of claims 35 to 38, wherein the subject has been treated with an anti-inflammatory agent or an immunosuppressant or a combination thereof, wherein the VEGF level in the subject's aqueous humor is above a threshold value, optionally the anti-inflammatory agent or immunosuppressant is selected from glucocorticoids, methotrexate, cyclosporin A, tacrolimus, azathioprine or a biologic targeting a pro-inflammatory molecule such as interleukin-6, interleukin-8 and tumor necrosis factor-α.

41. An anti-VEGF therapeutic for use according to any one of claims 35-38, wherein the subject has not been treated with an anti-inflammatory agent or an immunosuppressant or a combination thereof, wherein the subject's VEGF level in aqueous humor is above a threshold value, optionally the anti-inflammatory agent or immunosuppressant is selected from glucocorticoids, methotrexate, cyclosporin A, tacrolimus, azathioprine or a biologic targeting a pro-inflammatory molecule such as interleukin-6, interleukin-8 and tumor necrosis factor-α.

42. An anti-VEGF therapeutic for use according to any one of claims 35 to 41, wherein the cut-off value is between 30 pg / mL and 90 pg / mL.

43. An anti-VEGF therapeutic for use according to any one of claims 35-42, wherein the cut-off value is about 35 pg / mL, 40 pg / mL, 45 pg / mL, 50 pg / mL, 55 pg / mL, 60 pg / mL, 65 pg / mL, 70 pg / mL, 75 pg / mL, 80 pg / mL or 85 pg / mL.

44. An anti-VEGF therapeutic for use according to any one of claims 35-43, wherein the subject has an anatomical response to the anti-VEGF therapeutic as measured by a change in central macular thickness (CMT) or foveal thickness (CFT), or as measured by a relative change in central macular thickness (CMT), wherein the relative change in CMT is defined by the change in CMT divided by the baseline central macular thickness (bl-CMT).

45. An anti-VEGF therapeutic for use according to any one of claims 35-44, wherein the subject's baseline central macular thickness (bl-CMT) level is above the bl-CMT cut-off value, and wherein the bl-CMT cut-off value is between about 250 μm and about 600 μm.

46. ​​A method for predicting an anatomical response of an eye of a subject to treatment with an anti-VEGF therapeutic agent, the method comprising determining a VEGF level in aqueous humor collected from the subject, wherein a determined VEGF level above a cutoff value indicates that the eye has a good anatomical response to treatment with the anti-VEGF therapeutic agent, wherein a determined VEGF level not above the cutoff value indicates that the eye has a poor anatomical response to treatment with the anti-VEGF therapeutic agent, and wherein the cutoff value is between 10 pg / mL and 100 pg / mL.

47. The method of claim 46, wherein the anatomical response to the anti-VEGF therapeutic is measured by a change in central macular thickness (CMT) or foveal thickness (CFT), wherein CMT or CFT is defined as the perpendicular distance between the inner limiting membrane and the retinal pigment epithelium at the center of the fovea; or wherein the anatomical response to the anti-VEGF therapeutic is measured by a relative change in central macular thickness (CMT), defined as the change in CMT divided by the baseline central macular thickness (bl-CMT).

48. The method of claim 47, wherein a determined VEGF level above a cut-off value is predictive of a change in CFT of < -200 μm.

49. The method of claim 47, wherein a determined VEGF level below a cut-off value is predictive of a change in CFT > -200 μm.

50. The method of any one of claims 46-49, wherein the cutoff value is between 30 pg / mL and 90 pg / mL.

51. The method of any one of claims 46-50, wherein the cutoff value is about 35 pg / mL, 40 pg / mL, 45 pg / mL, 50 pg / mL, 55 pg / mL, 60 pg / mL, 65 pg / mL, 70 pg / mL, 75 pg / mL, 80 pg / mL, or 85 pg / mL.

52. The method of any one of claims 46-51, wherein a determined VEGF level above 90 pg / mL predicts a good anatomical response of the eye to treatment with the anti-VEGF therapeutic agent.

53. The method of any one of claims 46-52, wherein a determined VEGF level below 30 pg / mL predicts that the eye will have a poor anatomical response to treatment with the anti-VEGF therapeutic agent.

54. The method of any one of claims 46-53, wherein VEGF levels are determined by immunoassay, optionally an enzyme immunoassay.

55. The method of any one of claims 46-54, wherein the subject is in need of treatment for retinal vein occlusion (RVO).

56. The method of any one of claims 46-55, wherein the subject is in need of treatment for macular edema secondary to retinal vein occlusion (ME-RVO), optionally the retinal vein occlusion is central retinal vein occlusion (CRVO) and / or branch retinal vein occlusion (BRVO).

57. The method of any one of claims 46-56, further comprising administering to the subject intravitreally, suprachoroidally, or subretinally an anti-VEGF therapeutic or other anti-VEGF therapeutic when the determined VEGF level is above a cut-off value.

58. The method of claim 57, wherein the VEGF level determined is greater than 90 pg / mL.

59. The method of claim 58, comprising administering to the subject intravitreally, suprachoroidally, or subretinally an anti-VEGF therapeutic or other anti-VEGF therapeutic.

60. The method of any one of claims 46-56, further comprising administering to the subject a treatment other than the anti-VEGF therapeutic agent when the determined VEGF level is below a threshold value.

61. The method of claim 60, wherein the VEGF level determined is less than 30 pg / mL.

62. The method of claim 61, wherein the method does not administer to the subject intravitreal, suprachoroidal, or subretinal anti-VEGF therapeutics or other anti-VEGF therapeutics.

63. The method of any one of claims 46-62, wherein prior to the determining step, the subject has not been treated with the anti-VEGF therapeutic or another anti-VEGF therapeutic.

64. The method of any one of claims 46-62, wherein prior to the determining step, the subject has been treated with the anti-VEGF therapeutic or another anti-VEGF therapeutic.

65. The method according to any one of claims 46-64, wherein prior to the determining step, the subject has not received steroid therapy, the steroid can be selected as a corticosteroid, further selected as a glucocorticoid.

66. The method according to any one of claims 46-64, wherein prior to the determining step, the subject has been receiving steroid therapy, the steroid being selected as a corticosteroid, further selected as a glucocorticoid.

67. The method of any one of claims 46-66, wherein the subject is selected to initiate, continue or resume use of an anti-VEGF drug or other anti-VEGF drug, and / or to discontinue treatment with a steroid, which may be a corticosteroid, further selected as a glucocorticoid.

68. The method of any one of claims 46-66, wherein the subject chooses to discontinue use of an anti-VEGF drug or other anti-VEGF drug, and / or chooses to initiate, continue or resume treatment with a steroid, which may be a corticosteroid, further selected as a glucocorticoid.

69. The method of any one of claims 46-68, wherein the subject is a human.

70. The method of any one of claims 46-69, wherein the anti-VEGF therapeutic is formulated for intravitreal, suprachoroidal and / or subretinal administration, optionally further comprising mixing the anti-VEGF therapeutic with a pharmaceutically acceptable excipient or carrier.

71. The method of any one of claims 46-70, further comprising determining a level of bl-CMT in the subject, wherein a determined level of bl-CMT above a bl-CMT cutoff value indicates that the eye has a good anatomical response to treatment with the anti-VEGF therapeutic agent, wherein a determined level of bl-CMT not above a bl-CMT cutoff value indicates that the eye has a poor anatomical response to treatment with the anti-VEGF therapeutic agent, and wherein the bl-CMT cutoff value is between about 250 μm and about 600 μm.

72. A method for predicting anatomic response of an eye of a subject to treatment with an anti-VEGF therapeutic agent, the method comprising determining the level of VEGF in aqueous humor collected from the subject and the level of bl-CMT in the subject, wherein the determined VEGF level is above a VEGF cutoff value, and the determined bl-CMT level is above a bl-CMT cutoff value, indicating that the eye has a favorable anatomical response to treatment with the anti-VEGF therapeutic agent; and Wherein the determined VEGF level is not higher than the VEGF cut-off value or the determined bl-CMT level is not higher than the bl-CMT cut-off value, it indicates that the eye has a poor anatomical response to treatment with the anti-VEGF therapeutic agent.

73. The method of claim 72, wherein the VEGF cut-off value is between 10 pg / mL and 100 pg / mL.

74. The method of claim 72 or 73, wherein the bl-CMT threshold is between about 250 μm and about 600 μm.

75. The method of any one of claims 72-74, wherein the anatomical response to the anti-VEGF therapeutic is measured by changes in CMT, which is defined as the perpendicular distance between the internal limiting membrane and the retinal pigment epithelium at the center of the fovea.

76. The method of any one of claims 72-74, wherein the anatomical response to the anti-VEGF therapeutic is measured by a relative change in CMT, wherein the relative change in CMT is defined by the change in CMT divided by bl-CMT.

77. The method of any one of claims 72-76, wherein the determined VEGF level is above a VEGF cutoff of 10 pg / mL to 100 pg / mL, and the determined bl-CMT level is above a bl-CMT cutoff of about 250 μm to about 600 μm, indicating that the eye has a good anatomical response to treatment with the anti-VEGF therapeutic agent.

78. The method of any one of claims 72-77, wherein a determined VEGF level above a VEGF cut-off value and a determined bl-CMT level above a bl-CMT cut-off value is predictive of a change in CFT of <-200 μm.

79. The method of any one of claims 72-77, wherein a determined VEGF level below a VEGF cut-off value or a determined bl-CMT level below a bl-CMT cut-off value is predictive of a change in CFT of > -200 μm.

80. The method of any one of claims 72-79, wherein the VEGF cutoff is between 30 pg / mL and 90 pg / mL.

81. The method of any one of claims 72-80, wherein the VEGF cutoff is about 35 pg / mL, 40 pg / mL, 45 pg / mL, 50 pg / mL, 55 pg / mL, 60 pg / mL, 65 pg / mL, 70 pg / mL, 75 pg / mL, 80 pg / mL, or 85 pg / mL.

82. The method of any one of claims 72-81, wherein the bl-CMT threshold is between 250 μm and 600 μm.

83. The method of any one of claims 72-82, wherein the bl-CMT threshold is about 250 μm, 300 μm, 350 μm, 400 μm, 450 μm, 500 μm, 550 μm, or 600 μm.

84. The method of any one of claims 72-83, wherein VEGF levels are determined by immunoassay, optionally an enzyme immunoassay.

85. The method of any one of claims 72-84, wherein the subject is in need of treatment for retinal vein occlusion (RVO).

86. The method of any one of claims 72-85, wherein the subject is in need of treatment for macular edema secondary to retinal vein occlusion (ME-RVO), optionally the retinal vein occlusion is central retinal vein occlusion (CRVO) and / or branch retinal vein occlusion (BRVO).

87. The method of any one of claims 72-78 and 80-86, further comprising administering to the subject intravitreal, suprachoroidal, or subretinal an anti-VEGF therapeutic or other anti-VEGF therapeutic when the determined VEGF level is above a VEGF cutoff value and when the determined bl-CMT level is above a bl-CMT cutoff value.

88. The method of claim 87, wherein the VEGF level determined is greater than 90 pg / mL.

89. The method of claim 88, comprising administering to the subject intravitreally, suprachoroidally, or subretinally an anti-VEGF therapeutic or other anti-VEGF therapeutic.

90. The method of any one of claims 72-76 and 79-86, further comprising administering to the subject a treatment other than the anti-VEGF therapeutic agent when the determined VEGF level is below the VEGF cutoff value.

91. The method of claim 90, wherein the determined VEGF level is less than 30 pg / mL and the subject is not administered an intravitreal, suprachoroidal, or subretinal anti-VEGF therapeutic or other anti-VEGF therapeutic.

92. The method of claim 90 or 91, wherein the treatment other than the anti-VEGF therapeutic is a steroid, optionally a corticosteroid, further optionally a glucocorticoid.

93. The method of any one of claims 72-92, wherein prior to the determining step, the subject has or has not been treated with the anti-VEGF therapeutic or another anti-VEGF therapeutic.

94. The method according to any one of claims 72-93, wherein prior to the determining step, the subject has or has not been treated with steroids, the steroids may be selected as corticosteroids, further selected as glucocorticoids.

95. The method of any one of claims 72-92, wherein the subject has or has not been treated with an anti-inflammatory agent or an immunosuppressant, or a combination thereof.

96. The method of claim 95, wherein the anti-inflammatory agent or immunosuppressant is selected from glucocorticoids, methotrexate, cyclosporin A, tacrolimus, azathioprine, or a biologic targeting a pro-inflammatory molecule such as interleukin-6, interleukin-8, and tumor necrosis factor-α.

97. The method of any one of claims 72-96, wherein the subject is selected to initiate, continue or resume use of an anti-VEGF drug or other anti-VEGF drug, and / or to discontinue treatment with a steroid, which may be a corticosteroid, further selected as a glucocorticoid.

98. The method of any one of claims 72-96, wherein the subject is chosen to stop taking an anti-VEGF drug or other anti-VEGF drug, and / or is chosen to start, continue or resume treatment with a steroid, which may be a corticosteroid, further selected as a glucocorticoid.

99. The method of any one of claims 72-98, wherein the subject is a human.

100. The method of any one of claims 72-99, wherein the anti-VEGF therapeutic is formulated for intravitreal, suprachoroidal and / or subretinal administration, optionally further comprising mixing the anti-VEGF therapeutic with a pharmaceutically acceptable excipient or carrier.

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