Methods and applications for detecting genetic polymorphisms linked to age-related central nervous system conditions and drug response

Abstract

AMD is a complex, multifactorial disease lacking curative therapy. Understanding of AMD pathophysiology highlights mechanisms such as mitochondrial dysfunction, visual cycle defects, autophagy impairment, and unresolved inflammation and oxidative stress. However, current therapies only target select aspects of AMD pathology. As described herein, variants in the ELOVL2 gene are utilized to predict the progression of age-related central nervous system conditions, including AMD, and to assess susceptibility to treatment. This approach enables more effective early intervention and personalized therapeutic strategies.

Description

METHODS AND APPLICATIONS FOR DETECTING GENETIC POLYMORPHISMS LINKED TO AGE-RELATED CENTRAL NERVOUS SYSTEM CONDITIONS AND DRUG RESPONSECROSS-REFERENCE TO RELATED APPLICATIONS

[0001] This application claims benefit of U.S. Provisional Application No. 63 / 666,328 filed July 1 , 2024, the specification of which is incorporated herein in its entirety by reference.FIELD OF THE INVENTION

[0002] The present invention utilizes variants in the Elovl2 gene to predict the progression of age-related central nervous system conditions (e.g., age-related macular degeneration (AMD)) and the susceptibility to treatment.BACKGROUND OF THE INVENTION

[0003] The specific composition of lipids within membranes dictates their biophysical properties, such as diffusion, permeability, domain formation, and curvature generation. In the human retina, where lipids constitute nearly half of the tissue’s dry weight and are predominantly localized to cellular membranes, the composition of these lipid building blocks — defined by their chemical structures and relative abundance — is critical for maintaining membrane integrity and function. Notably, retinal tissue is particularly enriched in long- and very long-chain polyunsaturated fatty acids (LC-PUFAs and VLC-PUFAs, respectively), which are essential components of photoreceptor disc membranes. The lipidome, encompassing both cellular and intracellular membranes, undergoes significant changes throughout the lifespan, and age-related alterations in membrane lipid composition have been postulated as one of the hallmarks of aging. In the aged retina, levels of PUFAs decline significantly, a phenomenon that is further exacerbated in retinas affected by age-related macular degeneration (AMD). The depletion or reduced levels of LC- and VLC-PUFAs have similarly been implicated as a key feature of AMD pathogenesis.

[0004] Currently, there are no approved therapies capable of effectively halting or slowing the progression of age-related macular degeneration (AMD) during its early or intermediate stages. While significant efforts have been devoted to understanding the genetic underpinnings of AMD — including numerous genome-wide association studies — no mutations or variants in the ELOVL2 gene have been identified that are associated with an increased risk of developing the disease. As described herein, the present invention provides novel methods for distinguishing between early and intermediate stages of AMD with greater precision. Such diagnostic tools enable more accurate patient stratification, ensuring individuals receive stage-appropriate monitoring, interventions, and potential treatments as they become available, thereby improving clinical management and prognosis.BRIEF SUMMARY OF THE INVENTION

[0005] It is an objective of the present invention to provide compositions and methods that allow for the prognosis and treatment of age-related central nervous system conditions (e.g., age-related macular degeneration (AMD)), as specified in the independent claims. Embodiments of the invention are given in the dependent claims. Embodiments of the present invention can be freely combined with each other if they are not mutually exclusive.

[0006] The ELOVL2 gene (Elongation of Very Long Chain Fatty Acids-Like 2) encodes a transmembrane enzyme essential for the biosynthesis of long-chain (C22 and C24) n-3 and n-6 polyunsaturated fatty acids (PUFAs) (FIG. 1). In the retina, ELOVL2 is highly expressed in cone photoreceptors and, to a lesser extent, in rods and the retinal pigment epithelium (RPE). Functionally, ELOVL2 catalyzes the elongation of docosapentaenoic acid (DPA, 22:5n-3) to 24:5n-3, a critical precursor for both very long-chain PUFAs (VLC-PUFAs) and docosahexaenoic acid (DHA, 22:6n-3). Notably, the regulatory regions of ELOVL2 exhibit progressive, age-associated methylation, a phenomenon consistently observed across multiple tissues and species, including rodents and humans. This epigenetic modification corresponds with decreased ELOVL2 expression, and reduced expression or specific single-nucleotide polymorphisms (SNPs) within the ELOVL2 gene have been linked to lower DHA levels, a known risk factor for age-related macular degeneration (AMD). Supporting this association, lipidomic analyses have demonstrated that total lipid extracts from photoreceptors of AMD patients contain significantly less DHA compared to those from age-matched healthy individuals. Of note, the reproducible inverse relationship between levels of methylation of ELOVL2 promoter with age was shown to be one of the best DNA methylation markers correlated with the chronological age of the organisms.

[0007] The downregulation of ELOVL2 expression is increasingly recognized as an age-related phenomenon rather than one exclusive to age-related macular degeneration (AMD). The present invention relates to specific genetic variants that correlate with an increased risk of developing AMD. These polymorphisms influence the activity or expression of ELOVL2, providing insight into an individual’s susceptibility to the disease. The invention further encompasses nucleic acid molecules containing these polymorphisms, as well as tools and methods for detecting these genetic variations and applying this information to AMD risk assessment, diagnostics, and personalized treatment strategies.

[0008] Aging is one of the major risk factors for developing AMD. The Inventors recently initiated an investigation of changes in Elovl2 expression in the retina over the lifespan of the mouse. Elovl2 expression is highest at the peak of photoreceptor differentiation (post-natal day 4), remains high in young and mature adult animals (~6 months), but significantly decreases at oldage (18 months), while DNA methylation of Elovl2 regulatory elements increases with age. Age-related changes in Elovl2 expression and methylation correlate with the appearance of autofluorescent spots in the fundus and with a decline in visual function, as measured by electroretinogram (ERG) after dark adaptation. A homozygous mutant mouse, E / ov / 2C234W, lacking the ability to convert the C22 PUFA DPA to 24:5n-3 (Tetracosapentaenoic acid) was generated. LC-MS lipidomic analysis of the E / ov / 2C234Whomozygous mouse retinas demonstrated a buildup of a substrate of the ELOVL2 (FIG. 1), while the levels of primary products of ELOVL2 and DHA were significantly decreased, indicating a loss of ELOVL2-specific enzymatic activity. Other relevant features of Elovl2C2i / -'JJmutant mice include (1) an accelerated decrease in scotopic (low light) response amplitude, (2) an accumulation of sub-RPE deposits containing several protein markers found in human drusen and (3) thickening of Bruch’s membrane and sub-RPE structures as assessed by transmission electron microscopy (TEM). In summary, the present invention links reduced ELOVL2 activity with deficiencies in PUFA production and visual impairments in mouse models and, as such, to the Inventor’s knowledge, it is the first example of a causative role of DNA methylation marker in age-related functional change.

[0009] In some embodiments, the present invention may feature a method of treating an age-related central nervous system condition (e.g., AMD) in a subject in need thereof. In some embodiments, the method comprises determining whether the subject has a genetic polymorphism, e.g., a SNR in Elongation of Very Long Chain Fatty Acids-Like 2 (ELOVL2) by obtaining or having obtained a biological sample from the subject and performing or having performed a genotyping assay on the biological sample to determine if the subject has a genetic polymorphism in ELOVL2. The method may further comprise administering a treatment to the subject if the subject has genetic polymorphism in ELOVL2. Non-limiting examples of age-related central nervous system conditions include, but are not limited to, age-related macular degeneration (AMD), Alzheimer's disease (AD), Parkinson’s disease (PD), dementia, or the like.

[0010] In some embodiments, the genetic polymorphism is single-nucleotide polymorphism (SNP). In some embodiments, the SNP is in the fifth intron of ELOVL2 (e.g., SNP rs911196). In other embodiments, the SNP is in the first intron of ELOVL2 (e.g., SNP rs9468304).

[0011] The aforementioned method may be used for stratifying subjects diagnosed with early AMD to identify individuals with an increased likelihood of accelerated progression from early to intermediate AMD, and to determine a suitable therapeutic intervention, including the type and / or dosage of medication or other treatment, for reducing or preventing further progression of the disease..

[0012] In certain embodiments, the present invention provides methods for identifying subjects likely to respond to therapies aimed at improving lipid content or modulating ELOVL2 activity. Such therapies may include, but are not limited to, interventions that increase the expression or function of ELOVL2, promote ELOVL2 demethylation, or enhance lipid biosynthesis through supplementation or gene therapy. In some embodiments, the treatment improves lipid content in retinal cells, including through mechanisms that increase ELOVL2 expression, restore or enhance ELOVL2 function, or reverse age-associated methylation of the ELOVL2 gene. These approaches may be used alone or in combination to slow the progression of AMD, particularly in subjects identified as carrying one or more genetic polymorphisms in ELOVL2, and may be further utilized to stratify patients for clinical trials or guide personalized treatment strategies.

[0013] In some embodiments, the present invention provides a method for predicting the progression of age-related macular degeneration (AMD) in a subject in need thereof. The method comprises obtaining or having obtained a biological sample from a subject diagnosed with AMD, and performing or having performed a genotyping assay on the biological sample to determine whether the subject possesses a genetic polymorphism in the Elongation of Very Long Chain Fatty Acids-Like 2 (ELOVL2) gene. The presence of the genetic polymorphism in the ELOVL2 gene in the subject predicts a faster progression of AMD compared to a subject who does not possess the genetic polymorphism.

[0014] One of the unique and inventive technical features of the present invention is the utilization of variants, such as single-nucleotide polymorphisms (SNPs), in the ELOVL2 gene. Without wishing to limit the invention to any theory or mechanism, it is believed that the technical feature of the present invention advantageously provides methods for predicting the progression of age-related central nervous system conditions, such as age-related macular degeneration (AMD), and assessing susceptibility to treatment. None of the presently known prior references or works have the unique, inventive technical feature of the present invention.

[0015] Moreover, the prior references teach away from the present invention. For example, current AMD therapies primarily target VEGF and complement modulation, but these are often ineffective as increased complement factor indicates an advanced disease stage. In contrast, the present invention leverages variants, such as SNPs, in the ELOVL2 gene to identify early disease progression, enabling more effective therapies.

[0016] Furthermore, the inventive technical features of the present invention contributed to a surprising result. For example, the Inventors were surprisingly able to identify variants in ELOVL2 (e.g., SNPs) by searching for variants within the AMD population. This approach is rarely employed, as the usual method involves examining the differences in variants betweendisease and control populations.

[0017] Additionally, testing patients for the presence of these variants can aid in predicting the onset and progression of age-related macular degeneration, thereby assisting in treatment decisions. Moreover, individuals with AMD who possess these variants may be identified as optimal candidates for therapies aimed at modulating the expression or function of ELOVL2. Notably, this correlation has not been previously established.

[0018] Any feature or combination of features described herein are included within the scope of the present invention provided that the features included in any such combination are not mutually inconsistent as will be apparent from the context, this specification, and the knowledge of one of ordinary skill in the art. Additional advantages and aspects of the present invention are apparent in the following detailed description and claims.BRIEF DESCRIPTION OF THE SEVERAL VIEWS OF THE DRAWING(S)

[0019] The features and advantages of the present invention will become apparent from a consideration of the following detailed description presented in connection with the accompanying drawings in which:

[0020] FIG. 1 shows a schematic representation of the present invention. In some embodiments, ELOVL2 is a key enzyme in VLC-PUFA elongation pathway and DHA synthesis.

[0021] FIG. 2 shows a lolliplot of variants correlated to the age of onset of AMD in the ELOVL2 gene, relative to the gene structure. Top: The negative decadic P-Value of the correlation between the alleles and age of onset / diagnosis is depicted on the y-axis. The lead variant rs911196 denoted with an asterisk (*) and the correlated variants are coded according to their genetic correlation (R2) to the lead variant. Bottom: CpGs found in the ELOVL2 locus and their effect on aging expressed as the slope (i.e., higher methylation in those sites was correlated to older age). CpGs significantly positively correlated with the G allele at rs911196 causing earlier onset of AMD are denoted with an asterisk.

[0022] FIG. 3 shows the association of ELOVL2 variants and PUFA levels in plasma and colostrum. The ELOVL2 gene and its antisense transcript ELVOL2-AS are localed on chromosome 6 (hg19; chr6; 10,980,000-11 ,080,070). Vertical lines indicate the location of selected single nucleotide polymorphisms (SNPs) across the locus.DETAILED DESCRIPTION OF THE INVENTION

[0023] Disclosed are various peptides, solvents, solutions, carriers, and / or components to be used to prepare compositions to be used within the methods disclosed herein. Also disclosedare the various steps, elements, amounts, routes of administration, symptoms, and / or treatments that are used or observed when performing the disclosed methods, as well as the methods themselves. These and other materials, steps, and / or elements are disclosed herein, and it is understood that when combinations, subsets, interactions, groups, etc. of these materials are disclosed, that while specific reference of each various individual and collective combination and permutation of these compounds may not be explicitly disclosed, each is specifically contemplated and described herein. It is also to be understood that the terminology used herein is for the purpose of describing particular embodiments only and is not intended to be limiting.

[0024] Unless otherwise explained, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which a disclosed invention belongs. The singular terms "a," "an," and "the" include plural referents unless context clearly indicates otherwise. Similarly, the word "or" is intended to include "and" unless the context clearly indicates otherwise. The term "comprising" means that other elements can also be present in addition to the defined elements presented. The use of "comprising" indicates inclusion rather than limitation. Stated another way, the term "comprising" means "including principally, but not necessary solely". Furthermore, variation of the word "comprising", such as "comprise" and "comprises", have correspondingly the same meanings. In one respect, the technology described herein related to the herein described compositions, methods, and respective component(s) thereof, as essential to the invention, yet open to the inclusion of unspecified elements, essential or not ("comprising").

[0025] Suitable methods and materials for the practice and / or testing of embodiments of the disclosure are described below. Such methods and materials are illustrative only and are not intended to be limiting. Other methods and materials similar or equivalent to those described herein can be used. For example, conventional methods well known in the art to which the disclosure pertains are described in various general and more specific references, including, for example, Sambrook et al., Molecular Cloning: A Laboratory Manual, 2d ed., Cold Spring Harbor Laboratory Press, 1989; Sambrook et al., Molecular Cloning: A Laboratory Manual, 3d ed., Cold Spring Harbor Press, 2001 ; Ausubel et al., Current Protocols in Molecular Biology, Greene Publishing Associates, 1992 (and Supplements to 2000); Ausubel et al., Short Protocols in Molecular Biology: A Compendium of Methods from Current Protocols in Molecular Biology, 4th ed., Wiley & Sons, 1999; Harlow and Lane, Antibodies: A Laboratory Manual, Cold Spring Harbor Laboratory Press, 1990; and Harlow and Lane, Using Antibodies: A Laboratory Manual, Cold Spring Harbor Laboratory Press, 1999, Gene Expression Technology (Methods in Enzymology, Vol. 185, edited by D. Goeddel, 1991. Academic Press, San Diego, Calif.), "Guideto Protein Purification” in Methods in Enzymology (M. P. Deutshcer, ed., (1990) Academic Press, Inc.); PCR Protocols: A Guide to Methods and Applications (Innis, et al. 1990. Academic Press, San Diego, Calif.), Culture of Animal Cells: A Manual of Basic Technique, 2nd Ed. (R. I. Freshney. 1987. Liss, Inc. New York, N.Y.), Gene Transfer and Expression Protocols, pp. 109-128, ed. E. J. Murray, The Humana Press Inc., Clifton, N.J.), and the Ambion 1998 Catalog (Ambion, Austin, Tex.), the disclosures of which are incorporated in their entirety herein by reference.

[0026] All publications, patent applications, patents, and other references mentioned herein are incorporated by reference in their entirety for all purposes. In case of conflict, the present specification, including explanations of terms, will control.

[0027] Although methods and materials similar or equivalent to those described herein can be used to practice or test the disclosed technology, suitable methods and materials are described below. The materials, methods, and examples are illustrative only and not intended to be limiting.

[0028] As used herein, the terms “subject” and “patient” are used interchangeably. As used herein, a subject can be a mammal such as a non-primate (e.g., cows, pigs, horses, cats, dogs, rats, etc.) or a primate (e.g., monkey and human). In specific embodiments, the subject is a human. In one embodiment, the subject is a mammal (e.g., a human) having a disease, disorder, or condition described herein. In another embodiment, the subject is a mammal (e.g., a human) at risk of developing a disease, disorder, or condition described herein. In certain instances, the term patient refers to a human.

[0029] As used herein, the terms "treat," “treating,” or "treatment" refer to both therapeutic treatment and prophylactic or preventative measures, with the objective of preventing, reducing, slowing down (lessen), inhibiting, or eliminating an undesired physiological change, symptom, disease, or disorder. For example, the disease may be Age-Related Macular Degeneration (AMD). For purposes of this invention, beneficial or desired clinical results include, but are not limited to, alleviation of symptoms, diminishment of extent of disease, stabilized (i.e., not worsening) state of disease, delay or slowing of disease progression, amelioration or palliation of the disease state, and remission (whether partial or total), whether detectable or undetectable. "Treatment" can also mean prolonging survival as compared to expected survival if not receiving treatment. Those in need of treatment include those already with the condition or disorder as well as those prone to have the condition or disorder or those in which the condition or disorder is to be prevented or onset delayed. Optionally, the subject or patient may be identified (e.g., diagnosed) as one suffering from the disease or condition prior to administrationof the compositions of the invention. Subjects at risk for the disease can be identified by, for example, any or a combination of appropriate diagnostic or prognostic assays known in the art.

[0030] As used herein, “clinical improvement” may refer to a noticeable reduction in the symptoms of a disorder, or cessation thereof.

[0031] The terms “manage,” “managing,” and “management” refer to preventing or slowing the progression, spread, or worsening of a disease or disorder, or of one or more symptoms thereof. In certain cases, the beneficial effects that a subject derives from a prophylactic or therapeutic agent do not result in a cure of the disease or disorder.

[0032] The terms “regress,” “regressing,” and “regression” may refer to a decrease in the size of a tumor or in the extent of cancer in the body. In some embodiments, “regression” may refer to a decrease in severity of the disease. In some embodiments, regression may generally refer to lighter symptoms without the disease completely disappearing. In certain cases, the beneficial effects that a subject derives from a prophylactic or therapeutic agent do not result in a cure of the disease or disorder. In some embodiments, symptoms of the disease may return.

[0033] As used herein, a “biological sample” or “clinical sample” may be used interchangeably and may refer to any biological material taken from a subject. Non-limiting examples of a biological material to be taken for a biological sample may include but are not limited to, saliva, blood, or urine, buccal swabs, and nasopharyngeal specimens.

[0034] Referring now to FIGs. 1-3, the present invention features methods and compositions for utilizing variants in the Elovl2 gene to predict the progression of age-related central nervous system conditions (e.g., age-related macular degeneration (AMD)) and the susceptibility to treatment. Without wishing to limit the present invention to any theory or mechanism, it is believed that patients with certain variants as described herein may respond more favorably, potentially deriving greater benefit from the treatment.

[0035] The present invention may feature a method of treating an age-related central nervous system condition (e.g., AMD) in a subject in need thereof. In some embodiments, the method comprises determining whether the subject has a genetic polymorphism, e.g., a SNP, in Elongation of Very Long Chain Fatty Acids-Like 2 (ELOVL2) by obtaining or having obtained a biological sample from the subject and performing or having performed a genotyping assay on the biological sample to determine if the subject has a genetic polymorphism in ELOVL2. The method may further comprise administering a treatment to the subject if the subject has genetic polymorphism in ELOVL2. Non-limiting examples of age-related central nervous system conditions include, but are not limited to, age-related macular degeneration (AMD), Alzheimer'sdisease (AD), Parkinson’s disease (PD), dementia, or the like.

[0036] For example, in some embodiments, the present invention pertains to age-related macular degeneration (AMD) and its treatment, focusing on the effects of specific genetic variations in the human genome. These variations may be associated with AMD progression and the varying responses individuals have to AMD treatments, including preventative measures. Additionally, in some embodiments, the identified genetic polymorphisms could serve as targets for diagnostic tools and therapeutic agents and may be valuable for predicting AMD occurrence, severity, and recovery, as well as for assessing individual responses to AMD treatments. Additionally, these genetic variations might have applications in human identification. In some embodiments, the present invention provides methods, assays, kits, and reagents for detecting these genetic variations and their products.

[0037] In some embodimetns, the SNPs described herein may also be used to identify individuals most likely to respond to lipid- or ELOVL2-related therapies in clinical trials.

[0038] The methods described herein may also be used for stratifying subjects diagnosed with early AMD or intermediate AMD to determine a suitable therapeutic intervention, including the type and / or dosage of medication or other treatment, for reducing or preventing the progression of AMD. In certain embodiments, the present invention provides methods for identifying subjects likely to respond to therapies aimed at improving lipid content or modulating ELOVL2 activity. Such therapies may include, but are not limited to, interventions that increase the expression or function of ELOVL2, promote ELOVL2 demethylation, or enhance lipid biosynthesis through supplementation or gene therapy. In some embodiments, the treatment improves lipid content in retinal cells, including through mechanisms that increase ELOVL2 expression, restore or enhance ELOVL2 function, or reverse age-associated methylation of the ELOVL2 gene. These approaches may be used alone or in combination to slow the progression of AMD, particularly in subjects identified as carrying one or more genetic polymorphisms in ELOVL2, and may be further utilized to stratify patients for clinical trials or guide personalized treatment strategies.

[0039] In some embodiments, building on the identification of Single Nucleotide Polymorphisms (SNPs) associated with age-related macular degeneration (AMD), the present invention may encompass a comprehensive array of methods and applications. For example, the present invention may involve the detection of these genetic variations and extend to the design and development of reagents necessary for this purpose.

[0040] In certain embodiments, the present invention features novel SNPs within genetic sequences implicated in AMD and / or the response to AMD treatment. These SNPs may beidentified within isolated nucleic acid molecules, encompassing both DNA and RNA, which harbor these genetic variations. Furthermore, the present invention may include variant proteins encoded by these nucleic acid molecules, as well as antibodies targeting these variant proteins.

[0041] In some embodiments, the present invention goes beyond mere identification of the novel SNPs. Methods described herein may be provided for precisely detecting these SNPs within test samples (e.g., biological samples), facilitating personalized risk assessment for AMD progression and prognosis of its severity. Additionally, the present invention may provide potential methods for tailored treatment strategies based on an individual’s SNP profile, enabling the prediction of treatment responsiveness in AMD. By identifying specific genetic polymorphisms associated with AMD, the present invention enables the prediction of an individual’s likelihood of responding to particular therapeutic interventions, thereby facilitating more effective and personalized treatment decisions. Such personalized approaches may include, for example, lipid supplementation or statin therapy, to improve visual outcomes and slow disease progression.

[0042] In some embodiments, individuals with AMD who possess one or more variants described herein may be identified as optimal candidates for therapies targeting the modulation of ELOVL2 expression or function. Other potential treatments may include therapies targeting the demethylation of the regulatory region of ELOVL2. Potential therapies to improve lipid content may include ELOVL2 demethylation, ELOVL2 gene therapy, or lipid supplementation.

[0043] Furthermore, in some embodiments, the present invention may cover the identification of individuals predisposed to AMD or more prone to treatment effects based on the presence of one or more SNPs. The present invention may also encompass methods for screening candidate compounds for potential efficacy in treating disorders linked to variant genes or proteins implicated in AMD pathogenesis. For example, in certain embodiments, induced pluripotent stem cell-derived retinal pigment epithelium (iPSC-RPE) cells comprising one or more ELOVL2 polymorphisms may be used in high-throughput screening assays to identify compounds that modulate ELOVL2 activity, expression, or associated cellular processes. In some embodiments, animal models, such as Elovl2C234Wmice, may be utilized to evaluate the efficacy of such compounds in vivo. Any compounds identified through these methods may also be within the scope of the present invention.

[0044] In some embodiments, the utility of the SNPs described herein may extend to human identification applications, enhancing forensic and identity verification procedures. By leveraging these genetic markers, the invention offers a multifaceted approach to understanding, predicting progression, and treating AMD, catering to both research and clinical needs. io

[0045] In some embodiments, the present invention features a method of treating an age-related central nervous system condition in a subject in need thereof. The method may comprise determining whether the subject has a genetic polymorphism in Elongation of Very Long Chain Fatty Acids-Like 2 (ELOVL2) by obtaining or having obtained a biological sample from the subject and performing or having performed a genotyping assay on the biological sample to determine if the patient has a genetic polymorphism in ELOVL2. In some embodiments, if the subject has a genetic polymorphism in ELOVL2, a treatment is administered to the subject.

[0046] In other embodiments, the present invention features a method of predicting the progression of an age-related central nervous system condition in a subject in need thereof. The method may comprise determining whether the subject has a genetic polymorphism in Elongation of Very Long Chain Fatty Acids-Like 2 (ELOVL2) by obtaining or having obtained a biological sample from the subject and performing or having performed a genotyping assay on the biological sample to determine if the patient has a genetic polymorphism in ELOVL2. In some embodiments, the presence of the genetic polymorphism in ELOVL2 in the subject predicts early progression of an age-related central nervous system condition.

[0047] The present invention may also provide in vitro methods for determining the risk of progression of age-related macular degeneration (AMD). For example, the method may comprise obtaining a biological sample from a subject diagnosed with AMD and performing a genotyping assay on the biological sample to determine whether the subject possesses a genetic polymorphism in the ELOVL2 gene. The presence of the genetic polymorphism in ELOVL2 predicts an increased likelihood of accelerated progression of AMD in the subject. In additional embodiments, the present invention provides an in vitro method for screening lipid supplements for their potential to slow the progression of AMD. In some embodiments, the method comprises providing a cell culture comprising retinal pigment epithelial (RPE) cells or photoreceptor cells, contacting the cell culture with a candidate lipid supplement, and assessing one or more cellular or molecular markers indicative of AMD progression. A change in the one or more markers relative to a control identifies the lipid supplement as having potential to slow the progression of AMD.

[0048] Non-limiting examples of biological samples that may be used in accordance with the present invention include but are not limited to a skin sample, a blood sample, a saliva sample, a urine sample, a buccal swab, or a nasopharyngeal sample.

[0049] Non-limiting examples of age-related central nervous system conditions include but are not limited to age-related macular degeneration (AMD), Alzheimer's disease (AD), Parkinson’sdisease (PD), or the like.

[0050] In some embodiments, the genetic polymorphism is single-nucleotide polymorphism (SNP). In some embodiments, the SNP is in the fifth intron of ELOVL2 (e.g., SNP rs911196). In other embodiments, the SNP is in the first intron of ELOVL2 (e.g., SNP rs9468304). Other single-nucleotide polymorphisms (SNPs) relevant to the present invention may include, but are not limited to, cg01799681 , cg23642061 , cg16867657, cg1572722, rs9468304, rs8523, rs17606561 , rs3734398, rs2236212, rs116021127, rs2295602, rs3798712, rs3798713, rs16870891 , and rs3756963. In addition, other relevant SNPs may include rs3734397, rs953413, rs1570069, rs7744440, rs3756963, rs3734397, rs953413, rs3798719, rs3734398, rs2236212, rs3798713, rs9368564, and rs12662634. The present invention is not limited to the aforementioned SNPs.

[0051] In some embodiments, the treatment modulates either the expression of ELOVL2, the function of ELOVL2, or both the expression and function of ELOVL2.

[0052] In some embodiments, treatments may include lipid supplementation or gene therapy.

[0053] EXAMPLE

[0054] The following is a non-limiting example of the present invention. It is to be understood that said example is not intended to limit the present invention in any way. Equivalents or substitutes are within the scope of the present invention.

[0055] At the genetic level, several variants within the ELOVL2 locus have been described. Specifically, variants within noncoding regions (introns and regulatory elements) have been associated with n-3 PUFA levels in plasma and colostrum, as well as with a risk of coronary artery disease and Alzheimer’s disease. To date, despite numerous genome-wide studies, no ELOVL2 mutations or variants have been detected that correlate with the risk of AMD. Two possible explanations may account for this observation: (1) ELOVL2 is an essential gene for population survival, and heterozygosity of the gene in a mouse model causes infertility in C57BL / 6 mice. Therefore, variants that can be potentially correlated with the disease are rare and have yet to be discovered. (2) The downregulation of ELOVL2 expression is not exclusive to AMD but is associated with aging. In this scenario, potential variants affecting the activity or expression of ELOVL2 may correlate more with the onset of the disease rather than the disease itself. To address the latter possibility, genomic data was used from the International AMD Genomics Consortium and the UK Biobank and asked whether there are any variants in the ELOVL2 genomic region that modify the onset of the disease. With this approach, several non-coding variants were identified within the gene region of ELOVL2. While the associatedvariants were not found to influence ELOVL2 gene expression in the retina or brain, the G allele of the lead variant rs911196 increases methylation of cg01799681 , cg16867657, cg21572722, and cg23642061 in whole blood. Those CpG sites are located near the ELOVL2 promoter, and increased methylation at those sites is strongly associated with advanced age.

[0056] To investigate the association between genetic variants within the ELOVL2 gene (chr6: 10980992-11044624) and the age of onset of intermediate AMD, two independent cohorts were used: unrelated, European individuals with intermediate AMD from the International AMD Genomics Consortium (IAMGDC, n=2,407, mean age at diagnosis: 74.1 years), as well as incident AMD cases from the UK Biobank (n=1 ,309, mean age of onset: 62.8 years). In the IAMDGC cohort, linear regression was used with age of onset as the outcome and genotype as the exposure, adjusted for study, sex, and the first five principal components of ancestry. In the UK Biobank, the analyses were additionally adjusted by smoking status and BMI.

[0057] To study the role of ELOVL2 genetics in AMD risk, the correlation between those variants and age at disease onset or diagnosis was computed. First, the incident AMD cases were identified from the UK Biobank (the data was accessed and analyzed under the UK Biobank project ID 73446). Detailed ophthalmological grading is not available for the majority of the UK Biobank participants. Therefore, the general practice read codes were leveraged to ascertain incident AMD patients. In total, 1 ,309 unrelated European individuals developed any type of AMD in the UK Biobank after recruitment. In this case-only analysis, linear regression was used with age at onset as the outcome and each variant separately as exposure. The analyses were adjusted by sex, BMI, smoking status, and the first ten principal components of ancestry. To replicate the findings, all non-imputed variants were extracted from the Illumina HumanCoreExome in the genomic region of ELOVL2 (chr6: 10980992 - 11044624) for unrelated and European participants with intermediate AMD from a recent GWAS (n=2,407) conducted by the International AMD Genomics Consortium. Intermediate AMD was defined as individuals which had pigmentary changes in the RPE, were at least 50 years of age and did not exhibit signs of late stage AMD. Similar to the UK Biobank analyses, we used linear regression adjusted for study, sex, and the first five principal components of ancestry were used. The resulting slopes and standard errors from both cohorts were then meta-analyzed with the REML algorithm implemented in the rma function from the metafor package in R. The association results were plotted as a lolliplot, implemented in the track iewer package in R.

[0058] The strongest association was observed for rs911196 in the fifth intron of ELOVL2. The minor G allele (allele frequency 25% in Europeans) resulted in 4.7 months (95% Confidence Interval: 2.1 months - 7.3 months) earlier onset for intermediate AMD in our analyses (FIG. 2, p=0.0003). This effect was seen in both cohorts (IAMDGC: 5.7 months earlier onset, UKBiobank: 4.5 months earlier). The G allele of the correlated variant rs9468304 in the first intron also showed a significant association, however, the effect was weaker (31% allele frequency, 4.32 months earlier onset, p=0.0009). None of the remaining variants had a statistically significant correlation (p>0.05). Importantly, no such correlation was detected in ELOVL4 and ELOVL5 loci (data not shown).

[0059] As used herein, the term “about” refers to plus or minus 10% of the referenced number.

[0060] Although there has been shown and described the preferred embodiment of the present invention, it will be readily apparent to those skilled in the art that modifications may be made thereto which do not exceed the scope of the appended claims. Therefore, the scope of the invention is only to be limited by the following claims. In some embodiments, the figures presented in this patent application are drawn to scale, including the angles, ratios of dimensions, etc. In some embodiments, the figures are representative only and the claims are not limited by the dimensions of the figures. In some embodiments, descriptions of the inventions described herein using the phrase “comprising” includes embodiments that could be described as “consisting essentially of’ or “consisting of’, and as such the written description requirement for claiming one or more embodiments of the present invention using the phrase “consisting essentially of’ or “consisting of’ is met.

Claims

WHAT IS CLAIMED IS:1 . A method of treating an age-related central nervous system condition in a subject in need thereof, the method comprising a) determining whether the subject has a genetic polymorphism in Elongation ofVery Long Chain Fatty Acids-Like 2 (ELOVL2) by: i. obtaining or having obtained a biological sample from the subject; and ii. performing or having performed a genotyping assay on the biological sample to determine if the subject has a genetic polymorphism in ELOVL2; and b) if the subject has genetic polymorphism in ELOVL2, then administering a treatment to the subject.

2. The method of claim 1 , wherein the age-related central nervous system condition is age-related macular degeneration (AMD).

3. The method of claim 1 , wherein the age-related central nervous system condition is Alzheimer's disease.

4. The method of claim 1 , wherein the age-related central nervous system condition is Parkinson’s disease.

5. The method of claim 1 , wherein the age-related central nervous system condition is dementia.A method of treating age-related macular degeneration (AMD) in a subject in need thereof, the method comprising a) determining whether the subject has a genetic polymorphism in Elongation of Very Long Chain Fatty Acids-Like 2 (ELOVL2) by: i. obtaining or having obtained a biological sample from the subject; and ii. performing or having performed a genotyping assay on the biological sample to determine if the subject has a genetic polymorphism in ELOVL2; and b) if the subject has genetic polymorphism in ELOVL2, then administering a treatment to the subject.

7. The method of claim 6, wherein said method is for stratifying subjects with early AMD to identify individuals with an increased likelihood of accelerated progression from early to intermediate AMD, and determine appropriate medication and level of medication and / or intervention(s) to prevent progression of AMD.

8. The method of claim 6, wherein said method is for identifying subjects that will respond to a therapy aimed at improving lipid content, modulating ELOVL2 expression or function, or a combination thereof.

9. The method of any one of claims 1-8, wherein the biological sample is a blood sample, a saliva sample, a urine sample, a buccal swab, or a nasopharyngeal sample.

10. The method of any one of claims 1-9, wherein the genetic polymorphism is single-nucleotide polymorphisms (SNP).11 . The method of claim 10, wherein the SNP is in a fifth intron of ELOVL2.

12. The method of claim 10 or claim 11 , wherein the SNP is rs911196.

13. The method of claim 10, wherein the SNP is in a first intron of ELOVL2.

14. The method of claim 10 or claim 13, wherein the SNP is rs9468304.

15. The method of any one of claims 1-14, wherein the treatment improves lipid content.

16. The method of any one of claims 1-15, wherein the treatment modulates expression of ELOVL2, function of ELOVL2, or a combination thereof.

17. The method of claim 16, wherein the treatment increases the expression of ELOVL2, function of ELOVL2, or a combination thereof.

18. The method of claim 16, wherein the treatment causes ELOVL2 demethylation.

19. A method for predicting the progression of age-related macular degeneration (AMD) in a subject in need thereof, the method comprising: a) obtaining or having obtained a biological sample from a subject diagnosed with AMD; and b) performing or having performed a genotyping assay on the biological sample to determine whether the subject possesses a genetic polymorphism in the Elongation of Very Long Chain Fatty Acids-Like 2 (ELOVL2) gene; wherein the presence of the genetic polymorphism in ELOVL2 in the subject predicts faster progression of AMD compared to a subject without a genetic polymorphism.

20. The method of claim 19, wherein the biological sample is a blood sample, a saliva sample, a urine sample, a buccal swab, or a nasopharyngeal sample.

21. The method of claim 19 or claim 20 wherein the genetic polymorphism is single-nucleotide polymorphisms (SNP).

22. The method of claim 21 , wherein the SNP is in a fifth intron of ELOVL2.

23. The method of claim 21 or claim 22, wherein the SNP is rs911196.

24. The method of claim 21 , wherein the SNP is in a first intron of ELOVL2.

25. The method of claim 21 or claim 24, wherein the SNP is rs9468304.

26. The method of any one of claims 19-25, wherein the treatment improves lipid content.

27. The method of any one of claims 19-26, wherein the treatment modulates expression of ELOVL2, function of ELOVL2, or a combination thereof.

28. The method of claim 27, wherein the treatment increases the expression of ELOVL2, function of ELOVL2, or a combination thereof.

29. The method of claim 27, wherein the treatment causes ELOVL2 demethylation.

30. An in vitro method for determining the risk of the progression of age-related macular degeneration (AMD), the method comprising: a) obtaining a biological sample from a subject diagnosed with AMD; and b) performing a genotyping assay on the biological sample to determine if an Elongation of Very Long Chain Fatty Acids-Like 2 (ELOVL2) gene has a genetic polymorphism; wherein the presence of the genetic polymorphism in ELOVL2 in the subject predicts early progression of AMD.

31. The method of claim 30, wherein the biological sample is a blood sample, a saliva sample, a urine sample, a buccal swab, or a nasopharyngeal sample.

32. The method of claim 30 or claim 31 , wherein the genetic polymorphism is single-nucleotide polymorphisms (SNP).

33. The method of claim 32, wherein the SNP is in a fifth intron of ELOVL2.

34. The method of claim 32 or claim 33, wherein the SNP rs911196.

35. The method of claim 32, wherein the SNP is in a first intron of ELOVL2.

36. The method of claim 32 or claim 35, wherein the SNP rs9468304.

37. An in vitro method for screening lipid supplements for their potential to slow the progression of age-related macular degeneration (AMD), the method comprising: a) providing a cell culture comprising retinal pigment epithelial (RPE) cells or photoreceptor cells; b) contacting the cell culture with a candidate lipid supplement; and c) assessing one or more cellular or molecular markers indicative of AMD progression, wherein a change in the one or more markers relative to a control identifies the lipid supplement as having potential to slow the progression of AMD.

38. The method of claim 37, wherein the genetic polymorphism is single-nucleotide polymorphisms (SNP).

39. The method of claim 38, wherein the SNP is in a fifth intron of ELOVL2.

40. The method of claim 38 or claim 39, wherein the SNP rs911196.41 . The method of claim 38, wherein the SNP is in a first intron of ELOVL2.

42. The method of claim 38 or claim 41 , wherein the SNP rs9468304.

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