Treatment of lipid disorders and cardiovascular diseases with phospholipase a2 group XIIB (PLA2g12b) inhibitors

Abstract

The present disclosure relates generally to the treatment of subjects having a lipid disorder or a cardiovascular disease or at risk of developing a lipid disorder or a cardiovascular disease by administering a Phospholipase A2 Group XIIB (PLA2G12B) inhibitor to the subject.

Description

[0001] DOCKET NO.: 38120-4555 (11318WO01)

[0002] - 1 - Treatment Of Lipid Disorders And Cardiovascular Diseases With Phospholipase A2 Group XII B (PLA2G12B) Inhibitors

[0003] Field

[0004] The present disclosure relates generally to the treatment of subjects having a lipid disorder or a cardiovascular disease, or at risk of developing a lipid disorder or a cardiovascular disease, by administering a Phospholipase A2 Group XIIB (PLA2G12B) inhibitorto the subject, and methods of identifying subjects having an increased risk of developing a lipid disorder or a cardiovascular disease, or at risk of developing a lipid disorder or a cardiovascular disease.

[0005] Background

[0006] Lipid disorders encompass an array of conditions characterized by abnormal levels of lipids or fats in the blood. Such fats include low-density lipoproteins (LDLs), triglycerides, and high-density lipoproteins (HDLs). Typically, lipid disorders are the result of increased levels of LDLs, triglycerides, or both. Increased levels of LDLs, triglycerides, or both can result from a variety of causes including lifestyle factors, medical conditions, medication side effects, and genetics. Lifestyle factors that can increase the level of fats include an unhealthy diet high in fat, lack of exercise, and smoking. Medical conditions that can lead to increased fats include diabetes, hypothyroidism, metabolic syndrome, and kidney disease. Certain medications such as diuretics can increase fat levels as well. Underlying genetic factors can also lead to increased fat levels ultimately resulting in a lipid disorder. While some of the risk factors for lipid disorders, more specifically high LDL, are easier to attenuate, i.e. lifestyle factors, others present a challenge.

[0007] PLA2G12B is encoded by a 20 kb gene located at 10q22.1 and is present in 2 isoforms. PLA2G12B protein is 195 amino acids long and is a 22 kDa protein that is part of a family of enzymes which function as enzymes for glycolipid hydrolysis. PLA2G12B encodes a group 12B phospholipase A2 (PLA2) enzyme. This family of enzymes hydrolyze phospholipids into fatty acids and other lipophilic molecules. Phospholipases A2 include several families and the two most notable families are secreted (sPLA2) and cytosolic (sPLA2). Secreted PLA2 and cytosolic PLA2 require Ca2+for activity and play various roles in digestion, inflammation and cell signaling.

[0008] 118452910DOCKET NO.: 38120-4555 (11318WO01)

[0009] - 2 - Summary

[0010] The present disclosure provides methods of treating a subject having a lipid disorder or a cardiovascular disease, or at risk of developing a lipid disorder or a cardiovascular disease, the methods comprising administering a PLA2G12B inhibitor to the subject.

[0011] The present disclosure also provides methods of treating a subject having a lipid disorder or a cardiovascular disease, or at risk of developing a lipid disorder or a cardiovascular disease by administering a lipid disorder therapeutic agent or a cardiovascular disease therapeutic agent, the methods comprising: determining or having determined whether the subject has a PLA2G12B variant nucleic acid molecule, by: obtaining or having obtained a biological sample from the subject; and performing or having performed a sequence analysis on the biological sample to determine if the subject has a genotype comprising a PLA2G12B variant nucleic acid molecule; and administering or continuing to administer the lipid disorder therapeutic agent orthe cardiovascular disease therapeutic agent in an amountthat is the same as or less than a standard dosage amount and / or administering a PLA2G12B inhibitor to a subject that is PLA2G12B reference; administering or continuing to administer the lipid disorder therapeutic agent orthe cardiovascular disease therapeutic agent in an amountthat is the same as or less than a standard dosage amount and / or administering a PLA2G12B inhibitor to a subject that is heterozygous for the PLA2G12B variant nucleic acid molecule; or administering or continuing to administer the lipid disorder therapeutic agent or the cardiovascular disease therapeutic agent in a standard dosage amount to a subject that is homozygous for the PLA2G12B variant nucleic acid molecule; wherein the presence of a genotype having the PLA2G12B variant nucleic acid molecule indicates the subject has a decreased risk of developing a lipid disorder or a cardiovascular disease.

[0012] The present disclosure also provides methods of identifying a subject having an increased risk of developing a lipid disorder or a cardiovascular disease, the methods comprising: determining or having determined the presence or absence of a PLA2G12B variant nucleic acid molecule in a biological sample obtained from the subject; wherein: when the subject is PLA2G12B reference, then the subject has an increased risk of developing a lipid disorder or a cardiovascular disease; and when the subject is heterozygous or homozygous for the PLA2G12B variant nucleic acid molecule, then the subject has a decreased risk of developing a lipid disorder or a cardiovascular disease.

[0013] 118452910DOCKET NO.: 38120-4555 (11318WO01)

[0014] - 3 - The present disclosure also provides lipid disorder therapeutic agents or cardiovascular disease therapeutic agents for use in the treatment or prevention of a lipid disorder or a cardiovascular disease in a subject having a PLA2G12B variant nucleic acid molecule.

[0015] The present disclosure also provides PLA2G12B inhibitors for use in the treatment or prevention of a lipid disorder or a cardiovascular disease in a subject that is PLA2G12B reference or is heterozygous for a PLA2G12B variant nucleic acid molecule.

[0016] Description

[0017] Various terms relating to aspects of the present disclosure are used throughout the specification and claims. Such terms are to be given their ordinary meaning in the art, unless otherwise indicated. Other specifically defined terms are to be construed in a manner consistent with the definitions provided herein.

[0018] Unless otherwise expressly stated, it is in no way intended that any method or aspect set forth herein be construed as requiring that its steps be performed in a specific order.

[0019] Accordingly, where a method claim does not specifically state in the claims or descriptions that the steps are to be limited to a specific order, it is in no way intended that an order be inferred, in any respect. This holds for any possible non-expressed basis for interpretation, including matters of logic with respect to arrangement of steps or operational flow, plain meaning derived from grammatical organization or punctuation, or the number or type of aspects described in the specification.

[0020] As used herein, the singular forms "a," "an" and "the" include plural referents unless the context clearly dictates otherwise.

[0021] As used herein, the term "about" means that the recited numerical value is approximate and small variations would not significantly affect the practice of the disclosed embodiments. Where a numerical value is used, unless indicated otherwise by the context, the term "about" means the numerical value can vary by ±10% and remain within the scope of the disclosed embodiments.

[0022] As used herein, the term "comprising" may be replaced with "consisting" or "consisting essentially of' in particular embodiments as desired.

[0023] As used herein, the term "isolated", in regard to a nucleic acid molecule or a polypeptide, means that the nucleic acid molecule or polypeptide is in a condition other than its native environment, such as apart from blood and / or animal tissue. In some embodiments, an

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[0025] - 4-isolated nucleic acid molecule or polypeptide is substantially free of other nucleic acid molecules or other polypeptides, particularly other nucleic acid molecules or polypeptides of animal origin. In some embodiments, the nucleic acid molecule or polypeptide can be in a highly purified form, i.e., greater than 95% pure or greater than 99% pure. When used in this context, the term "isolated" does not exclude the presence of the same nucleic acid molecule or polypeptide in alternative physical forms, such as dimers or alternately phosphorylated or derivatized forms.

[0026] As used herein, the terms "nucleic acid", "nucleic acid molecule", "nucleic acid sequence", "polynucleotide", or "oligonucleotide" can comprise a polymeric form of nucleotides of any length, can comprise DNA and / or RNA, and can be single-stranded, doublestranded, or multiple stranded. One strand of a nucleic acid also refers to its complement.

[0027] As used herein, the term "subject" includes any animal, including mammals. Mammals include, but are not limited to, farm animals (such as, for example, horses, cows, and pigs), companion animals (such as, for example, dogs and cats), laboratory animals (such as, for example, mice, rats, and rabbits), and non-human primates. In some embodiments, the subject is a human. In some embodiments, the human is a patient underthe care of a physician.

[0028] It has been observed in accordance with the present disclosure that rare PLA2G12B variant nucleic acid molecules (whetherthese variants are homozygous or heterozygous in a particular subject) associate with a decreased risk of developing a lipid disorder, such as an increased low-density lipoprotein (LDL) level. It is believed that predicted loss-of-function PLA2G12B variant nucleic acid molecules have not been associated with a lipid disorder in humans. Therefore, subjects that are PLA2G12B reference or heterozygous for a PLA2G12B variant nucleic acid molecule may be treated with a PLA2G12B inhibitor such that a lipid disorder or a cardiovascular disease is inhibited or prevented, the symptoms thereof are reduced or prevented, and / or development of symptoms is repressed or prevented. It is also believed that such subjects having a lipid disorder or a cardiovascular disease may further be treated with one or more lipid disorder therapeutic agents or one or more cardiovascular disease therapeutic agents. In addition, the present disclosure provides methods of leveraging the presence or absence of PLA2G12B variant nucleic acid molecules in subjects to identify or stratify risk is such subjects of developing a lipid disorder or a cardiovascular disease, or to diagnose subjects as having an increased risk of developing a lipid disorder or a cardiovascular disease.

[0029] 118452910DOCKET NO.: 38120-4555 (11318WO01)

[0030] - 5 - For purposes of the present disclosure, any particular subject, such as a human, can be categorized as having one of three PLA2G12B genotypes: i) PLA2G12B reference; ii) heterozygous for a PLA2G12B variant nucleic acid molecule; or iii) homozygous for a PLA2G12B variant nucleic acid molecule. A subject is PLA2G12B reference when the subject does not have a copy of a PLA2G12B variant nucleic acid molecule. A subject is heterozygous for a PLA2G12B variant nucleic acid molecule when the subject has a single copy of a PLA2G12B variant nucleic acid molecule. A subject is homozygous for a PLA2G12B variant nucleic acid molecule when the subject has two copies of a PLA2G12B variant nucleic acid molecule.

[0031] PLA2G12B variant nucleic acid molecules can be any nucleic acid molecule (such as, a genomic nucleic acid molecule, an mRNA molecule, or a cDNA molecule produced from an mRNA molecule) encoding a PLA2G12B polypeptide having a partial loss-of-function, a complete loss-of-function, a predicted partial loss-of-function, or a predicted complete loss-of-function. In some embodiments, the PLA2G12B variant nucleic acid molecule is a missense variant nucleic acid molecule. In some embodiments, the PLA2G12B variant nucleic acid molecule comprises a variation in a coding region. In some embodiments, the PLA2G12B variant nucleic acid molecule does not comprise a variation in a non-coding region, except for splice acceptor regions (two bases before the start of any exon except the first). In some embodiments, the PLA2G12B variant nucleic acid molecule is a splice-site variant, a stop-gain variant, a start-loss variant, a stop-loss variant, a frameshift variant, an in-frame indel variant, or a variant that encodes a truncated PLA2G12B variant polypeptide. A subject who has a PLA2G12B polypeptide having a partial loss-of-function (or predicted partial loss-of-function) is hypomorphic for PLA2G12B. In some embodiments, the PLA2G12B variant nucleic acid molecule is any nucleic acid molecule resulting in decreased or aberrant expression of PLA2G12B mRNA or polypeptide. In some embodiments, the PLA2G12B variant nucleic acid molecule is associated with a reduced in vitro response to PLA2G12B ligands compared with reference PLA2G12B. In some embodiments, the PLA2G12B variant nucleic acid molecule is results or is predicted to result in a premature truncation of a PLA2G12B polypeptide compared to the reference PLA2G12B. In some embodiments, the PLA2G12B variant nucleic acid molecule is a variant that is predicted to be damaging to the protein function (and hence, in this case, protective to the human) by in vitro prediction algorithms such as Polyphen, SIFT, or similar algorithms. In some embodiments, the PLA2G12B variant nucleic acid molecule is a variant that causes or is predicted to cause a nonsynonymous amino-acid substitution in a PLA2G12B

[0032] 118452910DOCKET NO.: 38120-4555 (11318WO01)

[0033] - 6 -nucleic acid molecule and whose allele frequency is less than 1 / 100 alleles or less than 1 / 1000 alleles or less than 1 / 5000 alleles in the population from which the subject is selected. In some embodiments, the PLA2G12B variant nucleic acid molecule is any rare missense variant (allele frequency < 0.1%; or 1 in 1,000 alleles), or any splice-site, stop-gain, start-loss, stop-loss, frameshift, or in-frame indel, or other frameshift PLA2G12B variant.

[0034] In any of the embodiments described herein, the PLA2G12B variant genomic nucleic acid molecule can include one or more variations at any of the positions of chromosome 10 (i.e., positions 72,934,762-72,954,806) using the nucleotide sequence of the PLA2G12B reference genomic nucleic acid molecule in the GRCh38 / hg38 human genome assembly (see, ENST00000373032.4 annotated in the in the Ensembl database (URL: world wide web at "useast.ensembl.org / Homo_sapiens / Gene / Summary?g= ENSG00000138308;r=10:72934762-72954806;t=ENST00000373032)) as a reference sequence. The sequences provided in these transcripts for the PLA2G12B genomic nucleic acid molecule are only exemplary sequences. Other sequences for the PLA2G12B genomic nucleic acid molecule are also possible. In some embodiments, the PLA2G12B variant nucleic acid molecule is a PLA2G12B variant genomic nucleic acid molecule that comprises the genetic variation 10:72935723:AGG:A, 10:72941168:C:T, 10:72941227:A:T, 10:72941235:G:A, 10:72941336:T:C, 10:72942659:GGT:G, 10:72942688:G:T, 10:72942730:TG:T, 10:72954474:C:A, 10:72954480:CG:C, 10:72954481:G:A, 10:72954565:C:A, 10:72954604:C:A, 10:72954685:T:C, 10:72935708:AAC:A, 10:72941259:T:A, 10:72941302:G:T, 10:72942650:A:G, 10:72942742:T:A, 10:72954591:G:C, 10:72954601:C:A, or 10:72941311:C:CA, or is an mRNA molecule produced therefrom, or is a cDNA molecule produced from the mRNA molecule.

[0035] For subjects that are genotyped or determined to be PLA2G12B reference, such subjects have an increased risk of developing a lipid disorder or a cardiovascular disease. For subjects that are genotyped or determined to be either PLA2G12B reference or heterozygous for a PLA2G12B variant nucleic acid molecule, such subjects can be treated with a PLA2G12B inhibitor.

[0036] In any of the embodiments described herein, the subject in whom a lipid disorder or a cardiovascular disease is prevented by administering a PLA2G12B inhibitor can be anyone at risk for developing a lipid disorder or a cardiovascular disease including, but not limited to, subjects with a genetic predisposition for developing a lipid disorder or a cardiovascular disease. Additional risk factors for a lipid disorder or a cardiovascular disease include, but are

[0037] 118452910DOCKET NO.: 38120-4555 (11318WO01)

[0038] - 7 -not limited to, poor diet, obesity, lack of exercise, smoking, alcohol, age (over 40), race (African Americans typically have higher HDL and LDL cholesterol levels than Caucasians), gender (before the age of menopause, women have lower total cholesterol levels than men of the same age; after the age of menopause, a woman's LDL levels tend to rise), and familial hypercholesterolemia (FH). In some embodiments, administering a PLA2G12B inhibitor to a subject having a lipid disorder or a cardiovascular disease may be carried out to prevent development of another a lipid disorder or a cardiovascular disease in a subject who has already had a lipid disorder or a cardiovascular disease. In any of the embodiments described herein, the methods can be used to improve a lipid disorder or a cardiovascular disease.

[0039] In any of the embodiments described herein, the PLA2G12B predicted loss-of-function polypeptide can be any PLA2G12B polypeptide having a partial loss-of-function, a complete loss-of-function, a predicted partial loss-of-function, ora predicted complete loss-of-function.

[0040] Any one or more (i.e., any combination) of the PLA2G12B variant nucleic acid molecules described herein can be used within any of the methods described herein to determine whether a subject has an increased risk of developing a lipid disorder or a cardiovascular disease. The combinations of particular variants can form a mask used for statistical analysis of the particular correlation of PLA2G12B and an increased risk of developing a lipid disorder or a cardiovascular disease. In some embodiments, the mask used for statistical analysis of the particular correlation of PLA2G12B and an increased risk of developing a lipid disorder or a cardiovascular disease can exclude any one or more of these PLA2G12B variant nucleic acid molecules described herein.

[0041] In any of the embodiments described herein, the subject can have a lipid disorder. In any of the embodiments described herein, the subject can be at risk of developing a lipid disorder. In any of the embodiments described herein, the lipid disorder can be increased LDL (i.e., hypercholesterolemia, such as familial hypercholesterolemia), increased ApoB, and increased total cholesterol, or complications of increased LDL, complications of increased ApoB, and complications of increased total cholesterol: aneurysm, coronary artery disease, peripheral artery disease, familial hypercholesterolemia, heart failure and stroke. The lipid disorder can be a re-occurrence of aneurysm, coronary artery disease, peripheral artery disease, familial hypercholesterolemia, heart failure, and stroke. In any of the embodiments described herein, the lipid disorder can be increased LDL. In any of the embodiments described herein, the lipid disorder can be increased total cholesterol. In any of the embodiments described herein, the

[0042] 118452910DOCKET NO.: 38120-4555 (11318WO01)

[0043] - 8 -methods can be used to treat a complication or co-morbid ity of a lipid disorder, or reduce the risk of developing the same.

[0044] In any of the embodiments described herein, the subject can have a cardiovascular disease. In any of the embodiments described herein, the cardiovascular disease may be an aneurism (e.g., an abdominal aortic aneurysm, a thoracic aortic aneurysm, a cerebral aneurysm, a carotid aneurysm, a popliteal aneurysm, a mesenteric aneurysm, or a splenic aneurysm), heart failure (e.g., non-ischemic heart failure; and can be independent of any one or more of the ACEi pathway, ARB pathway, ARNi pathway, SGLT2i pathway, and beta blockade pathway), elevated level of coronary artery calcification, Type lid glycosylation (CDG-lld), elevated level of pericardial fat, coronary artery disease (CAD), myocardial infarction (Ml), peripheral artery disease (PAD), stroke, pulmonary embolism, deep vein thrombosis (DVT), bleeding diatheses, an atherothrombotic condition (e.g., elevated level of fibrinogen, a fibrinogen-mediated blood clot), aortic stenosis (AoS), or aortic valve stenosis. In some embodiments, the cardiovascular disease is an aneurism (e.g., an abdominal aortic aneurysm, a thoracic aortic aneurysm, a cerebral aneurysm, a carotid aneurysm, a popliteal aneurysm, a mesenteric aneurysm, or a splenic aneurysm). In some embodiments, the aneurism is an abdominal aortic aneurysm. In some embodiments, the aneurism is a thoracic aortic aneurysm. In some embodiments, the aneurism is a cerebral aneurysm. In some embodiments, the aneurism is a carotid aneurysm. In some embodiments, the aneurism is a popliteal aneurysm. In some embodiments, the aneurism is a mesenteric aneurysm. In some embodiments, the aneurism is a splenic aneurysm. In some embodiments, the cardiovascular disease is heart failure (e.g., non-ischemic heart failure; and can be independent of any one or more of the ACEi pathway, ARB pathway, ARNi pathway, SGLT2i pathway, and beta blockade pathway). In some embodiments, the heart failure is nonischemic heart failure. In some embodiments, the heart failure is independent of the ACEi pathway. In some embodiments, the heart failure is independent of the ARB pathway. In some embodiments, the heart failure is independent of the ARNi pathway. In some embodiments, the heart failure is independent of the SGLT2i pathway. In some embodiments, the heart failure is independent of the beta blockade pathway. In some embodiments, the cardiovascular disease is an elevated level of coronary artery calcification. In some embodiments, the cardiovascular disease is Type lid glycosylation (CDG-lld). In some embodiments, the cardiovascular disease is an elevated level of pericardial fat. In some embodiments, the cardiovascular disease is CAD. In some embodiments, the cardiovascular disease is Ml. In some

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[0046] - 9-embodiments, the cardiovascular disease is PAD. In some embodiments, the cardiovascular disease is stroke. In some embodiments, the cardiovascular disease is pulmonary embolism. In some embodiments, the cardiovascular disease is DVT. In some embodiments, the cardiovascular disease is a bleeding diatheses. In some embodiments, the cardiovascular disease is an atherothrombotic condition (e.g., elevated level of fibrinogen, a fibrinogen-mediated blood clot). In some embodiments, the cardiovascular disease is AoS. In some embodiments, the cardiovascular disease is aortic valve stenosis.

[0047] The present disclosure provides methods of treating a subject having a lipid disorder or a cardiovascular disease or at risk of developing a lipid disorder or a cardiovascular disease, the methods comprising administering a PLA2G12B inhibitor to the subject.

[0048] In some embodiments, the PLA2G12B inhibitor comprises an inhibitory nucleic acid molecule. Examples of inhibitory nucleic acid molecules include, but are not limited to, antisense nucleic acid molecules, small interfering RNAs (siRNAs), and short hairpin RNAs (shRNAs). Such inhibitory nucleic acid molecules can be designed to target any region of a PLA2G12B nucleic acid molecule. In some embodiments, the antisense RNA, siRNA, or shRNA hybridizes to a sequence within a PLA2G12B genomic nucleic acid molecule or mRNA molecule and decreases expression of the PLA2G12B polypeptide in a cell in the subject. In some embodiments, the PLA2G12B inhibitor comprises an antisense molecule that hybridizesto a PLA2G12B genomic nucleic acid molecule or mRNA molecule and decreases expression of the PLA2G12B polypeptide in a cell in the subject. In some embodiments, the PLA2G12B inhibitor comprises an siRNA that hybridizes to a PLA2G12B genomic nucleic acid molecule or mRNA molecule and decreases expression of the PLA2G12B polypeptide in a cell in the subject. In some embodiments, the PLA2G12B inhibitor comprises an shRNA that hybridizes to a PLA2G12B genomic nucleic acid molecule or mRNA molecule and decreases expression of the PLA2G12B polypeptide in a cell in the subject.

[0049] The inhibitory nucleic acid molecules can comprise RNA, DNA, or both RNA and DNA. The inhibitory nucleic acid molecules can also be linked or fused to a heterologous nucleic acid sequence, such as in a vector, or a heterologous label. For example, the inhibitory nucleic acid molecules can be within a vector or as an exogenous donor sequence comprising the inhibitory nucleic acid molecule and a heterologous nucleic acid sequence. The inhibitory nucleic acid molecules can also be linked or fused to a heterologous label. The label can be directly detectable (such as, for example, fluorophore) or indirectly detectable (such as, for example,

[0050] 118452910DOCKET NO.: 38120-4555 (11318WO01)

[0051] - 10-hapten, enzyme, or fluorophore quencher). Such labels can be detectable by spectroscopic, photochemical, biochemical, immunochemical, or chemical means. Such labels include, for example, radiolabels, pigments, dyes, chromogens, spin labels, and fluorescent labels. The label can also be, for example, a chemiluminescent substance; a metal-containing substance; or an enzyme, where there occurs an enzyme-dependent secondary generation of signal. The term "label" can also refer to a "tag" or hapten that can bind selectively to a conjugated molecule such that the conjugated molecule, when added subsequently along with a substrate, is used to generate a detectable signal. For example, biotin can be used as a tag along with an avidin or streptavidin conjugate of horseradish peroxidate (HRP) to bind to the tag, and examined using a calorimetric substrate (such as, for example, tetra methylbenzidine (TMB)) or a fluorogenic substrate to detect the presence of HRP. Exemplary labels that can be used as tags to facilitate purification include, but are not limited to, myc, HA, FLAG or3XFLAG, GXHis or polyhistidine, glutathione-S-transferase (GST), maltose binding protein, an epitope tag, or the Fc portion of immunoglobulin. Numerous labels include, for example, particles, fluorophores, haptens, enzymes and their calorimetric, fluorogenic and chemiluminescent substrates and other labels.

[0052] The inhibitory nucleic acid molecules can comprise, for example, nucleotides or nonnatural or modified nucleotides, such as nucleotide analogs or nucleotide substitutes. Such nucleotides include a nucleotide that contains a modified base, sugar, or phosphate group, or that incorporates a non-natural moiety in its structure. Examples of non-natural nucleotides include, but are not limited to, dideoxynucleotides, biotinylated, aminated, deaminated, alkylated, benzylated, and fluorophor-labeled nucleotides.

[0053] The inhibitory nucleic acid molecules can also comprise one or more nucleotide analogs or substitutions. A nucleotide analog is a nucleotide which contains a modification to either the base, sugar, or phosphate moieties. Modifications to the base moiety include, but are not limited to, natural and synthetic modifications of A, C, G, and T / U, as well as different purine or pyrimidine bases such as, for example, pseudouridine, u racil-5-y I, hypoxanthin-9-yl (I), and 2-aminoadenin-9-yl. Modified bases include, but are not limited to, 5-methylcytosine (5-me-C), 5-hydroxymethyl cytosine, xanthine, hypoxanthine, 2-aminoadenine, 6-methyl and other alkyl derivatives of adenine and guanine, 2-propyl and other alkyl derivatives of adenine and guanine, 2-thiouracil, 2-thiothymine and 2-thiocytosine, 5-halouracil and cytosine, 5-propynyl uracil and cytosine, 6-azo uracil, cytosine and thymine, 5-uracil (pseudouracil),

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[0055] - 11 - 4-thiouracil, 8-halo, 8-amino, 8-thiol, 8-thioalkyl, 8-hydroxyl and other 8-substituted adenines and guanines, 5-halo (such as, for example, 5-bromo), 5-trifluoromethyl and other 5-substituted uracils and cytosines, 7-methylguanine, 7-methyladenine, 8-azaguanine, 8-azaadenine, 7-deazaguanine, 7-deazaadenine, 3-deazaguanine, and 3-deazaadenine.

[0056] Nucleotide analogs can also include modifications of the sugar moiety. Modifications to the sugar moiety include, but are not limited to, natural modifications of the ribose and deoxy ribose as well as synthetic modifications. Sugar modifications include, but are not limited to, the following modifications at the 2' position: OH; F; O-, S-, or N-alkyl; O-, S-, or N-alkenyl; O-, S- or N-alkynyl; or O-a Iky l-O-al kyl, wherein the alkyl, alkenyl, and alkynyl may be substituted or unsubstituted Ci-walkyl or C2-ioalkenyl, and C2-ioalkynyl. Exemplary 2' sugar modifications also include, but are not limited to, -O[(CH2)nO]mCH3, -O(CH2)nOCH3, -O(CH2)nNH2, -O(CH2)nCH3, -O(CH2)n-ONH2, and -O(CH2)nON[(CH2)nCH3)]2, where n and m, independently, are from 1 to about 10. Other modifications at the 2' position include, but are not limited to, Ci-walkyl, substituted lower alkyl, alkaryl, aralkyl, O-alkaryl or O-aralkyl, SH, SCH3, OCN, Cl, Br, CN, CF3, OCF3, SOCH3, SO2CH3, ONO2, NO2, N3, NH2, heterocycloalkyl, heterocycloalkaryl, aminoalkylamino, polyalkylamino, substituted silyl, an RNA cleaving group, a reporter group, an intercalator, a group for improving the pharmacokinetic properties of an oligonucleotide, or a group for improving the pharmacodynamic properties of an oligonucleotide, and other substituents having similar properties. Similar modifications may also be made at other positions on the sugar, particularly the 3' position of the sugar on the 3' terminal nucleotide or in 2'-5' linked oligonucleotides and the 5' position of 5' terminal nucleotide. Modified sugars can also include those that contain modifications at the bridging ring oxygen, such as CH2and S. Nucleotide sugar analogs can also have sugar mimetics, such as cyclobutyl moieties in place of the pentofu ranosyl sugar.

[0057] Nucleotide analogs can also be modified at the phosphate moiety. Modified phosphate moieties include, but are not limited to, those that can be modified so that the linkage between two nucleotides contains a phosphorothioate, chiral phosphorothioate, phosphorodithioate, phosphotriester, aminoalkylphosphotriester, methyl and other alkyl phosphonates including 3'-alkylene phosphonate and chiral phosphonates, phosphinates, phosphoramidates including 3'-amino phosphoramidate and aminoalkylphosphoramidates, thionophosphoramidates, thionoalkylphosphonates, thionoalkylphosphotriesters, and boranophosphates. These phosphate or modified phosphate linkage between two nucleotides can be through a 3'-5'

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[0059] - 12 -linkage or a 2'-5' linkage, and the linkage can contain inverted polarity such as 3'-5' to 5'-3' or 2' -5' to 5'-2'. Various salts, mixed salts, and free acid forms are also included. Nucleotide substitutes also include peptide nucleic acids (PNAs).

[0060] In some embodiments, the antisense nucleic acid molecules are gapmers, whereby the first one to seven nucleotides at the 5' and 3' ends each have 2'-methoxyethyl (2'-MOE) modifications. In some embodiments, the first five nucleotides at the 5' and 3' ends each have 2'-MOE modifications. In some embodiments, the first one to seven nucleotides at the 5' and 3' ends are RNA nucleotides. In some embodiments, the first five nucleotides at the 5' and 3' ends are RNA nucleotides. In some embodiments, each of the backbone linkages between the nucleotides is a phosphorothioate linkage.

[0061] In some embodiments, the siRNA molecules have termini modifications. In some embodiments, the 5' end of the antisense strand is phosphorylated. In some embodiments, 5'-phosphate analogs that cannot be hydrolyzed, such as 5'-(E)-vinyl-phosphonate are used.

[0062] In some embodiments, the siRNA molecules have backbone modifications. In some embodiments, the modified phosphodiester groups that link consecutive ribose nucleosides have been shown to enhance the stability and in vivo bioavailability of siRNAs The non-ester groups (-OH, =0) of the phosphodiester linkage can be replaced with sulfur, boron, or acetate to give phosphorothioate, boranophosphate, and phosphonoacetate linkages. In addition, substituting the phosphodiester group with a phosphotriester can facilitate cellular uptake of siRNAs and retention on serum components by eliminating their negative charge. In some embodiments, the siRNA molecules have sugar modifications. In some embodiments, the sugars are deprotonated (reaction catalyzed by exo- and endonucleases) whereby the

[0063] 2'-hydroxyl can act as a nucleophile and attack the adjacent phosphorous in the phosphodiester bond. Such alternatives include 2'-O-methyl, 2'-O-methoxyethyl, and 2'-fluoro modifications.

[0064] In some embodiments, the siRNA molecules have base modifications. In some embodiments, the bases can be substituted with modified bases such as pseudouridine, 5'-methylcytidine, N6-methyladenosine, inosine, and N7-methylguanosine.

[0065] In some embodiments, the siRNA molecules are conjugated to lipids. Lipids can be conjugated to the 5' or 3' termini of siRNA to improve their in vivo bioavailability by allowing them to associate with serum lipoproteins. Representative lipids include, but are not limited to, cholesterol and vitamin E, and fatty acids, such as palmitate and tocopherol.

[0066] In some embodiments, a representative siRNA has the following formula:

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[0068] - 13 - Sense: mN*mN* / i2FN / mN / i2FN / mN / i2FN / mN / i2FN / mN / i2FN / mN / i2FN / mN / i2FN / mN / i2FN / *mN* / 32FN /

[0069] Antisense: / 52FN / * / i2FN / *mN / i2FN / mN / i2FN / mN / i2FN / mN / i2FN / mN / i2FN / mN / i2FN / mN / i2FN / mN / i2FN / mN*N*N

[0070] wherein: "N" is the base; "2F" is a 2'-F modification; "m" is a 2'-O-methyl modification, "I" is an internal base; and is a phosphorothioate backbone linkage.

[0071] In any of the embodiments described herein, the inhibitory nucleic acid molecules may be administered, for example, as one to two hour i.v. infusions or s.c. injections. In any of the embodiments described herein, the inhibitory nucleic acid molecules may be administered at dose levels that range from about 50 mg to about 900 mg, from about 100 mg to about 800 mg, from about 150 mg to about 700 mg, or from about 175 to about 640 mg (2.5 to 9.14 mg / kg; 92.5 to 338 mg / m2- based on an assumption of a body weight of 70 kg and a conversion of mg / kg to mg / m2dose levels based on a mg / kg dose multiplier value of 37 for humans).

[0072] The present disclosure also provides vectors comprising any one or more of the inhibitory nucleic acid molecules. In some embodiments, the vectors comprise any one or more of the inhibitory nucleic acid molecules and a heterologous nucleic acid. The vectors can be viral or nonviral vectors capable of transporting a nucleic acid molecule. In some embodiments, the vector is a plasmid or cosmid (such as, for example, a circular double-stranded DNA into which additional DNA segments can be ligated). In some embodiments, the vector is a viral vector, wherein additional DNA segments can be ligated into the viral genome. Expression vectors include, but are not limited to, plasmids, cosmids, retroviruses, adenoviruses, adeno-associated viruses (AAV), plant viruses such as cauliflower mosaic virus and tobacco mosaic virus, yeast artificial chromosomes (YACs), Epstein-Barr (EBV)-derived episomes, and other expression vectors known in the art.

[0073] The present disclosure also provides compositions comprising any one or more of the inhibitory nucleic acid molecules. In some embodiments, the composition is a pharmaceutical composition. In some embodiments, the compositions comprise a carrier and / or excipient. Examples of carriers include, but are not limited to, poly(lactic acid) (PLA) microspheres, poly(D,L-lactic-coglycolic-acid) (PLGA) microspheres, liposomes, micelles, inverse micelles, lipid cochleates, and lipid microtubules. A carrier may comprise a buffered salt solution such as PBS, HBSS, etc.

[0074] In some embodiments, the PLA2G12B inhibitor comprises a nuclease agent that

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[0076] - 14-induces one or more nicks or double-strand breaks at a recognition sequence(s) or a DNA-binding protein that bindsto a recognition sequence within a PLA2G12B genomic nucleic acid molecule. The recognition sequence can be located within a coding region of the PLA2G12B gene, or within regulatory regions that influence the expression of the gene. A recognition sequence of the DNA-binding protein or nuclease agent can be located in an intron, an exon, a promoter, an enhancer, a regulatory region, or any non-protein coding region. The recognition sequence can include or be proximate to the start codon of the PLA2G12B gene. For example, the recognition sequence can be located about 10, about 20, about 30, about 40, about 50, about 100, about 200, about 300, about 400, about 500, or about 1,000 nucleotides from the start codon. As another example, two or more nuclease agents can be used, each targeting a nuclease recognition sequence including or proximate to the start codon. As another example, two nuclease agents can be used, one targeting a nuclease recognition sequence including or proximate to the start codon, and one targeting a nuclease recognition sequence including or proximate to the stop codon, wherein cleavage by the nuclease agents can result in deletion of the coding region between the two nuclease recognition sequences. Any nuclease agent that induces a nick or double-strand break into a desired recognition sequence can be used in the methods and compositions disclosed herein. Any DNA-binding protein that binds to a desired recognition sequence can be used in the methods and compositions disclosed herein.

[0077] Suitable nuclease agents and DNA-binding proteins for use herein include, but are not limited to, zinc finger protein or zinc finger nuclease (ZFN) pair, Transcription Activator-Like Effector (TALE) protein or Transcription Activator-Like Effector Nuclease (TALEN), or Clustered Regularly Interspersed Short Palindromic Repeats (CRISPR) / CRISPR-associated (Cas) systems. The length of the recognition sequence can vary, and includes, for example, recognition sequences that are about 30-36 bp for a zinc finger protein or ZFN pair, about 15-18 bp for each ZFN, about 36 bp for a TALE protein or TALEN, and about 20 bp for a CRISPR / Cas guide RNA.

[0078] In some embodiments, CRISPR / Cas systems can be used to modify a PLA2G12B genomic nucleic acid molecule within a cell. The methods and compositions disclosed herein can employ CRISPR-Cas systems by utilizing CRISPR complexes (comprising a guide RNA (gRNA) complexed with a Cas protein) for site-directed cleavage of PLA2G12B nucleic acid molecules.

[0079] Cas proteins generally comprise at least one RNA recognition or binding domain that can interact with gRNAs. Cas proteins can also comprise nuclease domains (such as, for example, DNase or RNase domains), DNA binding domains, helicase domains, protein-protein

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[0081] - 15 -interaction domains, dimerization domains, and other domains. Suitable Cas proteins include, for example, a wild type Cas9 protein and a wild type Cpfl protein (such as, for example, FnCpfl). A Cas protein can have full cleavage activity to create a double-strand break in a PLA2G12B genomic nucleic acid molecule or it can be a nickase that creates a single-strand break in a PLA2G12B genomic nucleic acid molecule. Additional examples of Cas proteins include, but are not limited to, Casl, CaslB, Cas2, Cas3, Cas4, Cas5, Cas5e (CasD), Cas6, Cas6e, CasSf, Cas7, Cas8al, Cas8a2, Cas8b, Cas8c, Cas9 (Csnl or Csxl2), CaslO, CaslOd, CasF, CasG, CasH, Csyl, Csy2, Csy3, Csel (CasA), Cse2 (CasB), Cse3 (CasE), Cse4 (CasC), Cscl, Csc2, Csa5, Csn2, Csm2, Csm3, Csm4, Csm5, Csm6, Cmrl , Cmr3, Cmr4, Cmr5, Cmr6, Csbl, Csb2, Csb3, Csxl7, Csxl4, CsxlO, CsxlG, CsaX, Csx3, Csxl, Csxl5, Csfl, Csf2, Csf3, Csf4, and Cul966, and homologs or modified versions thereof. In some embodiments, a Cas system, such as Casl2a, can have multiple gRNAs encoded into a single crRNA. Cas proteins can also be operably linked to heterologous polypeptides as fusion proteins. For example, a Cas protein can be fused to a cleavage domain, an epigenetic modification domain, a transcriptional activation domain, or a transcriptional repressor domain. Cas proteins can be provided in any form. For example, a Cas protein can be provided in the form of a protein, such as a Cas protein complexed with a gRNA. Alternately, a Cas protein can be provided in the form of a nucleic acid molecule encoding the Cas protein, such as an RNA or DNA.

[0082] In some embodiments, targeted genetic modifications of PLA2G12B genomic nucleic acid molecules can be generated by contacting a cell with a Cas protein and one or more gRNAs that hybridize to one or more gRNA recognition sequences within a target genomic locus in the PLA2G12B genomic nucleic acid molecule. The gRNA recognition sequence can include or be proximate to the start codon of a PLA2G12B genomic nucleic acid molecule orthe stop codon of a PLA2G12B genomic nucleic acid molecule. For example, the gRNA recognition sequence can be located from about 10, from about 20, from about 30, from about 40, from about 50, from about 100, from about 200, from about 300, from about 400, from about 500, or from about 1,000 nucleotides of the start codon orthe stop codon.

[0083] The gRNA recognition sequences within a target genomic locus in a PLA2G12B genomic nucleic acid molecule are located near a Protospacer Adjacent Motif (PAM) sequence, which is a 2-6 base pair DNA sequence immediately following the DNA sequence targeted by the Cas9 nuclease. The canonical PAM is the sequence 5'-NGG-3' where "N" is any nucleobase followed by two guanine ("G") nucleobases. gRNAs can transport Cas9 to anywhere in the genome for

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[0085] - 16 -gene editing, but no editing can occur at any site other than one at which Cas9 recognizes PAM. In addition, 5'-NGA-3' can be a highly efficient non-canonical PAM for human cells. Generally, the PAM is about 2-6 nucleotides downstream of the DNA sequence targeted by the gRNA. The PAM can flank the gRNA recognition sequence. In some embodiments, the gRNA recognition sequence can be flanked on the 3' end by the PAM. In some embodiments, the gRNA recognition sequence can be flanked on the 5' end by the PAM. For example, the cleavage site of Cas proteins can be about 1 to about 10, about 2 to about 5 base pairs, or three base pairs upstream or downstream of the PAM sequence. In some embodiments (such as when Cas9 from S. pyogenes or a closely related Cas9 is used), the PAM sequence of the non-complementary strand can be 5'-NGG-3', where N is any DNA nucleotide and is immediately 3' of the gRNA recognition sequence of the non-complementary strand of the target DNA. As such, the PAM sequence of the complementary strand would be 5'-CCN-3', where N is any DNA nucleotide and is immediately 5' of the gRNA recognition sequence of the complementary strand of the target DNA.

[0086] A gRNA is an RNA molecule that binds to a Cas protein and targets the Cas protein to a specific location within a PLA2G12B genomic nucleic acid molecule. An exemplary gRNA is a gRNA effective to direct a Cas enzyme to bind to or cleave a PLA2G12B genomic nucleic acid molecule, wherein the gRNA comprises a DNA-targeting segment that hybridizes to a gRNA recognition sequence within the PLA2G12B genomic nucleic acid molecule. Exemplary gRNAs comprise a DNA-targeting segment that hybridizes to a gRNA recognition sequence present within a PLA2G12B genomic nucleic acid molecule that includes or is proximate to the start codon or the stop codon. For example, a gRNA can be selected such that it hybridizes to a gRNA recognition sequence that is located from about 5, from about 10, from about 15, from about 20, from about 25, from about 30, from about 35, from about 40, from about 45, from about 50, from about 100, from about 200, from about 300, from about 400, from about 500, or from about 1,000 nucleotides of the start codon or located from about 5, from about 10, from about 15, from about 20, from about 25, from about 30, from about 35, from about 40, from about 45, from about 50, from about 100, from about 200, from about 300, from about 400, from about 500, or from about 1,000 nucleotides of the stop codon. Suitable gRNAs can comprise from about 17 to about 25 nucleotides, from about 17 to about 23 nucleotides, from about 18 to about 22 nucleotides, or from about 19 to about 21 nucleotides. In some embodiments, the gRNAs can comprise 20 nucleotides.

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[0088] - 17 - The Cas protein and the gRNA form a complex, and the Cas protein cleaves the PLA2G12B genomic nucleic acid molecule. The Cas protein can cleave the nucleic acid molecule at a site within or outside of the nucleic acid sequence present in the PLA2G12B genomic nucleic acid molecule to which the DNA-targeting segment of a gRNA will bind. For example, formation of a CRISPR complex (comprising a gRNA hybridized to a gRNA recognition sequence and complexed with a Cas protein) can result in cleavage of one or both strands in or near (such as, for example, within 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 20, 50, or more base pairs from) the nucleic acid sequence present in the PLA2G12B genomic nucleic acid molecule to which a DNA-targeting segment of a gRNA will bind.

[0089] Such methods can result, for example, in a PLA2G12B genomic nucleic acid molecule in which a region of the PLA2G12B genomic nucleic acid molecule is disrupted, the start codon is disrupted, the stop codon is disrupted, or the coding sequence is disrupted or deleted.

[0090] Optionally, the cell can be further contacted with one or more additional gRNAs that hybridize to additional gRNA recognition sequences within the target genomic locus in the PLA2G12B genomic nucleic acid molecule. By contactingthe cell with one or more additional gRNAs (such as, for example, a second gRNA that hybridizes to a second gRNA recognition sequence), cleavage by the Cas protein can create two or more double-strand breaks or two or more single-strand breaks.

[0091] In any of the methods of treatment or prevention described herein, the subject being treated can comprise a PLA2G12B variant nucleic acid molecule. In some embodiments, the methods of treatment or prevention further comprise detecting the presence or absence of a PLA2G12B variant nucleic acid molecule in a biological sample from the subject. In some embodiments, the subject being treated is heterozygous for the PLA2G12B variant nucleic acid molecule. In some embodiments, the subject being treated is PLA2G12B reference. In some embodiments, the subject being treated is PLA2G12B reference. The PLA2G12B variant nucleic acid molecule can be any of the PLA2G12B variant nucleic acid molecules disclosed herein. In some embodiments, the PLA2G12B variant nucleic acid molecule is a PLA2G12B variant genomic nucleic acid molecule that comprises the genetic variation 10:72935723:AGG:A, 10:72941168:C:T, 10:72941227:A:T, 10:72941235:G:A, 10:72941336:T:C, 10:72942659:GGT:G, 10:72942688:G:T, 10:72942730:TG:T, 10:72954474:C:A, 10:72954480:CG:C, 10:72954481:G:A, 10:72954565:C:A, 10:72954604:C:A, 10:72954685:T:C, 10:72935708:AAC:A, 10:72941259:T:A, 10:72941302:G:T, 10:72942650:A:G, 10:72942742:T:A, 10:72954591:G:C, 10:72954601:C:A, or

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[0093] - 18 - 10:72941311:C:CA, or is an mRNA molecule produced therefrom, or is a cDNA molecule produced from the mRNA molecule.

[0094] In some embodiments, the PLA2G12B variant nucleic acid molecule is a PLA2G12B variant genomic nucleic acid molecule that comprises the genetic variation 10:72935723:AGG:A, or is an mRNA molecule produced therefrom, or is a cDNA molecule produced from the mRNA molecule. In some embodiments, the PLA2G12B variant nucleic acid molecule is a PLA2G12B variant genomic nucleic acid molecule that comprises the genetic variation 10:72941168:C:T, or is an mRNA molecule produced therefrom, or is a cDNA molecule produced from the mRNA molecule. In some embodiments, the PLA2G12B variant nucleic acid molecule is a PLA2G12B variant genomic nucleic acid molecule that comprises the genetic variation 10:72941227:A:T, or is an mRNA molecule produced therefrom, or is a cDNA molecule produced from the mRNA molecule. In some embodiments, the PLA2G12B variant nucleic acid molecule is a PLA2G12B variant genomic nucleic acid molecule that comprises the genetic variation 10:72941235:G:A, or is an mRNA molecule produced therefrom, or is a cDNA molecule produced from the mRNA molecule. In some embodiments, the PLA2G12B variant nucleic acid molecule is a PLA2G12B variant genomic nucleic acid molecule that comprises the genetic variation 10:72941336:T:C, or is an mRNA molecule produced therefrom, or is a cDNA molecule produced from the mRNA molecule. In some embodiments, the PLA2G12B variant nucleic acid molecule is a PLA2G12B variant genomic nucleic acid molecule that comprises the genetic variation 10:72942659:GGT:G, or is an mRNA molecule produced therefrom, or is a cDNA molecule produced from the mRNA molecule. In some embodiments, the PLA2G12B variant nucleic acid molecule is a PLA2G12B variant genomic nucleic acid molecule that comprises the genetic variation 10:72942688:G:T, or is an mRNA molecule produced therefrom, or is a cDNA molecule produced from the mRNA molecule. In some embodiments, the PLA2G12B variant nucleic acid molecule is a PLA2G12B variant genomic nucleic acid molecule that comprises the genetic variation 10:72942730:TG:T, or is an mRNA molecule produced therefrom, or is a cDNA molecule produced from the mRNA molecule. In some embodiments, the PLA2G12B variant nucleic acid molecule is a PLA2G12B variant genomic nucleic acid molecule that comprises the genetic variation 10:72954474:C:A, or is an mRNA molecule produced therefrom, or is a cDNA molecule produced from the mRNA molecule. In some embodiments, the PLA2G12B variant nucleic acid molecule is a PLA2G12B variant genomic nucleic acid molecule that comprises the genetic variation 10:72954480:CG:C, or is an mRNA molecule produced therefrom, or is a cDNA

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[0096] - 19-molecule produced from the mRNA molecule. In some embodiments, the PLA2G12B variant nucleic acid molecule is a PLA2G12B variant genomic nucleic acid molecule that comprises the genetic variation 10:72954481:G:A, or is an mRNA molecule produced therefrom, or is a cDNA molecule produced from the mRNA molecule. In some embodiments, the PLA2G12B variant nucleic acid molecule is a PLA2G12B variant genomic nucleic acid molecule that comprises the genetic variation 10:72954565:C:A, or is an mRNA molecule produced therefrom, or is a cDNA molecule produced from the mRNA molecule. In some embodiments, the PLA2G12B variant nucleic acid molecule is a PLA2G12B variant genomic nucleic acid molecule that comprises the genetic variation 10:72954604:C:A, or is an mRNA molecule produced therefrom, or is a cDNA molecule produced from the mRNA molecule. In some embodiments, the PLA2G12B variant nucleic acid molecule is a PLA2G12B variant genomic nucleic acid molecule that comprises the genetic variation 10:72954685:T:C, or is an mRNA molecule produced therefrom, or is a cDNA molecule produced from the mRNA molecule. In some embodiments, the PLA2G12B variant nucleic acid molecule is a PLA2G12B variant genomic nucleic acid molecule that comprises the genetic variation 10:72935708:AAC:A, or is an mRNA molecule produced therefrom, or is a cDNA molecule produced from the mRNA molecule. In some embodiments, the PLA2G12B variant nucleic acid molecule is a PLA2G12B variant genomic nucleic acid molecule that comprises the genetic variation 10:72941259:T:A, or is an mRNA molecule produced therefrom, or is a cDNA molecule produced from the mRNA molecule. In some embodiments, the PLA2G12B variant nucleic acid molecule is a PLA2G12B variant genomic nucleic acid molecule that comprises the genetic variation 10:72941302:G:T, or is an mRNA molecule produced therefrom, or is a cDNA molecule produced from the mRNA molecule. In some embodiments, the PLA2G12B variant nucleic acid molecule is a PLA2G12B variant genomic nucleic acid molecule that comprises the genetic variation 10:72942650:A:G, or is an mRNA molecule produced therefrom, or is a cDNA molecule produced from the mRNA molecule. In some embodiments, the PLA2G12B variant nucleic acid molecule is a PLA2G12B variant genomic nucleic acid moleculethat comprises the genetic variation 10:72942742:T:A, or is an mRNA molecule produced therefrom, or is a cDNA molecule produced from the mRNA molecule. In some embodiments, the PLA2G12B variant nucleic acid molecule is a PLA2G12B variant genomic nucleic acid molecule that comprises the genetic variation 10:72954591:G:C, or is an mRNA molecule produced therefrom, or is a cDNA molecule produced from the mRNA molecule. In some embodiments, the PLA2G12B variant nucleic acid molecule is a PLA2G12B

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[0098] - 20 -variant genomic nucleic acid molecule that comprises the genetic variation 10:72954601:C:A, or is an mRNA molecule produced therefrom, or is a cDNA molecule produced from the mRNA molecule. In some embodiments, the PLA2G12B variant nucleic acid molecule is a PLA2G12B variant genomic nucleic acid molecule that comprises the genetic variation 10:72941311:C:CA, or is an mRNA molecule produced therefrom, or is a cDNA molecule produced from the mRNA molecule.

[0099] The present disclosure also provides methods of treating a subject with one or more lipid disorder therapeutic agents or one or more cardiovascular disease therapeutic agents that treat or inhibit a lipid disorder or a cardiovascular disease, wherein the subject has a lipid disorder or a cardiovascular disease or is at risk of developing a lipid disorder or a cardiovascular disease. The methods comprise determining whether the subject has a PLA2G12B variant nucleic acid molecule by obtaining or having obtained a biological sample from the subject and performing or having performed a sequence analysis on the biological sample to determine if the subject has a genotype comprising the PLA2G12B variant nucleic acid molecule. In embodiments where the subject is PLA2G12B reference, the methods further comprise administering or continuing to administer the lipid disorder therapeutic agent or the cardiovascular disease therapeutic agent in an amount that is the same as or less than a standard dosage amount and / or administering a PLA2G12B inhibitorto the subject. In embodiments where the subject is heterozygous for the PLA2G12B variant nucleic acid molecule, the methods further comprise administering or continuing to administer the lipid disorder therapeutic agent or the cardiovascular disease therapeutic agent in an amount that is the same as or less than a standard dosage amount and / or administering a PLA2G12B inhibitor to the subject. In embodiments where the subject is homozygous for the PLA2G12B variant nucleic acid molecule, the methods further comprise administering or continuing to administer the lipid disorder therapeutic agent or the cardiovascular disease therapeutic agent in a standard dosage amount. The presence of a genotype having the PLA2G12B variant nucleic acid molecule indicates the subject has a decreased risk of developing a lipid disorder or a cardiovascular disease. In some embodiments, the subject is PLA2G12B reference. In some embodiments, the subject is heterozygous for a PLA2G12B variant nucleic acid molecule. In some embodiments, the subject is homozygous for a PLA2G12B variant nucleic acid molecule. In any of the embodiments described herein, the PLA2G12B inhibitor is an example of a lipid disorder therapeutic agent or a cardiovascular disease therapeutic agent. In some

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[0101] - 21 -embodiments, the PLA2G12B variant nucleic acid molecule is a PLA2G12B variant genomic nucleic acid molecule that comprises any of the genetic variants described herein, or is an mRNA molecule produced therefrom, or is a cDNA molecule produced from the mRNA molecule.

[0102] For subjects that are genotyped or determined to be either PLA2G12B reference or heterozygous for a PLA2G12B variant nucleic acid molecule, such subjects can be administered a PLA2G12B inhibitor, as described herein.

[0103] Detecting the presence or absence of a PLA2G12B variant nucleic acid molecule in a biological sample from a subject and / or determining whether a subject has a PLA2G12B variant nucleic acid molecule can be carried out by any of the methods described herein. In some embodiments, these methods can be carried out in vitro. In some embodiments, these methods can be carried out in situ. In some embodiments, these methods can be carried out in vivo. In any of these embodiments, the nucleic acid molecule can be present within a cell obtained from the subject.

[0104] In some embodiments, the treatment or prevention methods comprise detecting the presence or absence of a PLA2G12B variant polypeptide in a biological sample from the subject. In some embodiments, when the subject does not have a PLA2G12B variant polypeptide, the subject is administered a lipid disorder therapeutic agent or a cardiovascular disease therapeutic agent in an amount that is the same as or less than a standard dosage amount. In some embodiments, when the subject has a PLA2G12B variant polypeptide, the subject is administered a lipid disorder therapeutic agent or a cardiovascular disease therapeutic agent in a standard dosage amount.

[0105] The present disclosure also provides methods of treating a subject with a lipid disorder therapeutic agent or a cardiovascular disease therapeutic agent, wherein the subject has a lipid disorder or a cardiovascular disease or is at risk of developing a lipid disorder or a cardiovascular disease. The methods comprise determining whether the subject has a PLA2G12B variant polypeptide by obtaining or having obtained a biological sample from the subject and performing or having performed an assay on the biological sample to determine if the subject has a PLA2G12B variant polypeptide. When the subject does not have a PLA2G12B variant polypeptide, the lipid disorder therapeutic agent or the cardiovascular disease therapeutic agent is administered in an amount that is the same as or less than a standard dosage amount and / or a PLA2G12B inhibitor is administered to the subject. When the subject

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[0107] - 22 -has a PLA2G12B variant polypeptide, the lipid disorder therapeutic agent or the cardiovascular disease therapeutic agent is administered in a standard dosage amount and / or a PLA2G12B inhibitor is administered to the subject. The presence of a PLA2G12B variant polypeptide indicates the subject has a decreased risk of developing a lipid disorder or a cardiovascular disease. In some embodiments, the subject has a PLA2G12B variant polypeptide. In some embodiments, the subject does not have a PLA2G12B variant polypeptide.

[0108] The present disclosure also provides methods of preventing a subject from developing a lipid disorder or a cardiovascular disease by administering a lipid disorder therapeutic agent or a cardiovascular disease therapeutic agent. In some embodiments, the method comprises determining whether the subject has a PLA2G12B variant polypeptide by obtaining or having obtained a biological sample from the subject and performing or having performed an assay on the biological sample to determine if the subject has a PLA2G12B variant polypeptide. When the subject does not have a PLA2G12B variant polypeptide, the lipid disorder therapeutic agent or the cardiovascular disease therapeutic agent is administered in an amount that is the same as or less than a standard dosage amount and / or a PLA2G12B inhibitor is administered to the subject. When the subject has a PLA2G12B variant polypeptide, the lipid disorder therapeutic agent or the cardiovascular disease therapeutic agent is administered in a standard dosage amount and / or a PLA2G12B inhibitor is administered to the subject. The presence of a PLA2G12B variant polypeptide indicates the subject has a decreased risk of developing a lipid disorder or a cardiovascular disease. In some embodiments, the subject has a PLA2G12B variant polypeptide. In some embodiments, the subject does not have a PLA2G12B variant polypeptide.

[0109] Detecting the presence or absence of a PLA2G12B variant polypeptide in a biological sample from a subject and / or determining whether a subject has a PLA2G12B variant polypeptide can be carried out by any of the methods described herein. In some embodiments, these methods can be carried out in vitro. In some embodiments, these methods can be carried out in situ. In some embodiments, these methods can be carried out in vivo. In any of these embodiments, the polypeptide can be present within a cell obtained from the subject.

[0110] In some embodiments, the PLA2G12B inhibitor is a small molecule. In some embodiments, the small molecule is low molecular weight (< 900 daltons) organic compound.

[0111] In some embodiments, the PLA2G12B inhibitor comprises an antibody, or antigenbinding fragment thereof. In some embodiments, the antibody, or antigen-binding fragment thereof, binds specifically to human PLA2G12B. In some embodiments, the antibody is a fully

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[0113] - 23 -human monoclonal antibody (mAb), or antigen-binding fragment thereof, that specifically binds and neutralizes, inhibits, blocks, abrogates, reduces, or interferes with, at least one activity of PLA2G12B, in particular, human PLA2G12B. In some embodiments, an antibody or fragment thereof can neutralize, inhibit, block, abrogate, reduce, or interfere with, an activity of PLA2G12B by binding to an epitope of PLA2G12B that is directly involved in the targeted activity of PLA2G12B. In some embodiments, an antibody or fragment thereof can neutralize, inhibit, block, abrogate, reduce, or interfere with, an activity of PLA2G12B by binding to an epitope of PLA2G12B that is not directly involved in the targeted activity of PLA2G12B, but the antibody or fragment binding thereto sterica I ly or conformationally inhibits, blocks, abrogates, reduces, or interferes with, the targeted activity of PLA2G12B. In some embodiments, an antibody or fragment thereof binds to an epitope of PLA2G12B that is not directly involved in the targeted activity of PLA2G12B (i.e., a non-blocking antibody), but the antibody or fragment binding thereto results in the enhancement of the clearance of PLA2G12B from the circulation, compared to the clearance of PLA2G12B in the absence of the antibody orfragment thereof, thereby indirectly inhibiting, blocking, abrogating, reducing, or interfering with, an activity of PLA2G12B. Clearance of PLA2G12B from the circulation can be particularly enhanced by combining two or more different non-blocking antibodies that do not compete with one another for specific binding to PLA2G12B. The antibodies can be full-length (for example, an IgGl or lgG4 antibody) or may comprise only an antigen-binding portion (for example, a Fab, F(a b')2 or scFv fragment), and may be modified to affect functionality, e.g., to eliminate residual effector functions (Reddy et aL, J. Immunol., 2000, 164, 1925-1933).

[0114] In some embodiments, the antibody or antigen-binding fragment thereof specifically binds to PLA2G12B with an equilibrium dissociation constant (KD) of about 7 nM or less, about 6 nM or less, about 5 nM or less, about 4 nM or less, about 3 nM or less, about 2 nM or less, or about 1 nM or less, as measured by surface plasmon resonance assay (for example, BIACORE™). In some embodiments, the antibody exhibits a KD of about 800 pM or less, about 700 pM or less; about 600 pM or less; about 500 pM or less; about 400 pM or less; about 300 pM or less; about 200 pM or less; about 100 pM or less; or about 50 pM or less.

[0115] In some embodiments, the anti-PLA2G12B antibodies have a modified glycosylation pattern. In some applications, modification to remove undesirable glycosylation sites may be useful, or e.g., removal of a fucose moiety to increase antibody dependent cellular cytotoxicity (ADCC) function (see, Shield et al., J. Biol. Chem., 2002, 277, 26733). In other applications,

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[0117] - 24-removal of N-glycosylation site may reduce undesirable immune reactions against the therapeutic antibodies or increase affinities of the antibodies. In yet other applications, modification of galactosylation can be made in order to modify complement dependent cytotoxicity (CDC).

[0118] The present disclosure also provides compositions comprising a combination of an antibody or antigen-binding fragment thereof and a lipid disorder therapeutic agent or a cardiovascular disease therapeutic agent.

[0119] In some embodiments, the lipid disorder therapeutic agent orthe cardiovascular disease therapeutic agent include, but are not limited to, those disclosed herein. In some embodiments, the lipid disorder therapeutic agent orthe cardiovascular disease therapeutic agent can be combined with a PLA2G12B inhibitor. The lipid disorder therapeutic agent orthe cardiovascular disease therapeutic agent may be delayed or avoided altogether by treatment with a PLA2G12B inhibitor as described herein.

[0120] Numerous suitable lipid disordertherapeutic agents for low HDL-C levels (hypoalphalipoproteinemia) include, but are not limited to, niacin, a fibrate (e.g., gemfibrozil (Lopid), fenofibrate, clofibrate, and ciprofibrate), or a statin (e.g., atorvastatin, pravastatin, fluvastatin, lovastatin, simvastatin, and cerivastatin).

[0121] Lipid disordertherapeutic agents for treating increased LDL include, but are not limited to, statins (e.g., atorvastatin, pravastatin, fluvastatin, lovastatin, simvastatin, and cerivastatin) and proprotein convertase subtilisin / kexin type 9 (PCSK9) inhibitors (e.g., alirocumab (Praluent®) and evolocumab (Repatha®)).

[0122] Lipid disordertherapeutic agents for treating chronic kidney disease include, but are not limited to, blood pressure medicines, such as an ACE inhibitors or angiotensin II receptor blockers (ARB), and statins, such as those recited herein.

[0123] Lipid disordertherapeutic agents for treating chronic peripheral venous insufficiency include, but are not limited to, diuretics, vasodilators, such as oxpentifylline, or anticoagulants, such as warfarin, rivaroxaban, and dabigatran.

[0124] Lipid disordertherapeutic agents for treating pure hyperglyceridemia include, but are not limited to, fibric acid derivatives, such as gemfibrozil (Lopid) and fenofibrate, niacin, and omega-3 fatty acids.

[0125] Lipid disordertherapeutic agents for treating type 1 diabetes mellitus include, but are not limited to, alpha-glucosidase inhibitors, such as acarbose or miglitol; biguanides, such as

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[0127] - 25 -metformins; dopamine agonists such as bromocriptine; DPP-4 inhibitors, such as alogliptins and linagliptins; glucagon-like peptides such as albiglutide, dulaglutide, exenatide, and liraglutide; SGLT 2 inhibitors, such as dapagliflozins, canagliflozins, and empagliflozins; sulfonylureas, such as glimepirides, gliclazide, glipizides, glyburides, chlorpropamide, tolazamide, or tolbutamide; and thiazolidinediones, such as rosiglitazone, pioglitazone, 5-((4-(2-(methyl-2-pyridinylamino)ethoxy)phenyl)methyl)-2,4-thiazolidinedione, troglitazone, ciglitazone, WAY-120,744, englitazone, AD 5075, and darglitazone.

[0128] Lipid disorder therapeutic agents for treating combined systolic (congestive) and diastolic (congestive) heart failure include, but are not limited to, ACE inhibitors, such as captopril, enalapril, lisinopril, benazepril, and ramipril; beta blockers such as epinephrine and digoxin; and diuretics, such as furosemide, hydrochlorothiazide, hydrochlorothiazide, bumetanide, torsemide, spironolactone, and metolazone.

[0129] Lipid disorder therapeutic agents for treating rheumatic tricuspid valve diseases and heart failure include, but are not limited to, diuretics, such as furosemide, hydrochlorothiazide, hydrochlorothiazide, bumetanide, torsemide, spironolactone, and metolazone; and arrhythmia medications, such as calcium channel blockers (such as amlodipine, diltiazem, felodipine, isradipine, nicardipine, nifedipine, nisoldipine, and verapamil), beta blockers (such as epinephrine), and anticoagulants (such as warfarin, rivaroxaban, and dabigatran).

[0130] Therapeutic agents for treating a coagulation condition include, but are not limited to heparin, warfarin, rivaroxaban, dabigatran, apixaban, edoxaban, enoxaparin, fondaparinux, dalteparin, bivalirudin, argatroban, or antithrombin III.

[0131] Therapeutic agents for treating CAD include, but are not limited to a cholesterol-mod ifying medication (such as, for example, a statin, niacin, a fibrate, or a bile acid sequestra nt), aspirin, a beta blocker, nitroglycerin, an angiotensin-converting enzyme (ACE) inhibitor, and / or an angiotensin II receptor blocker (ARB).

[0132] Therapeutic agents for treating heart failure include, but are not limited to ANGPTL3 inhibitors. In some embodiments, the ANGPTL3 inhibitor is (12mer-)heparin or CAT-2003. In some embodiments, the ANGPTL3 inhibitor is a vaccine. In some embodiments, the vaccine comprises a peptide corresponding to the LPL inhibitory domain of ANGPTL3. In some embodiments, the vaccine comprises a peptide having an amino acid sequence comprising amino acids 32 to 41 of ANGPTL3 (i.e., EPKSRFAMLD). In some embodiments, the ANGPTL3 inhibitor is an antibody, or antigen-binding fragment thereof. Exemplary antibodies, and

[0133] 118452910DOCKET NO.: 38120-4555 (11318WO01)

[0134] - 26 -fragments thereof, are disclosed in PCT Publication WO 2020 / 243031, which is incorporated herein by reference in its entirety.

[0135] Therapeutic agents for treating an aneurysm include, but are not limited to acetaminophen, a calcium channel blocker (such as, for example, verapamil and diltiazem), nimodipine, a vasodilator, levetiracetam, phenytoin, valproic acid, metoprolol, a beta blocker (such as, for example, metoprolol, esmolol, and la beta lol), an angiotensin II receptor blocker, including losartan, valsartan, and olmesartan, a statin, including atorvastatin, lovastatin, simvastatin, a nitroprusside, an adrenergic modifier, an angiotensin-converting enzyme (ACE) inhibitor, a direct renin inhibitor, a direct vasodilator, and a diuretic. Additional aneurysm therapies include any therapy used to reduce or manage aneurysm risk factors. In some embodiments, the aneurysm therapy is surgical clipping to close off an aneurysm, endovascular aortic aneurysm repair, flow diverter, an intraluminal flow disrupter, coiling, coiling in combination with stenting, different combinations of various devices to destroy the aneurysm from inside the blood vessel, open-chest surgery, aortic root surgery, angioplasty, ventricular or lumbar draining catheters, shunt surgery, rehabilitative therapy, exercise. In some embodiments, the therapeutic agent for treating an aneurysm is an APOC3 inhibitor. In some embodiments, the APOC3 inhibitor comprises an antibody, or antigen-binding fragment thereof.

[0136] Additional cardiovascular disease therapeutic agents include, but are not limited to: 1) 3-hydroxy-3-methylglutaryl-coenzyme A (HMG-CoA) reductase inhibitors, such as cerivastatin, atorvastatin, simvastatin, pitavastatin, rosuvastatin, fluvastatin, lovastatin, pravastatin, and the like; 2) inhibitors of cholesterol uptake and / or bile acid re-absorption; 3) niacin, which increases lipoprotein catabolism; 4) fibrates or amphipathic carboxylic acids, which reduce low-density lipoprotein (LDL) level, improve high-density lipoprotein (HDL) and TG levels, and reduce the number of non-fatal heart attacks; 5) activators of the LXR transcription factor that plays a role in cholesterol elimination such as 22-hydroxycholesterol, or fixed combinations such as ezetimibe plus simvastatin; a statin with a bile resin (e.g., cholestyramine, colestipol, colesevelam), a fixed combination of niacin plus a statin (e.g., niacin with lovastatin); or with other lipid lowering agents such as omega-3-fatty acid ethyl esters (for example, omacor); 6) an angiotensin-converting enzyme (ACE) inhibitor (ACE i), such as, for example, benazepril, captopril, enalapril, fosinopril, lisinopril, moexipril, perindopril, quinapril, ramipril, and trandolapril, or any combination thereof; 7) an angiotensin receptor blocker (ARB), such as, for

[0137] 118452910DOCKET NO.: 38120-4555 (11318WO01)

[0138] - 27 -example, candesartan, irbesartan, olmesartan, losartan, valsartan, telmisartan, and eprosartan, or any combination thereof; and 8) an angiotensin receptor / neprilysin inhibitor (ARNi), such as, for example, sacubitril / valsartan.

[0139] In some embodiments, the dose ofthe lipid disorder therapeutic agent orthe cardiovascular disease therapeutic agent can be decreased by about 10%, by about 20%, by about 30%, by about 40%, by about 50%, by about 60%, by about 70%, by about 80%, or by about 90% for subjects that are PLA2G12B reference or heterozygous for a PLA2G12B variant nucleic acid molecule (i.e., a less than the standard dosage amount) compared to subjects that are homozygous for a PLA2G12B variant nucleic acid molecule (who may receive standard dosage amount). In some embodiments, the dose ofthe lipid disordertherapeutic agent orthe cardiovascular disease therapeutic agent can be decreased by about 10%, by about 20%, by about 30%, by about 40%, or by about 50%. In addition, subjects that are PLA2G12B reference or heterozygous for a PLA2G12B variant nucleic acid molecule can be administered the lipid disordertherapeutic agent orthe cardiovascular disease therapeutic agent less frequently compared to subjects that are homozygous for a PLA2G12B variant nucleic acid molecule.

[0140] Administration ofthe lipid disordertherapeutic agents orthe cardiovascular disease therapeutic agents and / or PLA2G12B inhibitors can be repeated, for example, after one day, two days, three days, five days, one week, two weeks, three weeks, one month, five weeks, six weeks, seven weeks, eight weeks, two months, or three months. The repeated administration can be at the same dose or at a different dose. The administration can be repeated once, twice, three times, four times, five times, six times, seven times, eight times, nine times, ten times, or more. For example, according to certain dosage regimens a subject can receive therapy for a prolonged period of time such as, for example, 6 months, 1 year, or more.

[0141] Administration ofthe lipid disordertherapeutic agents orthe cardiovascular disease therapeutic agents and / or PLA2G12B inhibitors can occur by any suitable route including, but not limited to, parenteral, intravenous, oral, subcutaneous, intra-arterial, intracranial, intrathecal, intraperitoneal, topical, intranasal, or intramuscular. Pharmaceutical compositions for administration are desirably sterile and substantially isotonic and manufactured under GMP conditions. Pharmaceutical compositions can be provided in unit dosage form (i.e., the dosage for a single administration). Pharmaceutical compositions can be formulated using one or more physiologically and pharmaceutically acceptable carriers, diluents, excipients, or auxiliaries. The formulation depends on the route of administration chosen. The term "pharmaceutically

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[0143] - 28 -acceptable" means that the carrier, diluent, excipient, or auxiliary is compatible with the other ingredients of the formulation and not substantially deleterious to the recipient thereof. The terms "treat", "treating", and "treatment" and "prevent", "preventing", and "prevention" as used herein, refer to eliciting the desired biological response, such as a therapeutic and prophylactic effect, respectively. In some embodiments, a therapeutic effect comprises one or more of a decrease / reduction in a lipid disorder or a cardiovascular disease, a decrease / reduction in the severity of a lipid disorder or a cardiovascular disease (such as, for example, a reduction or inhibition of development of a lipid disorder or a cardiovascular disease), a decrease / reduction in symptoms and disease-related effects, delaying the onset of symptoms and disease-related effects, reducing the severity of symptoms of disease-related effects, reducing the number of symptoms and disease-related effects, reducing the latency of symptoms and disease-related effects, an amelioration of symptoms and disease-related effects, reducing secondary symptoms, reducing secondary infections, preventing relapse to a lipid disorder or a cardiovascular disease, decreasing the number or frequency of relapse episodes, increasing latency between symptomatic episodes, increasing time to sustained progression, speeding recovery, or increasing efficacy of or decreasing resistance to alternative therapeutics, and / or an increased survival time of the affected host animal, following administration of the agent or composition comprisingthe agent. A prophylactic effect may comprise a complete or partial avoidance / inhibition or a delay of a lipid disorder or a cardiovascular disease development / progression (such as, for example, a complete or partial avoidance / inhibition or a delay), and an increased survival time of the affected host animal, following administration of a therapeutic protocol. Treatment of a lipid disorder or a cardiovascular disease encompasses the treatment of a subject already diagnosed as having any form of a lipid disorder or a cardiovascular disease at any clinical stage or manifestation, the delay of the onset or evolution or aggravation or deterioration of the symptoms or signs of a lipid disorder or a cardiovascular disease, or preventing and / or reducing the severity of a lipid disorder or a cardiovascular disease.

[0144] In some embodiments, the PLA2G12B inhibitor and the lipid disordertherapeutic agent or the cardiovascular disease therapeutic agent are disposed within a pharmaceutical composition. In some embodiments, the PLA2G12B inhibitor is disposed within a first pharmaceutical composition and the lipid disordertherapeutic agent or the cardiovascular disease therapeutic agent is disposed within a second pharmaceutical composition. In some

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[0146] - 29-embodiments, the first pharmaceutical composition and the second pharmaceutical composition are administered simultaneously. In some embodiments, the first pharmaceutical composition is administered before the second pharmaceutical composition. In some embodiments, the first pharmaceutical composition is administered after the second pharmaceutical composition.

[0147] In some embodiments, the PLA2G12B predicted loss-of-function variant nucleic acid molecule comprises a splice-site variant, a stop-gain variant, a start-loss variant, a stop-loss variant, a frameshift variant, a missense variant, an in-frame indel variant, and / or a variant that encodes a truncated PLA2G12B variant polypeptide.

[0148] The present disclosure also provides methods of identifying a subject having an increased risk of developing a lipid disorder or a cardiovascular disease. In some embodiments, the method comprises determining or having determined in a biological sample obtained from the subject the presence or absence of a PLA2G12B variant nucleic acid molecule (such as a genomic nucleic acid molecule, mRNA molecule, and / or cDNA molecule). When the subject lacks a PLA2G12B variant nucleic acid molecule (i.e., the subject is genotypically categorized as PLA2G12B reference), then the subject has an increased risk of developing a lipid disorder or a cardiovascular disease. When the subject has a PLA2G12B variant nucleic acid molecule (i.e., the subject is heterozygous or homozygous for a PLA2G12B variant nucleic acid molecule), then the subject has a decreased risk of developing a lipid disorder or a cardiovascular disease. In some embodiments, the PLA2G12B variant nucleic acid molecule is a PLA2G12B variant genomic nucleic acid molecule that comprises the genetic variation 10:72935723:AGG:A, 10:72941168:C:T, 10:72941227:A:T, 10:72941235:G:A, 10:72941336:T:C, 10:72942659:GGT:G, 10:72942688:G:T, 10:72942730:TG:T, 10:72954474:C:A, 10:72954480:CG:C, 10:72954481:G:A, 10:72954565:C:A, 10:72954604:C:A, 10:72954685:T:C, 10:72935708:AAC:A, 10:72941259:T:A, 10:72941302:G:T, 10:72942650:A:G, 10:72942742:T:A, 10:72954591:G:C, 10:72954601:C:A, or 10:72941311:C:CA, or is an mRNA molecule produced therefrom, or is a cDNA molecule produced from the mRNA molecule.

[0149] In some embodiments, the PLA2G12B variant nucleic acid molecule is a PLA2G12B variant genomic nucleic acid molecule that comprises the genetic variation 10:72935723:AGG:A, or is an mRNA molecule produced therefrom, or is a cDNA molecule produced from the mRNA molecule. In some embodiments, the PLA2G12B variant nucleic acid molecule is a PLA2G12B variant genomic nucleic acid molecule that comprises the genetic

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[0151] - 30 -variation 10:72941168:C:T, or is an mRNA molecule produced therefrom, or is a cDNA molecule produced from the mRNA molecule. In some embodiments, the PLA2G12B variant nucleic acid molecule is a PLA2G12B variant genomic nucleic acid molecule that comprises the genetic variation 10:72941227:A:T, or is an mRNA molecule produced therefrom, or is a cDNA molecule produced from the mRNA molecule. In some embodiments, the PLA2G12B variant nucleic acid molecule is a PLA2G12B variant genomic nucleic acid molecule that comprises the genetic variation 10:72941235:G:A, or is an mRNA molecule produced therefrom, or is a cDNA molecule produced from the mRNA molecule. In some embodiments, the PLA2G12B variant nucleic acid molecule is a PLA2G12B variant genomic nucleic acid molecule that comprises the genetic variation 10:72941336:T:C, or is an mRNA molecule produced therefrom, or is a cDNA molecule produced from the mRNA molecule. In some embodiments, the PLA2G12B variant nucleic acid molecule is a PLA2G12B variant genomic nucleic acid molecule that comprises the genetic variation 10:72942659:GGT:G, or is an mRNA molecule produced therefrom, or is a cDNA molecule produced from the mRNA molecule. In some embodiments, the PLA2G12B variant nucleic acid molecule is a PLA2G12B variant genomic nucleic acid molecule that comprises the genetic variation 10:72942688:G:T, or is an mRNA molecule produced therefrom, or is a cDNA molecule produced from the mRNA molecule. In some embodiments, the PLA2G12B variant nucleic acid molecule is a PLA2G12B variant genomic nucleic acid molecule that comprises the genetic variation 10:72942730:TG:T, or is an mRNA molecule produced therefrom, or is a cDNA molecule produced from the mRNA molecule. In some embodiments, the PLA2G12B variant nucleic acid molecule is a PLA2G12B variant genomic nucleic acid molecule that comprises the genetic variation 10:72954474:C:A, or is an mRNA molecule produced therefrom, or is a cDNA molecule produced from the mRNA molecule. In some embodiments, the PLA2G12B variant nucleic acid molecule is a PLA2G12B variant genomic nucleic acid molecule that comprises the genetic variation 10:72954480:CG:C, or is an mRNA molecule produced therefrom, or is a cDNA molecule produced from the mRNA molecule. In some embodiments, the PLA2G12B variant nucleic acid molecule is a PLA2G12B variant genomic nucleic acid molecule that comprises the genetic variation 10:72954481:G:A, or is an mRNA molecule produced therefrom, or is a cDNA molecule produced from the mRNA molecule. In some embodiments, the PLA2G12B variant nucleic acid molecule is a PLA2G12B variant genomic nucleic acid molecule that comprises the genetic variation 10:72954565:C:A, or is an mRNA molecule produced therefrom, or is a cDNA molecule produced from the mRNA molecule. In some embodiments, the PLA2G12B variant

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[0153] - 31 -nucleic acid molecule is a PLA2G12B variant genomic nucleic acid molecule that comprises the genetic variation 10:72954604:C:A, or is an mRNA molecule produced therefrom, or is a cDNA molecule produced from the mRNA molecule. In some embodiments, the PLA2G12B variant nucleic acid molecule is a PLA2G12B variant genomic nucleic acid molecule that comprises the genetic variation 10:72954685:T:C, or is an mRNA molecule produced therefrom, or is a cDNA molecule produced from the mRNA molecule. In some embodiments, the PLA2G12B variant nucleic acid molecule is a PLA2G12B variant genomic nucleic acid molecule that comprises the genetic variation 10:72935708:AAC:A, or is an mRNA molecule produced therefrom, or is a cDNA molecule produced from the mRNA molecule. In some embodiments, the PLA2G12B variant nucleic acid molecule is a PLA2G12B variant genomic nucleic acid molecule that comprises the genetic variation 10:72941259:T:A, or is an mRNA molecule produced therefrom, or is a cDNA molecule produced from the mRNA molecule. In some embodiments, the PLA2G12B variant nucleic acid molecule is a PLA2G12B variant genomic nucleic acid molecule that comprises the genetic variation 10:72941302:G:T, or is an mRNA molecule produced therefrom, or is a cDNA molecule produced from the mRNA molecule. In some embodiments, the PLA2G12B variant nucleic acid molecule is a PLA2G12B variant genomic nucleic acid molecule that comprises the genetic variation 10:72942650:A:G, or is an mRNA molecule produced therefrom, or is a cDNA molecule produced from the mRNA molecule. In some embodiments, the PLA2G12B variant nucleic acid molecule is a PLA2G12B variant genomic nucleic acid moleculethat comprises the genetic variation 10:72942742:T:A, or is an mRNA molecule produced therefrom, or is a cDNA molecule produced from the mRNA molecule. In some embodiments, the PLA2G12B variant nucleic acid molecule is a PLA2G12B variant genomic nucleic acid molecule that comprises the genetic variation 10:72954591:G:C, or is an mRNA molecule produced therefrom, or is a cDNA molecule produced from the mRNA molecule. In some embodiments, the PLA2G12B variant nucleic acid molecule is a PLA2G12B variant genomic nucleic acid molecule that comprises the genetic variation 10:72954601:C:A, or is an mRNA molecule produced therefrom, or is a cDNA molecule produced from the mRNA molecule. In some embodiments, the PLA2G12B variant nucleic acid molecule is a PLA2G12B variant genomic nucleic acid molecule that comprises the genetic variation 10:72941311:C:CA, or is an mRNA molecule produced therefrom, or is a cDNA molecule produced from the mRNA molecule.

[0154] 118452910DOCKET NO.: 38120-4555 (11318WO01)

[0155] - 32 - Having a single copy of a PLA2G12B variant nucleic acid molecule is more protective of a subject from developing a lipid disorder or a cardiovascular disease than having no copies of a PLA2G12B variant nucleic acid molecule. Without intending to be limited to any particular theory or mechanism of action, it is believed that a single copy of a PLA2G12B variant nucleic acid molecule (i.e., heterozygous for a PLA2G12B variant nucleic acid molecule) is protective of a subject from developing a lipid disorder or a cardiovascular disease and it is also believed that having two copies of a PLA2G12B variant nucleic acid molecule (i.e., homozygous for a PLA2G12B variant nucleic acid molecule) may be more protective of a subject from developing a lipid disorder or a cardiovascular disease, relative to a subject with a single copy. Thus, in some embodiments, a single copy of a PLA2G12B variant nucleic acid molecule may not be completely protective, but instead, may be partially or incompletely protective of a subject from developing a lipid disorder or a cardiovascular disease. While not desiring to be bound by any particular theory, there may be additional factors or molecules involved in the development of a lipid disorder or a cardiovascular disease that are still present in a subject having a single copy of a PLA2G12B variant nucleic acid molecule, thus resulting in less than complete protection from the development of a lipid disorder or a cardiovascular disease.

[0156] Determining whether a subject has a PLA2G12B variant nucleic acid molecule in a biological sample from a subject and / or determining whether a subject has a PLA2G12B variant nucleic acid molecule can be carried out by any of the methods described herein. In some embodiments, these methods can be carried out in vitro. In some embodiments, these methods can be carried out in situ. In some embodiments, these methods can be carried out in vivo. In any of these embodiments, the nucleic acid molecule can be present within a cell obtained from the subject.

[0157] In some embodiments, when a subject is identified as having an increased risk of developing a lipid disorder or a cardiovascular disease, the subject is administered a lipid disorder therapeutic agent or a cardiovascular disease therapeutic agent and / or a PLA2G12B inhibitor, as described herein. For example, when the subject is PLA2G12B reference, and therefore has an increased risk of developing a lipid disorder or a cardiovascular disease, the subject can be administered a lipid disorder therapeutic agent or a cardiovascular disease therapeutic agent and / or administered a PLA2G12B inhibitor. In some embodiments, when the subject is heterozygous for a PLA2G12B variant nucleic acid molecule, the subject is administered the lipid disorder therapeutic agent or the cardiovascular disease therapeutic

[0158] 118452910DOCKET NO.: 38120-4555 (11318WO01)

[0159] - 33 -agent in an amount that is the same as or less than a standard dosage amount or a lipid disorder or a cardiovascular disease therapy, and / or is administered a PLA2G12B inhibitor. In some embodiments, when the subject is homozygous for a PLA2G12B variant nucleic acid molecule, the subject is administered a lipid disordertherapeutic agent or a cardiovascular disease therapeutic agent in a standard dosage amount. In some embodiments, the subject is PLA2G12B reference. In some embodiments, the subject is heterozygous for a PLA2G12B variant nucleic acid molecule. In some embodiments, the subject is homozygous for a PLA2G12B variant nucleic acid molecule.

[0160] The present disclosure also provides methods of determining a subject's aggregate burden, or risk score, of having two or more PLA2G12B variant nucleic acid molecules, and / or two or more PLA2G12B variant polypeptides associated with a decreased risk of developing a lipid disorder or a cardiovascular disease. The aggregate burden is the sum of two or more genetic variants that can be carried out in an association analysis with a lipid disorder or a cardiovascular disease. In some embodiments, the subject is homozygous for one or more PLA2G12B variant nucleic acid molecules associated with a decreased risk of developing a lipid disorder or a cardiovascular disease. In some embodiments, the subject is heterozygous for one or more PLA2G12B variant nucleic acid molecules associated with a decreased risk of developing a lipid disorder or a cardiovascular disease. When the subject has a lower aggregate burden, the subject has an increased risk of developing a lipid disorder or a cardiovascular disease and the subject is administered or continued to be administered the lipid disorder therapeutic agent orthe cardiovascular disease therapeutic agent in an amountthat is the same as or less than a standard dosage amount and / or a PLA2G12B inhibitor. When the subject has a greater aggregate burden, the subject has a decreased risk of developing a lipid disorder or a cardiovascular disease and the subject is administered or continued to be administered the lipid disorder therapeutic agent or the cardiovascular disease therapeutic agent in a standard dosage amount. The greater the aggregate burden, the lower the risk of developing a lipid disorder or a cardiovascular disease.

[0161] In some embodiments, a subject's aggregate burden of having any two or more PLA2G12B variant nucleic acid molecules represents a weighted sum of a plurality of any of the PLA2G12B variant nucleic acid molecules. In some embodiments, the aggregate burden is calculated using at least about 2, at least about 3, at least about 4, at least about 5, at least about 10, at least about 20, at least about 30, at least about 40, at least about 50, at least about

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[0163] - 34- 60, at least about 70, at least about 80, at least about 100, at least about 120, at least about 150, at least about 200, at least about 250, at least about 300, at least about 400, at least about 500, at least about 1,000, at least about 10,000, at least about 100,000, or at least about or more than 1,000,000 genetic variants present in or around (up to 10 Mb) the PLA2G12B gene, where the genetic burden is the number of alleles multiplied by the association estimate with a lipid disorder or a cardiovascular disease or related outcome for each allele (e.g., a weighted polygenic burden score). In some embodiments, when the subject has an aggregate burden above a desired threshold score, the subject has a decreased risk of developing a lipid disorder or a cardiovascular disease. In some embodiments, when the subject has an aggregate burden below a desired threshold score, the subject has an increased risk of developing a lipid disorder or a cardiovascular disease.

[0164] In some embodiments, the aggregate burden may be divided into quintiles, e.g., top quintile, second quintile, intermediate quintile, fourth quintile, and bottom quintile, wherein the top quintile of aggregate burden corresponds to the lowest risk group and the bottom quintile of aggregate burden corresponds to the highest risk group. In some embodiments, a subject having a greater aggregate burden comprises the highest weighted aggregate burdens, including, but not limited to the top 10%, top 20%, top 30%, top 40%, or top 50% of aggregate burdens from a subject population. In some embodiments, the genetic variants comprise the genetic variants having association with a lipid disorder or a cardiovascular disease in the top 10%, top 20%, top 30%, top 40%, or top 50% of p-value range for the association. In some embodiments, each of the identified genetic variants comprise the genetic variants having association with a lipid disorder or a cardiovascular disease with p-value of no more than about 10-2, about 10-3, about 10'4, about 10'5, about 10'6, about 10'7, about 10'8, about 10'9, about 10"10, about 10-11, about 1012, about 1013, about 1014, about or 1015. In some embodiments, the identified genetic variants comprise the genetic variants having association with a lipid disorder or a cardiovascular disease with p-value of less than 5 x 10'8. In some embodiments, the identified genetic variants comprise genetic variants having association with a lipid disorder or a cardiovascular disease in high-risk subjects as compared to the rest of the reference population with odds ratio (OR) about 1.5 or greater, about 1.75 or greater, about 2.0 or greater, or about 2.25 or greater forthe top 20% of the distribution; or about 1.5 or greater, about 1.75 or greater, about 2.0 or greater, about 2.25 or greater, about 2.5 or greater, or about 2.75 or greater. In some embodiments, the odds ratio (OR) may range from about 1.0 to

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[0166] - 35 -about 1.5, from about 1.5 to about 2.0, from about 2.0 to about 2.5, from about 2.5 to about 3.0, from about 3.0 to about 3.5, from about 3.5 to about 4.0, from about 4.0 to about 4.5, from about 4.5 to about 5.0, from about 5.0 to about 5.5, from about 5.5 to about 6.0, from about 6.0 to about 6.5, from about 6.5 to about 7.0, or greater than 7.0. In some embodiments, high-risk subjects comprise subjects having aggregate burdens in the bottom decile, quintile, or tertile in a reference population. The threshold of the aggregate burden is determined on the basis of the nature of the intended practical application and the risk difference that would be considered meaningful forthat practical application.

[0167] In embodiments where the aggregate burden is determined for PLA2G12B genetic variants associated with a lipid disorder or a cardiovascular disease, then the aggregate burden represents a subject's risk score for developing a lipid disorder or a cardiovascular disease. In some embodiments, the aggregate burden or risk score includes any of the PLA2G12B variant genomic nucleic acid molecules described herein. In some embodiments, a subject's aggregate burden can be determined for PLA2G12B genetic variants associated with a lipid disorder or a cardiovascular disease in combination with additional genetic variants for other genes also associated with a lipid disorder or a cardiovascular disease to produce a polygenic risk score (PRS) for developing a lipid disorder or a cardiovascular disease. In some embodiments, the PRS includes any of the PLA2G12B variant genomic nucleic acid molecules described herein.

[0168] The present disclosure also provides methods of detecting the presence or absence of a PLA2G12B variant nucleic acid molecule (i.e., a genomic nucleic acid molecule, an mRNA molecule, or a cDNA molecule produced from an mRNA molecule) in a biological sample from a subject. It is understood that gene sequences within a population and mRNA molecules encoded by such genes can vary due to polymorphisms such as single-nucleotide polymorphisms.

[0169] The biological sample can be derived from any cell, tissue, or biological fluid from the subject. The biological sample may comprise any clinically relevant tissue, such as a bone marrow sample, a tumor biopsy, a fine needle aspirate, or a sample of bodily fluid, such as blood, gingival crevicular fluid, plasma, serum, lymph, ascitic fluid, cystic fluid, or urine. In some cases, the sample comprises a buccal swab. The biological sample used in the methods disclosed herein can vary based on the assay format, nature of the detection method, and the tissues, cells, or extracts that are used as the sample. A biological sample can be processed differently depending on the assay being employed. For example, when detecting any

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[0171] - 36 - PLA2G12B variant nucleic acid molecule, preliminary processing designed to isolate or enrich the biological sample forthe genomic DNA can be employed. A variety of techniques may be used for this purpose. When detecting the level of any PLA2G12B variant nucleic acid molecule, different techniques can be used enrich the biological sample with mRNA molecules. Various methods to detect the presence or level of an mRNA molecule or the presence of a particular variant genomic DNA locus can be used.

[0172] In some embodiments, detecting a PLA2G12B variant nucleic acid molecule in a subject comprises performing a sequence analysis on a biological sample obtained from the subject to determine whether a PLA2G12B genomic nucleic acid molecule in the biological sample, and / or a PLA2G12B mRNA molecule in the biological sample, and / or a PLA2G12B cDNA molecule produced from an mRNA molecule in the biological sample, comprises one or more variations that cause a loss-of-fu notion (partial or complete) or are predicted to cause a loss-of-function (partial or complete). In some embodiments, the methods detect any of the PLA2G12B variant nucleic acid molecules described herein.

[0173] In some embodiments, the methods of detecting the presence or absence of a PLA2G12B variant nucleic acid molecule (such as, for example, a genomic nucleic acid molecule, an mRNA molecule, and / or a cDNA molecule produced from an mRNA molecule) in a subject, comprise performing an assay on a biological sample obtained from the subject. The assay determines whether a nucleic acid molecule in the biological sample comprises a particular nucleotide sequence.

[0174] In some embodiments, the biological sample comprises a cell or cell lysate. Such methods can further comprise, for example, obtaining a biological sample from the subject comprising a PLA2G12B genomic nucleic acid molecule or mRNA molecule, and if mRNA, optionally reverse transcribing the mRNA into cDNA. Such assays can comprise, for example determining the identity of these positions of the particular PLA2G12B nucleic acid molecule. In some embodiments, the method is an in vitro method.

[0175] In some embodiments, the determining step, detecting step, or sequence analysis comprises sequencing at least a portion of the nucleotide sequence of the PLA2G12B genomic nucleic acid molecule, the PLA2G12B mRNA molecule, orthe PLA2G12B cDNA molecule in the biological sample, wherein the sequenced portion comprises one or more variations that cause a loss-of-function (partial or complete) or are predicted to cause a loss-of-function (partial or complete).

[0176] 118452910DOCKET NO.: 38120-4555 (11318WO01)

[0177] - 37 - In some embodiments, the assay comprises sequencingthe entire nucleic acid molecule. In some embodiments, only a PLA2G12B genomic nucleic acid molecule is analyzed. In some embodiments, only a PLA2G12B mRNA is analyzed. In some embodiments, only a PLA2G12B cDNA obtained from the PLA2G12B mRNA is analyzed.

[0178] Alteration-specific polymerase chain reaction techniques can be used to detect mutations such as SNPs in a nucleic acid sequence. Alteration-specific primers can be used because the DNA polymerase will not extend when a mismatch with the template is present.

[0179] In some embodiments, the nucleic acid molecule in the sample is mRNA and the mRNA is reverse-transcribed into a cDNA prior to the amplifying step. In some embodiments, the nucleic acid molecule is present within a cell obtained from the subject.

[0180] In some embodiments, the assay comprises contacting the biological sample with a primer or probe, such as an alteration-specific primer or alteration-specific probe, that specifically hybridizes to a PLA2G12B variant genomic sequence, variant mRNA sequence, or variant cDNA sequence and not the corresponding PLA2G12B reference sequence under stringent conditions and determining whether hybridization has occurred.

[0181] In some embodiments, the determining step, detecting step, or sequence analysis comprises: a) amplifying at least a portion of the PLA2G12B nucleic acid molecule that encodes the PLA2G12B polypeptide; b) labeling the amplified nucleic acid molecule with a detectable label; c) contacting the labeled nucleic acid molecule with a support comprising an alterationspecific probe; and d) detecting the detectable label.

[0182] In some embodiments, the assay comprises RNA sequencing (RNA-Seq). In some embodiments, the assays also comprise reverse transcribing mRNA into cDNA, such as by the reverse transcriptase polymerase chain reaction (RT-PCR).

[0183] In some embodiments, the methods utilize probes and primers of sufficient nucleotide length to bind to the target nucleotide sequence and specifically detect and / or identify a polynucleotide comprising a PLA2G12B variant genomic nucleic acid molecule, variant mRNA molecule, or variant cDNA molecule. The hybridization conditions or reaction conditions can be determined by the operator to achieve this result. The nucleotide length may be any length that is sufficient for use in a detection method of choice, including any assay described or exemplified herein. Such probes and primers can hybridize specifically to a target nucleotide sequence under high stringency hybridization conditions. Probes and primers may have complete nucleotide sequence identity of contiguous nucleotides within the target nucleotide

[0184] 118452910DOCKET NO.: 38120-4555 (11318WO01)

[0185] - 38 -sequence, although probes differing from the target nucleotide sequence and that retain the ability to specifically detect and / or identify a target nucleotide sequence may be designed by conventional methods. Probes and primers can have about 80%, about 85%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, about 99%, or 100% sequence identity or complementarity with the nucleotide sequence of the target nucleic acid molecule.

[0186] Illustrative examples of nucleic acid sequencing techniques include, but are not limited to, chain terminator (Sanger) sequencing and dye terminator sequencing. Other methods involve nucleic acid hybridization methods other than sequencing, including using labeled primers or probes directed against purified DNA, amplified DNA, and fixed cell preparations (fluorescence in situ hybridization (FISH)). In some methods, a target nucleic acid molecule may be amplified prior to or simultaneous with detection. Illustrative examples of nucleic acid amplification techniques include, but are not limited to, polymerase chain reaction (PCR), ligase chain reaction (LCR), strand displacement amplification (SDA), and nucleic acid sequence based amplification (NASBA). Other methods include, but are not limited to, ligase chain reaction, strand displacement amplification, and thermophilic SDA (tSDA).

[0187] In hybridization techniques, stringent conditions can be employed such that a probe or primer will specifically hybridize to its target. In some embodiments, a polynucleotide primer or probe under stringent conditions will hybridize to its target sequence to a detectably greater degree than to other non-target sequences, such as, at least 2-fold, at least 3-fold, at least 4-fold, or more over background, including over 10-fold over background. In some embodiments, a polynucleotide primer or probe under stringent conditions will hybridize to its target nucleotide sequence to a detectably greater degree than to other nucleotide sequences by at least 2-fold. In some embodiments, a polynucleotide primer or probe under stringent conditions will hybridize to its target nucleotide sequence to a detectably greater degree than to other nucleotide sequences by at least 3-fold. In some embodiments, a polynucleotide primer or probe under stringent conditions will hybridize to its target nucleotide sequence to a detectably greater degree than to other nucleotide sequences by at least 4-fold. In some embodiments, a polynucleotide primer or probe under stringent conditions will hybridize to its target nucleotide sequence to a detectably greater degree than to other nucleotide sequences by over 10-fold over background. Stringent conditions are sequence-dependent and will be different in different circumstances.

[0188] 118452910DOCKET NO.: 38120-4555 (11318WO01)

[0189] - 39- Appropriate stringency conditions which promote DNA hybridization, for example, 6X sodium chloride / sodium citrate (SSC) at about 45°C., followed by a wash of 2X SSC at 50°C, are known or can be found in Current Protocols in Molecular Biology, John Wiley & Sons, N.Y. (1989), 6.3.1-6.3.6. Typically, stringent conditions for hybridization and detection will be those in which the salt concentration is less than about 1.5 M Na+ion, typically about 0.01 to 1.0 M Na+ion concentration (or other salts) at pH 7.0 to 8.3 and the temperature is at least about 30°C for short probes (such as, for example, 10 to 50 nucleotides) and at least about 60°C for longer probes (such as, for example, greater than 50 nucleotides). Stringent conditions may also be achieved with the addition of destabilizing agents such as formamide. Optionally, wash buffers may comprise about 0.1% to about 1% SDS. Duration of hybridization is generally less than about 24 hours, usually about 4 to about 12 hours. The duration of the wash time will be at least a length of time sufficient to reach equilibrium.

[0190] In some embodiments, such isolated nucleic acid molecules comprise or consist of at least about 5, at least about 8, at least about 10, at least about 11, at least about 12, at least about 13, at least about 14, at least about 15, at least about 16, at least about 17, at least about 18, at least about 19, at least about 20, at least about 21, at least about 22, at least about 23, at least about 24, at least about 25, at least about 30, at least about 35, at least about 40, at least about 45, at least about 50, at least about 55, at least about 60, at least about 65, at least about 70, at least about 75, at least about 80, at least about 85, at least about 90, at least about 95, at least about 100, at least about 200, at least about 300, at least about 400, at least about 500, at least about 600, at least about 700, at least about 800, at least about 900, at least about 1000, at least about 2000, at least about 3000, at least about 4000, or at least about 5000 nucleotides. In some embodiments, such isolated nucleic acid molecules comprise or consist of at least about 5, at least about 8, at least about 10, at least about 11, at least about 12, at least about 13, at least about 14, at least about 15, at least about 16, at least about 17, at least about 18, at least about 19, at least about 20, at least about 21, at least about 22, at least about 23, at least about 24, or at least about 25 nucleotides. In some embodiments, the isolated nucleic acid molecules comprise or consist of at least about 18 nucleotides. In some embodiments, the isolated nucleic acid molecules comprise or consists of at least about 15 nucleotides. In some embodiments, the isolated nucleic acid molecules consist of or comprise from about 10 to about 35, from about 10 to about 30, from about 10 to about 25, from about 12 to about 30, from about 12 to about 28, from about 12 to about 24, from about 15 to about 30, from about

[0191] 118452910DOCKET NO.: 38120-4555 (11318WO01)

[0192] - 40 - 15 to about 25, from about 18 to about 30, from about 18 to about 25, from about 18 to about 24, or from about 18 to about 22 nucleotides. In some embodiments, the isolated nucleic acid molecules consist of or comprise from about 18 to about 30 nucleotides. In some embodiments, the isolated nucleic acid molecules comprise or consist of at least about 15 nucleotides to at least about 35 nucleotides.

[0193] In some embodiments, such isolated nucleic acid molecules hybridize to PLA2G12B variant nucleic acid molecules (such as genomic nucleic acid molecules, mRNA molecules, and / or cDNA molecules) under stringent conditions. Such nucleic acid molecules can be used, for example, as probes, primers, alteration-specific probes, or alteration-specific primers as described or exemplified herein, and include, without limitation primers, probes, antisense RNAs, shRNAs, and siRNAs, each of which is described in more detail elsewhere herein and can be used in any of the methods described herein.

[0194] In some embodiments, the isolated nucleic acid molecules hybridize to at least about 15 contiguous nucleotides of a nucleic acid molecule that is at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or 100% identical to PLA2G12B variant nucleic acid molecules. In some embodiments, the isolated nucleic acid molecules consist of or comprise from about 15 to about 100 nucleotides, or from about 15 to about 35 nucleotides. In some embodiments, the isolated nucleic acid molecules consist of or comprise from about 15 to about 100 nucleotides. In some embodiments, the isolated nucleic acid molecules consist of or comprise from about 15 to about 35 nucleotides.

[0195] In some embodiments, the alteration-specific probes and alteration-specific primers comprise DNA. In some embodiments, the alteration-specific probes and alteration-specific primers comprise RNA.

[0196] In some embodiments, the probes and primers described herein (including alterationspecific probes and alteration-specific primers) have a nucleotide sequence that specifically hybridizes to any of the nucleic acid molecules disclosed herein, or the complement thereof. In some embodiments, the probes and primers specifically hybridize to any of the nucleic acid molecules disclosed herein under stringent conditions.

[0197] In some embodiments, the primers, including alteration-specific primers, can be used in second generation sequencing or high throughput sequencing. In some instances, the primers, including alteration-specific primers, can be modified. In particular, the primers can

[0198] 118452910DOCKET NO.: 38120-4555 (11318WO01)

[0199] - 41 -comprise various modifications that are used at different steps of, for example, Massive Parallel Signature Sequencing (MPSS), Polony sequencing, and 454 Pyrosequencing. Modified primers can be used at several steps of the process, including biotinylated primers in the cloning step and fluorescently labeled primers used at the bead loading step and detection step. Polony sequencing is generally performed using a paired-end tags library wherein each molecule of DNA template is about 135 bp in length. Biotinylated primers are used at the bead loading step and emulsion PCR. Fluorescently labeled degenerate nonamer oligonucleotides are used at the detection step. An adaptor can contain a 5'-biotin tag for immobilization of the DNA library onto streptavidin-coated beads.

[0200] The probes and primers described herein can be used to detect a nucleotide variation within any of the PLA2G12B variant nucleic acid molecules disclosed herein. The primers described herein can be used to amplify any PLA2G12B variant nucleic acid molecule, or a fragment thereof.

[0201] In the context of the disclosure "specifically hybridizes" means that the probe or primer (such as, for example, the alteration-specific probe or alteration-specific primer) does not hybridize to a nucleic acid sequence encoding a PLA2G12B reference genomic nucleic acid molecule, a PLA2G12B reference mRNA molecule, and / or a PLA2G12B reference cDNA molecule.

[0202] In some embodiments, the probes (such as, for example, an alteration-specific probe) comprise a label. In some embodiments, the label is a fluorescent label, a radiolabel, or biotin.

[0203] The present disclosure also provides supports comprising a substrate to which any one or more of the probes disclosed herein is attached. Solid supports are solid-state substrates or supports with which molecules, such as any of the probes disclosed herein, can be associated. A form of solid support is an array. Another form of solid support is an array detector. An array detector is a solid support to which multiple different probes have been coupled in an array, grid, or other organized pattern. A form for a solid-state substrate is a microtiter dish, such as a standard 96-well type. In some embodiments, a multiwell glass slide can be employed that normally contains one array per well.

[0204] The genomic nucleic acid molecules, mRNA molecules, and cDNA molecules can be from any organism. For example, the genomic nucleic acid molecules, mRNA molecules, and cDNA molecules can be human or an ortholog from another organism, such as a non-human

[0205] 118452910DOCKET NO.: 38120-4555 (11318WO01)

[0206] - 42 -mammal, a rodent, a mouse, or a rat. It is understood that gene sequences within a population can vary due to polymorphisms such as single-nucleotide polymorphisms.

[0207] Also provided herein are functional polynucleotides that can interact with the disclosed nucleic acid molecules. Examples of functional polynucleotides include, but are not limited to, antisense molecules, aptamers, ribozymes, triplex forming molecules, and external guide sequences. The functional polynucleotides can act as effectors, inhibitors, modulators, and stimulators of a specific activity possessed by a target molecule, or the functional polynucleotides can possess a de novo activity independent of any other molecules.

[0208] The isolated nucleic acid molecules disclosed herein can comprise RNA, DNA, or both RNA and DNA. The isolated nucleic acid molecules can also be linked or fused to a heterologous nucleic acid sequence, such as in a vector, or a heterologous label. For example, the isolated nucleic acid molecules disclosed herein can be within a vector or as an exogenous donor sequence comprising the isolated nucleic acid molecule and a heterologous nucleic acid sequence. The isolated nucleic acid molecules can also be linked or fused to a heterologous label. The label can be directly detectable (such as, for example, fluorophore) or indirectly detectable (such as, for example, hapten, enzyme, or fluorophore quencher). Such labels can be detectable by spectroscopic, photochemical, biochemical, immunochemical, or chemical means. Such labels include, for example, radiolabels, pigments, dyes, chromogens, spin labels, and fluorescent labels. The label can also be, for example, a chemiluminescent substance; a metal-containing substance; or an enzyme, where there occurs an enzyme-dependent secondary generation of signal. The term "label" can also refer to a "tag" or hapten that can bind selectively to a conjugated molecule such that the conjugated molecule, when added subsequently along with a substrate, is used to generate a detectable signal. For example, biotin can be used as a tag along with an avidin or streptavidin conjugate of horseradish peroxidate (HRP) to bind to the tag, and examined using a calorimetric substrate (such as, for example, tetramethylbenzidine (TMB)) or a fluorogenic substrate to detect the presence of HRP. Exemplary labels that can be used as tags to facilitate purification include, but are not limited to, myc, HA, FLAG or 3XFLAG, 6Xhis or polyhistidine, glutathione-S-transferase (GST), maltose binding protein, an epitope tag, or the Fc portion of immunoglobulin. Numerous labels include, for example, particles, fluorophores, haptens, enzymes and their calorimetric, fluorogenic and chemiluminescent substrates and other labels.

[0209] 118452910DOCKET NO.: 38120-4555 (11318WO01)

[0210] - 43 - Percent identity (or percent complementarity) between particular stretches of nucleotide sequences within nucleic acid molecules or amino acid sequences within polypeptides can be determined routinely using BLAST programs (basic local alignment search tools) and PowerBLAST programs (Altschul et al., J. Mol. Biol., 1990, 215, 403-410; Zhang and Madden, Genome Res., 1997, 7, 649-656) or by using the Gap program (Wisconsin Sequence Analysis Package, Version 8 for Unix, Genetics Computer Group, University Research Park, Madison Wis.), using default settings, which uses the algorithm of Smith and Waterman (Adv. Appl. Math., 1981, 2, 482-489). Herein, if reference is made to percent sequence identity, the higher percentages of sequence identity are preferred over the lower ones.

[0211] The present disclosure also provides lipid disorder therapeutic agents or cardiovascular disease therapeutic agents for use in the treatment or prevention of a lipid disorder or a cardiovascular disease in a subject having a PLA2G12B variant nucleic acid molecule. Any of the lipid disorder therapeutic agents or the cardiovascular disease therapeutic agents described herein can be used in these uses. Any of the PLA2G12B variant nucleic acid molecules disclosed herein can be used in these uses.

[0212] The present disclosure also provides uses of lipid disorder therapeutic agents or cardiovascular disease therapeutic agents for use in the preparation of a medicament for treating or preventing a lipid disorder or a cardiovascular disease in a subject having a PLA2G12B variant nucleic acid molecule. Any of the lipid disorder therapeutic agents orthe cardiovascular disease therapeutic agents described herein can be used in these uses. Any of the PLA2G12B variant nucleic acid molecules disclosed herein can be used in these uses.

[0213] The present disclosure also provides PLA2G12B inhibitors for use in the treatment or prevention of a lipid disorder or a cardiovascular disease in a subject that is PLA2G12B reference or is heterozygous for a PLA2G12B variant nucleic acid molecule. Any of the PLA2G12B inhibitors described herein can be used in these uses. Any of the PLA2G12B variant nucleic acid molecules disclosed herein can be used in these uses.

[0214] The present disclosure also provides PLA2G12B inhibitors in the preparation of a medicament for treating or preventing a lipid disorder or a cardiovascular disease in a subject that is PLA2G12B reference or is heterozygous for a PLA2G12B variant nucleic acid molecule. Any of the PLA2G12B inhibitors described herein can be used in these uses. Any of the PLA2G12B variant nucleic acid molecules disclosed herein can be used in these uses.

[0215] 118452910DOCKET NO.: 38120-4555 (11318WO01)

[0216] - 44- In some embodiments, the PLA2G12B inhibitor and the lipid disordertherapeutic agent or the cardiovascular disease therapeutic agent are disposed within a pharmaceutical composition. In some embodiments, the PLA2G12B inhibitor is disposed within a first pharmaceutical composition and the lipid disordertherapeutic agent or the cardiovascular disease therapeutic agent is disposed within a second pharmaceutical composition. In some embodiments, the first pharmaceutical composition and the second pharmaceutical composition are administered simultaneously. In some embodiments, the first pharmaceutical composition is administered before the second pharmaceutical composition. In some embodiments, the first pharmaceutical composition is administered after the second pharmaceutical composition.

[0217] In some embodiments, the PLA2G12B predicted loss-of-function variant nucleic acid molecule comprises a splice-site variant, a stop-gain variant, a start-loss variant, a stop-loss variant, a frameshift variant, an in-frame indel variant, and / or a variant that encodes a truncated PLA2G12B predicted loss-of-function polypeptide. In some embodiments, the PLA2G12B variant nucleic acid molecule is a PLA2G12B variant genomic nucleic acid molecule that comprises the genetic variation 10:72935723:AGG:A, 10:72941168:C:T, 10:72941227:A:T, 10:72941235:G:A, 10:72941336:T:C, 10:72942659:GGT:G, 10:72942688:G:T, 10:72942730:TG:T, 10:72954474:C:A, 10:72954480:CG:C, 10:72954481:G:A, 10:72954565:C:A, 10:72954604:C:A, 10:72954685:T:C, 10:72935708:AAC:A, 10:72941259:T:A, 10:72941302:G:T, 10:72942650:A:G, 10:72942742:T:A, 10:72954591:G:C, 10:72954601:C:A, or 10:72941311:C:CA, or is an mRNA molecule produced therefrom, or is a cDNA molecule produced from the mRNA molecule.

[0218] In some embodiments, the PLA2G12B variant nucleic acid molecule is a PLA2G12B variant genomic nucleic acid molecule that comprises the genetic variation 10:72935723:AGG:A, or is an mRNA molecule produced therefrom, or is a cDNA molecule produced from the mRNA molecule. In some embodiments, the PLA2G12B variant nucleic acid molecule is a PLA2G12B variant genomic nucleic acid molecule that comprises the genetic variation 10:72941168:C:T, or is an mRNA molecule produced therefrom, or is a cDNA molecule produced from the mRNA molecule. In some embodiments, the PLA2G12B variant nucleic acid molecule is a PLA2G12B variant genomic nucleic acid molecule that comprises the genetic variation 10:72941227:A:T, or is an mRNA molecule produced therefrom, or is a cDNA molecule produced from the mRNA molecule. In some embodiments, the PLA2G12B variant nucleic acid molecule is a PLA2G12B variant genomic nucleic acid molecule that comprises the genetic

[0219] 118452910DOCKET NO.: 38120-4555 (11318WO01)

[0220] - 45 -variation 10:72941235:G:A, or is an mRNA molecule produced therefrom, or is a cDNA molecule produced from the mRNA molecule. In some embodiments, the PLA2G12B variant nucleic acid molecule is a PLA2G12B variant genomic nucleic acid molecule that comprises the genetic variation 10:72941336:T:C, or is an mRNA molecule produced therefrom, or is a cDNA molecule produced from the mRNA molecule. In some embodiments, the PLA2G12B variant nucleic acid molecule is a PLA2G12B variant genomic nucleic acid molecule that comprises the genetic variation 10:72942659:GGT:G, or is an mRNA molecule produced therefrom, or is a cDNA molecule produced from the mRNA molecule. In some embodiments, the PLA2G12B variant nucleic acid molecule is a PLA2G12B variant genomic nucleic acid molecule that comprises the genetic variation 10:72942688:G:T, or is an mRNA molecule produced therefrom, or is a cDNA molecule produced from the mRNA molecule. In some embodiments, the PLA2G12B variant nucleic acid molecule is a PLA2G12B variant genomic nucleic acid molecule that comprises the genetic variation 10:72942730:TG:T, or is an mRNA molecule produced therefrom, or is a cDNA molecule produced from the mRNA molecule. In some embodiments, the PLA2G12B variant nucleic acid molecule is a PLA2G12B variant genomic nucleic acid molecule that comprises the genetic variation 10:72954474:C:A, or is an mRNA molecule produced therefrom, or is a cDNA molecule produced from the mRNA molecule. In some embodiments, the PLA2G12B variant nucleic acid molecule is a PLA2G12B variant genomic nucleic acid molecule that comprises the genetic variation 10:72954480:CG:C, or is an mRNA molecule produced therefrom, or is a cDNA molecule produced from the mRNA molecule. In some embodiments, the PLA2G12B variant nucleic acid molecule is a PLA2G12B variant genomic nucleic acid molecule that comprises the genetic variation 10:72954481:G:A, or is an mRNA molecule produced therefrom, or is a cDNA molecule produced from the mRNA molecule. In some embodiments, the PLA2G12B variant nucleic acid molecule is a PLA2G12B variant genomic nucleic acid molecule that comprises the genetic variation 10:72954565:C:A, or is an mRNA molecule produced therefrom, or is a cDNA molecule produced from the mRNA molecule. In some embodiments, the PLA2G12B variant nucleic acid molecule is a PLA2G12B variant genomic nucleic acid molecule that comprises the genetic variation 10:72954604:C:A, or is an mRNA molecule produced therefrom, or is a cDNA molecule produced from the mRNA molecule. In some embodiments, the PLA2G12B variant nucleic acid molecule is a PLA2G12B variant genomic nucleic acid molecule that comprises the genetic variation 10:72954685:T:C, or is an mRNA molecule produced therefrom, or is a cDNA molecule produced from the mRNA molecule. In some embodiments, the PLA2G12B variant

[0221] 118452910DOCKET NO.: 38120-4555 (11318WO01)

[0222] - 46 -nucleic acid molecule is a PLA2G12B variant genomic nucleic acid molecule that comprises the genetic variation 10:72935708:AAC:A, or is an mRNA molecule produced therefrom, or is a cDNA molecule produced from the mRNA molecule. In some embodiments, the PLA2G12B variant nucleic acid molecule is a PLA2G12B variant genomic nucleic acid molecule that comprises the genetic variation 10:72941259:T:A, or is an mRNA molecule produced therefrom, or is a cDNA molecule produced from the mRNA molecule. In some embodiments, the PLA2G12B variant nucleic acid molecule is a PLA2G12B variant genomic nucleic acid molecule that comprises the genetic variation 10:72941302:G:T, or is an mRNA molecule produced therefrom, or is a cDNA molecule produced from the mRNA molecule. In some embodiments, the PLA2G12B variant nucleic acid molecule is a PLA2G12B variant genomic nucleic acid molecule that comprises the genetic variation 10:72942650:A:G, or is an mRNA molecule produced therefrom, or is a cDNA molecule produced from the mRNA molecule. In some embodiments, the PLA2G12B variant nucleic acid molecule is a PLA2G12B variant genomic nucleic acid moleculethat comprises the genetic variation 10:72942742:T:A, or is an mRNA molecule produced therefrom, or is a cDNA molecule produced from the mRNA molecule. In some embodiments, the PLA2G12B variant nucleic acid molecule is a PLA2G12B variant genomic nucleic acid molecule that comprises the genetic variation 10:72954591:G:C, or is an mRNA molecule produced therefrom, or is a cDNA molecule produced from the mRNA molecule. In some embodiments, the PLA2G12B variant nucleic acid molecule is a PLA2G12B variant genomic nucleic acid molecule that comprises the genetic variation 10:72954601:C:A, or is an mRNA molecule produced therefrom, or is a cDNA molecule produced from the mRNA molecule. In some embodiments, the PLA2G12B variant nucleic acid molecule is a PLA2G12B variant genomic nucleic acid molecule that comprises the genetic variation 10:72941311:C:CA, or is an mRNA molecule produced therefrom, or is a cDNA molecule produced from the mRNA molecule.

[0223] All patent documents, websites, other publications, accession numbers and the like cited above or below are incorporated by reference in their entirety for all purposes to the same extent as if each individual item were specifically and individually indicated to be so incorporated by reference. If different versions of a sequence are associated with an accession number at different times, the version associated with the accession number at the effective filing date of this application is meant. The effective filing date means the earlier of the actual filing date or filing date of a priority application referring to the accession number if applicable.

[0224] 118452910DOCKET NO.: 38120-4555 (11318WO01)

[0225] - 47 - Likewise, if different versions of a publication, website or the like are published at different times, the version most recently published at the effective filing date of the application is meant unless otherwise indicated. Any feature, step, element, embodiment, or aspect of the present disclosure can be used in combination with any other feature, step, element, embodiment, or aspect unless specifically indicated otherwise. Although the present disclosure has been described in some detail by way of illustration and example for purposes of clarity and understanding, it will be apparent that certain changes and modifications may be practiced within the scope of the appended claims.

[0226] The following examples are provided to describe the embodiments in greater detail. They are intended to illustrate, not to limit, the claimed embodiments. The following examples provide those of ordinary skill in the art with a disclosure and description of how the compounds, compositions, articles, devices and / or methods described herein are made and evaluated and are intended to be purely exemplary and are not intended to limit the scope of any claims. Efforts have been made to ensure accuracy with respect to numbers (such as, for example, amounts, temperature, etc.), but some errors and deviations may be accounted for. Unless indicated otherwise, parts are parts by weight, temperature is in °C or is at ambient temperature, and pressure is at or near atmospheric.

[0227] Examples

[0228] Example 1: General Methods

[0229] Participating cohorts

[0230] Genetic association studies were performed in individuals of African or Admixed African, Admixed American, European, and East and South Asian ancestry from the United Kingdom Biobank (UKB) cohort (Bycroft et al., Nature, 2018, 562, 203-209; and Van Hout et al., Nature, 2020, 586, 749-756), the MyCode Community Health Initiative cohort from the Geisinger Health System (GHS) (Carey et al., Genet Med., 2016, 18, 906-13), the University of Pennsylvania Medicine BioBank (PMBB), the Malmo Diet and Cancer Study (MDCS) (Berglund et al., J. Int. Med., 1993, 233, 45-51) and Mount Sinai's BioMe Personalized Medicine Cohort (SINAI) (Gottesman et al., Genet. Med., 2013, 15, 761-771).

[0231] The UKB is a population-based cohort study of people aged between 40 and 69 years recruited through 22 testing centers in the UK between 2006-2010. Exome sequencing and phenotype data were available for 463,300 individuals. The GHS MyCode study is a health

[0232] 118452910DOCKET NO.: 38120-4555 (11318WO01)

[0233] - 48 -system-based cohort of patients from Central and Eastern Pennsylvania (USA) recruited in 2007-2019. A total of 132,812 participants from GHS with available whole-exome sequencing and phenotype data were included. Mount Sinai's BioMe Personalized Medicine Cohort (SINAI) is an electronic health record-linked clinical care cohort, of which 21,591 individuals had available phenotypes and exome sequencing data and were included in analysis. The Malmo Diet and Cancer Study (MDCS) included 5,116 individuals with available phenotype and exome sequencing data. The University of Pennsylvania Medicine BioBank (PMBB) included a total of 24,834 individuals with available phenotype and exome sequencing data.

[0234] Phenotype definitions

[0235] Lipid levels were measured during a visit to the assessment center for UK Biobank participants or were extracted from electronic health record datasets for participants from other contributing cohorts. Direct LDL measurements were available for UK biobank participants. LDL levels were estimated using the Friedewald equation for the other participating individuals. Individuals known to be on lipid lowering medication had their pretreatment LDL and Apolipoprotein B levels estimated using a correction factor for individuals known to be on lipid lowering medication.

[0236] Example 2: Loss of Function of the Gene Encoding PLA2G12B is Associated with Lower Levels of LDL Cholesterol

[0237] To identify genetic factors influencing levels of LDL cholesterol, a major cause of cardiovascular disease, imputed genotype and exome sequencing data were analyzed for association with lipid levels in up to 626,062 participants from the Geisinger Heath System MyCode Community Health Initiative study (GHS), Malmo Diet and Cancer Study (MDCS), Mount Sinai's BioMe personalized Medicine Cohort (SINAI), the UK Biobank (UKB), and the University of Pennsylvania Penn Medicine Biobank (UPENN-PMBB). Associations with lower LDL and HDL cholesterol and lower apolipoprotein B levels were identified among individuals with rare predicted loss of function (pLOF) variants in PLA2G12B (see, Table 1). A total of 25 predicted loss of function variants contributed to the associations (see, Table 2).

[0238] 118452910DOCKET NO.: 38120-4555 (11318WO01)

[0239] - 49- Table 1: Rare predicted loss of function variants in PLA2G12B are associated with lower levels of LDL cholesterol

[0240]

[0241] Effect sizes are given in standard deviation (SD) units. RR: count of homozygous individuals for the reference allele, RA: count of heterozygous individuals carrying one pLOF variant, AA: count of homozygous individuals carrying pLOF variants in both PLA2G12B alleles.

[0242] Table 2: Predicted loss of function variants in PLA2G12B identified by whole exome sequencing

[0243] > > >

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[0245]

[0246] 118452910DOCKET NO.: 38120-4555 (11318WO01)

[0247] - 50 -

[0248] > > > > >

[0249] > > > > >

[0250]

[0251] Various modifications of the described subject matter, in addition to those described herein, will be apparent to those skilled in the art from the foregoing description. Such modifications are also intended to fall within the scope of the appended claims. Each reference (including, but not limited to, journal articles, U.S. and non-U. S. patents, patent application publications, international patent application publications, gene bank accession numbers, and the like) cited in the present application is incorporated herein by reference in its entirety and for all purposes.

[0252] 118452910

Claims

DOCKET NO.: 38120-4555 (11318WO01)- 51 - What is Claimed is:

1. A method of treating a subject having a lipid disorder or a cardiovascular disease, or at risk of developing a lipid disorder or a cardiovascular disease, the method comprising administering a Phospholipase A2 Group XI IB (PLA2G12B) inhibitor to the subject.

2. The method of claim 1, wherein the lipid disorder is increased low-density lipoprotein (LDL) and / or increased total cholesterol.

3. The method of claim 1 or claim 2, wherein the PLA2G12B inhibitor comprises an inhibitory nucleic acid molecule that hybridizes to a PLA2G12B nucleic acid molecule.

4. The method of claim 3, wherein the inhibitory nucleic acid molecule comprises an antisense nucleic acid molecule, a small interfering RNA (siRNA), and / or a short hairpin RNA (shRNA).

5. The method of claim 4, wherein the inhibitory nucleic acid molecule comprises an siRNA.

6. The method of claim 4, wherein the inhibitory nucleic acid molecule comprises an antisense nucleic acid molecule.

7. The method of any one of claims 1 to 6, wherein the subject is also administered a lipid disorder therapeutic agent or a cardiovascular disease therapeutic agent.

8. The method of claim 7, wherein the PLA2G12B inhibitor and the lipid disorder therapeutic agent or the cardiovascular disease therapeutic agent are disposed within the same pharmaceutical composition.

9. The method of claim 7, wherein the PLA2G12B inhibitor is disposed within a first pharmaceutical composition and the lipid disorder therapeutic agent or the cardiovascular disease therapeutic agent is disposed within a second pharmaceutical composition.

10. The method of claim 9, wherein the first pharmaceutical composition and the second pharmaceutical composition are administered simultaneously.

11. The method of claim 9, wherein the first pharmaceutical composition is administered before the second pharmaceutical composition.

12. The method of claim 9, wherein the first pharmaceutical composition is administered after the second pharmaceutical composition.

13. The method of any one of claims 1 to 12, further comprising detecting the presence or absence of a PLA2G12B variant nucleic acid molecule in a biological sample from the subject.118452910DOCKET NO.: 38120-4555 (11318WO01)- 52 - 14. The method of claim 13, wherein the PLA2G12B variant nucleic acid molecule comprises a splice-site variant, a stop-gain variant, a start-loss variant, a stop-loss variant, a frameshift variant, a missense variant, an in-frame indel variant, and / or a variant that encodes a truncated PLA2G12B variant polypeptide.

15. The method of claim 13 or claim 14, wherein the PLA2G12B variant nucleic acid molecule comprises the genetic variation 10:72935723:AGG:A, 10:72941168:C:T, 10:72941227:A:T, 10:72941235:G:A, 10:72941336:T:C, 10:72942659:GGT:G, 10:72942688:G:T, 10:72942730:TG:T, 10:72954474:C:A, 10:72954480:CG:C, 10:72954481 :G: A, 10:72954565:C:A, 10:72954604:C:A, 10:72954685:T:C, 10:72935708:AAC:A, 10:72941259:T:A, 10:72941302:G:T, 10:72942650:A:G, 10:72942742:T:A, 10:72954591:G:C, 10:72954601:C:A, or 10:72941311:C:CA.

16. A method of treating a subject having a lipid disorder or a cardiovascular disease or at risk of developing a lipid disorder or a cardiovascular disease by administering a lipid disorder therapeutic agent or a cardiovascular disease therapeutic agent, the method comprising:determining or having determined whether the subject has a Phospholipase A2 Group XIIB (PLA2G12B) variant nucleic acid molecule, by:obtaining or having obtained a biological sample from the subject; andperforming or having performed a sequence analysis on the biological sample to determine if the subject has a genotype comprising a PLA2G12B variant nucleic acid molecule; andadministering or continuing to administer the lipid disorder therapeutic agent or the cardiovascular disease therapeutic agent in an amount that is the same as or less than a standard dosage amount and / or administering a PLA2G12B inhibitorto a subject that is PLA2G12B reference;administering or continuing to administerthe lipid disordertherapeutic agent orthe cardiovascular disease therapeutic agent in an amount that is the same as or less than a standard dosage amount and / or administering a PLA2G12B inhibitorto a subject that is heterozygous for the PLA2G12B variant nucleic acid molecule; oradministering or continuing to administerthe lipid disordertherapeutic agent orthe cardiovascular disease therapeutic agent in a standard dosage amount to a subject that is homozygous for the PLA2G12B variant nucleic acid molecule;118452910DOCKET NO.: 38120-4555 (11318WO01)- 53 - wherein the presence of a genotype having the PLA2G12B variant nucleic acid molecule indicates the subject has a decreased risk of developing a lipid disorder or a cardiovascular disease.

17. The method of claim 16, wherein the lipid disorder is increased low-density lipoprotein (LDL) and / or increased total cholesterol.

18. The method of claim 16 or claim 17, wherein the PLA2G12B inhibitor comprises an inhibitory nucleic acid molecule that hybridizes to a PLA2G12B nucleic acid molecule.

19. The method of claim 18, wherein the inhibitory nucleic acid molecule comprises an antisense nucleic acid molecule, a small interfering RNA (siRNA), and / or a short hairpin RNA (shRNA).

20. The method of claim 19, wherein the inhibitory nucleic acid molecule comprises an siRNA.

21. The method of claim 19, wherein the inhibitory nucleic acid molecule comprises an antisense nucleic acid molecule.

22. The method of any one of claims 16 to 21, wherein the PLA2G12B inhibitor and the lipid disorder therapeutic agent or the cardiovascular disease therapeutic agent are disposed within the same pharmaceutical composition.

23. The method of any one of claims 16 to 21, wherein the PLA2G12B inhibitor is disposed within a first pharmaceutical composition and the lipid disorder therapeutic agent or the cardiovascular disease therapeutic agent is disposed within a second pharmaceutical composition.

24. The method of claim 23, wherein the first pharmaceutical composition and the second pharmaceutical composition are administered simultaneously.

25. The method of claim 23, wherein the first pharmaceutical composition is administered before the second pharmaceutical composition.

26. The method of claim 23, wherein the first pharmaceutical composition is administered after the second pharmaceutical composition.

27. The method of any one of claims 16 to 26, wherein the subject is heterozygous forthe PLA2G12B variant nucleic acid molecule, and the subject is administered or continued to be administered the lipid disorder therapeutic agent or the cardiovascular disease therapeutic agent in an amount that is the same as or less than a standard dosage amount and a PLA2G12B inhibitor.118452910DOCKET NO.: 38120-4555 (11318WO01)- 54- 28. The method of any one of claims 16 to 27, wherein the PLA2G12B variant nucleic acid molecule comprises a splice-site variant, a stop-gain variant, a start-loss variant, a stop-loss variant, a frameshift variant, a missense variant, an in-frame indel variant, and / or a variant that encodes a truncated PLA2G12B variant polypeptide.

29. The method of any one of claims 16 to 27, wherein the PLA2G12B variant nucleic acid molecule comprises the genetic variation 10:72935723:AGG:A, 10:72941168:C:T, 10:72941227:A:T, 10:72941235:G:A, 10:72941336:T:C, 10:72942659:GGT:G, 10:72942688:G:T, 10:72942730:TG:T, 10:72954474:C:A, 10:72954480:CG:C, 10:72954481 :G: A, 10:72954565:C:A, 10:72954604:C:A, 10:72954685:T:C, 10:72935708:AAC:A, 10:72941259:T:A, 10:72941302:G:T, 10:72942650:A:G, 10:72942742:T:A, 10:72954591:G:C, 10:72954601:C:A, or 10:72941311:C:CA.

30. A method of identifying a subject having an increased risk of developing a lipid disorder or a cardiovascular disease, the method comprising:determining or having determined the presence or absence of a PLA2G12B variant nucleic acid molecule in a biological sample obtained from the subject;wherein:when the subject is PLA2G12B reference, then the subject has an increased risk of developing a lipid disorder or a cardiovascular disease; and when the subject is heterozygous or homozygous for the PLA2G12B variant nucleic acid molecule, then the subject has a decreased risk of developing a lipid disorder or a cardiovascular disease.

31. The method of claim 30, wherein the PLA2G12B variant nucleic acid molecule is a splice-site variant, a stop-gain variant, a start-loss variant, a stop-loss variant, a frameshift variant, a missense variant, an in-frame indel variant, or a variant that encodes a truncated PLA2G12B variant polypeptide.

32. The method of claim 30 or claim 31, wherein the PLA2G12B variant nucleic acid molecule comprises the genetic variation 10:72935723:AGG:A, 10:72941168:C:T, 10:72941227:A:T, 10:72941235:G:A, 10:72941336:T:C, 10:72942659:GGT:G, 10:72942688:G:T, 10:72942730:TG:T, 10:72954474:C:A, 10:72954480:CG:C, 10:72954481 :G: A, 10:72954565:C:A, 10:72954604:C:A, 10:72954685:T:C, 10:72935708:AAC:A, 10:72941259:T:A, 10:72941302:G:T, 10:72942650:A:G, 10:72942742:T:A, 10:72954591:G:C, 10:72954601:C:A, or 10:72941311:C:CA.118452910DOCKET NO.: 38120-4555 (11318WO01)- 55 - 33. The method of any one of claims 30 to 32, further comprising administering a lipid disorder therapeutic agent or a cardiovascular disease therapeutic agent in a standard dosage amount and / or administering a PLA2G12B inhibitor to a subject that is PLA2G12B reference.

34. The method of any one of claims 30 to 32, further comprising administering a lipid disorder therapeutic agent or a cardiovascular disease therapeutic agent in an amount that is the same as or less than a standard dosage amount and / or administering a PLA2G12B inhibitor to a subject that is heterozygous for a PLA2G12B variant nucleic acid molecule.

35. The method of claim 33 or claim 34, wherein the PLA2G12B inhibitor comprises an inhibitory nucleic acid molecule that hybridizes to a PLA2G12B nucleic acid molecule.

36. The method of claim 35, wherein the inhibitory nucleic acid molecule comprises an antisense nucleic acid molecule, a small interfering RNA (siRNA), and / or a short hairpin RNA (shRNA).

37. The method of claim 36, wherein the inhibitory nucleic acid molecule comprises an siRNA.

38. The method of claim 36, wherein the inhibitory nucleic acid molecule comprises an antisense nucleic acid molecule.

39. The method of any one of claims 30 to 38, wherein the PLA2G12B inhibitor and the lipid disorder therapeutic agent or the cardiovascular disease therapeutic agent are disposed within the same pharmaceutical composition.

40. The method of any one of claims 30 to 38, wherein the PLA2G12B inhibitor is disposed within a first pharmaceutical composition and the lipid disorder therapeutic agent or the cardiovascular disease therapeutic agent is disposed within a second pharmaceutical composition.

41. The method of claim 40, wherein the first pharmaceutical composition and the second pharmaceutical composition are administered simultaneously.

42. The method of claim 40, wherein the first pharmaceutical composition is administered before the second pharmaceutical composition.

43. The method of claim 40, wherein the first pharmaceutical composition is administered after the second pharmaceutical composition.

44. The method of any one of claims 30 to 43, wherein the subject is PLA2G12B reference, and the subject is administered or continued to be administered the lipid disorder therapeutic118452910DOCKET NO.: 38120-4555 (11318WO01)- 56 -agent or the cardiovascular disease therapeutic agent in an amount that is the same as or less than a standard dosage amount, and the subject is administered the PLA2G12B inhibitor.

45. The method of any one of claims 30 to 44, wherein the subject is heterozygous forthe PLA2G12B variant nucleic acid molecule, and the subject is administered or continued to be administered the lipid disorder therapeutic agent or the cardiovascular disease therapeutic agent in an amount that is the same as or less than a standard dosage amount, and the subject is administered the PLA2G12B inhibitor.

46. A lipid disorder therapeutic agent or a cardiovascular disease therapeutic agent for use in the treatment or prevention of a lipid disorder or a cardiovascular disease in a subject having a PLA2G12B variant nucleic acid molecule.

47. The lipid disorder therapeutic agent or the cardiovascular disease therapeutic agent of claim 46, wherein the PLA2G12B variant nucleic acid molecule is a splice-site variant, a stopgain variant, a start-loss variant, a stop-loss variant, a frameshift variant, a missense variant, an in-frame indel variant, or a variant that encodes a truncated PLA2G12B variant polypeptide.

48. The lipid disorder therapeutic agent or the cardiovascular disease therapeutic agent of claim 46 or claim 47, wherein the PLA2G12B variant nucleic acid molecule comprises the genetic variation in 10:72935723:AGG:A, 10:72941168:C:T, 10:72941227:A:T, 10:72941235:G:A, 10:72941336:T:C, 10:72942659:GGT:G, 10:72942688:G:T, 10:72942730:TG:T, 10:72954474:C:A, 10:72954480:CG:C, 10:72954481:G:A, 10:72954565:C:A, 10:72954604:C:A, 10:72954685:T:C, 10:72935708:AAC:A, 10:72941259:T:A, 10:72941302:G:T, 10:72942650:A:G, 10:72942742:T:A, 10:72954591:G:C, 10:72954601:C:A, or 10:72941311:C:CA.

49. A PLA2G12B inhibitor for use in the treatment or prevention of a lipid disorder or a cardiovascular disease in a subject that is PLA2G12B reference or is heterozygous for a PLA2G12B variant nucleic acid molecule.

50. The PLA2G12B inhibitor of claim 49, wherein the PLA2G12B variant nucleic acid molecule is a splice-site variant, a stop-gain variant, a start-loss variant, a stop-loss variant, a frameshift variant, a missense variant, an in-frame indel variant, or a variant that encodes a truncated PLA2G12B variant polypeptide.

51. The PLA2G12B inhibitor of claim 49 or claim 50, wherein the PLA2G12B variant nucleic acid molecule comprises the genetic variation 10:72935723:AGG:A, 10:72941168:C:T, 10:72941227:A:T, 10:72941235:G:A, 10:72941336:T:C, 10:72942659:GGT:G, 10:72942688:G:T, 10:72942730:TG:T, 10:72954474:C:A, 10:72954480:CG:C, 10:72954481 :G: A, 10:72954565:C:A,118452910DOCKET NO.: 38120-4555 (11318WO01)- 57 - 10:72954604:C:A, 10:72954685:T:C, 10:72935708:AAC:A, 10:72941259:T:A, 10:72941302:G:T, 10:72942650:A:G, 10:72942742:T:A, 10:72954591:G:C, 10:72954601:C:A, or 10:72941311:C:CA.

52. The PLA2G12B inhibitor of any one of claims 49 to 51, wherein the PLA2G12B inhibitor comprises an inhibitory nucleic acid molecule that hybridizesto a PLA2G12B nucleic acid molecule.

53. The PLA2G12B inhibitor of claim 52, wherein the inhibitory nucleic acid molecule comprises an antisense nucleic acid molecule, a small interfering RNA (siRNA), and / or a short hairpin RNA (shRNA).

54. The PLA2G12B inhibitor of claim 53, wherein the inhibitory nucleic acid molecule comprises an siRNA.

55. The PLA2G12B inhibitor of claim 53, wherein the inhibitory nucleic acid molecule comprises an antisense nucleic acid molecule.118452910

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