Modulating neuroinflammation

Abstract

To provide compositions and methods for treating neuroinflammation and neurodegeneration and / or reducing risk of development or progression of neuroinflammation and neurodegeneration.SOLUTION: The present invention provides a method comprising administration, to a subject in need thereof, of therapeutically effective amounts of one or more of: (i) HSP60 or HSP27, or active fragments thereof; and / or (ii) IGFBPL or active fragments thereof.SELECTED DRAWING: None

Description

Technical Field

[0001] This application claims the benefit of U.S. Provisional Patent Application No. 62 / 813,556, filed Mar. 4, 2019. The entire content of the above patent document is hereby incorporated by reference.

[0002] This invention was made with government support under grant number EY025259 awarded by the National Institutes of Health. The government has certain rights in this invention.

[0003] Compositions and methods for treating neuroinflammation and neurodegeneration and / or reducing the risk of onset or progression of neuroinflammation and neurodegeneration, for example, HSP60 and / or HSP27 for nasal administration, and for example, IGFBPL1 for nasal, systemic, or intravitreal administration, are described herein.

Background Art

[0004] Glaucoma is a leading cause of blindness and remains an unsolved global medical problem.

Summary of the Invention

[0005] Methods are provided for treating neuroinflammation and neurodegeneration and / or reducing the risk of onset or progression of neuroinflammation and neurodegeneration in a subject in need thereof. The methods include administering to the subject a therapeutically effective amount of (i) HSP60 and / or HSP27, or an active fragment thereof, and / or (ii) IGFBPL or an active fragment thereof.

[0006] ​​​​​​​​​​In some embodiments, a therapeutically effective amount is sufficient to reduce inflammation and neuronal cell death in a subject.

[0007] In some embodiments, the method includes the nasal administration of HSP60 and / or HSP27 as well as the systemic (e.g., nasal or oral) or intravitreal administration of IGFBPL.

[0008] In some embodiments, the intravitreal administration of IGFBPL includes an intravitreal injection.

[0009] In some embodiments, the subject has glaucoma, autism, multiple sclerosis, Alzheimer's disease, Parkinson's disease, ischemic retinopathy, age-related macular degeneration, seizures, ischemic and traumatic optic neuropathy, or diabetic retinopathy.

[0010] In some embodiments, the method reduces inflammation and neuronal cell death in the eye of the subject.

[0011] In some embodiments, the method reduces inflammation and neuronal cell death in the brain or spinal cord of the subject.

[0012] Also provided herein is a kit comprising a composition comprising HSP60 and / or HSP 27 for use in the methods described herein, and a composition comprising IGFBPL1.

[0013] Further provided herein is a composition comprising HSP60 and / or HSP27 for use in a method for treating and / or reducing the risk of onset or progression of neuroinflammation and neurodegeneration, and / or a composition comprising IGFBPL1.

[0014] ​​​​​​In some embodiments, HSP60 and / or HSP27 are formulated for nasal administration , and IGFBPL1 is formulated for administration to the eye, e.g., intravitreal administration. In some embodiments , HSP60 and / or HSP27 are formulated for nasal administration and IGFBPL1 is formulated for administration systemically, e.g., to the eye or for nasal administration. In some embodiments, HSP60 and / or HSP27 are formulated for nasal administration and IGFBPL1 is formulated for nasal administration . In some embodiments, one, two, or all three of HSP60 and / or HSP27 and IGFB PL1 are formulated together in a single composition for nasal administration .

[0015] Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. Methods and materials are described herein for use in the present invention, and other suitable methods and materials known in the art can also be used . The materials, methods, and examples are illustrative only and not intended to be limiting. All publications , patent applications, patents, sequences, database entries, and other references mentioned herein are incorporated by reference in their entirety. In case of conflict, the present specification, including definitions , will control .

[0016] Other features and advantages of the present invention will become apparent from the following detailed description and the drawings, and from the scope of the claims

[0017] The patent or application file contains at least one drawing executed in color. The A copy of this patent or patent application publication with drawings (multiple possible) is provided by the Patent Office upon payment of the claims and the necessary fees. This is provided by the Patent Office upon payment of the claims and the necessary fees.

Brief Description of the Drawings

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Mode for Carrying Out the Invention

[0019] Methods for treating neuroinflammation and neurodegeneration and for reducing the risk of the onset or progression of neuroinflammation and neurodegeneration are described herein. The methods include (i) administration of HSP60 or HSP27, or an active fragment thereof, and / or (ii) administration of IGFB PL or an active fragment thereof. Use of the methods can reduce inflammation and neuronal death in the eye as well as in other locations including the CNS and PNS. The methods can include, for example, nasal administration of HSP60 or HSP27 for the treatment of glaucoma, and nasal, systemic, or intraocular administration of IGFBPL1, for example, intravitreal injection or including topical ocular administration. ​​​

[0020] HSP60 (HSPD1, Heat shock protein family D (Hsp60) member 1 ) New evidence implicating an autoimmune mechanism in glaucoma is emerging, but its relative etiological importance has not yet been proven. As shown herein, neuronal and visual loss in glaucoma, and other immune-related conditions, are associated with existing memory T cells that have been primed by exposure to bacterial HSP60 early in life. Elucidating the association of the immune mechanism in glaucoma and its progression to neurodegeneration with commensal microorganisms provides a basis for new diagnostic approaches.

[0021] The amino acid sequence of human hsp60 is provided by GenBank accession number NP_002147.2, and the amino acid sequence of bacterial hsp60 is provided by GenBank accession number WP_000729117.1, and these amino acid sequences are incorporated herein by reference. HSP60 is preferably formulated for nasal administration to induce tolerance to HSP60. Alternatively, oral or subcutaneous administration can be used. See also WO2012118863.

[0022] HSP27 (Heat shock protein family B (small) member 1 (HSPB1)) HSP27, also known as HSPB1, is shown herein to directly induce a pro-inflammatory response in HMC3 cells. The amino acid sequence of human HSP27 is provided by GenBank accession number NP_001531.1, and this amino acid sequence is incorporated herein by reference. HSP27 is formulated for nasal administration to induce tolerance to HSP27. ​​​​​​​​​​It is preferably administered parenterally. Alternatively, oral or subcutaneous administration can be used. W See also O2012118863.

[0023] IGFBPL (insulin-like growth factor-binding protein-like 1) Insulin growth factor-binding protein-like 1 (IGFBPL1) regulates the survival of neonatal mouse retinal ganglion cells (RGCs) via the insulin-like growth factor 1-mediated signaling pathway and promotes neurite outgrowth (Guo et al., Sci Rep. 2018 Feb 1;8(1):2054). As shown in this specification, IGFBPL1 actively suppresses microglia in adult animals / animals after the neonatal period. The amino acid sequence of human IGFBPL1 is provided by GenBank accession number NP_001007564, and this amino acid sequence is incorporated herein by reference. See also WO2012118796.

[0024] Methods of treatment and prevention By administering the compositions described herein to a subject, disorders associated with abnormal or unwanted immune responses, such as neuroinflammation or neurodegenerative disorders associated with excessive or abnormal activation of microglia, can be treated or prevented. Examples of such disorders include non-arteritic ischemic optic neuropathy (NAION), autism, multiple sclerosis, Alzheimer's disease, Parkinson's disease, ischemic retinopathy, glaucoma, age-related macular degeneration, stroke, ischemic and traumatic optic neuropathies, as well as diabetic retinopathy, but are not limited thereto; in some embodiments, the disease is associated with vision loss and / or increased intraocular pressure. Using the methods, subjects having those diseases can be treated, e.g., to reverse vision loss and nerve cell loss associated with those diseases ​​​​​It is possible to reduce or treat these. US8 / 198,284; WO / 2 017 / 213504; WO2012118863; and WO2012118796 are also referred to. All of the above patent documents are hereby incorporated by reference into this specification.

[0025] The methods of treatment or prevention described herein can include, for example, administering to a subject a sufficient transnasal or subcutaneous HSP60 or HSP27 composition to stimulate the mucosal immune system for ten minutes. In some embodiments, the method includes administering a sufficient transnasal HSP60 or HSP27 composition to increase the level of regulatory T cells by, for example, about 50%, 75%, 100%, 200%, 300% or more above baseline In some embodiments, the method includes administering a transnasal or subcutaneous HSP60 or HSP27 composition and / or an IGFBPL1 composition in an amount sufficient to effect an improvement in one or more clinical markers of vision loss (e.g., decreased visual acuity) or disability ; for example, in multiple sclerosis, such markers can include gadolinium-enhanced lesions visualized by MRI, or the Fazekas, or Barkhof MRI criteria, or the McDonald diagnostic criteria of the patient The IGFBPL1 composition can be administered transnasally, systemically, or intravitreally, for example, by eye drops or intravitreal administration

[0026] In some embodiments, the treatment is administered to a subject diagnosed with a disorder associated with microglial activation; such diagnosis is made by a skilled practitioner using known methods and conventional techniques In some embodiments, the method includes administering a transnasal or subcutaneous HSP60 or HSP27 composition and / or an IGFBPL1 composition in an amount sufficient to effect an improvement in one or more clinical markers of vision loss (e.g., decreased visual acuity) or disability ; for example, in multiple sclerosis, such markers can include gadolinium-enhanced lesions visualized by MRI, or the Fazekas, or Barkhof MRI criteria, or the McDonald diagnostic criteria of the patient ; for example, in multiple sclerosis, such markers can include gadolinium-enhanced lesions visualized by MRI, or the Fazekas, or Barkhof MRI criteria, or the McDonald diagnostic criteria of the patient

[0027] ​​​​​​It can be performed. In some embodiments, the method has a disorder associated with microglial activation comprising diagnosing or identifying or selecting a subject, or identifying or selecting a subject based on the presence or diagnosis of a disorder associated with microglial activation. In some embodiments, the subject is an adult, e.g., a human at least 18 years old, or a postnatal human, e.g., at least 6 months old or 1 year old.

[0028] Pharmaceutical Compositions and Methods of Administration The methods described herein include the use of a pharmaceutical composition comprising one or more of HSP60, HSP27, or IGF BPL1 as an active ingredient.

[0029] The pharmaceutical composition typically comprises a pharmaceutically acceptable carrier. As used herein , the term "pharmaceutically acceptable carrier" includes saline, solvents, dispersion media, coatings, antibacterial and antifungal agents, isotonic and absorption delaying agents, and the like, which are compatible with pharmaceutical administration.

[0030] The pharmaceutical composition is typically formulated to be compatible with its intended route of administration.

[0031] Examples of routes of administration include systemic parenteral administration, e.g., intravenous, intraperitoneal, intradermal, or subcutaneous; topical administration to the eye, e.g., topical, intravitreal, intraocular, orbital, periorbital, subconjunctival, subretinal, sub-Tenon's capsule or trans-scleral; and systemic oral administration. In some embodiments, intravitreal administration or, in particular, administration by eye drops, ointments, creams, gels, or lotions may be used. In some embodiments, the composition is administered systemically, e.g., orally; in a preferred embodiment, the composition is administered, e.g., topically (eye drops, lotion, or ointment). and is administered to the eye by topical, subconjunctival, or local injection, for example, periorbitally or intravitreally; see, for example, Gaudana et al., AAPS J. 12(3):348-360 (2010); Fischer et al., Eur J Ophthalm ol. 21 Suppl 6:S20-6 (2011). Administration may be provided as a regular bolus (e.g., intravitreally or intravenously) or as a continuous infusion from an internal reservoir (e.g., from an implantable tablet placed in an intraocular or extraocular location; see U.S. Patent Nos. 5,443,505 and 5,76 6,242) or from an external reservoir (e.g., from an intravenous drip bag or a contact lens sustained release formulation system). The composition may be administered locally, for example, by continuous release from a sustained release drug delivery device fixed to the inner wall of the eye or via targeted transscleral controlled release into the choroid (see, for example, PCT / US0 0 / 00207, PCT / US02 / 14279, PCT / US2004 / 004625 , Ambati et al. (2000) Invest. Ophthalmol. Vis. Sci. 41:1181-1185, and Ambati et al (2000) Invest. Ophthalmol. Vis. Sci. 41:1186-1191). Various devices suitable for local administration of drugs into the eye are known in the art; see, for example, U.S. Patent Nos. 6,251,090, 6,299,895, 6,416, 777, 6,413,540, and 6,375,972, as well as PCT / US00 / 28187.

[0032] A pharmaceutical composition is typically formulated to be compatible with its intended route of administration. Examples of routes of administration include systemic (e.g., parenteral, nasal, subcutaneous, and oral) and topical (ophthalmic, e.g., intravitreal or topical ocular) administration. Thus, compositions containing the compositions described herein, such as in formulations for ophthalmic administration, including, for example, microcapsules, microemulsions, or nanoparticles, in the form of, for example, eye drops, lotions, creams, are also within the scope of the present disclosure. Methods of formulating pharmaceutical compositions suitable for ophthalmic delivery are known in the art, see, for example, Losa et al., Pharmaceutical Research 10:1(80 - 87 (1993); Gasco et al., J. Pharma Biomed Anal., 7(4):433 - 439 (1989); Fischer et al., Eur J Ophthalmol. 21 Suppl 6:S20 - 6 (2011); and Tangri and Khurana, Intl J Res Pharma Biomed Sci., 2(4):1541 - 1442 (2011). Examples of routes of administration include systemic (e.g., parenteral, nasal, subcutaneous, and oral) and topical (ophthalmic, e.g., intravitreal or topical ocular) administration. Thus, compositions containing the compositions described herein, such as in formulations for ophthalmic administration, including, for example, microcapsules, microemulsions, or nanoparticles, in the form of, for example, eye drops, lotions, creams, are also within the scope of the present disclosure. Methods of formulating pharmaceutical compositions suitable for ophthalmic delivery are known in the art, see, for example, Losa et al., Pharmaceutical Research 10:1(80 - 87 (1993); Gasco et al., J. Pharma Biomed Anal., 7(4):433 - 439 (1989); Fischer et al., Eur J Ophthalmol. 21 Suppl 6:S20 - 6 (2011); and Tangri and Khurana, Intl J Res Pharma Biomed Sci., 2(4):1541 - 1442 (2011). General methods of formulating suitable pharmaceutical compositions are known in the art, see, for example, Remington: The Science and Practice of Pharmacy, 21st ed., 2005; and the books in the series Drugs and the Pharmaceutical Sciences: a Series of Textbooks and Monographs (Dekker, NY). For example, solutions or suspensions used for parenteral, intradermal, or subcutaneous application may contain the following components: water for injection, saline, fixed oils, polyethylene glycols, glycerin, propylene glycol or other solvents; antibacterial agents such as benzyl alcohol or methyl paraben; antioxidants such as ascorbic acid or sodium bisulfite; chelating agents such as ethylenediaminetetraacetic acid; buffers such as

[0033] buffers such as acetate, citrate or phosphate, and agents for adjusting tonicity such as sodium chloride or dextrose. Parenteral formulations may be presented in unit - dose or multi - dose containers, for example, sealed ampoules and vials, and may be stored in a freeze - dried (lyophilized) condition requiring only the addition of the sterile liquid carrier, for example, water for injection, immediately prior to use. Sterile injectable solutions are prepared by incorporating the active compound in the required amount in Sterile dilute solvents such as ethylene glycol, glycerin, propylene glycol or other synthetic solvents. Antibacterial agents such as benzyl alcohol or methylparaben; ascorbic acid or Antioxidants such as sodium bisulfate; chelating agents such as ethylenediaminetetraacetic acid; acetic acid; Buffers such as citrate or phosphate and osmosides such as sodium chloride or glucose It is possible to include drugs for adjusting the pressure. The pH can be adjusted with an acid such as hydrochloric acid or sodium hydroxide. Parenteral formulations can be prepared using glass or plastic. Can be packaged in plastic ampoules, disposable syringes or multi-dose vials It is Noh.

[0034] Pharmaceutical compositions suitable for injectable use include sterile aqueous solutions (where water soluble) or dispersions and and sterile powders for the extemporaneous preparation of sterile injectable solutions or dispersions. For intravenous administration, suitable carriers include physiological saline, bacteriostatic water, Cremophor EL™ (BAS F, Parsippany, NJ) or phosphate-buffered saline (PBS). In this case, the composition must be sterile and fluid enough to exist easy syringability. The composition should be stable under the conditions of manufacture and storage and should be free from bacteria and fungi. The carrier must be protected against the contaminating action of microorganisms such as water, ethanol, etc. alcohols, polyols (e.g., glycerol, propylene glycol, and liquid polyethylene) The solvent or dispersion medium may contain ethylene glycol, etc., and suitable mixtures thereof. Proper fluidity can be achieved, for example, in the case of dispersions, by the use of a coating such as lecithin. This can be achieved by maintaining the required particle size and by using surfactants. Yes. Prevention of microbial activity can be achieved by various antibacterial and antifungal agents such as parabens, chlorobutanol, phenol, ascorbic acid, thimerosal and the like. In many cases, it is preferable to include in the composition tonicity agents such as saccharides, polyhydric alcohols such as mannitol and sorbitol, and sodium chloride. Long-term absorption of the injectable composition can be brought about by including in the composition agents that delay absorption such as aluminum monostearate and gelatin.

[0035] Sterile injectable solutions can be prepared by incorporating the active compound in the required amounts into a suitable solvent, optionally together with one or a combination of the ingredients listed above, and subsequently filtering and sterilizing. Generally, dispersions are prepared by incorporating the active compound into a sterile vehicle containing a basic dispersion medium and the required other ingredients from those listed above. In the case of sterile powders for the preparation of sterile injectable solutions, the preferred methods of preparation are vacuum drying and freeze-drying, by which powders of the active ingredient and any additional desired ingredients are obtained from

[0036] sterile-filtered solutions thereof beforehand. Oral compositions generally include an inert diluent or an edible carrier. For the purpose of oral therapeutic administration, the active compound can be incorporated with excipients and used in the form of tablets, troches, or capsules, such as gelatin capsules. Oral compositions can also be prepared using a fluid carrier for use as a mouthwash. Pharmaceutically compatible The pessary, troche, etc. are composed of the following ingredients, or compounds with similar properties: crystalline cellulose, Binders such as tragacanth gum or gelatin; excipients such as starch or lactose; Disintegrants such as alginic acid, Primogel, or corn starch; lubricants such as magnesium stearate or Sterotes; Glidants such as colloidal silicon dioxide; sweeteners such as sucrose or saccharin; or flavoring agents such as peppermint, methyl salicylate, or orange flavor, and can contain any of them.

[0037] For administration by inhalation, the compound can be delivered in the form of an aerosol spray from a pressurized container or dispenser containing a suitable propellant, such as a gas like carbon dioxide, or a nebulizer. Such methods include those described in U.S. Patent No. 6,468,798.

[0038] Systemic administration of the therapeutic compounds described herein is also possible by transmucosal or transdermal means. For transmucosal or transdermal administration, suitable penetration enhancers are used in the formulation for the barrier to be traversed. Such penetration enhancers are generally known in the art and include, for example, surfactants, bile salts, and fusidic acid derivatives for transmucosal administration. Transmucosal administration can be achieved through the use of nasal sprays or suppositories. For transdermal administration, the active compound is formulated into ointments, plasters, gels, or creams generally known in the art.

[0039] The pharmaceutical composition can be a suppository (e.g., with conventional suppository bases such as cocoa butter and other glycerides) It is also possible to prepare it in the form of a retention enema for co - administration or rectal delivery.

[0040] In one embodiment, the therapeutic compound is prepared together with a carrier that protects the therapeutic compound from rapid disappearance from the body, such as controlled - release formulations including implant tablets and microcapsule delivery systems. It can be prepared with biodegradable, biocompatible polymers such as ethylene vinyl acetate, polyanhydrides, polyglycolic acid, collagen, polyorthoesters, and polylactic acid. Such formulations can be prepared using standard techniques or, for example, purchased from Alza Corporation and Nova Pharmaceuticals, Inc. Liposome suspensions (including liposomes targeted to cells selected with monoclonal antibodies against cell antigens) can also be used as pharmaceutically acceptable carriers. These compounds can be prepared according to methods known to those skilled in the art, such as those described in U.S. Patent No. 4,522,811.

[0041] The pharmaceutical composition can be included together with instructions for administration in a container, pack, or dispenser.

[0042] Kit Kits for use in the methods described herein are also provided herein. For example, the kit can include, for example, a composition containing HSP60 or HSP27 for nasal administration, and a composition containing IGF - FBPL1 for nasal, systemic (e.g., oral) or ocular (e.g., topical ocular or intravitreal) administration. Instructions for use can also be included in the kit.

Examples

[0043] The present invention will be further described in the following examples, which do not limit the scope of the present invention described in the claims.

[0044] [Example 1] Immune tolerance to HSP60 attenuates neurodegeneration in a mouse model of glaucoma Primary open-angle glaucoma (POAG), the world's leading cause of blindness, is a disease that damages the optic nerve. POAG has been mainly associated with high intraocular pressure (IOP), but treatments that affect IOP do not completely prevent vision loss and blindness. 1,2 .

[0045] Elevated intraocular pressure (IOP) induces a T cell-mediated autoimmune response against HSP60, and in mice raised without a microbiota, HSP-specific T cell responses and neuronal loss disappear after elevated IOP. 3~9 . In this example, it was tested whether induction of tolerance to HSP60 attenuates glaucoma damage.

[0046] Immune tolerance to HSP60 was induced in male and female C57BL / 6J mice, 6 - 8 weeks old, by administering low-dose HSP60 intranasally (2 μM HSP60, daily for 7 days). Control mice were treated with saline. Glaucoma was induced by injecting microbeads (MB) into the anterior chamber twice and maintaining elevated IOP for 8 weeks. IOP was monitored weekly. Visual function was evaluated by optokinetic response (OMR) and scotopic threshold response of electroretinogram (pSTR). Mice were sacrificed at 2, 4, and 8 weeks. The immune response and T cell tolerance to HSP60 were analyzed by fluorescence-activated cell sorting (FACS). ​​ was performed. Glaucoma nerve damage was quantified by counting retinal ganglion cells (RGCs) and axons. was performed.

[0047] By 4 weeks, the MB-injected eyes showed an IOP of 20.1 ± 0.56 mmHg or higher compared to 11.6 ± 0.21 mmHg in the contralateral non-injected eyes (Figure 1). As shown by FACS analysis, nasal administration of low-dose HSP60 induced immune tolerance and increased the level of Tregs (Figure 2). No significant difference in IOP levels was observed between HSP60-treated mice and saline-treated mice. Treatment with HSP60 did not change visual acuity (VA), contrast sensitivity (CS), or pSTR compared to saline-treated mice prior to MB injection. However, when evaluated by OMR, HSP60-treated mice showed significantly higher VA and CS than saline-treated mice at all time points after MB injection (Figure 3A - C). Consistent with this, RGC function evaluated by pSTR was also significantly improved in HSP60-treated mice compared to saline-treated non-immune-tolerant mice at all time points after MB injection (Figure 4A - B). As shown in Figure 5, these results indicate that immune tolerance to HSP60 attenuates glaucoma RGC loss and functional loss in mice. (Figure 3A - C). Consistently, RGC function evaluated by pSTR was also significantly improved in HSP60-treated mice compared to saline-treated non-immune-tolerant mice at all time points after MB injection (Figure 4A - B).

[0048] As shown in Figure 5, these results indicate that immune tolerance to HSP60 attenuates glaucoma RGC loss and functional loss in mice.

[0049] [Example 2] Role of insulin-like growth factor-binding protein-like protein 1 in microglia in adult mice Insulin-like growth factor-binding protein-like protein 1 (IGFBPL1) plays a crucial role in promoting axonal outgrowth and survival of retinal ganglion cells (RGCs) during development. ​ (Guo et al., Sci Rep. 2018 Feb 1;8(1):2054). This requires the presence of insulin-like growth factor 1 (IGF1) and is mediated through the IGF1 receptor (IGF1R). Mature human RGCs are known to express low or undetectable levels of IGF1R , so in this example, it was investigated whether IGFBPL1 supports RGC survival after adult retinal injury.

[0050] The expression of IGF1, IGFBPL1, and IGF1R in the adult retina was examined in retinal whole-mounts using immunohistochemistry . Retinal whole-mounts were double-immunolabeled with primary antibodies against the RGC marker Brn -3 or the microglial marker Iba-1 to identify the cell-type specific expression of IGF1, IGF 1R, and IGFBPL1.

[0051] Purified microglia and RGCs isolated from 5-day-old Cx3CR1 / GFP mice were co-cultured in the presence or absence of LPS and / or IGFBPL1 and IGF1. Neuronal survival was determined using a LIVE / DEAD cell viability kit, and the percentage of surviving R GCs was quantified using Image J software.

[0052] According to the results, in the adult mouse retina, IGF1, IGF1R, and IGFBPL1 were shown to be expressed by microglia but not by RGCs (Figs. 6A - B). Adding IGF1 and / or IGFBPL1 to purified RGC cultures did not promote neuronal survival, but LPS stimulated microglial activation and significantly increased RGC death compared to control cultures. Caused death (P<0.05). IGF1 and / or IGFBPL1 significantly reduced neuronal death in LPS-treated microglia-induced RGC death in microglia-RGC co-cultures (see Fig. 12). In the glaucoma model, intravitreal injection of IGFBPL1 on days 3 and 10 after IOP elevation protected RGCs against elevated IOP-induced damage and prevented loss of RGC function and vision (Figs. 7A-D). IGFBPL1 suppressed microglial activation in glaucomatous retina (Figs. 8A-B), and also inhibited microglial activation, reactive gliosis, and production of pro-inflammatory cytokines (Fig. 9). As shown in Figs. 10A-B, when IGFBPL1 was deficient, microglial activation occurred in the adult retina and the levels of pro-inflammatory cytokines increased. Fig. 11 shows that RGC loss and functional deficits in IGFBPL1-deficient mice were rescued by intravitreal injection of IGFBPL1.

[0053] In the glaucoma model, intravitreal injection of IGFBPL1 on days 3 and 10 after IOP elevation protected RGCs against elevated IOP-induced damage and prevented loss of RGC function and vision (Figs. 7A-D). IGFBPL1 suppressed microglial activation in glaucomatous retina (Figs. 8A-B), and also inhibited microglial activation, reactive gliosis, and production of pro-inflammatory cytokines (Fig. 9). As shown in Figs. 10A-B, when IGFBPL1 was deficient, microglial activation occurred in the adult retina and the levels of pro-inflammatory cytokines increased. Fig. 11 shows that RGC loss and functional deficits in IGFBPL1-deficient mice were rescued by intravitreal injection of IGFBPL1.

[0054] This study reveals that in adult mice, IGFBPL1 is expressed rather by microglia than by RGCs. IGFBPL1 exerts a neuroprotective effect by acting on microglia. These results suggest that IGFBPL1 protects against neuronal death by regulating neuroinflammation.

[0055] [Example 3] In IGFBPL1 mice with ocular hypertension, IGFBPL1 protected against neuronal and - / - vision loss Insulin-like growth factor-binding protein-like 1 (IGFBPL1) is an insulin-like growth factor ​​​​​​​​​Survival and neurite outgrowth of neonatal mouse retinal ganglion cells (RGCs) regulated via the 1 - mediator signaling pathway are promoted (Guo et al., Sci Rep. 2018 Feb 1;8(1):2054). Newborn IGFBPL1 - deficient (IGFBPL1 ) mice had approximately 20% fewer RGCs compared to wild - type control mice. This study investigated the role of the IGF - / - BPL1 protein in neuronal cell survival and visual performance in adult mice with or without elevated intraocular pressure (IOP). The RGC density of 1, 2, and 7 - month - old IGFBPL1 mice was determined by Brn3a immunolabeling in retinal flat mounts. To induce an increase in IOP, 2 microliters of polystyrene microbeads (MB; 5×10 - / - / ml) were injected into the anterior chamber of one eye of adult male and female IGFBPL1 mice. Recombinant IGFBPL1 protein or sterile saline as a control was administered by intravitreal injection 3, 7, and 17 days after MB injection. Four weeks after MB injection, two researchers recorded the contrast sensitivity and optomotor response of visual acuity in mice in a masked manner. The mice were sacrificed, and the retinas were flat - mounted and processed for Brn3a immunolabeling to reveal surviving RGCs. Student's t - test was used for statistical analysis.

[0056] According to the data, it was revealed that in IGFBPL1 - / - mice, RGCs were progressively lost in the absence of IGFBPL1. In adult IGFBPL1 mice, IO The RGC density of 1, 2, and 7 - month - old IGFBPL1 6 mice was determined by Brn3a immunolabeling in retinal flat mounts. To induce an increase in IOP, 2 microliters of polystyrene microbeads (MB; 5×10 / ml) were injected into the anterior chamber of one eye of adult male and female IGFBPL1 - / - mice. Recombinant IGFBPL1 protein or sterile saline as a control was administered by intravitreal injection 3, 7, and 17 days after MB injection. Four weeks after MB injection, two researchers recorded the contrast sensitivity and optomotor response of visual acuity in mice in a masked manner. The mice were sacrificed, and the retinas were flat - mounted and processed for Brn3a immunolabeling to reveal surviving RGCs. Student's t - test was used for statistical analysis. protein or sterile saline as a control was administered by intravitreal injection 3, 7, and 17 days after MB injection. Four weeks after MB injection, two researchers recorded the contrast sensitivity and optomotor response of visual acuity in mice in a masked manner. The mice were sacrificed, and the retinas were flat - mounted and processed for Brn3a immunolabeling to reveal surviving RGCs. Student's t - test was used for statistical analysis. Four weeks after MB injection, two researchers recorded the contrast sensitivity and optomotor response of visual acuity in mice in a masked manner. The mice were sacrificed, and the retinas were flat - mounted and processed for Brn3a immunolabeling to reveal surviving RGCs. Student's t - test was used for statistical analysis. The mice were sacrificed, and the retinas were flat - mounted and processed for Brn3a immunolabeling to reveal surviving RGCs. Student's t - test was used for statistical analysis. The mice were sacrificed, and the retinas were flat - mounted and processed for Brn3a immunolabeling to reveal surviving RGCs. Student's t - test was used for statistical analysis. Student's t - test was used for statistical analysis.

[0057] According to the data, in IGFBPL1 - / - mice, it was revealed that RGCs were progressively lost in the absence of IGFBPL1. In adult IGFBPL1 mice, IO - / - mice, IO A transient increase in P led to a significant decrease in visual performance and RGC loss. IGFBP Treatment with L1 improved visual function (P<0.05) and RGC survival (P<0.05) in wild-type and IGFBPL1 - / - mice with glaucoma. Furthermore, retinal ischemic reperfusion injury was induced unilaterally in mice, and saline (isotype) or IGFBPL1 (BPL

[0058] 1) was intravitreally injected on day 1 (early ) or day 10 (late) after injury. Mice were sacrificed 4 weeks after injury, and the RGC density was quantified . RGC function was evaluated by pSTR amplitude at 2 and 4 weeks after injury (before sacrifice), and visual contrast sensitivity (CS) and visual acuity (VA) were measured using the optokinetic response (OKR) assay . As shown in Figure 13, after ischemic injury, RGC density was significantly increased, and pSTR amplitude as well as CS and VA values were improved in IGF FBPL1-treated mice compared to the saline-treated group. IGFBPL1 was known to be strongly expressed in the embryonic retina and barely detectable in the adult retina. These results indicate that in IGFB

[0059] PL1-deficient mice, lack of IGFB PL1 during embryogenesis induces progressive degeneration of RGCs. Administration of IGFBPL1 protected mice with glaucoma from RGC loss and vision loss. Collectively - / - speaking, IGFBPL1 is an important neuroprotective agent in retinas undergoing progressive degeneration, such as in glaucoma.

[0060] [Example 4] Heat shock proteins 27 and 60 directly activate human microglia​​​​ Glaucoma has autoimmune components caused by CD4+ T cells pre-stimulated by commensal bacteria. These CD4+ T cells enter the retina and cross-react with heat shock protein (HSP)-expressing neurons via the mechanism of molecular mimicry. As shown herein, microglial activation is the cause of the immune response and retinal degeneration in glaucoma. Considering the limited availability of primary human microglia, the immortalized human microglial cell clone 3 cell line (HMC3) is useful for examining microglial behavior under pathological conditions. To test whether HSP27 and HSP60 can induce microglial activation, cytokine expression and morphological changes were examined in HMC3. Other known inflammation promoters were used as positive controls. Glaucoma has autoimmune components caused by CD4+ T cells pre-stimulated by commensal bacteria. These CD4+ T cells enter the retina and cross-react with heat shock protein (HSP)-expressing neurons via the mechanism of molecular mimicry. As shown herein, microglial activation is the cause of the immune response and retinal degeneration in glaucoma. Considering the limited availability of primary human microglia, the immortalized human microglial cell clone 3 cell line (HMC3) is useful for examining microglial behavior under pathological conditions. To test whether HSP27 and HSP60 can induce microglial activation, cytokine expression and morphological changes were examined in HMC3. Other known inflammation promoters were used as positive controls. Glaucoma has autoimmune components caused by CD4+ T cells pre-stimulated by commensal bacteria. These CD4+ T cells enter the retina and cross-react with heat shock protein (HSP)-expressing neurons via the mechanism of molecular mimicry. As shown herein, microglial activation is the cause of the immune response and retinal degeneration in glaucoma. Considering the limited availability of primary human microglia, the immortalized human microglial cell clone 3 cell line (HMC3) is useful for examining microglial behavior under pathological conditions. To test whether HSP27 and HSP60 can induce microglial activation, cytokine expression and morphological changes were examined in HMC3. Other known inflammation promoters were used as positive controls. Glaucoma has autoimmune components caused by CD4+ T cells pre-stimulated by commensal bacteria. These CD4+ T cells enter the retina and cross-react with heat shock protein (HSP)-expressing neurons via the mechanism of molecular mimicry. As shown herein, microglial activation is the cause of the immune response and retinal degeneration in glaucoma. Considering the limited availability of primary human microglia, the immortalized human microglial cell clone 3 cell line (HMC3) is useful for examining microglial behavior under pathological conditions. To test whether HSP27 and HSP60 can induce microglial activation, cytokine expression and morphological changes were examined in HMC3. Other known inflammation promoters were used as positive controls. Glaucoma has autoimmune components caused by CD4+ T cells pre-stimulated by commensal bacteria. These CD4+ T cells enter the retina and cross-react with heat shock protein (HSP)-expressing neurons via the mechanism of molecular mimicry. As shown herein, microglial activation is the cause of the immune response and retinal degeneration in glaucoma. Considering the limited availability of primary human microglia, the immortalized human microglial cell clone 3 cell line (HMC3) is useful for examining microglial behavior under pathological conditions. To test whether HSP27 and HSP60 can induce microglial activation, cytokine expression and morphological changes were examined in HMC3. Other known inflammation promoters were used as positive controls. Glaucoma has autoimmune components caused by CD4+ T cells pre-stimulated by commensal bacteria. These CD4+ T cells enter the retina and cross-react with heat shock protein (HSP)-expressing neurons via the mechanism of molecular mimicry. As shown herein, microglial activation is the cause of the immune response and retinal degeneration in glaucoma. Considering the limited availability of primary human microglia, the immortalized human microglial cell clone 3 cell line (HMC3) is useful for examining microglial behavior under pathological conditions. To test whether HSP27 and HSP60 can induce microglial activation, cytokine expression and morphological changes were examined in HMC3. Other known inflammation promoters were used as positive controls. Glaucoma has autoimmune components caused by CD4+ T cells pre-stimulated by commensal bacteria. These CD4+ T cells enter the retina and cross-react with heat shock protein (HSP)-expressing neurons via the mechanism of molecular mimicry. As shown herein, microglial activation is the cause of the immune response and retinal degeneration in glaucoma. Considering the limited availability of primary human microglia, the immortalized human microglial cell clone 3 cell line (HMC3) is useful for examining microglial behavior under pathological conditions. To test whether HSP27 and HSP60 can induce microglial activation, cytokine expression and morphological changes were examined in HMC3. Other known inflammation promoters were used as positive controls. Glaucoma has autoimmune components caused by CD4+ T cells pre-stimulated by commensal bacteria. These CD4+ T cells enter the retina and cross-react with heat shock protein (HSP)-expressing neurons via the mechanism of molecular mimicry. As shown herein, microglial activation is the cause of the immune response and retinal degeneration in glaucoma. Considering the limited availability of primary human microglia, the immortalized human microglial cell clone 3 cell line (HMC3) is useful for examining microglial behavior under pathological conditions. To test whether HSP27 and HSP60 can induce microglial activation, cytokine expression and morphological changes were examined in HMC3. Other known inflammation promoters were used as positive controls. Glaucoma has autoimmune components caused by CD4+ T cells pre-stimulated by commensal bacteria. These CD4+ T cells enter the retina and cross-react with heat shock protein (HSP)-expressing neurons via the mechanism of molecular mimicry. As shown herein, microglial activation is the cause of the immune response and retinal degeneration in glaucoma. Considering the limited availability of primary human microglia, the immortalized human microglial cell clone 3 cell line (HMC3) is useful for examining microglial behavior under pathological conditions. To test whether HSP27 and HSP60 can induce microglial activation, cytokine expression and morphological changes were examined in HMC3. Other known inflammation promoters were used as positive controls.

[0061] Method: The HMC3 cell line (ATCC) was cultured in EMEM medium and challenged with 10 μg / ml of HSP27, 10 μg / ml of HSP60, 200 ng / ml of LPS, or 100 ng / ml of LPS with or without 5 mM of ATP for an additional 30 minutes. Cells that received only the medium were used as controls. After 24 hours, the RNA of HMC3 cells was collected using the ZYMO Research Quick-RNA Microprep kit, and RNA reverse transcription was performed using PrimeScript™ RT Master Mix. Sybr green RT-PCR mixtures containing different primers and cDNA samples were subjected to PCR using an EP realplex real-time PCR device. The relative fold change in mRNA transcripts was presented and compared with the control group. Additionally, low-density HMC3 cell cultures were prepared, and images of cell morphology were taken for LPS, HSP27, or HS Method: The HMC3 cell line (ATCC) was cultured in EMEM medium and challenged with 10 μg / ml of HSP27, 10 μg / ml of HSP60, 200 ng / ml of LPS, or 100 ng / ml of LPS with or without 5 mM of ATP for an additional 30 minutes. Cells that received only the medium were used as controls. After 24 hours, the RNA of HMC3 cells was collected using the ZYMO Research Quick-RNA Microprep kit, and RNA reverse transcription was performed using PrimeScript™ RT Master Mix. Sybr green RT-PCR mixtures containing different primers and cDNA samples were subjected to PCR using an EP realplex real-time PCR device. The relative fold change in mRNA transcripts was presented and compared with the control group. Additionally, low-density HMC3 cell cultures were prepared, and images of cell morphology were taken for LPS, HSP27, or HS Method: The HMC3 cell line (ATCC) was cultured in EMEM medium and challenged with 10 μg / ml of HSP27, 10 μg / ml of HSP60, 200 ng / ml of LPS, or 100 ng / ml of LPS with or without 5 mM of ATP for an additional 30 minutes. Cells that received only the medium were used as controls. After 24 hours, the RNA of HMC3 cells was collected using the ZYMO Research Quick-RNA Microprep kit, and RNA reverse transcription was performed using PrimeScript™ RT Master Mix. Sybr green RT-PCR mixtures containing different primers and cDNA samples were subjected to PCR using an EP realplex real-time PCR device. The relative fold change in mRNA transcripts was presented and compared with the control group. Additionally, low-density HMC3 cell cultures were prepared, and images of cell morphology were taken for LPS, HSP27, or HS Method: The HMC3 cell line (ATCC) was cultured in EMEM medium and challenged with 10 μg / ml of HSP27, 10 μg / ml of HSP60, 200 ng / ml of LPS, or 100 ng / ml of LPS with or without 5 mM of ATP for an additional 30 minutes. Cells that received only the medium were used as controls. After 24 hours, the RNA of HMC3 cells was collected using the ZYMO Research Quick-RNA Microprep kit, and RNA reverse transcription was performed using PrimeScript™ RT Master Mix. Sybr green RT-PCR mixtures containing different primers and cDNA samples were subjected to PCR using an EP realplex real-time PCR device. The relative fold change in mRNA transcripts was presented and compared with the control group. Additionally, low-density HMC3 cell cultures were prepared, and images of cell morphology were taken for LPS, HSP27, or HS Method: The HMC3 cell line (ATCC) was cultured in EMEM medium and challenged with 10 μg / ml of HSP27, 10 μg / ml of HSP60, 200 ng / ml of LPS, or 100 ng / ml of LPS with or without 5 mM of ATP for an additional 30 minutes. Cells that received only the medium were used as controls. After 24 hours, the RNA of HMC3 cells was collected using the ZYMO Research Quick-RNA Microprep kit, and RNA reverse transcription was performed using PrimeScript™ RT Master Mix. Sybr green RT-PCR mixtures containing different primers and cDNA samples were subjected to PCR using an EP realplex real-time PCR device. The relative fold change in mRNA transcripts was presented and compared with the control group. Additionally, low-density HMC3 cell cultures were prepared, and images of cell morphology were taken for LPS, HSP27, or HS Method: The HMC3 cell line (ATCC) was cultured in EMEM medium and challenged with 10 μg / ml of HSP27, 10 μg / ml of HSP60, 200 ng / ml of LPS, or 100 ng / ml of LPS with or without 5 mM of ATP for an additional 30 minutes. Cells that received only the medium were used as controls. After 24 hours, the RNA of HMC3 cells was collected using the ZYMO Research Quick-RNA Microprep kit, and RNA reverse transcription was performed using PrimeScript™ RT Master Mix. Sybr green RT-PCR mixtures containing different primers and cDNA samples were subjected to PCR using an EP realplex real-time PCR device. The relative fold change in mRNA transcripts was presented and compared with the control group. Additionally, low-density HMC3 cell cultures were prepared, and images of cell morphology were taken for LPS, HSP27, or HS Method: The HMC3 cell line (ATCC) was cultured in EMEM medium and challenged with 10 μg / ml of HSP27, 10 μg / ml of HSP60, 200 ng / ml of LPS, or 100 ng / ml of LPS with or without 5 mM of ATP for an additional 30 minutes. Cells that received only the medium were used as controls. After 24 hours, the RNA of HMC3 cells was collected using the ZYMO Research Quick-RNA Microprep kit, and RNA reverse transcription was performed using PrimeScript™ RT Master Mix. Sybr green RT-PCR mixtures containing different primers and cDNA samples were subjected to PCR using an EP realplex real-time PCR device. The relative fold change in mRNA transcripts was presented and compared with the control group. Additionally, low-density HMC3 cell cultures were prepared, and images of cell morphology were taken for LPS, HSP27, or HS Method: The HMC3 cell line (ATCC) was cultured in EMEM medium and challenged with 10 μg / ml of HSP27, 10 μg / ml of HSP60, 200 ng / ml of LPS, or 100 ng / ml of LPS with or without 5 mM of ATP for an additional 30 minutes. Cells that received only the medium were used as controls. After 24 hours, the RNA of HMC3 cells was collected using the ZYMO Research Quick-RNA Microprep kit, and RNA reverse transcription was performed using PrimeScript™ RT Master Mix. Sybr green RT-PCR mixtures containing different primers and cDNA samples were subjected to PCR using an EP realplex real-time PCR device. The relative fold change in mRNA transcripts was presented and compared with the control group. Additionally, low-density HMC3 cell cultures were prepared, and images of cell morphology were taken for LPS, HSP27, or HS Method: The HMC3 cell line (ATCC) was cultured in EMEM medium and challenged with 10 μg / ml of HSP27, 10 μg / ml of HSP60, 200 ng / ml of LPS, or 100 ng / ml of LPS with or without 5 mM of ATP for an additional 30 minutes. Cells that received only the medium were used as controls. After 24 hours, the RNA of HMC3 cells was collected using the ZYMO Research Quick-RNA Microprep kit, and RNA reverse transcription was performed using PrimeScript™ RT Master Mix. Sybr green RT-PCR mixtures containing different primers and cDNA samples were subjected to PCR using an EP realplex real-time PCR device. The relative fold change in mRNA transcripts was presented and compared with the control group. Additionally, low-density HMC3 cell cultures were prepared, and images of cell morphology were taken for LPS, HSP27, or HS Method: The HMC3 cell line (ATCC) was cultured in EMEM medium and challenged with 10 μg / ml of HSP27, 10 μg / ml of HSP60, 200 ng / ml of LPS, or 100 ng / ml of LPS with or without 5 mM of ATP for an additional 30 minutes. Cells that received only the medium were used as controls. After 24 hours, the RNA of HMC3 cells was collected using the ZYMO Research Quick-RNA Microprep kit, and RNA reverse transcription was performed using PrimeScript™ RT Master Mix. Sybr green RT-PCR mixtures containing different primers and cDNA samples were subjected to PCR using an EP realplex real-time PCR device. The relative fold change in mRNA transcripts was presented and compared with the control group. Additionally, low-density HMC3 cell cultures were prepared, and images of cell morphology were taken for LPS, HSP27, or HS Recordings were made 24 hours after P60 treatment and morphological changes were quantified.

[0062] Results: Data show that LPS with or without ATP inhibited IFNγ and other cytokines in HMC3. It induced an increase in the expression of all pro-inflammatory cytokines, including HSP27 and HSP60. It was shown that HMC3 could be activated to express higher levels of IFNγ and TNFα. Quantification of cell morphology revealed that LPS, HSP27 and HS were significantly increased in the cytoplasmic endothelial cells compared to the vehicle control group. In the group stimulated by P60, the dendrites were shortened and the round cell body size was enlarged. It was shown that (P<0.05).

[0063] Conclusion: This study demonstrates that HMC3 cells respond to known pro-inflammatory agents in a manner similar to primary microglia. We found that HSP27 and HSP60 promote inflammation in HMC3 cells. These results suggest that HSPs can directly induce microglial responses and may be an early cause of glaucoma-associated immune responses. This supports the idea that it may induce cell activation.

[0064] [Example 5] Investigating the spatiotemporal dynamics of microglia / macrophage polarization following ischemia / reperfusion in the retina Background: Microglia / macrophages are diverse and diverse under various microenvironmental stimuli and disease processes. Microglial phenotype changes during ischemia / reperfusion (I / R) The spatiotemporal patterns of microglia / macrophage polarization after I / R remain unclear. By identifying these pathways, our understanding of post-I / R injury and recovery has improved. There is a possibility that this will happen.

[0065] Method: The eye was connected to a saline reservoir to maintain an intraocular pressure of 110 mmHg for 60 minutes. I / R was induced in rats by cannulation with a 30-gauge needle. The retinas of rats were collected on the 1st, 2nd, 7th, and 14th days after surgery. To characterize the phenotypic changes of retinal cells including microglia and infiltrating macrophages, flow cytometry, reverse transcriptase polymerase chain reaction, Western blot, and immunohistochemical staining

[0066] for M1 and M2 markers were performed. + CD45 high Results: According to the flow cytometry results, there was a significant increase in CD11b - CD45 high , probably macrophages and / or activated microglia, as early as 12 hours after I / R, followed by an increase in CD11b - CD45 high lymphocytes on the 1st day, both peaking on the 7th day. A rapid increase in both CD16 + Iba1 + (M1 marker) cells and Ym - 1 + I ba1 + (M2 marker) cells was found in the retina on the 1st and 2nd days after I / R. These cells showed round bodies with rare short dendrites and were distributed from the inner granular layer to the ganglion cell layer from the 1st day to the

[0067] 7th day. Conclusion: I / R induced an initial response of microglia / macrophages activated in both M1 and M2 types, resulting in a gradual increase in lymphocytes. Therefore, microglia / macrophages may play a dominant role

[0068] [Example 6] Targeting HSPs: Immune tolerance to HSP60 attenuates neurodegeneration in glaucoma Objective: Previous studies have demonstrated that bacterially primed T cell-mediated autoimmune mechanisms are involved in the pathogenesis of glaucoma. It has been suggested that heat shock proteins (HSPs) play a role in pathogenic autoantibodies. We have demonstrated that immune tolerance to bacterial HSP60 is involved in glaucoma. Induction of tolerance may block such pathogenic immune responses and attenuate neuronal loss. It was assumed that:

[0069] Methods: Adult C57BL / 6J mice received low doses of recombinant bacterial HSP60 in the nostrils daily for 7 days. , ovalbumin (OVA) or saline (both as controls). Elevated OP was induced unilaterally by injection of polystyrene microbeads (MB) into the anterior chamber. Visual and retinal function was assessed by optokinetic response (OMR) and electroretinogram positive scotopic threshold potential (p The mice were sacrificed 2, 4 and 8 weeks after MB injection. T cell responses to HSP60 were assessed by ear DTH (delayed type hypersensitive response) assay and flow cytometry. Glaucomatous nerve damage was analyzed by catalysis of retinal ganglion cells (RGCs) and axons. The data were quantified by counting.

[0070] Results: Nasal administration of low doses of HSP60 enhanced immunity as indicated by reduced DTH responses. Induce immune tolerance and increase levels of regulatory T cells as seen by flow cytometry analysis. MB-injected eyes had a 2 ± 2 mmHg increase compared with 12 ± 2 mmHg in the contralateral non-injected eyes. An IOP level of 5±3 mmHg was maintained. Treatment with HSP60, OVA and saline No significant differences in IOP levels were observed between mice treated with HSP60 or OVA. Treatment at did not change the baseline levels of visual acuity (VA), contrast sensitivity (CS), or pSTR compared to untreated or saline-treated mice prior to MB injection. However, after MB injection, VA and CS evaluated by OMR were significantly better in HSP60-treated mice compared to saline or OVA-treated mice at all time points. Consistently, both RGC function and RGC count evaluated by pSTR amplitude were significantly higher in HSP60-treated mice compared to mice treated with saline or OVA after MB injection. Conclusion: Intranasal administration of multiple low doses of bacterial HSP60 induced immune tolerance and attenuated RGC loss and functional deterioration in a mouse model of MB-induced glaucoma. These results suggest an attractive antigen-specific treatment strategy for preventing vision loss in glaucoma. Such research helps the inventors' understanding of the etiology of neurodegenerative disorders and may also provide innovative therapeutic interventions for the treatment of neurodegeneration affecting other parts of the central nervous system.

[0071] References

[0072]

Table 1

[0073] Other Embodiments The present invention has been described in conjunction with its detailed description, but it should be understood that the foregoing description is intended to illustrate and not limit the scope of the present invention, which is limited by the appended claims. Other aspects, advantages, and modifications are within the scope of the following claims. ​

Claims

1. Treating neuroinflammation and neurodegeneration and / or preventing the onset of neuroinflammation and neurodegeneration or reducing the risk of progression of a pulmonary hypertension comprising: (i) administering to the patient HSP60 or HSP2 7, or an active fragment thereof, and / or (ii) IGFBPL or an active fragment thereof. administering to a subject in need thereof a therapeutically effective amount of one or more of The method.

2. The therapeutically effective amount is sufficient to reduce inflammation and neuronal cell death in the subject. The method according to claim 1 .

3. The method includes administering HSP60 intranasally or subcutaneously and administering IGFBPL systemically or ocularly. The method according to claim 1 or 2.

4. The method of claim 3 , wherein the ocular administration of IGFBPL comprises intravitreal injection.

5. The subject is suffering from non-arteritic ischemic optic neuropathy (NAION), glaucoma, autism, multiple sclerosis. Alzheimer's disease, Parkinson's disease, ischemic retinopathy, age-related macular degeneration, stroke, ischemic and traumatic optic neuropathy, or diabetic retinopathy.

6. The method according to any one of claims 1 to 5, wherein the method reduces inflammation and neuronal death in the eye of the subject. method.

7. HSP60 and / or HSPs for use in the methods described herein 27, and a kit comprising a composition comprising IGFBPL1.

8. Treating neuroinflammation and neurodegeneration and / or preventing the onset of neuroinflammation and neurodegeneration or for use in a method for reducing the risk of progression of HSP60 and / or A composition comprising HSP27, and / or a composition comprising IGFBPL1.

9. The HSP60 and / or HSP27 are formulated for nasal administration, and the IGFBP The kit or kit of claim 7, wherein L1 is formulated for ocular, e.g., intravitreal, administration. A composition for use according to claim 8.

10. The HSP60 and / or HSP27 are formulated for nasal or subcutaneous administration, The kit of claim 7 or the kit of claim 8, wherein IGFBPL1 is formulated for systemic administration. A composition for the described uses.

11. The HSP60 and / or HSP27 are formulated for nasal or subcutaneous administration, The kit according to claim 7 or the kit according to claim 8, wherein IGFBPL1 is formulated for nasal administration. A composition for the described uses.

12. one of said HSP60 and / or HSP27 and said IGFBPL1, wherein two or all three are formulated together in a single composition for nasal or subcutaneous administration. A kit or composition for use according to claim 11.

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