Compositions and methods for regulating cardiac angiogenesis post ischemia

Abstract

Described herein are compositions (e.g., an inhibitory nucleic acid molecule) for reducing expression of decorin (DCN) and methods thereof for (i) treating a medical condition resulting from a myocardial infarction (Ml), and / or (ii) promoting angiogenesis in a subject. The inhibitory nucleic acid molecule may be a small interfering RNA (siRNA), a double-stranded RNA (dsRNA), an anti-sense oligonucleotide (ASO), a microRNA (miRNA), or a short hairpin RNA (shRNA)), or a gapmeR described herein, or a composition (e.g., pharmaceutical composition) thereof. Advantageously, the composition described herein provides therapeutic effects (e.g., angiogenesis) for cardiac tissue following myocardial infarction (Ml).

Description

[0001] COMPOSITIONS AND METHODS FOR REGULATING CARDIAC ANGIOGENESIS POST ISCHEMIA

[0002] CROSS-REFERENCE TO RELATED APPLICATIONS

[0003] This application claims benefit of U.S. Provisional Application No. 63 / 626,184, filed on January 29, 2024, which is incorporated herein by reference in its entirety for any purpose.

[0004] SEQUENCE LISTING

[0005] The instant application contains a Sequence Listing which has been submitted electronically in XML format and is hereby incorporated by reference in its entirety. Said XML copy, created on January 27, 2025, is named “51665-004WO2_Sequence_Listing_1_27_25.xml” and is 26,689 bytes in size.

[0006] TECHNICAL FIELD

[0007] This disclosure relates to inhibitory nucleic acid molecules useful for targeting Decorin (DCN), and methods of using such inhibitory nucleic acid molecules for (i) treating medical complications resulting from a myocardial infarction (Ml), and / or (ii) promoting angiogenesis in a subject after the subject has suffered an Ml.

[0008] BACKGROUND

[0009] Over 800,000 people experience a myocardial infarction (Ml) or heart attack annually in the US. Ml occurs when occlusion of a coronary artery prevents blood from perfusing the myocardium, creating an infarct. Angiogenesis around the infarct is crucial to limiting damage and recovery of cardiac function. Patients with diabetes are more likely to suffer from myocardial injury compared to those without diabetes. For patients with diabetes, longevity and quality of life suffer considerably post-MI, in part due to impaired angiogenesis and tissue repair. Moreover, elevated blood glucose level (hyperglycemia) can alter endothelial cell (EC) metabolism, causing endothelial dysfunction. Chronic exposure to high glucose levels has been shown to induce oxidative stress in vascular ECs via mitochondrial reactive oxygen species (ROS) production, which makes them more susceptible to apoptosis. These metabolic perturbations can negatively impact normal cell signaling pathways during angiogenesis. There remains an unmet need for therapeutics that can rescue post-ischemic angiogenesis in these patients. The role of microRNAs (miRNAs) in angiogenesis is relatively unexplored.

[0010] SUMMARY OF THE INVENTION

[0011] In a first aspect, the invention provides an inhibitory nucleic acid molecule comprising sufficient complementarity to a target nucleic acid molecule, wherein (i) the inhibitory nucleic acid molecule is at least 15 nucleotides in length, and (ii) the target nucleic acid molecule comprises a nucleotide sequence having at least 80% sequence identity to SEQ ID NO: 14.

[0012] In some embodiments, the target nucleic acid molecule comprises a nucleotide sequence having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to SEQ ID NO: 14.

[0013] In some embodiments, the target nucleic acid molecule comprises the nucleotide sequence of

[0014] SEQ ID NO: 14, or a splice variant thereof. In some embodiments, the inhibitory nucleic acid molecule is 15 to 6,850 nucleotides in length (e.g., 50 to 6,850 nucleotides in length, 100 to 6,850 nucleotides in length, 250 to 6,850 nucleotides in length, 500 to 6,850 nucleotides in length, 750 to 6,850 nucleotides in length, 1 ,000 to 6,850 nucleotides in length, 1 ,500 to 6,850 2,000 to 6,850, 4,000 to 6,850, 5,000 to 6,850, or 6,000 to 6,850 nucleotides in length).

[0015] In some embodiments, the inhibitory nucleic acid molecule is 15 to 49 nucleotides in length (e.g., 15, 16, 17, 18, 19, 20, 21 , 22, 23, 24, 25, 26, 27, 28, 29, 30, 31 , 32, 33, 34, 35, 36, 37, 38, 39, 40, 41 , 42,

[0016] 43, 44, 45, 46, 47, 48, or 49 nucleotides in length), 50 to 99 nucleotides in length (e.g., 50, 51 , 52, 53, 54,

[0017] 55, 56, 57, 58, 59, 60, 61 , 62, 63, 64, 65, 66, 67, 68, 69, 70, 71 , 72, 73, 74, 75, 76, 77, 78, 79, 80, 81 , 82,

[0018] 83, 84, 85, 86, 87, 88, 89, 90, 91 , 92, 93, 94, 95, 96, 97, 98, or 99 nucleotides in length), or 100 to 6,850 nucleotides in length (e.g., 100, 125, 150, 175, 200, 225, 250, 275, 300, 325, 350, 375, 400, 425, 450, 475, 500, 525, 550, 575, 600, 625, 650, 675, 700, 725, 750, 775, 800, 825, 850, 875, 900, 925, 950, 975, 1000, 1 ,025, 1 ,050, 1 ,075, 1 ,100, 1 ,125, 1 ,150, 1 ,175, 1 ,200, 1 ,225, 1 ,250, 1 ,275, 1 ,300, 1 ,325, 1 ,350, 1 ,375, 1 ,400, 1 ,425, 1 ,450, 1 ,475, 1 ,500, 1 ,525, 1 ,550, 1 ,575, 1 ,600, 1 ,625, 1 ,650, 1 ,675, 1 ,700, 1 ,725, 1 ,750, 1 ,775, 1 ,800, 1 ,900, 2,000, 2,500, 3,000, 3,500, 4,000, 4,500, 5,000, 5,500, 6,000, or 6,500 nucleotides in length).

[0019] In some embodiments, the inhibitory nucleic acid molecule is 20 to 28 nucleotides in length (e.g., 20, 21 , 22, 23, 24, 25, 26, 27, or 28 nucleotides in length).

[0020] In some embodiments, the inhibitory nucleic acid molecule is 18 nucleotides in length. In some embodiments, the inhibitory nucleic acid molecule is 19 nucleotides in length. In some embodiments, the inhibitory nucleic acid molecule is 20 nucleotides in length. In some embodiments, the inhibitory nucleic acid molecule is 21 nucleotides in length. In some embodiments, the inhibitory nucleic acid molecule is 22 nucleotides in length. In some embodiments, the inhibitory nucleic acid molecule is 23 nucleotides in length. In some embodiments, the inhibitory nucleic acid molecule is 24 nucleotides in length. In some embodiments, the inhibitory nucleic acid molecule is 25 nucleotides in length.

[0021] In some embodiments, the inhibitory nucleic acid molecule is 23 or 25 nucleotides in length.

[0022] In some embodiments, the inhibitory nucleic acid molecule comprises at least 85% (e.g., 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) complementarity to the target nucleic acid molecule, or splice variant thereof.

[0023] In some embodiments, the inhibitory nucleic acid molecule includes at least 90% (e.g., 90% 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) complementarity to the target nucleic acid molecule, or splice variant thereof.

[0024] In some embodiments, the inhibitory nucleic acid molecule includes at least 95% (e.g., 95%, 96%, 97%, 98%, 99%, or 100%) complementarity to the target nucleic acid molecule, or splice variant thereof.

[0025] In some embodiments, the inhibitory nucleic acid molecule is complementary to the target nucleic acid molecule, or splice variant thereof.

[0026] In some embodiments, the inhibitory nucleic acid molecule further includes a modification.

[0027] In some embodiments, the modification includes: (a) a non-natural or modified nucleoside or nucleotide; and / or (b) a covalently or non-covalently conjugated moiety. In some embodiments: (a) the non-natural or modified nucleoside or nucleotide is selected from the group consisting of: a locked nucleic acid (LNA), a 2'-O-methyl (2'-0-Me) modified nucleoside, a phosphorothioate (PS) bond between nucleosides, and a 2'-fluoro (2’-F) modified nucleoside; and / or (b) the covalently or non-covalently conjugated moiety is selected from the group consisting of: a targeting moiety, a hydrophobic moiety, a cell penetrating peptide, or a polymer.

[0028] In some embodiments, the targeting moiety is vascular cell adhesion protein 1 (VCAM1). In some embodiments, the targeting moiety is arginylglycylaspartic acid (RGD),

[0029] In some embodiments, the inhibitory nucleic acid molecule is selected from the group consisting of: a small interfering RNA (siRNA), a double-stranded RNA (dsRNA), a microRNA (miRNA), a short hairpin RNA (shRNA), an anti-sense oligonucleotide (ASO), and a gapmeR.

[0030] In some embodiments, the inhibitory nucleic acid molecule is a dsRNA.

[0031] In some embodiments, the inhibitory nucleic acid molecule is a miRNA.

[0032] In some embodiments, the inhibitory nucleic acid molecule is an shRNA.

[0033] In some embodiments, the inhibitory nucleic acid molecule is an ASO.

[0034] In some embodiments, the inhibitory nucleic acid molecule is a gapmeR.

[0035] In some embodiments, the inhibitory nucleic acid molecule is an siRNA.

[0036] In some embodiments, the siRNA comprises an antisense strand comprising at least 88% (e.g., 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) sequence identity to any one of SEQ ID NOs: 1 -6.

[0037] In some embodiments, the antisense strand comprises at least 92% (e.g., 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) sequence identity to any one of SEQ ID NOs: 1 -6.

[0038] In some embodiments, the antisense strand comprises at least 96% (e.g., 97%, 98%, 99%, or 100%) sequence identity to any one of SEQ ID NOs: 1 -6.

[0039] In some embodiments, the antisense strand comprises the nucleotide sequence of any one of SEQ ID NOs: 1 -6.

[0040] In some embodiments, the siRNA further comprises a sense strand comprising at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to any one of SEQ ID NOs: 7-12.

[0041] In some embodiments, the siRNA includes: (a) the antisense strand of SEQ ID NO: 1 and the sense strand of SEQ ID NO: 7; (b) the antisense strand of SEQ ID NO: 2 and the sense strand of SEQ ID NO: 8; (c) the antisense strand of SEQ ID NO: 3 and the sense strand of SEQ ID NO: 9; (d) the antisense strand of SEQ ID NO: 4 and the sense strand of SEQ ID NO: 10; (e) the antisense strand of SEQ ID NO: 5 and the sense strand of SEQ ID NO: 11 ; or (f) the antisense strand of SEQ ID NO: 6 and the sense strand of SEQ ID NO: 12.

[0042] In some embodiments, the siRNA contains 3’ overhangs selected from the group consisting of: (i) a single uracil overhang at one or more 3’ ends of the siRNA; (ii) a double uracil overhang at one or more 3’ ends of the siRNA; (iii) a single thymine overhang at one or more 3’ ends of the siRNA; (iv) a double thymine overhang at one or more 3’ ends of the siRNA; or (v) a single cytosine and single thymine overhang at one or more 3’ ends of the siRNA.

[0043] In some embodiments, the siRNA targets the nucleotide sequence of any one of SEQ ID NOs: In some embodiments, the inhibitory nucleic acid molecule is a miRNA.

[0044] In some embodiments, the miRNA comprises a modification selected from: (a) a non-natural or modified nucleoside or nucleotide; and / or (b) a covalently or non-covalently conjugated moiety.

[0045] In some embodiments: (a) the non-natural or modified nucleoside or nucleotide is selected from the group consisting of: a locked nucleic acid (LNA), a 2'-O-methyl (2'-0-Me) modified nucleoside, a phosphorothioate (PS) bond between nucleosides, and a 2'-fluoro (2’-F) modified nucleoside; and / or (b) the covalently or non-covalently conjugated moiety is selected from the group consisting of: a targeting moiety, a hydrophobic moiety, a cell penetrating peptide, or a polymer.

[0046] In some embodiments, the targeting moiety is vascular cell adhesion protein 1 (VCAM1). In some embodiments, the targeting moiety is arginylglycylaspartic acid (RGD).

[0047] In some embodiments, the miRNA comprises a nucleotide sequence comprising at least 86% (e.g., 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) sequence identity to SEQ ID NO: 13.

[0048] In some embodiments, the miRNA comprises a nucleotide sequence comprising at least 91% (e.g., 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) sequence identity to SEQ ID NO: 13.

[0049] In some embodiments, the miRNA comprises a nucleotide sequence comprising at least 95% (e.g., 95%, 96%, 97%, 98%, 99%, or 100%) sequence identity to SEQ ID NO: 13.

[0050] In some embodiments, the miRNA comprises the nucleotide sequence of SEQ ID NO: 13.

[0051] In some embodiments, the miRNA is miR-342-3p.

[0052] In some embodiments, the inhibitory nucleic acid molecule is formulated in a delivery vehicle.

[0053] In some embodiments, the delivery vehicle is selected from the group consisting of: a vector, a plasmid, a micelle, a liposome, an exosome, and a lipid nano particle (LNP).

[0054] In some embodiments, the vector is a viral vector.

[0055] In some embodiments, the viral vector is an adeno-associated viral (AAV) vector.

[0056] In some embodiments, the AAV vector is selected from the group consisting of: AAV1 , AAV2, AAV3, AAV4, AAV5, AAV6, AAV7, AAV8, AAV9, AAV10, AAV11 , and AAV12.

[0057] In some embodiments of any of the foregoing aspects, the inhibitory nucleic acid molecule is formulated as a pharmaceutical composition.

[0058] In some embodiments, the pharmaceutical composition includes a pharmaceutically acceptable excipient, diluent, and / or carrier.

[0059] In a second aspect, the invention provides a method of treating or reducing the likelihood of a medical complication resulting from a myocardial infarction (Ml) in a subject, the method comprising administering the inhibitory nucleic acid molecule of the first aspect.

[0060] In some embodiments, the medical complication is an arrhythmic-related complication, an ischemic-related complication, a mechanical-related complication, an inflammatory-related complication, and / or a systemic complication.

[0061] In some embodiments: (a) the arrhythmic-related complication is a heart block, an atrial arrhythmia, and / or a ventricular arrhythmia; (b) the ischemic-related complication is reinfarction, periinfarct ischemia, and / or an infarct extension; (c) the mechanical-related complication is a mitral valve rupture or tear, a chordae rupture or tear, a ventricular septal defect (VSD), a ventricular free wall rupture, a cardiac tamponade, and / or an aneurysm; (d) the inflammatory-related complication is pericarditis and / or Dressier syndrome; and / or (e) the systemic complication is cardiogenic shock, cardiomyopathy, heart failure, embolic stroke, systemic embolism, and / or a lower extremity embolism.

[0062] In a third aspect, the invention provides a method of promoting angiogenesis in a subject, the method comprising administering the inhibitory nucleic acid molecule of the first aspect.

[0063] In some embodiments of any of the foregoing aspects, the subject has previously experienced a myocardial infarction.

[0064] In some embodiments of any of the foregoing aspects, the subject has an ischemic injury.

[0065] In some embodiments of any of the foregoing aspects, the subject has a cardiovascular disease.

[0066] In some embodiments, the cardiovascular disease is coronary artery disease, peripheral artery disease, cardiomyopathy, or stroke.

[0067] In some embodiments of any of the foregoing aspects, the subject has a metabolic disorder, or the subject is at risk of developing the metabolic disorder.

[0068] In some embodiments, the metabolic disorder is diabetes.

[0069] In some embodiments of any of the foregoing aspects, the subject at risk of developing diabetes is prediabetic and / or has one or more of the following: (a) hyperglycemia; (b) glucose resistance; (c) insulin resistance; (d) hyperlipidemia; and (e) has a family history of diabetes.

[0070] In some embodiments of any of the foregoing aspects, the inhibitory nucleic acid molecule is administered to the subject intravenously, intraperitoneally, subcutaneously, intraarticularly, or intramuscularly.

[0071] In some embodiments of any of the foregoing aspects, the inhibitory nucleic acid molecule is delivered to the coronary endothelium, remote zone of the heart, and / or border zone of the heart.

[0072] In some embodiments of any of the foregoing aspects, the inhibitory nucleic acid molecule is delivered to an endothelial cell, a cardiomyocyte, a fibroblast, a vascular smooth muscle cell, and / or a leukocyte.

[0073] In some embodiments of the second or third aspect, the further comprising administering an additional therapeutic agent.

[0074] In some embodiments, the additional therapeutic agent is a statin or hepatocyte growth factor (HGF).

[0075] Other features and advantages of the invention will be apparent from the following description of the preferred embodiments thereof, and from the claims.

[0076] BRIEF DESCRIPTION OF THE DRAWINGS

[0077] The accompanying drawings are included to illustrate embodiments of the disclosure and further an understanding of its implementations.

[0078] FIG. 1A-FIG. 1G demonstrate that miR-342-3p is a proangiogenic miRNA. FIG. 1A displays the expression kinetics of miR-342-3pin cardiac endothelial cells (ECs) post-myocardial infarction (Ml) between chow and high fat sucrose containing (HFSC) diet groups. Dominant phases for inflammation and angiogenesis are indicated. FIG. 1B is a representative image of a spheroid sprouting assay following transfection of ECs with miR-342-3p mimics. These data demonstrate that miR-342-3p promoted spheroid sprouting while its inhibition elicited angiostatic responses. FIG. 1C are graphs quantifying the number of sprouts after the transfection of ECs with miR-342-3p mimics. This data demonstrates that miR-342-3p promoted spheroid sprouting while its inhibition elicited angiostatic responses. Two-tailed t-tests, p<0.05. FIG. 1D is a representative image of a scratch closure assay following transfection of ECs with miR-342-3p mimics. These data demonstrate that miR-342-3p accelerated scratch closure while its inhibition elicited angiostatic responses. FIG. 1E are graphs quantifying the percent scratch area after the transfection of ECs with miR-342-3p mimics. These data demonstrate that miR-342-3p accelerated scratch closure while its inhibition elicited angiostatic responses. FIG. 1F are graphs quantifying the area under the curve of a scratch area assay after the transfection of ECs with miR-342-3p mimics. These data demonstrate that miR-342-3p accelerated scratch closure while its inhibition elicited angiostatic responses. Two-tailed t-tests, p<0.05. FIG. 1G are graphs quantifying a BrdU incorporation assay following transfection of ECs with miR-342-3p mimics. These data demonstrate that miR-342-3p increased EC proliferation while its inhibition elicited angiostatic responses. Two-tailed t-tests, p<0.05.

[0079] FIG. 2A-FIG. 2H show miR-342-3p’s targets and associated signaling pathways. FIG. 2A is a Venn diagram illustrating the identification of decorin (DCN) as one of several targets of miR-342-3p. FIG. 2B is a graph quantifying DCN transcripts following miR-342-3p overexpression. These data demonstrate that overexpression miR-342-3p suppresses DCN at the transcriptional level in ECs. Two-tailed t-tests, p<0.05. FIG. 2C are a Western blot (WB) and a graph quantifying DCN protein following miR-342-3p overexpression. These data demonstrate that overexpression miR-342-3p suppresses DCN at the translational level in ECs. Two-tailed t-tests, p<0.05. FIG. 2D shows DCN’s 3’ UTR and the seed sequence for miR-342-3p (upper) (SEQ ID NOs: 21 -23), along with bar graphs (lower) quantifying DCN 3’UTR transcriptional activity, which was reduced significantly in the presence of miR-342-3p vs control. Mutated DCN 3’UTR is not targeted. Two-tailed t-tests, p<0.05. FIG. 2E is a chord plot showing top up- regulated and top down-regulated genes, as well as their associated pathways in miR-342-3p overexpressing ECs vs control. RNA sequencing analysis implicated hepatocyte growth factor (HGF)- mesenchymal-epithelial transition factor (MET) signaling as one of the top pathways upregulated by miR- 342-3p (box). FIG. 2F are graphs showing that miR-342-3p overexpression induces MET mRNA expression (upper graph) and protein expression (lower graph). Two-tailed t-tests, p<0.05. FIG. 2G is a WB showing that miR-342-3p overexpression induces MET protein expression. FIG. 2H is a graph showing that miR-342-3p induces HGF mRNA concordantly. HGF is the canonical ligand of the MET receptor. Two-tailed t-tests, p<0.05.

[0080] FIG. 3A-FIG. 3F demonstrate the effect of hypoxia and diabetogenic stimuli on miR-342-3p, DCN and MET. FIG. 3A is a graph showing that hypoxia induces miR-342-3p expression under control conditions, but high glucose conditions blunted this induction. One-way analysis of variance (ANOVA), p<0.05. FIG. 3B is a graph showing that DCN is suppressed by hypoxia and is induced by glucose conditions. One-way ANOVA, p<0.05. FIG. 3C is a graph showing that MET expression mirrors that of miR-342-3p; it is induced by hypoxia but abrogated by glucose. One-way ANOVA, p<0.05. FIG. 3D is a graph showing that hypoxia induces miR-342-3p expression under control conditions, but palmitate treatment blunts this induction. One-way ANOVA, p<0.05. FIG. 3E is a graph showing that DCN is suppressed by hypoxia and is induced by palmitate treatment. One-way ANOVA, p<0.05. FIG. 3F is a graph showing that MET expression mirrors that of miR-342-3p; it is induced by hypoxia but abrogated by palmitate treatment. One-way ANOVA, p<0.05. FIG. 4A-FIG. 41 demonstrate that miR-342-3p rescues cardiac function and promotes angiogenesis in vivo. FIG. 4A is a schematic of the experimental procedure in which miR-342-3p or control mimic was injected in the border zone (BZ) of mice following 45 minutes of ischemia. FIG. 4B are representative mode images of non-specific control (NS) vs miR-342-3p treated hearts. FIG. 4C is a graph quantifying the ejection fraction (EF) and fractional shortening (FS) in miR-342-3p-injected hearts, which had significantly higher EF and FS at day 14 when compared to control. Two-way ANOVA, two- tailed t-tests, p<0.05. FIG. 4D is a graph of a quantitative pCR (qPCR) validation of miR-342-3p overexpression in the BZ at day 14. Two-way ANOVA, two-tailed t-tests, p<0.05. FIG. 4E is a graph showing that DCN mRNA was suppressed in the miR-342-3p injected hearts vs control. Two-way ANOVA, two-tailed t-tests, p<0.05. FIG. 4F is a graph showing that MET mRNA was significantly upregulated in the miR-342-3p treated group at day 14. Two-way ANOVA, two-tailed t-tests, p<0.05. FIG. 4G are a representative immunofluorescence images showing proliferating (ki67+) endothelial cells in BZs of control or miR-342-3p mimic injected hearts. FIG. 4H is a graph quantifying capillary density, which was higher in miR-342-3p hearts vs. control, respectively. Two-way ANOVA, two-tailed t-tests, p<0.05. FIG. 41 is a graph quantifying ki67+ EC count, which was higher in miR-342-3p hearts vs. control, respectively. Two-way ANOVA, two-tailed t-tests, p<0.05.

[0081] Fig. 5A-FIG. 5H are a series of graphs demonstrating that miR-342-3p induces EC proliferation via HGF-MET signaling. For all graphs: one-way ANOVA, two-way ANOVA. p<0.05. FIG. 5A is a graph of BrdU incorporation assays showing inhibition of the MET receptor by PHA-665752 attenuates EC proliferation induced by miR-342-3p. FIG. 5B is a graph showing that the addition of exogenous recombinant DCN (rDCN) protein neutralized miR-342-3p-induced EC proliferation, mirroring results from FIG. 5A. FIG. 5C is a graph showing that PHA-665752 blunted EC proliferation, even in the presence of HGF. FIG. 5D is a graph showing that rDCN blunted EC proliferation, even in the presence of HGF. FIG. 5E is a graph quantifying a BrdU assay performed under mannitol or glucose conditions in the absence of HGF. While miR-342-3p induced EC proliferation, its induction was attenuated by a MET inhibitor. FIG. 5F is a graph quantifying a BrdU assay performed under mannitol or glucose conditions in the presence of HGF. While miR-342-3p induced EC proliferation, its induction was attenuated by a MET inhibitor. FIG. 5G is a graph showing that exogenous rDCN counteracts the proliferative effects of miR- 342-3p in the absence of HGF. FIG. 5H is a graph showing that exogenous rDCN counteracts the proliferative effects of miR-342-3p in the presence of HGF.

[0082] FIG. 6A-FIG. 60 demonstrate that DCN knockdown (KD) phenocopied miR-342-3p overexpression (OE). FIG. 6A is a WB showing the induction of MET and repression of DCN by miR-342- 3p. FIG. 6B is a graph showing the densitometry of relative MET levels from FIG. 6A. Two-way ANOVA, two-tailed t-tests, p<0.05. FIG. 6C is a graph showing the densitometry of relative DCN levels from FIG. 6A. Two-way ANOVA, two-tailed t-tests, p<0.05. FIG. 6D is a WB showing that siRNA-mediated KD of DCN enhanced MET protein while suppressing DCN; these results phenocopy the results from FIG. 6A- FIG. 6C. FIG. 6E is a quantification of MET proteins from FIG. 6D. Two-way ANOVA, two-tailed t-tests, p<0.05. FIG. 6F is a quantification of DCN protein from FIG. 6D. Two-way ANOVA, two-tailed t-tests, p<0.05. FIG. 6G is a graph showing that the KD of DCN induced EC proliferation under normal and high glucose conditions. Two-way ANOVA, two-tailed t-tests, p<0.05. FIG. 6H is a panel of representative endothelial spheroid images showing robust sprouting in the miR-342-3p OE group under both high mannitol and glucose conditions. FIG. 61 is a panel of representative endothelial spheroid images showing that DCN KD also promoted sprouting under mannitol and glucose conditions. FIG. 6J is a panel of representative endothelial spheroid images showing that the addition of exogenous rDCN attenuates miR-342-induced sprouting. FIG. 6K is a panel of representative endothelial spheroid images showing that supplementation with rDCN abrogates siDCN-mediated sprouting, echoing results from FIG. 6J. FIG. 6L is a quantification of sprout numbers from FIG. 6H. Two-way ANOVA, two-tailed t-tests, p<0.05. FIG. 6M is a quantification of sprout numbers from FIG. 61. Two-way ANOVA, two-tailed t-tests, p<0.05. FIG. 6N is a quantification of sprout numbers from FIG. 6J. Two-way ANOVA, two-tailed t-tests, p<0.05. FIG. 60 is a quantification of sprout numbers from FIG. 6K. Two-way ANOVA, two-tailed t-tests, p<0.05.

[0083] FIG. 7 is a graphical summary of miR-342-3p-induced angiogenesis. Under physiologic conditions, hypoxia induces miR-342-3p expression, inhibiting DCN. miR-342-3p induced HGF-MET signaling precipitates sprouting angiogenesis. Diabetes associated stimuli, such as high glucose or fatty acid levels, prevents hypoxia-induced expression of miR-342- 3p, causing an overabundance of DCN protein which can then be secreted and bind to the MET receptor. DCN-MET binding leads to internalization and degradation of the MET receptor. This blockade of the HGF-MET pathway impairs angiogenesis.

[0084] FIG. 8A-FIG. 8B demonstrate that DCN protein expression is impacted by diabetes-associated stimuli. FIG. 8A is a WB blot and quantification thereof showing the suppression of DCN due to hypoxia in mannitol treated ECs. High glucose treatment significantly elevates DCN levels under normoxia and hypoxia vs mannitol control. Two-way ANOVA p<0.05. FIG. 8B is a WB and quantification thereof showing that hypoxia suppressed DCN protein expression in a time-dependent manner vs normoxia when treated with BSA. Palmitate treatment significantly upregulates DCN protein and keeps it elevated even under hypoxic conditions Two-way ANOVA p<0.05.

[0085] FIG. 9A-FIG. 9E demonstrate that miR-342-3p acts via the HGF-MET signaling pathway in both mice and humans. For all graphs: one-way and two-way ANOVA p<0.05. FIG. 9A is a quantification of a BrdU assay showing that EC proliferation is induced by HGF (50 ng / mL) supplementation in HUVECs transfected with either control or miR-342 mimic. FIG. 9B is a graph showing that the addition of PHA- 665752 blunted EC proliferation caused by miR-342 under normal (mannitol) conditions. HUVECs proliferated less under high glucose conditions even in the absence of the inhibitor. Addition of the inhibitor did not synergistically reduce proliferation. miR-342 OE induced EC proliferation albeit to a smaller extent. FIG. 9C is a graph showing that supplementation with rDCN attenuated proliferation in a manner similar to that of the MET inhibitor. FIG. 9D is a quantification of a BrdU assay showing similar results FIG. 9B except with added HGF. FIG. 9E is a graph showing that rDCN acts similarly to the conditions set forth in FIG. 9C, even in the presence of added HGF.

[0086] FIG. 10A-FIG. 10D are dose-responses of PHA-665752 and DCN using the scratch closure assay. FIG. 10A is a dose curve showing scratch closure rates in response to vehicle (DMSO) or increasing concentrations of PHA-665752 - a MET inhibitor. FIG. 10B is an area under curve (AUC) analysis from FIG. 10A showing significantly higher AUC, or slower scratch closure, in ECs treated with 100 nM of the inhibitor. One-way ANOVA p<0.05. FIG. 10C is a dose curve showing scratch closure rates in response to increasing concentrations of rDCN protein. FIG. 10D is an AUC analysis from FIG. 10C showing effective inhibition of scratch closure at 25 nM dose. One-way ANOVA p<0.05. FIG. 11 A displays the expression kinetics of DCN mRNA in cardiac ECs post-MI.

[0087] FIG. 11B displays the expression kinetics of MET mRNA in cardiac ECs post-MI.

[0088] FIG. 11C is a panel of representative light-sheet images in which infarct size and capillary density was quantified.

[0089] DEFINTIONS

[0090] Unless otherwise defined herein, scientific, and technical terms used herein have the meanings that are commonly understood by those of ordinary skill in the art. In the event of any latent ambiguity, definitions provided herein take precedent over any dictionary or extrinsic definition. Unless otherwise required by context, singular terms shall include pluralities and plural terms shall include the singular. The use of "or" means "and / or" unless stated otherwise. The use of the term "including," as well as other forms, such as "includes" and "included," is not limiting.

[0091] As used herein, the term "about," as applied to one or more values of interest, refers to a value that falls within 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1%, or less in either direction (greater than or less than) of a stated reference value, unless otherwise stated or otherwise evident from the context (except where such number would exceed 100% of a possible value).

[0092] As used herein, “administration” refers to providing or giving a subject a therapeutic agent by any effective route. Exemplary routes of administration are described herein below.

[0093] As used herein, the term “administered in combination” or “combined administration” means that two or more agents are administered to a subject at the same time or within an interval such that there may be an overlap of an effect of each agent on the patient. In some embodiments, they are administered within about 60, 30, 15, 10, 5, or 1 minute of one another. In some embodiments, the administrations of the agents are spaced sufficiently closely together such that a combinatorial (e.g., a synergistic) effect is achieved.

[0094] As used herein, the term "auxiliary moiety" refers to any moiety, including, but not limited to, a small molecule, a peptide, a carbohydrate, a neutral organic polymer, a positively charged polymer, a therapeutic agent, a targeting moiety, an endosomal escape moiety, and any combination thereof, which can be conjugated to a nucleic acid molecule. In some embodiments, an "auxiliary moiety" is linked to an inhibitory nucleic acid molecule disclosed herein by forming one or more covalent or non-covalent bonds with one or more conjugating groups attached to a phosphate linkage, a phosphorothioate linkage, a 5' positions of a nucleotide sugar, or any portion of a nucleobase. One skilled in the art will readily understand appropriate points of attachment of a particular auxiliary moiety to a nucleic acid molecule.

[0095] As used herein, “delivery vehicle” refers to any substance (e.g., molecule, peptide, conjugate, and construct) that facilitates, at least in part, the in vivo delivery of a nucleic acid molecule to targeted cells.

[0096] As used herein, the terms “effective amount,” “therapeutically effective amount,” and a “sufficient amount” of a composition described herein refer to a quantity sufficient to, when administered to the subject, effect beneficial or desired results; as such, an “effective amount” or synonym thereto depends upon the context in which it is being applied. For example, in the context of decreasing decorin (DCN), it is an amount of the composition sufficient to achieve a treatment response as compared to the response obtained without administration of the composition. The amount of a given composition described herein that will correspond to such an amount will vary depending upon various factors, such as the given agent, the pharmaceutical compositions, the route of administration, the type of disease or disorder, the identity of the subject (e.g., age, sex, weight) or host being treated, and the like, but can nevertheless be routinely determined by one skilled in the art.

[0097] As used herein, a “formulation” includes at least an inhibitory nucleic acid molecule and a delivery vehicle.

[0098] As used herein, the term “in vitro” refers to events that occur in an artificial environment, e.g., in a test tube or reaction vessel, in cell culture, in a Petri dish, etc., rather than within an organism (e.g., animal, plant, or microbe).

[0099] As used herein, the term “in vivo” refers to events that occur within an organism (e.g., animal, plant, or microbe or cell or tissue thereof).

[0100] As used herein, the term “inhibitory nucleic acid molecule” refers to a nucleic acid molecule that has sufficient complementarity to bind to a target nucleic acid molecule to inhibit expression of a product (e.g., a mRNA) encoded by the target nucleic acid molecule. Exemplary inhibitory nucleic acid molecules are anti-sense oligonucleotides (ASOs), small interfering RNA (siRNAs), short hairpin RNA (shRNAs), double stranded RNAs (dsRNAs), and microRNA (miRNAs). Inhibitory nucleic acid molecules may reduce the target’s expression by 10% or more (e.g., 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%, 98%, 99% or more). In one embodiment, the target nucleic acid molecule encodes DCN.

[0101] As used herein the term “modified” refers to a changed state or structure of a nucleic acid molecule described herein. Molecules may be modified in many ways including chemically, structurally, and functionally. In one embodiment, the inhibitory nucleic acid molecules of the present invention are modified by the introduction of non-natural nucleosides and / or nucleotides. In other embodiments, the inhibitory nucleic acid molecules of the present invention are modified by conjugation of an auxiliary moiety.

[0102] As used herein, the term “pharmaceutical composition” refers to a mixture containing a therapeutic agent, optionally in combination with one or more pharmaceutically acceptable excipients, diluents, and / or carriers, to be administered to a subject, such as a mammal, e.g., a human, in order to prevent, treat or control a particular disease or condition affecting or that may affect the subject.

[0103] As used herein, the term “pharmaceutically acceptable” refers to those compounds, materials, compositions, and / or dosage forms which are, within the scope of sound medical judgment, suitable for use in contact with the tissues a subject, such as a mammal (e.g., a human) without excessive toxicity, irritation, allergic response, or other problem or complication, commensurate with a reasonable benefit / risk ratio.

[0104] “Percent (%) sequence identity” with respect to a reference polynucleotide or polypeptide sequence is defined as the percentage of nucleic acids or amino acids in a candidate sequence that are identical to the nucleic acids or amino acids in the reference polynucleotide or polypeptide sequence, after aligning the sequences and introducing gaps, if necessary, to achieve the maximum percent sequence identity. Alignment for purposes of determining percent nucleic acid or amino acid sequence identity can be achieved in various ways that are within the capabilities of one of skill in the art, for example, using publicly available computer software such as BLAST, BLAST-2, or Megalign software. Those skilled in the art can determine appropriate parameters for aligning sequences, including any algorithms needed to achieve maximal alignment over the full length of the sequences being compared. For example, percent sequence identity values may be generated using the sequence comparison computer program BLAST. As an illustration, the percent sequence identity of a given nucleic acid or amino acid sequence, A, to, with, or against a given nucleic acid or amino acid sequence, B, (which can alternatively be phrased as a given nucleic acid or amino acid sequence, A that has a certain percent sequence identity to, with, or against a given nucleic acid or amino acid sequence, B) is calculated as follows:

[0105] 100 multiplied by (the fraction X / Y) where X is the number of nucleotides or amino acids scored as identical matches by a sequence alignment program (e.g., BLAST) in that program’s alignment of A and B, and where Y is the total number of nucleic acids in B. It will be appreciated that where the length of nucleic acid or amino acid sequence A is not equal to the length of nucleic acid or amino acid sequence B, the percent sequence identity of A to B will not equal the percent sequence identity of B to A.

[0106] As used herein, an inhibitory nucleic acid molecule (e.g., an siRNA, a dsRNA, a miRNA, a shRNA, an ASO, or a gapmeR) having “sufficient complementarity” to a target nucleic acid molecule (e.g., a target mRNA, e.g., DCN) means that the inhibitory nucleic acid molecule includes a nucleotide sequence capable of hybridizing to, and triggering the destruction of, the target nucleic acid molecule (e.g., by RISC-mediated cleavage or Rnase H-mediated cleavage of the target nucleic acid molecule). The inhibitory nucleic acid molecule can be designed such that every nucleotide is complementary to a nucleotide in the target nucleic acid molecule. Alternatively, mismatched nucleotides may be introduced so long as there remains hybridization and destruction of the target nucleic acid molecule.

[0107] As used herein, the term “therapeutic agent” refers to any agent that, when administered to a subject, has a therapeutic, diagnostic, and / or prophylactic effect and / or elicits a desired biological and / or pharmacological effect.

[0108] As used herein, “treatment” and “treating” in reference to a disease or condition, refer to an approach for obtaining beneficial or desired results, e.g., clinical results. Beneficial or desired results can include, but are not limited to, alleviation or amelioration of one or more symptoms or conditions; diminishment of extent of disease or condition; stabilized (i.e., not worsening) state of disease, disorder, or condition; preventing spread of disease or condition; delay or slowing the progress of the disease or condition; amelioration or palliation of the disease or condition; and remission (whether partial or total), whether detectable or undetectable. “Treatment” can also mean prolonging survival as compared to expected survival if not receiving treatment. Those in need of treatment include those already with the condition or disorder, as well as those prone to have the condition or disorder or those in which the condition or disorder is to be prevented.

[0109] As used herein, the term “vector” is considered a replicon, such as plasmid, phage, viral construct or cosmid, to which another nucleic acid (e.g., DNA or RNA) segment may be attached. Vectors are used to transduce and express the nucleic acid segment in cells.

[0110] DETAILED DESCRIPTION

[0111] Described herein are compositions (e.g., an inhibitory nucleic acid molecule) for reducing expression of a target nucleic acid molecule (e.g., an mRNA molecule encoding decorin (DCN), e.g., SEQ ID NO: 14) and methods thereof for (i) treating a medical condition resulting from a myocardial infarction (Ml), and / or (ii) promoting angiogenesis in a subject. The inhibitory nucleic acid molecule may be a small interfering RNA (siRNA), a double-stranded RNA (dsRNA), an anti-sense oligonucleotide (ASO), a microRNA (miRNA), or a short hairpin RNA (shRNA)), or a gapmeR described herein, or a composition (e.g., pharmaceutical composition) thereof. Advantageously, the composition described herein provides therapeutic effects (e.g., angiogenesis) for cardiac tissue following myocardial infarction (Ml).

[0112] Inhibitory Nucleic Acid Molecules

[0113] Exemplary inhibitory nucleic acid molecules of the disclosure are siRNAs, dsRNAs, ASOs, miRNAs, gapmeRs, and shRNAs; however, any nucleic acid molecule capable of reducing DCN (e.g., SEQ ID NO: 14), or a variant thereof, is envisioned for use of the methods described herein. In some instances, the inhibitory nucleic acid molecules of the disclosure may be referred to as RNA inhibitory (RNAi) molecules.

[0114] For any of the inhibitory nucleic acid molecules described herein (e.g., siRNA, dsRNA, miRNA, shRNA, ASO, gapmeR, or other inhibitory nucleic acid molecules capable of reducing expression of a target gene) the inhibitory nucleic acid molecule contains at least some sequence complementarity to the nucleotide sequence of SEQ ID NO: 14 (or a variant thereof, e.g., see Table 3). In some embodiments, the inhibitory nucleic acid molecule comprises or consists of a sequence complementary to at least 15 contiguous nucleotides set forth within SEQ ID NO: 14 (or a variant thereof, e.g., see Table 3). In some embodiments, the inhibitory nucleic acid molecule comprises or consists of a sequence complementary to at least 16 contiguous nucleotides set forth within SEQ ID NO: 14 (or a variant thereof, e.g., see Table 3). In some embodiments, the inhibitory nucleic acid molecule comprises or consists of a sequence complementary to at least 17 contiguous nucleotides set forth within SEQ ID NO: 14 (or a variant thereof, e.g., see Table 3). In some embodiments, the inhibitory nucleic acid molecule comprises or consists of a sequence complementary to at least 18 contiguous nucleotides set forth within SEQ ID NO: 14 (or a variant thereof, e.g., see Table 3). In some embodiments, the inhibitory nucleic acid molecule comprises or consists of a sequence complementary to at least 19 contiguous nucleotides set forth within SEQ ID NO: 14 (or a variant thereof, e.g., see Table 3). In some embodiments, the inhibitory nucleic acid molecule comprises or consists of a sequence complementary to at least 20 contiguous nucleotides set forth within SEQ ID NO: 14 (or a variant thereof, e.g., see Table 3). In some embodiments, the inhibitory nucleic acid molecule comprises or consists of a sequence complementary to at least 21 contiguous nucleotides set forth within SEQ ID NO: 14 (or a variant thereof, e.g., see Table 3). In some embodiments, the inhibitory nucleic acid molecule comprises or consists of a sequence complementary to at least 22 contiguous nucleotides set forth within SEQ ID NO: 14 (or a variant thereof, e.g., see Table 3). In some embodiments, the inhibitory nucleic acid molecule comprises or consists of a sequence complementary to at least 23 contiguous nucleotides set forth within SEQ ID NO: 14 (or a variant thereof, e.g., see Table 3). In some embodiments, the inhibitory nucleic acid molecule comprises or consists of a sequence complementary to at least 24 contiguous nucleotides set forth within SEQ ID NO: 14 (or a variant thereof, e.g., see Table 3). In some embodiments, the inhibitory nucleic acid molecule comprises or consists of a sequence complementary to at least 25 contiguous nucleotides set forth within SEQ ID NOs: 16. In some embodiments, the inhibitory nucleic acid molecule comprises or consists of a sequence complementary to at least 26 contiguous nucleotides set forth within SEQ ID NO: 14 (or a variant thereof, e.g., see Table 3). In some embodiments, the inhibitory nucleic acid molecule comprises or consists of a sequence complementary to at least 27 contiguous nucleotides set forth within SEQ ID NO: 14 (or a variant thereof, e.g., see Table 3). In some embodiments, the inhibitory nucleic acid molecule comprises or consists of a sequence complementary to at least 28 contiguous nucleotides set forth within SEQ ID NO: 14 (or a variant thereof, e.g., see Table 3). In some embodiments, the inhibitory nucleic acid molecule comprises or consists of a sequence complementary to at least 29 contiguous nucleotides set forth within SEQ ID NO: 14 (or a variant thereof, e.g., see Table 3). In some embodiments, the inhibitory nucleic acid molecule comprises or consists of a sequence complementary to at least 30 contiguous nucleotides set forth within SEQ ID NO: 14 (or a variant thereof, e.g., see Table 3).

[0115] For any of the inhibitory nucleic acid molecules described herein (e.g., siRNA, dsRNA, miRNA, shRNA, ASO, gapmeR, or other inhibitory nucleic acid molecules capable of reducing expression of a target nucleic acid) the inhibitory nucleic acid molecule comprises or consists of a sequence complementary to at least 15 to 6850 contiguous nucleotides (e.g., 15 to 49, 20 to 28, 23-25, 50 to 99, 100 to 200, 150 to 300, 200 to 400, 300 to 700, 500 to 1000, 1000 to 5000, 100 to 6850, 16 to 30, 17 to 30, 18 to 30, 19 to 30, 20 to 30, 21 to 30, 22 to 30, 23 to 30, 24 to 30, 25 to 30, 36 to 30, 27 to 30, 28 to 30, or 29 to 30 contiguous nucleotide) set forth within SEQ ID NO: 14 (or a variant thereof, e.g., see Table 3).

[0116] In some embodiments, the inhibitory nucleic acid is an siRNA targeting DON (e.g., SEQ ID NO: 14), or a variant thereof. In some embodiments, the inhibitory nucleic acid is an dsRNA targeting DON (e.g., SEQ ID NO: 14), or a variant thereof. In some embodiments, the inhibitory nucleic acid is an ASO targeting DON (e.g., SEQ ID NO: 14), or a variant thereof. In some embodiments, the inhibitory nucleic acid is a gapmeR targeting DON (e.g., SEQ ID NO: 14), or a variant thereof. In some embodiments, the inhibitory nucleic acid is a miRNA targeting DON (e.g., SEQ ID NO: 14), or a variant thereof. In some embodiments, the inhibitory nucleic acid is an shRNA targeting DON (e.g., SEQ ID NO: 14), or a variant thereof. Each of these modalities is described further below. small interfering RNA (siRNA) siRNAs of the disclosure are single-stranded (ss) or double-stranded (ds) nucleic acid molecules made of DNA, RNA, or both DNA and RNA (e.g., a chimeric) that are complementary to a target gene of interest and prevent translation of the target’s mRNA into a protein. Once an siRNA molecule enters a cell, it is incorporated into an RNA-induced silencing complex (RISC). Upon siRNA hybridization to a target mRNA, the RISC complex will cleave the target mRNA, thereby inactivating the target mRNA, resulting in reduced mRNA and protein levels of the target.

[0117] In some embodiments, siRNAs of the disclosure may include a nucleotide sequence of about 10 to about 30 nucleotides in length (e.g., 9, about 10, about 11 , about 12, about 13, about 14, about 15, about 16, about 17, about 18, about 19, about 20, about 21 , about 22, about 23, about 24, about 25, about 26, about 27, about 28, about 29, about 30, or 31 nucleotides in length).

[0118] In some embodiments, siRNAs of the disclosure may include a nucleotide sequence of 10 to 30 nucleotides in length (e.g., 10 to 30, 11 to 30, 12 to 30, 13 to 30, 14 to 30, 15 to 30, 16 to 30, 17 to 30, 18 to 30, 19 to 30, 20 to 30, 21 to 30, 22 to 30, 23 to 30, 24 to 30, 25 to 30, 26 to 30, 27 to 30, 28 to 30, or 29 to 30 nucleotides in length, e.g., 10, 11 , 12, 13, 14, 15, 16, 17, 18, 19, 20, 21 , 22, 23, 24, 25, 26, 27, 28, 29, or 30 nucleotides in length).

[0119] In some embodiments, the siRNA includes a sequence complementary at least 15 to 30 contiguous nucleotides (e.g., 15 to 25, 20 to 28, 23 to 25, 25 to 28, 16 to 30, 17 to 30, 18 to 30, 19 to 30, 20 to 30, 21 to 30, 22 to 30, 23 to 30, 24 to 30, 25 to 30, 36 to 30, 27 to 30, 28 to 30, or 29 to 30 contiguous nucleotides, e.g., 15, 16, 17, 18, 19, 20, 21 , 22, 23, 24, 25, 26, 27, 28, 29, or 30 contiguous nucleotides) set forth within SEQ ID NO: 14 (or a variant thereof, e.g., see Table 3).

[0120] It is within the scope of the disclosure that any length, known and previously unknown in the art, may be employed for the current invention.

[0121] In some embodiments, the siRNA contains an antisense strand. In some embodiments, lengths for an antisense strand of the siRNA molecules of the present disclosure is between 10 and 30 nucleotides (e.g., 10 nucleotides, 11 nucleotides, 12 nucleotides, 13 nucleotides, 14 nucleotides, 15 nucleotides, 16 nucleotides, 17 nucleotides, 18 nucleotides, 19 nucleotides, 20 nucleotides, 21 nucleotides, 22 nucleotides, 23 nucleotides, 24 nucleotides, 25 nucleotides, 26 nucleotides, 27 nucleotides, 28 nucleotides, 29 nucleotides, or 30 nucleotides), between 15 and 25 nucleotides (e.g., 15 nucleotides, 16 nucleotides, 17 nucleotides, 18 nucleotides, 19 nucleotides, 20 nucleotides, 21 nucleotides, 22 nucleotides, 23 nucleotides, 24 nucleotides, or 25 nucleotides), between 20 and 28 nucleotides (e.g., 20 nucleotides, 21 nucleotides, 22 nucleotides, 23 nucleotides, 24 nucleotides, 25 nucleotides, 26 nucleotides, 27 nucleotides, or 28 nucleotides), between 23 and 25 nucleotides (e.g., 23, 24, or 25 nucleotides), or between 18 and 23 nucleotides (e.g., 18 nucleotides, 19 nucleotides, 20 nucleotides, 21 nucleotides, 22 nucleotides, or 23 nucleotides). In some embodiments, the antisense strand is 17 nucleotides. In some embodiments, the antisense strand is 18 nucleotides. In some embodiments, the antisense strand is 19 nucleotides. In some embodiments, the antisense strand is 20 nucleotides. In some embodiments, the antisense strand is 21 nucleotides. In some embodiments, the antisense strand is 22 nucleotides. In some embodiments, the antisense strand is 23 nucleotides. In some embodiments, the antisense strand is 24 nucleotides. In some embodiments, the antisense strand is 25 nucleotides. In some embodiments, the antisense strand is 26 nucleotides. In some embodiments, the antisense strand is 27 nucleotides. In some embodiments, the antisense strand is 28 nucleotides. In some embodiments, the antisense strand is 29 nucleotides. In some embodiments, the antisense strand is 30 nucleotides.

[0122] In some embodiments, the siRNA contains a sense strand. In some embodiments, the sense strand of the siRNA molecules of the present disclosure is between 10 and 30 nucleotides (e.g., 10 nucleotides, 11 nucleotides, 12 nucleotides, 13 nucleotides, 14 nucleotides, 15 nucleotides, 16 nucleotides, 17 nucleotides, 18 nucleotides, 19 nucleotides, 20 nucleotides, 21 nucleotides, 22 nucleotides, 23 nucleotides, 24 nucleotides, 25 nucleotides, 26 nucleotides, 27 nucleotides, 28 nucleotides, 29 nucleotides, or 30 nucleotides), between 15 and 25 nucleotides (e.g., 15 nucleotides, 16 nucleotides, 17 nucleotides, 18 nucleotides, 19 nucleotides, 20 nucleotides, 21 nucleotides, 22 nucleotides, 23 nucleotides, 24 nucleotides, or 25 nucleotides), between 20 and 28 nucleotides (e.g., 20 nucleotides, 21 nucleotides, 22 nucleotides, 23 nucleotides, 24 nucleotides, 25 nucleotides, 26 nucleotides, 27 nucleotides, or 28 nucleotides), between 23 and 25 nucleotides (e.g., 23, 24, or 25 nucleotides), or between 18 and 23 nucleotides (e.g., 18 nucleotides, 19 nucleotides, 20 nucleotides, 21 nucleotides, 22 nucleotides, or 23 nucleotides). In some embodiments, the sense strand is 15 nucleotides. In some embodiments, the sense strand is 16 nucleotides. In some embodiments, the sense strand is 17 nucleotides. In some embodiments, the sense strand is 18 nucleotides. In some embodiments, the sense strand is 19 nucleotides. In some embodiments, the sense strand is 20 nucleotides. In some embodiments, the sense strand is 21 nucleotides. In some embodiments, the sense strand is 22 nucleotides. In some embodiments, the sense strand is 23 nucleotides. In some embodiments, the sense strand is 24 nucleotides. In some embodiments, the sense strand is 25 nucleotides. In some embodiments, the sense strand is 26 nucleotides. In some embodiments, the sense strand is 27 nucleotides. In some embodiments, the sense strand is 28 nucleotides. In some embodiments, the sense strand is 29 nucleotides. In some embodiments, the sense strand is 30 nucleotides.

[0123] In some embodiments, the sense and antisense strands of an siRNA molecule of the disclosure are completely complementary. In some embodiments, the sense and antisense strands of an siRNA molecule of the disclosure are completely complementary to the extent that their lengths overlap with one another. Depending on the sequence of the first and second strand, complementarity need not be complete or perfect, which means that the first and second strand are not 100% base-paired due to mismatches. One or more mismatches may be present within the ds siRNA without impacting the siRNA’s ability to reduced expression of a target gene of interest.

[0124] The nucleotide sequence of an siRNA of the disclosure may contain sufficient complementarity to a portion of a target gene of interest (e.g., DCN (e.g., SEQ ID NO: 14), or a variant thereof, e.g., see Table 3) such that the siRNA can hybridize with the target gene of interest. In some embodiments, the siRNA is at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% complementary to the target gene of interest (e.g., DCN (e.g., SEQ ID NO: 14), or a variant thereof, e.g., see Table 3), or a portion thereof. In some embodiments, the siRNA is complementary to the target gene of interest (e.g., DCN (e.g., SEQ ID NO: 14), or a variant thereof, e.g., see Table 3), or a portion thereof.

[0125] In some embodiments, the nucleotide sequence of the siRNA may contain sufficient complementarity to an exon sequence of a target gene of interest (e.g., an exon of DCN (e.g., SEQ ID NO: 14), or a variant thereof). In some embodiments, the nucleotide sequence of the siRNA may contain sufficient complementarity to an intron sequence of a target gene of interest (e.g., an intron of DCN (e.g., SEQ ID NO: 14), or a variant thereof). In some embodiments, the siRNA of the disclosure may contain sufficient complementarity to a pre-mRNA transcript or an mRNA transcript encoding DCN (e.g., SEQ ID NO: 14), or a variant thereof. For example, the siRNA may hybridize to a target sequence of any one of SEQ ID NOs: 15-20 (e.g., see Table 2). The target gene of interest may be DCN (e.g., SEQ ID NO: 14, or a splice variant thereof, e.g., see Table 3).

[0126] In some embodiments, the siRNAs described herein have 0-7 nucleotide 3’ overhangs or 0-4 nucleotide 5’ overhangs. In some embodiments, the siRNA molecule has a single uracil (e.g., U) overhang at each 3’ end of the siRNA. In some embodiments, the siRNA molecule has a double uracil (e.g., UU) overhang at each 3’ end of the siRNA. In some embodiments, the siRNA molecule has a single thymine (e.g., T) overhang at each 3’ end of the siRNA. In some embodiments, the siRNA molecule has a double thymine (e.g., TT) overhang at each 3’ end of the siRNA. In some embodiments, the siRNA molecule has a cytosine and thymine (e.g., CT) overhang at each 3’ end of the siRNA.

[0127] Different siRNAs can be combined for decreasing mRNA expression of DCN (e.g., SEQ ID NO: 14), or a variant thereof (e.g., see Table 3). A combination of two siRNAs may be used in a method of the invention, such as two different siRNAs, three different siRNAs, four different siRNAs, or five different siRNAs targeting the same gene of interest (e.g., DCN (e.g., SEQ ID NO: 14), or a variant thereof). In some embodiments, the siRNA sequence may contain at least 85%, 86%, 87%, 88%, 89%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% sequence identity to any one or more of SEQ ID NOs: 1 -6 (e.g., see Table 1 ). In some embodiments, the siRNA sequence may contain the sequence of any one or more of SEQ ID NOs: 1 -6 (e.g., see Table 1 ).

[0128] In some embodiments, the siRNA contains at least 15 contiguous nucleotides set forth within any one of SEQ ID NOs: 1 -6 (e.g., see Table 1 ). In some embodiments, the siRNA contains at least 16 contiguous nucleotides set forth within any one of SEQ ID NOs: 1 -6 (e.g., see Table 1 ). In some embodiments, the siRNA contains at least 17 contiguous nucleotides set forth within any one of SEQ ID NOs: 1 -6 (e.g., see Table 1 ). In some embodiments, the siRNA contains at least 18 contiguous nucleotides set forth within any one of SEQ ID NOs: 1 -6 (e.g., see Table 1 ). In some embodiments, the siRNA contains at least 19 contiguous nucleotides set forth within any one of SEQ ID NOs: 1 -6 (e.g., see Table 1 ). In some embodiments, the siRNA contains at least 20 contiguous nucleotides set forth within any one of SEQ ID NOs: 1 -6 (e.g., see Table 1 ). In some embodiments, the siRNA contains 21 contiguous nucleotides set forth within any one of SEQ ID NOs: 1 -6 (e.g., see Table 1 ). In some embodiments, the siRNA contains 22 contiguous nucleotides set forth within any one of SEQ ID NOs: 1 -6 (e.g., see Table 1 ). In some embodiments, the siRNA contains 23 contiguous nucleotides set forth within any one of SEQ ID NOs: 1 -6 (e.g., see Table 1 ). In some embodiments, the siRNA contains 24 contiguous nucleotides set forth within any one of SEQ ID NOs: 1 -6 (e.g., see Table 1 ). In some embodiments, the siRNA contains 25 contiguous nucleotides set forth within any one of SEQ ID NOs: 1 -6 (e.g., see Table 1 ).

[0129] In any of the foregoing embodiments, the siRNA further contains the sequence of any one of SEQ ID NOs: 7-12. Table 1 below provides the antisense and sense strands of exemplary siRNA sequences of the invention.

[0130] TABLE 1. EXEMPLARY siRNA SEQUENCES A = adenine; C = cytosine; G = guanine; U = uracil.

[0131] In some embodiments, the siRNA of the disclosure may target a nucleotide sequence of any one of SEQ ID NOs: 14-20 (e.g., see Table 2), or a complementary sequence thereof, or variant thereof with at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% thereto.

[0132] TABLE 2. TARGET SEQUENCES

[0133] In some embodiments, the siRNA comprises a sequence complementary to at least 15 contiguous nucleotides set forth within SEQ ID NO: 14 (or a variant thereof, e.g., see Table 3). In some embodiments, the siRNA comprises a sequence complementary to at least 16 contiguous nucleotides set forth within SEQ ID NO: 14 (or a variant thereof, e.g., see Table 3). In some embodiments, the siRNA comprises a sequence complementary to at least 17 contiguous nucleotides set forth within SEQ ID NO: 14 (or a variant thereof, e.g., see Table 3). In some embodiments, the siRNA comprises a sequence complementary to at least 18 contiguous nucleotides set forth within SEQ ID NO: 14 (or a variant thereof, e.g., see Table 3). In some embodiments, the siRNA comprises a sequence complementary to at least 19 contiguous nucleotides set forth within SEQ ID NO: 14 (or a variant thereof, e.g., see Table 3). In some embodiments, the siRNA comprises a sequence complementary to at least 20 contiguous nucleotides set forth within SEQ ID NO: 14 (or a variant thereof, e.g., see Table 3). In some embodiments, the siRNA comprises a sequence complementary to at least 21 contiguous nucleotides set forth within SEQ ID NO: 14 (or a variant thereof, e.g., see Table 3). In some embodiments, the siRNA comprises a sequence complementary to at least 22 contiguous nucleotides set forth within SEQ ID NO: 14 (or a variant thereof, e.g., see Table 3). In some embodiments, the siRNA comprises a sequence complementary to at least 23 contiguous nucleotides set forth within SEQ ID NO: 14 (or a variant thereof, e.g., see Table 3). In some embodiments, the siRNA comprises a sequence complementary to at least 24 contiguous nucleotides set forth within SEQ ID NO: 14 (or a variant thereof, e.g., see Table 3). In some embodiments, the siRNA comprises a sequence complementary to at least 25 contiguous nucleotides set forth within SEQ ID NO: 14 (or a variant thereof, e.g., see Table 3). In some embodiments, the siRNA comprises a sequence complementary to at least 26 contiguous nucleotides set forth within SEQ ID NO: 14 (or a variant thereof, e.g., see Table 3). In some embodiments, the siRNA comprises a sequence complementary to at least 27 contiguous nucleotides set forth within SEQ ID NO: 14 (or a variant thereof, e.g., see Table 3). In some embodiments, the siRNA comprises a sequence complementary to at least 28 contiguous nucleotides set forth within SEQ ID NO: 14 (or a variant thereof, e.g., see Table 3). In some embodiments, the siRNA comprises a sequence complementary to at least 29 contiguous nucleotides set forth within SEQ ID NO: 14 (or a variant thereof, e.g., see Table 3). In some embodiments, the siRNA comprises a sequence complementary to at least 30 contiguous nucleotides set forth within SEQ ID NO: 14 (or a variant thereof, e.g., see Table 3). The nucleotide sequence of SEQ ID NO: 14 is set forth in Table 2.

[0134] TABLE 3. DCN SPLICE VARIANTS

[0135] Double-stranded RNA (ds RNA) dsRNAs of the disclosure are ds nucleic acid molecules made of DNA, RNA, or both DNA and RNA (e.g., a chimeric) that are complementary to a target gene of interest and prevent translation of the target’s mRNA into a protein. Typically, dsRNAs are longer than an siRNA and are processed within a cell to form an siRNA molecule. The siRNA is then incorporated into an RNA-induced silencing complex (RISC). Upon siRNA hybridization to a target mRNA, the RISC complex will cleave the target mRNA, thereby inactivating the target mRNA, resulting in reduced mRNA and protein levels of the target.

[0136] In some embodiments, dsRNAs of the disclosure may include a sense strand and an antisense strand, each containing a nucleotide sequence of about 25 to about 5000 nucleotides in length, or longer (e.g., 20, about 25, about 30, about 35, about 40, about 45, about 50, about 55, about 60, about 65, about 70, about 75, about 80, about 85, about 90, about 95, about 100, about 105, about 110, about 115, about 120, about 125, about 130, about 135, about 140, about 145, about 150, about 155, about 160, about

[0137] 165, about 170, about 175, about 180, about 185, about 190, about 195, about 200, about 210, about

[0138] 220, about 230, about 240, about 250, about 260, about 270, about 280, about 290, about 300, about

[0139] 310, about 320, about 330, about 340, about 350, about 360, about 370, about 380, about 380, about

[0140] 400, about 425, about 450, about 475, about 500, about 525, about 550, about 575, about 600, about

[0141] 625, about 650, about 675, about 700, about 725, about 750, about 775, about 800, about 825, about

[0142] 850, about 875, about 900, about 925, about 950, about 975, about 1000, about 1100, about 1200, about 1300, about 1400, about 1500, about 1600, about 1700, about 1800, about 1900, about 2000, about

[0143] 2200, about 2400, about 2600, about 2800, about 3000, about 3250, about 3500, about 3750, about

[0144] 4000, about 4250, about 4500, about 4750, about 5000, about 6000, or about 6850 nucleotides in length).

[0145] In some embodiments, dsRNAs of the disclosure may include a sense strand and an antisense strand, each containing a nucleotide sequence of 25 to 5000 nucleotides in length, or longer (e.g., 25 to 5000, 50 to 5000, 75 to 5000, 100 to 5000, 125 to 5000, 150 to 5000, 175 to 5000, 200 to 5000, 225 to 5000, 250 to 5000, 275 to 5000, 300 to 5000, 325 to 5000, 350 to 5000, 375 to 5000, 400 to 5000, 425 to

[0146] 5000, 450 to 5000, 475 to 5000, 500 to 5000, 550 to 5000, 600 to 5000, 650 to 5000, 700 to 5000, 750 to

[0147] 5000, 800 to 5000, 850 to 5000, 900 to 5000, 950 to 5000, 1000 to 5000, 1050 to 5000, 1100 to 5000, 1150 to 5000, 1200 to 5000, 1300 to 5000, 1400 to 5000, 1500 to 5000, 1600 to 5000, 1700 to 5000,

[0148] 1800 to 5000, 1900 to 5000, 2000 to 5000, 2100 to 5000, 2200 to 5000, 2300 to 5000, 2400 to 5000,

[0149] 2500 to 5000, 3000 to 5000, 3500 to 5000, 4000 to 5000, or 4500 to 5000 nucleotides in length, e.g., 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100, 105, 110, 115, 120, 125, 130, 135, 140, 145,

[0150] 150, 155, 160, 165, 170, 175, 180, 185, 190, 195, 200, 210, 220, 230, 240, 250, 260, 270, 280, 290, 300,

[0151] 310, 320, 330, 340, 350, 360, 370, 380, 380, 400, 425, 450, 475, 500, 525, 550, 575, 600, 625, 650, 675,

[0152] 700, 725, 750, 775, 800, 825, 850, 875, 900, 925, 950, 975, 1000, 1100, 1200, 1300, 1400, 1500, 1600,

[0153] 1700, 1800, 1900, 2000, 2200, 2400, 2600, 2800, 3000, 3250, 3500, 3750, 4000, 4250, 4500, 4750, or 5000 nucleotides in length).

[0154] In some embodiments, the dsRNA includes a sequence complementary at least 25 to 6850 contiguous nucleotides (e.g., 25 to 6850, 50 to 6850, 75 to 6850, 100 to 6850, 125 to 6850, 150 to 6850, 175 to 6850, 200 to 6850, 225 to 6850, 250 to 6850, 275 to 6850, 300 to 6850, 325 to 6850, 350 to 6850, 375 to 6850, 400 to 6850, 425 to 6850, 450 to 6850, 475 to 6850, 500 to 6850, 550 to 6850, 600 to 6850, 650 to 6850, 700 to 6850, 750 to 6850, 800 to 6850, 850 to 6850, 900 to 6850, 950 to 6850, 1000 to 6850, 1050 to 6850, 1100 to 6850, 1150 to 6850, 1200 to 6850, 1300 to 6850, 1400 to 6850, 1500 to 6850, 1600 to 6850, 1700 to 6850, 1800 to 6850, 1900 to 6850, 2000 to 6850, 2100 to 6850, 2200 to 6850, 2300 to 6850, 2400 to 6850, 2500 to 6850, 3000 to 6850, 3500 to 6850, 4000 to 6850, 4500 to 6850, 5000 to 6850, 5500 to 6850, 6000 to 6850, or 6500 to 6850 contiguous nucleotides, e.g., 25, 50, 75, 100, 125, 150, 175, 200, 225, 250, 275, 300, 325, 350, 375, 400, 425, 450, 475, 500, 550, 600, 650, 700, 750, 800, 850, 900, 950, 1000, 1100, 1200, 1300, 1400, 1500, 1600, 1700, 1800, 1900, 2000, 2100, 2200, 2300, 2400, 2500, 3000, 3500, 4000, 4500, 5000, 5500, 6000, 6500, or 6850 contiguous nucleotides) set forth within SEQ ID NO: 14 (or a variant thereof, e.g., see Table 3). It is within the scope of the disclosure that any length, known and previously unknown in the art, may be employed for the current invention.

[0155] In some embodiments, the sense and antisense strands of an dsRNA molecule of the disclosure are completely complementary. In some embodiments, the sense and antisense strands of an dsRNA molecule of the disclosure are completely complementary to the extent that their lengths overlap with one another. Depending on the sequence of the first and second strand, complementarity need not be complete or perfect, which means that the first and second strand are not 100% base-paired due to mismatches. One or more mismatches may be present within the ds dsRNA without impacting the dsRNA’s ability to reduced expression of a target gene of interest.

[0156] The nucleotide sequence of an dsRNA of the disclosure may contain sufficient complementarity to a portion of a target gene of interest (e.g., DCN (e.g., SEQ ID NO: 14), or a variant thereof) such that the dsRNA can hybridize with the target gene of interest. In some embodiments, the dsRNA is at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% complementary to the target gene of interest (e.g., DCN (e.g., SEQ ID NO: 14), or a variant thereof), or a portion thereof. In some embodiments, the dsRNA is complementary to the target gene of interest (e.g., DCN (e.g., SEQ ID NO: 14), or a variant thereof), or a portion thereof.

[0157] In some embodiments, the nucleotide sequence of the dsRNA may contain sufficient complementarity to an exon sequence of a target gene of interest (e.g., an exon of DCN (e.g., SEQ ID NO: 14), or a variant thereof, e.g., see Table 3). In some embodiments, the nucleotide sequence of the dsRNA may contain sufficient complementarity to an intron sequence of a target gene of interest (e.g., an intron of DCN (e.g., SEQ ID NO: 14), or a variant thereof, e.g., see Table 3). In some embodiments, the dsRNA of the disclosure may contain sufficient complementarity to a pre-mRNA transcript or an mRNA transcript encoding DCN (e.g., SEQ ID NO: 14), or a variant thereof, e.g., see Table 3. The target sequence may be any one of SEQ ID NOs: 15-20 (e.g., see Table 2).

[0158] Different dsRNAs can be combined for decreasing the protein expression of a target gene of interest (e.g., DCN (e.g., SEQ ID NO: 14), or a variant thereof). A combination of two dsRNAs may be used in a method of the invention, such as two different dsRNAs, three different dsRNAs, four different dsRNAs, or five different dsRNAs targeting the same gene of interest (e.g., DCN (e.g., SEQ ID NO: 14), or a variant thereof. micro RNA (miRNA) miRNAs of the disclosure are single stranded (ss) nucleic acid molecules made of DNA, RNA, or both DNA and RNA (e.g., a chimeric) that are complementary to a target gene of interest and prevent translation of the target’s mRNA into a protein. Once a miRNA molecule enters a cell, it is incorporated into a RNA-induced silencing complex (RISC). Upon miRNA hybridization to a target mRNA, the RISC complex will cleave the target mRNA, thereby inactivating the target mRNA, resulting in reduced mRNA and protein levels of the target.

[0159] In some embodiments, miRNAs of the disclosure may include a nucleotide sequence of about 6 to about 30 nucleotides in length (e.g., 5, 6, 7, 8, 9, 10, 11 , 12, 13, 14, 15, 16, 17, 18, 19, 20, 21 , 22, 23, 24, 25, 26, 27, 28, 29, 30, or 31 nucleotides in length). In some embodiments, miRNAs of the disclosure may include a nucleotide sequence of 6 to 30 nucleotides in length (e.g., 6 to 30, 7 to 30, 8 to 30, 9 to 30, 10 to 30, 11 to 30, 12 to 30, 13 to 30, 14 to 30, 15 to 30, 16 to 30, 17 to 30, 18 to 30, 19 to 30, 20 to 30, 21 to 30, 22 to 30, 23 to 30, 24 to 30, 25 to 30, 26 to 30, 27 to 30, 28 to 30, or 29 to 30 nucleotides in length, e.g., 6, 7, 8, 9, 10, 11 , 12, 13, 14, 15, 16, 17, 18, 19, 20, 21 , 22, 23, 24, 25, 26, 27, 28, 29, or 30 nucleotides in length).

[0160] In some embodiments, the miRNA comprises a sequence complementary at least 6 to 30 contiguous nucleotides (e.g., 6 to 30, 7 to 30, 8 to 30, 9 to 30, 10 to 30, 11 to 30, 12 to 30, 13 to 30, 14 to 30, 15 to 30, 16 to 30, 17 to 30, 18 to 30, 19 to 30, 20 to 30, 21 to 30, 22 to 30, 23 to 30, 24 to 30, 25 to 30, 26 to 30, 27 to 30, 28 to 30, or 29 to 30 contiguous nucleotides, e.g., 6, 7, 8, 9, 10, 11 , 12, 13, 14, 15, 16, 17, 18, 19, 20, 21 , 22, 23, 24, 25, 26, 27, 28, 29, or 30 contiguous nucleotides) set forth within SEQ ID NO: 14 (or a variant thereof, e.g., see Table 3).

[0161] It is within the scope of the disclosure that any length, known and previously unknown in the art, may be employed for the current invention.

[0162] The nucleotide sequence of the miRNA may contain sufficient complementarity to a portion of a target gene of interest (e.g., DCN (e.g., SEQ ID NO: 14), or a variant thereof) such that the miRNA can hybridize with the target gene of interest. In some embodiments, the miRNA is at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% complementary to the target gene of interest (e.g., DCN (e.g., SEQ ID NO: 14), or a variant thereof), or a portion thereof. In some embodiments, the miRNA is complementary to the target gene of interest (e.g., DCN (e.g., SEQ ID NO: 14), or a variant thereof), or a portion thereof.

[0163] In some embodiments, the nucleotide sequence of the miRNA may contain sufficient complementarity to an exon sequence of a target gene of interest (e.g., an exon of DCN (e.g., SEQ ID NO: 14), or a variant thereof). In some embodiments, the nucleotide sequence of the miRNA may contain sufficient complementarity to an intron sequence of a target gene of interest (e.g., an intron of DCN (e.g., SEQ ID NO: 14), or a variant thereof). In some embodiments, the miRNA of the disclosure may contain sufficient complementarity to a pre-mRNA transcript or an mRNA transcript encoding DCN (e.g., SEQ ID NO: 14), or a variant thereof. The target sequence may be any one of SEQ ID NOs: 15-20 (e.g., see Table 2).

[0164] In some embodiments, the nucleotide sequence of the miRNA includes at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to SEQ ID NO: 13. The miRNA may further include a modification described herein (e.g., a non-natural or modified nucleoside or nucleotide, and / or a covalently or non- covalently conjugated moiety). In some embodiments, the miRNA is miR-342-3p (e.g., SEQ ID NO: 13).

[0165] Different miRNAs can be combined for decreasing the protein expression of a target gene of interest (e.g., DCN (e.g., SEQ ID NO: 14), or a variant thereof). A combination of two or more miRNAs may be used in a method of the invention, such as two different miRNAs, three different miRNAs, four different miRNAs, or five different miRNAs targeting the same target gene of interest (e.g., DCN (e.g., SEQ ID NO: 14), or a variant thereof, e.g., see Table 3). In some embodiments, at least one of the miRNAs is miR-342-3p (e.g., SEQ ID NO: 13), or a variant thereof having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity thereto. short hairpin RNA (shRNA) shRNAs of the disclosure are ss or ds nucleic acid molecules made of DNA, RNA, or both DNA and RNA (e.g., a chimeric) that are complementary to a target gene of interest and prevent translation of the target’s mRNA into a protein. Once a shRNA molecule enters a cell, it is incorporated into a RNA- induced silencing complex (RISC). Upon shRNA hybridization to a target mRNA, the RISC complex will cleave the target mRNA, thereby inactivating the target mRNA, resulting in reduced mRNA and protein levels of the target.

[0166] In some embodiments, shRNAs of the disclosure may include a nucleotide sequence of about 50 to about 100 nucleotides in length (e.g., 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100, 105, or 110 nucleotides in length).

[0167] In some embodiments, shRNAs of the disclosure may include a nucleotide sequence of 50 to 100 nucleotides in length (e.g., 50 to 100, 55 to 100, 60 to 100, 65 to 100, 70 to 100, 75 to 100, 80 to 100, 85 to 100, 90 to 100, or 95 to 100 nucleotides in length, e.g., 50, 51 , 52, 53, 54, 55, 56, 57, 58, 59, 60, 61 , 62, 63, 64, 65, 66, 67, 68, 69, 70, 71 , 72, 73, 74, 75, 76, 77, 78, 79, 80, 81 , 82, 83, 84, 85, 86, 87, 88, 89, 90, 91 , 92, 93, 94, 95, 96, 97, 98, 99, or 100 nucleotides in length). shRNAs of the disclosure contain a variable hairpin loop structure and a stem sequence. In some embodiments the stem sequence may be 10 to 50 nucleotides in length (e.g., 10 to 50, 11 to 50, 12 to 50, 13 to 50, 14 to 50, 15 to 50, 16 to 50, 17 to 50, 18 to 50, 19 to 50, 20 to 50, 21 to 50, 22 to 50, 23 to 50,

[0168] 24 to 50, 25 to 50, 26 to 50, 27 to 50, 28 to 50, 29 to 50, 30 to 50, 31 to 50, 32 to 50, 33 to 50, 34 to 50,

[0169] 35 to 50, 36 to 50, 37 to 50, 38 to 50, 39 to 50, 40 to 50, 41 to 50, 42 to 50, 43 to 50, 44 to 50, 45 to 50,

[0170] 46 to 50, 47 to 50, 48 to 50, or 49 to 50 nucleotides in length, e.g., 10, 11 , 12, 13, 14, 15, 16, 17, 18, 19,

[0171] 20, 21 , 22, 23, 24, 25, 26, 27, 28, 29, or 30, 31 , 32, 33, 34, 35, 36, 37, 38, 39, 40, 41 , 42, 43, 44, 45, 46, 47, 48, 49, or 50 nucleotides in length). In some embodiments, the hairpin size is between 4 to 50 nucleotides in length (e.g., 4 to 50, 5 to 50, 6 to 50, 7 to 50, 8 to 50, 9 to 50, 10 to 50, 11 to 50, 12 to 50, 13 to 50, 14 to 50, 15 to 50, 16 to 50, 17 to 50, 18 to 50, 19 to 50, 20 to 50, 21 to 50, 22 to 50, 23 to 50,

[0172] 24 to 50, 25 to 50, 26 to 50, 27 to 50, 28 to 50, 29 to 50, 30 to 50, 31 to 50, 32 to 50, 33 to 50, 34 to 50,

[0173] 35 to 50, 36 to 50, 37 to 50, 38 to 50, 39 to 50, 40 to 50, 41 to 50, 42 to 50, 43 to 50, 44 to 50, 45 to 50,

[0174] 46 to 50, 47 to 50, 48 to 50, or 49 to 50 nucleotides in length, e.g., 4, 5, 6, 7, 8, 9, 10, 11 , 12, 13, 14, 15,

[0175] 16, 17, 18, 19, 20, 21 , 22, 23, 24, 25, 26, 27, 28, 29, 30, 31 , 32, 33, 34, 35, 36, 37, 38, 39, 40, 41 , 42, 43, 44, 45, 46, 47, 48, 49, or 50 nucleotides in length), although the loop size may be larger without significantly affecting silencing activity. shRNA molecules of the disclosure may contain mismatches, for example G-U mismatches between two strands of the shRNA stem without decreasing potency. In some embodiments, shRNAs are designed to include one or several G-U pairings in the hairpin stem to stabilize hairpins during propagation in bacteria, for example.

[0176] In some embodiments, the shRNA includes a sequence (e.g., a stem sequence) complementary at least 10 to 50 contiguous nucleotides (e.g., 10 to 50, 11 to 50, 12 to 50, 13 to 50, 14 to 50, 15 to 50, 16 to 50, 17 to 50, 18 to 50, 19 to 50, 20 to 50, 21 to 50, 22 to 50, 23 to 50, 24 to 50, 25 to 50, 26 to 50, 27 to

[0177] 50, 28 to 50, 29 to 50, 30 to 50, 31 to 50, 32 to 50, 33 to 50, 34 to 50, 35 to 50, 36 to 50, 37 to 50, 38 to

[0178] 50, 39 to 50, 40 to 50, 41 to 50, 42 to 50, 43 to 50, 44 to 50, 45 to 50, 46 to 50, 47 to 50, 48 to 50, or 49 to

[0179] 50 contiguous nucleotides, e.g., 10, 11 , 12, 13, 14, 15, 16, 17, 18, 19, 20, 21 , 22, 23, 24, 25, 26, 27, 28, 29, 30, 31 , 32, 33, 34, 35, 36, 37, 38, 39, 40, 41 , 42, 43, 44, 45, 46, 47, 48, 49, or 50 contiguous nucleotides) set forth within SEQ ID NO: 14 (or a variant thereof, e.g., see Table 3).

[0180] It is within the scope of the disclosure that any length, known and previously unknown in the art, may be employed for the current invention.

[0181] The nucleotide sequence of the shRNA may contain sufficient complementarity to a portion of a target gene of interest (e.g., DCN (e.g., SEQ ID NO: 14), or a variant thereof) such that the shRNA can hybridize with the target gene of interest. In some embodiments, the shRNA is at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% complementary to the target gene of interest (e.g., DCN (e.g., SEQ ID NO: 14), or a variant thereof), or a portion thereof. In some embodiments, the shRNA is complementary to the target gene of interest (e.g., DCN (e.g., SEQ ID NO: 14), or a variant thereof), or a portion thereof.

[0182] In some embodiments, the nucleotide sequence of the shRNA may contain sufficient complementarity to an exon sequence of a target gene of interest (e.g., an exon of DCN (e.g., SEQ ID NO: 14), or a variant thereof). In some embodiments, the nucleotide sequence of the shRNA may contain sufficient complementarity to an intron sequence of a target gene of interest (e.g., an intron of DCN (e.g., SEQ ID NO: 14), or a variant thereof). In some embodiments, the shRNA of the disclosure may contain sufficient complementarity to a pre-mRNA transcript or an mRNA transcript encoding DCN (e.g., SEQ ID NO: 14), or a variant thereof. The target sequence may be any one of SEQ ID NOs: 15-20 (e.g., see Table 2).

[0183] Different shRNAs can be combined for decreasing the protein expression of a target gene of interest (e.g., DCN (e.g., SEQ ID NO: 14), or a variant thereof). A combination of two or more shRNAs may be used in a method of the invention, such as two different shRNAs, three different shRNAs, four different shRNAs, or five different shRNAs targeting the same gene of interest (e.g., DCN (e.g., SEQ ID NO: 14), or a variant thereof).

[0184] Anti-Sense Oligonucleotide (ASO)

[0185] ASOs of the disclosure are single (ss) nucleic acid molecules made of DNA, RNA, or both DNA and RNA (e.g., a chimeric) that are complementary to a target gene of interest and prevent translation of the target’s mRNA into a protein. Upon hybridization to a target mRNA, RNase H will degrade the mRNA by hydrolyzation, resulting in reduced mRNA and protein levels of the target.

[0186] In some embodiments, ASOs of the disclosure may include a nucleotide sequence of about 12 to about 50 nucleotides in length (e.g., 11 , 12, 13, 14, 15, 16, 17, 18, 19, 20, 21 , 22, 23, 24, 25, 26, 27, 28, 29, 30, 31 , 32, 33, 34, 35, 36, 37, 38, 39, 40, 41 , 42, 43, 44, 45, 46, 47, 48, 49, 50, or 51 nucleotides in length).

[0187] In some embodiments, ASOs of the disclosure may include a nucleotide sequence of 12 to 50 nucleotides in length (e.g., 12 to 50, 13 to 50, 14 to 50, 15 to 50, 16 to 50, 17 to 50, 18 to 50, 19 to 50, 20 to 50, 21 to 50, 22 to 50, 23 to 50, 24 to 50, 25 to 50, 26 to 50, 27 to 50, 28 to 50, 29 to 50, 30 to 50, 31 to

[0188] 50, 32 to 50, 33 to 50, 34 to 50, 35 to 50, 36 to 50, 37 to 50, 38 to 50, 39 to 50, 40 to 50, 41 to 50, 42 to

[0189] 50, 43 to 50, 44 to 50, 45 to 50, 46 to 50, 47 to 50, 48 to 50, or 49 to 50 nucleotides in length, e.g., 12, 13,

[0190] 14, 15, 16, 17, 18, 19, 20, 21 , 22, 23, 24, 25, 26, 27, 28, 29, 30, 31 , 32, 33, 34, 35, 36, 37, 38, 39, 40, 41 ,

[0191] 42, 43, 44, 45, 46, 47, 48, 49, or 50 nucleotides in length). In some embodiments, the ASO includes a sequence complementary at least 12 to 50 contiguous nucleotides (e.g., 12 to 50, 13 to 50, 14 to 50, 15 to 50, 16 to 50, 17 to 50, 18 to 50, 19 to 50, 20 to 50, 21 to 50, 22 to 50, 23 to 50, 24 to 50, 25 to 50, 26 to 50, 27 to 50, 28 to 50, 29 to 50, 30 to 50,

[0192] 31 to 50, 32 to 50, 33 to 50, 34 to 50, 35 to 50, 36 to 50, 37 to 50, 38 to 50, 39 to 50, 40 to 50, 41 to 50,

[0193] 42 to 50, 43 to 50, 44 to 50, 45 to 50, 46 to 50, 47 to 50, 48 to 50, or 49 to 50 contiguous nucleotides, e.g., 12, 13, 14, 15, 16, 17, 18, 19, 20, 21 , 22, 23, 24, 25, 26, 27, 28, 29, 30, 31 , 32, 33, 34, 35, 36, 37,

[0194] 38, 39, 40, 41 , 42, 43, 44, 45, 46, 47, 48, 49, or 50 contiguous nucleotides) set forth within SEQ ID NO: 14 (or a variant thereof, e.g., see Table 3).

[0195] It is within the scope of the disclosure that any length, known and previously unknown in the art, may be employed for the current invention.

[0196] The nucleotide sequence of the ASO may contain sufficient complementarity to a portion of a target gene of interest (e.g., DON (e.g., SEQ ID NO: 14), or a variant thereof) such that the ASO can hybridize with the target gene of interest. In some embodiments, the ASO is at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% complementary to the target gene of interest (e.g., DON (e.g., SEQ ID NO: 14), or a variant thereof), or a portion thereof. In some embodiments, the ASO is complementary to the target gene of interest (e.g., DON (e.g., SEQ ID NO: 14), or a variant thereof), or a portion thereof.

[0197] In some embodiments, the nucleotide sequence of the ASO may contain sufficient complementarity to an exon sequence of a target gene of interest (e.g., an exon of DON (e.g., SEQ ID NO: 14), or a variant thereof). In some embodiments, the nucleotide sequence of the ASO may contain sufficient complementarity to an intron sequence of a target gene of interest (e.g., an intron of DON (e.g., SEQ ID NO: 14), or a variant thereof). In some embodiments, the ASO of the disclosure may contain sufficient complementarity to a pre-mRNA transcript or an mRNA transcript encoding DON (e.g., SEQ ID NO: 14), or a variant thereof. The target sequence may be any one of SEQ ID NOs: 15-20 (e.g., see Table 2).

[0198] Different ASOs can be combined for decreasing the protein expression of a target gene of interest (e.g., DON (e.g., SEQ ID NO: 14), or a variant thereof, e.g., see Table 3). A combination of two ASOs may be used in a method of the invention, such as two different ASOs, three different ASOs, four different ASOs, or five different ASOs targeting the same gene of interest (e.g., DON (e.g., SEQ ID NO: 14), or a variant thereof, e.g., see Table 3).

[0199] GapmeR

[0200] GapmeRs of the disclosure are single (ss) nucleic acid molecules made of DNA and RNA with the central 8-10 nucleotide of the gapmeR being DNA that is complementary to a target gene of interest, which prevent translation of the target’s mRNA into a protein. Upon hybridization to a target mRNA, RNase H will degrade the mRNA by hydrolyzation, resulting in reduced mRNA and protein levels of the target.

[0201] In some embodiments, gapmeRs of the disclosure may include a nucleotide sequence of about 12 to about 50 nucleotides in length (e.g., 11 , 12, 13, 14, 15, 16, 17, 18, 19, 20, 21 , 22, 23, 24, 25, 26, 27, 28, 29, 30, 31 , 32, 33, 34, 35, 36, 37, 38, 39, 40, 41 , 42, 43, 44, 45, 46, 47, 48, 49, 50, or 51 nucleotides in length). In some embodiments, gapmeRs of the disclosure may include a nucleotide sequence of 12 to 50 nucleotides in length (e.g., 12 to 50, 13 to 50, 14 to 50, 15 to 50, 16 to 50, 17 to 50, 18 to 50, 19 to 50, 20 to 50, 21 to 50, 22 to 50, 23 to 50, 24 to 50, 25 to 50, 26 to 50, 27 to 50, 28 to 50, 29 to 50, 30 to 50, 31 to 50, 32 to 50, 33 to 50, 34 to 50, 35 to 50, 36 to 50, 37 to 50, 38 to 50, 39 to 50, 40 to 50, 41 to 50, 42 to 50, 43 to 50, 44 to 50, 45 to 50, 46 to 50, 47 to 50, 48 to 50, or 49 to 50 nucleotides in length, e.g., 12, 13, 14, 15, 16, 17, 18, 19, 20, 21 , 22, 23, 24, 25, 26, 27, 28, 29, 30, 31 , 32, 33, 34, 35, 36, 37, 38, 39, 40, 41 , 42, 43, 44, 45, 46, 47, 48, 49, or 50 nucleotides in length).

[0202] In some embodiments, gapmeRs of the disclosure may include a nucleotide sequence of 8 to 9, 8 to 10, or 9 to 10 (e.g., 8, 9, or 10) internal DNA nucleotides.

[0203] In some embodiments, the gapmeR includes a sequence complementary at least 12 to 50 contiguous nucleotides (e.g., 12 to 50, 13 to 50, 14 to 50, 15 to 50, 16 to 50, 17 to 50, 18 to 50, 19 to 50, 20 to 50, 21 to 50, 22 to 50, 23 to 50, 24 to 50, 25 to 50, 26 to 50, 27 to 50, 28 to 50, 29 to 50, 30 to 50,

[0204] 31 to 50, 32 to 50, 33 to 50, 34 to 50, 35 to 50, 36 to 50, 37 to 50, 38 to 50, 39 to 50, 40 to 50, 41 to 50,

[0205] 42 to 50, 43 to 50, 44 to 50, 45 to 50, 46 to 50, 47 to 50, 48 to 50, or 49 to 50 contiguous nucleotides, e.g., 12, 13, 14, 15, 16, 17, 18, 19, 20, 21 , 22, 23, 24, 25, 26, 27, 28, 29, 30, 31 , 32, 33, 34, 35, 36, 37,

[0206] 38, 39, 40, 41 , 42, 43, 44, 45, 46, 47, 48, 49, or 50 contiguous nucleotides) set forth within SEQ ID NO: 14 (or a variant thereof, e.g., see Table 3).

[0207] It is within the scope of the disclosure that any length, known and previously unknown in the art, may be employed for the current invention.

[0208] The nucleotide sequence of the gapmeR may contain sufficient complementarity to a portion of a target gene of interest (e.g., DCN (e.g., SEQ ID NO: 14), or a variant thereof) such that the gapmeR can hybridize with the target gene of interest. In some embodiments, the gapmeR is at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% complementary to the target gene of interest (e.g., DCN (e.g., SEQ ID NO: 14), or a variant thereof), or a portion thereof. In some embodiments, the gapmeR is complementary to the target gene of interest (e.g., DCN (e.g., SEQ ID NO: 14), or a variant thereof), or a portion thereof.

[0209] In some embodiments, the nucleotide sequence of the gapmeR may contain sufficient complementarity to an exon sequence of a target gene of interest (e.g., an exon of DCN (e.g., SEQ ID NO: 14), or a variant thereof). In some embodiments, the nucleotide sequence of the gapmeR may contain sufficient complementarity to an intron sequence of a target gene of interest (e.g., an intron of DCN (e.g., SEQ ID NO: 14), or a variant thereof). In some embodiments, the gapmeR of the disclosure may contain sufficient complementarity to a pre-mRNA transcript or an mRNA transcript encoding DCN (e.g., SEQ ID NO: 14), or a variant thereof. The target sequence may be any one of SEQ ID NOs: 15-20 (e.g., see Table 2).

[0210] Different gapmeRs can be combined for decreasing the protein expression of a target gene of interest (e.g., DCN (e.g., SEQ ID NO: 14), or a variant thereof). A combination of two gapmeRs may be used in a method of the invention, such as two different gapmeRs, three different gapmeRs, four different gapmeRs, or five different gapmeRs targeting the same gene of interest (e.g., DCN (e.g., SEQ ID NO: 14), or a variant thereof). Modifications to the Inhibitory Nucleic Acid Molecules

[0211] It is contemplated that any of the inhibitory nucleic acid molecules disclosed herein may be used in the methods disclosed herein in an unmodified or in a modified form. Unmodified inhibitory nucleic acid molecules contain nucleobases that include the purine bases adenine (A) and guanine (G), and the pyrimidine bases thymine (T), cytosine (C), and uracil (U). Modified nucleic acid molecules are described in more detail below.

[0212] Modifications may be achieved by systematically adding or removing linked nucleosides to generate longer or shorter sequences.

[0213] Modifications may be achieved by incorporating, for example, one or more alternative nucleosides, alternative 2’ sugar moieties, and / or alternative internucleoside linkages, which are described further below. Typically, these types of modifications are introduced to optimize the molecule’s efficacy or biophysical properties (e.g., increasing serum stability or circulating half-life, increasing thermal stability, enhancing transmembrane delivery, reduce immunogenicity, and / or targeting to a particular location or cell type).

[0214] Modification may further be achieved by covalently or non-covalently conjugating a moiety (e.g., a targeting moiety, a hydrophobic moiety, a cell penetrating peptide, or a polymer) to the 5’ end and / or 3’ end of the inhibitory nucleic acid molecule, as described in more detail below. In some embodiments, the targeting moiety is vascular ceil adhesion protein 1 (VCAM1). In some embodiments, the targeting moiety is arginylglycylaspartic acid (RGD).

[0215] Nucleoside Modifications

[0216] Modification of the inhibitory nucleic acid molecules described herein include one or more of the following nucleoside modifications: 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 (-C=C-CH3) uracil and cytosine and other alkynyl derivatives of pyrimidine bases, 6-azo uracil, cytosine and thymine, 5-uracil (pseudouracil), 4-thiouracil, 8-halo, 8-amino, 8-thiol, 8- thioalkyl, 8-hydroxyl and other 8-substituted adenines and guanines, 5-halo particularly 5-bromo, 5- trifluoromethyl and other 5-substituted uracils and cytosines, 7-methylguanine and 7-methyladenine, 2-F- adenine, 2-amino-adenine, 8-azaguanine and 8-azaadenine, 7-deazaguanine and 7-deazaadenine, and / or 3-deazaguanine and 3-deazaadenine. The inhibitory nucleic acid molecules may also include nucleobases in which the purine or pyrimidine base is replaced with other heterocycles, for example 7- deaza-adenine, 7-deazaguanosine, 2-aminopyridine, and / or 2-pyridone. Further modification of the inhibitory nucleic acid molecules described herein may include nucleobases disclosed in US 3,687,808; Kroschwitz, J. I., ed. The Concise Encyclopedia of Polymer Science and Engineering, New York, John Wiley & Sons, 1990, pp. 858-859; Englisch et al., Angewandte Chemie, International Edition 30:613, 1991 ; and Sanghvi, Y.S., Chapter 16, Antisense Research and Applications, CRC Press, Gait, M.J. ed., 1993, pp. 289-302. Sugar Modifications

[0217] Modifications of the inhibitory nucleic acid molecules described herein may also include one or more of the following 2’ sugar modifications: 2’-O-methyl (2’-0-Me), 2'-methoxyethoxy (2'-O- CH2CH2OCH3, also known as 2'-O-(2-methoxyethyl) or 2'-MOE), 2'-dimethylaminooxyethoxy, i.e. , a O(CH2)2ON(CH3)2 group, also known as 2'-DMAOE, and / or 2'-dimethylaminoethoxyethoxy (also known in the art as 2'-O-dimethylamino-ethoxy-ethyl or 2'-DMAEOE), i.e., 2'-O-CH2OCH2N(CH3)2. Other possible 2'-modifications that can modify the inhibitory nucleic acid molecules described herein include all possible orientations of OH; F; O-, S-, or N-alkyl; O-, S-, or N-alkenyl; O-, S- or N-alkynyl; or O-alkyl-O- alkyl, wherein the alkyl, alkenyl and alkynyl may be substituted or unsubstituted C1 to C10 alkyl or C2 to C10 alkenyl and alkynyl. Other potential sugar substituent groups include, e.g., aminopropoxy (- OCH2CH2CH2NH2), allyl (-CH2-CH=CH2), -O-allyl (-O-CH2-CH=CH2) and fluoro (F). 2'-sugar substituent groups may be in the arabino (up) position or ribo (down) position. In some embodiments, the 2'-arabino modification is 2'-F. Similar modifications may also be made at other positions on the interfering RNA molecule, particularly the 3' position of the sugar on the 3' terminal nucleoside or in 2'-5' linked oligonucleotides and the 5' position of 5' terminal nucleotide. Oligonucleotides may also have sugar mimetics such as cyclobutyl moieties in place of the pentofuranosyl sugar.

[0218] Internucleoside Linkage Modifications

[0219] Modifications of the inhibitory nucleic acid molecules described herein may include one or more of the following internucleoside modifications: phosphorothioates, phosphorodithioates, phosphotriesters, aminoalkylphosphotriesters, methyl and other alkyl phosphonates including 3'-alkylene phosphonates, 5'- alkylene phosphonates, phosphinates, phosphoramidates including 3'-amino phosphoramidate and aminoalkylphosphoramidates, thionophosphoramidates, thionoalkylphosphonates, thionoalkylphosphotriesters, selenophosphates, and boranophosphates having normal 3'-5' linkages, 2'-5' linked analogs of these, and those having inverted polarity wherein one or more internucleotide linkages is a 3' to 3', 5' to 5' or 2' to 2' linkage.

[0220] Conjugates

[0221] Any of the inhibitory nucleic acid molecules described herein may be modified via the addition of an auxiliary moiety, e.g., a cell penetrating peptide (CPP), a polymer, a hydrophobic moiety, or a targeting moiety. The auxiliary moiety may be present as a 5’ terminal modification (e.g., covalently bonded to a 5’- terminal nucleoside), a 3’ terminal modification (e.g., covalently bonded to a 3’-terminal nucleoside), or an internucleoside linkage (e.g., covalently bonded to phosphate or phosphorothioate in an internucleoside linkage).

[0222] CPPs are known in the art (e.g., TAT or Arg8) (Snyder and Dowdy, 2005, Expert Opin. Drug Deliv. 2, 43-51 ). Specific examples of CPPs are provided in WO2011157713, which is incorporated herein by reference in its entirety.

[0223] Inhibitory nucleic acid molecules of the disclosure may include covalently attached neutral polymer-based auxiliary moieties. Neutral polymers include poly(C1 -6 alkylene oxide), e.g., polyethylene glycol) and polypropylene glycol) and copolymers thereof, e.g., di- and triblock copolymers. An inhibitory nucleic acid molecule containing a hydrophobic moiety may exhibit superior cellular uptake, as compared to an inhibitory nucleic acid molecule lacking the hydrophobic moiety. A hydrophobic moiety is a monovalent group (e.g., a bile acid (e.g., cholic acid, taurocholic acid, deoxycholic acid, oleyl lithocholic acid, or oleoyl cholenic acid), glycolipid, phospholipid, sphingolipid, isoprenoid, vitamin, saturated fatty acid, unsaturated fatty acid, fatty acid ester, triglyceride, pyrene, porphyrine, texaphyrine, adamantine, acridine, biotin, coumarin, fluorescein, rhodamine, Texas-Red, digoxygenin, dimethoxytrityl, t-butydimethylsilyl, t-butyldiphenylsilyl, cyanine dye (e.g., Cy3 or Cy5), Hoechst 33258 dye, psoralen, or ibuprofen) covalently linked to the nucleic acid backbone (e.g., 5’- terminus) of the inhibitory nucleic acid molecule.

[0224] A targeting moiety is selected based on its ability to target oligonucleotides of the invention to a desired or selected cell population that expresses the corresponding binding partner (e.g., either the corresponding receptor or ligand) for the selected targeting moiety. For example, an oligonucleotide of the invention could be targeted to hepatocytes expressing asialoglycoprotein receptor (ASGP-R) by selecting a targeting moiety containing N-acetylgalactosamine (GalNAc).

[0225] In some embodiments, the targeting moiety is vascular cell adhesion protein 1 (VCAM1). In some embodiments, the targeting moiety is arginylglycylaspartic acid (RGD).

[0226] A targeting moiety may include one or more ligands (e.g., 1 to 9 ligands, 1 to 6 ligands, 1 to 3 ligands, 3 ligands, or 1 ligand). The ligand may target a cell expressing asialoglycoprotein receptor (ASGP-R), IgA receptor, HDL receptor, LDL receptor, or transferrin receptor. Non-limiting examples of the ligands include N-acetylgalactosamine (e.g., a triantennary N-acetylgalactosamine), glycyrrhetinic acid, glycyrrhizin, lactobionic acid, lactoferrin, IgA, or a bile acid (e.g., lithocholyltaurine or taurocholic acid).

[0227] The ligand may be a small molecule, e.g., a small molecule targeting a cell expressing asialoglycoprotein receptor (ASGP-R). A non-limiting example of a small molecule targeting an asialoglycoprotein receptor is N-acetylgalactosamine. Alternatively, the ligand can be an antibody or an antigen-binding fragment or an engineered derivative thereof (e.g., Fcab or a fusion protein (e.g., scFv)).

[0228] Preparation of Inhibitory Nucleic Acid Molecules

[0229] Inhibitory nucleic acid molecules of the disclosure may be prepared using techniques and methods known in the art for the oligonucleotide synthesis. For example, inhibitory nucleic acid molecules of the disclosure may be prepared using a phosphoramidite-based synthesis cycle. This synthesis cycle includes the steps of (1 ) de-blocking a 5’-protected nucleotide to produce a 5’-deblocked nucleotide, (2) coupling the 5’-deblocked nucleotide with a 5’-protected nucleoside phosphoramidite to produce nucleosides linked through a phosphite, (3) repeating steps (1 ) and (2) one or more times as needed, (4) capping the 5’-terminus, and (5) oxidation or sulfurization of internucleoside phosphites. The reagents and reaction conditions useful for the oligonucleotide synthesis are known in the art.

[0230] The inhibitory nucleic acid molecules disclosed herein may be linked to solid support as a result of solid-phase synthesis. Cleavable solid supports that may be used are known in the art. Non-limiting examples of the solid support include, e.g., controlled pore glass or macroporous polystyrene bonded to a strand through a cleavable linker (e.g., succinate-based linker) known in the art (e.g., UnyLinkerTM). A nucleic acid linked to solid support may be removed from the solid support by cleaving the linker connecting a nucleic acid and solid support. Compositions

[0231] The inhibitory nucleic acid molecules described herein may be formulated into various compositions (e.g., a pharmaceutical composition) for administration to a subject in a biologically compatible form suitable for administration in vivo. For example, the inhibitory nucleic acid molecules described herein (e.g., the siRNA molecules of SEQ ID NOs: 1 -12, or variants thereof) may be administered in a suitable diluent, carrier, or excipient, and may further contain a preservative, e.g., to prevent the growth of microorganisms. Conventional procedures and ingredients for the selection and preparation of suitable compositions are described, for example, in Remington, J.P. The Science and Practice of Pharmacy, Easton, PA. Mack Publishers, 2012, 22nded. And in The United States Pharmacopeial Convention, The National Formulary, United States Pharmacopeial, 2015, USP 38 NF 33).

[0232] Although the descriptions of pharmaceutical compositions provided herein are principally directed to pharmaceutical compositions which are suitable for administration to humans, it will be understood by the skilled artisan that such compositions are generally suitable for administration to any other animal, e.g., to non-human animals, e.g. non-human mammals. Modification of pharmaceutical compositions suitable for administration to humans to render the compositions suitable for administration to various animals is well understood, and the ordinarily skilled veterinary pharmacologist can design and / or perform such modification with merely ordinary, if any, experimentation. Subjects to which administration of the pharmaceutical compositions is contemplated include, but are not limited to, humans and / or other primates and mammals.

[0233] Compositions containing the inhibitory nucleic acids described herein may further include a second therapeutic agent (e.g., a nucleic acid molecule to be expressed within a cell, a polypeptide, or a drug). For example, a second therapeutic agent may be a blood pressure medication, an antiinflammatory medication (e.g., a steroid or colchicine), or immunosuppressive agent. In some embodiments, the second therapeutic agent is a statin. Non-limiting examples of second therapeutic agents are a statin (e.g., atorvastatin), a proprotein convertase subtilisin / kexin type 9 (PCSK9) inhibitor (e.g., an siRNA or monoclonal antibody targeting PCSK9), hepatocyte growth factor (HGF), inclisiran (e.g., LEQVIO™), or ezetimibe (e.g., ZETIA™).

[0234] In some embodiments, the second therapeutic agent (e.g., statin) is administered in combination with an inhibitory nucleic acid molecule of the disclosure. In some embodiments, the subject is orally administered a statin. In some embodiments, the subject is administered a statin daily.

[0235] Methods of Treatment

[0236] The disclosure provides methods of (i) treating a medical condition resulting from a myocardial infarction (Ml), and / or (ii) promoting angiogenesis in a subject. In some embodiments, the method includes the step of administering to a subject an inhibitory nucleic acid molecule described herein, wherein the inhibitory nucleic acid molecule targets DCN (e.g., SEQ ID NO: 14), or a variant thereof (e.g., see Table 3). In some embodiments, the method includes the step of administering to a subject an siRNA described herein (e.g., any one or more of SEQ ID NOs: 1 -12, or a variant thereof), wherein the siRNA targets DCN (e.g., SEQ ID NO: 14), or a variant thereof (e.g., see Table 3). In some embodiments, the method includes the step of administering to a subject an dsRNA described herein, wherein the dsRNA targets DCN (e.g., SEQ ID NO: 14), or a variant thereof (e.g., see Table 3). In some embodiments, the method includes the step of administering to a subject an ASO described herein, wherein the ASO targets DCN (e.g., SEQ ID NO: 14), or a variant thereof (e.g., see Table 3). In some embodiments, the method includes the step of administering to a subject a gapmeR described herein, wherein the gapmeR targets DCN (e.g., SEQ ID NO: 14), or a variant thereof (e.g., see Table 3). In some embodiments, the method includes the step of administering to a subject an miRNA described herein, wherein the miRNA targets DCN (e.g., SEQ ID NO: 14), or a variant thereof (e.g., see Table 3). In some embodiments, the method includes the step of administering to a subject an shRNA described herein, wherein the shRNA targets DCN (e.g., SEQ ID NO: 14), or a variant thereof (e.g., see Table 3).

[0237] In some embodiments, the medical complication is an arrhythmic-related complication (e.g., a heart block, an atrial arrhythmia, and / or a ventricular arrhythmia), an ischemic-related complication (e.g., reinfarction, peri-infarct ischemia, and / or an infarct extension), a mechanical-related complication (e.g., a mitral valve rupture or tear, a chordae rupture or tear, a ventricular septal defect (VSD), a ventricular free wall rupture, a cardiac tamponade, and / or an aneurysm), an inflammatory-related complication (e.g., pericarditis and / or Dressier syndrome), and / or a systemic complication (e.g., cardiogenic shock, cardiomyopathy, heart failure, an embolic stroke, a systemic embolism, and / or a lower extremity embolism).

[0238] Any of the methods can administer a composition (e.g., a pharmaceutical composition) or delivery vehicle (e.g., a vector or nanoparticle) that contains or expresses any of the inhibitory nucleic acid molecules described herein (e.g., siRNA, dsRNA, miRNA, shRNA, ASO, or gapmeR).

[0239] In some embodiments, the methods of (i) treating a medical condition resulting from a myocardial infarction (Ml), and / or (ii) promoting angiogenesis in a subject further includes administering a second therapeutic agent (e.g., a nucleic acid molecule to be expressed within a cell, a polypeptide, or a drug). For example, a second therapeutic agent may be a blood pressure medication, an anti-inflammatory medication (e.g., a steroid or colchicine), or immunosuppressive agent. In some embodiments, the second therapeutic agent is a statin. Non-limiting examples of second therapeutic agents are a statin (e.g., atorvastatin), a proprotein convertase subtilisin / kexin type 9 (PCSK9) inhibitor (e.g., an siRNA or monoclonal antibody targeting PCSK9), hepatocyte growth factor (HGF), inclisiran (e.g., LEQVIO™), or ezetimibe (e.g., ZETIA™). In some embodiments, the additional therapeutic agent can be administered prior to, subsequent to, or concurrently with an inhibitory nucleic acid described herein.

[0240] Delivery Vehicle

[0241] The inhibitory nucleic acid molecule of the disclosure may be delivered to a subject (e.g., a human) using any suitable delivery vehicle. For example, a delivery vehicle for any of the inhibitory nucleic acid molecules described herein may be a vector, plasmid, or nano particle, (e.g., a micelle, a liposome, an exosome, or a lipid nano particle (LNP)).

[0242] The inhibitory nucleic acid molecule of the disclosure and compositions thereof may be delivered to a subject via a vector (e.g., a viral vector). Any suitable viral vector system can be used including, e.g., adenoviruses (e.g., Ad2, Ad5, Ad9, Ad15, Ad17, Ad19, Ad20, Ad22, Ad26, Ad27, Ad28, Ad30, or Ad39), rhabdoviruses (e.g., vesicular stomatitis virus), retroviruses, adeno-associated vectors (AAV), poxviruses, herpes viral vectors, and Sindbis viral vectors. For example, the vector may be an AAV vAAV1 , AAV2, AAV3, AAV4, AAV5, AAV6, AAV7, AAV8, AAV9, AAV10, AAV11 , or AAV12 vector.

[0243] The inhibitory nucleic acid molecule of the disclosure and compositions thereof may be delivered to a subject via liposomes. Liposomes are artificially-prepared vesicles which may primarily be composed of a lipid bilayer and may be used as a delivery vehicle for the administration of the inhibitory nucleic acids described herein, and compositions thereof. Liposomes can be of different sizes such as, but not limited to, a multilamellar vesicle (MLV) which may be hundreds of nanometers in diameter and may contain a series of concentric bilayers separated by narrow aqueous compartments, a small unicellular vesicle (SUV) which may be smaller than 50 nm in diameter, and a large unilamellar vesicle (LUV) which may be between 50 and 500 nm in diameter. Liposome design may include, but is not limited to, opsonins or ligands in order to improve the attachment of liposomes to unhealthy tissue or to activate events such as, but not limited to, endocytosis. Liposomes may contain a low or a high pH in order to improve the delivery of the pharmaceutical composition.

[0244] The inhibitory nucleic acid molecule of the disclosure and compositions thereof may be delivered to a subject via exosomes. Exosomes produced from cells can be collected from cell culture medium by any suitable method. Typically, a preparation of exosomes can be prepared from cell culture or tissue supernatant by centrifugation, filtration or combinations of these methods. For example, using standard methods, exosomes can be prepared by differential centrifugation, that is low speed (<20000 g) centrifugation to pellet larger particles followed by high speed (>100000 g) centrifugation to pellet exosomes, size filtration with appropriate filters (for example, 0.22 micrometer filter), gradient ultracentrifugation (for example, with sucrose gradient) or a combination of these methods.

[0245] The inhibitory nucleic acid molecules of the disclosure, and compositions thereof, may be delivered to a subject via LNPs. For example, the inhibitory nucleic acid molecules (e.g., siRNA, dsRNA, miRNA, shRNA, ASO, or gapmeR) may be formulated in a lipid nanoparticle such as those described in International Publication No. WO2012170930, herein incorporated by reference in its entirety. As a nonlimiting example, LNP formulations may contain cationic lipids, distearoylphosphatidylcholine (DSPC), cholesterol, polyethylene glycol (PEG), R-3-[(w-methoxy polyethylene glycol)2000)carbamoyl)]-1 ,2- dimyristyloxl-propyl-3-amine (PEG-c-DOMG), distearoyl-rac-glycerol (DSG) and / or dimethylaminobutanoate (DMA). As a non-limiting example, 1 -5% of the lipid molar ratio of PEG-c-DOMG as compared to the cationic lipid, DSPC and cholesterol. In another embodiment the PEG-c-DOMG may be replaced with a PEG lipid such as, but not limited to, PEG-DSG (1 ,2-Distearoyl-sn-glycerol, methoxypoly ethylene glycol) or PEG-DPG (1 ,2-Dipalmitoyl-sn-glycerol, methoxypolyethylene glycol). The cationic lipid may be selected from any lipid known in the art such as, but not limited to, (6Z,9Z,28Z,31Z)- heptatriacont-6,9,28,31 -tetraene-19-yl 4-(dimethylamino)butanoate (DLin-MC3-DMA), 1 ,2-dil inoleyloxy- n,n-dimethyl-3-aminopropane (DLin-DMA), C 12-200, and N,N-dimethyl-2,2-di-(9Z,12Z)-9,12- octadecadien-1 -yl-1 ,3-dioxolane-4-ethanamine (DLin-KC2-DMA).

[0246] Exemplary commercial reagents useful for lipid-based delivery of inhibitory nucleic acid molecules including, but not limited to, TransIT-TKO™ (Mirus, Catalog No. MIR 2150), Transmessenger™ (Qiagen, Catalog No. 301525), Oligofectamine™ and Lipofectamine™ (Invitrogen, Catalog No. MIR 12252-011 and Catalog No. 13778-075), siPORT™ (Ambion, Catalog No. 1631 ), and DharmaFECT™ (Fisher Scientific, Catalog No. T-2001 -01 ). Subject

[0247] The subject to be treated may have previously experienced a myocardial infarction (Ml). The subject to be treated may have an ischemic injury, which may have occurred as a result of the Ml. The subject to be treated may have a cardiovascular disease, including, but not limited to, coronary artery disease, peripheral artery disease. The subject may also have, or be at risk of developing, a stroke. Additionally, the subject to be treated may have a metabolic disorder, or is at risk of developing a metabolic disorder, such as diabetes. Subjects at risk of developing diabetes may be prediabetic and / or have experienced one or more of the following risk factors: hyperglycemia, glucose resistance, insulin resistance, hyperlipidemia, or has a family history of diabetes.

[0248] The inhibitory nucleic acid molecules of the disclosure, and compositions thereof, may be delivered to the subject’s coronary endothelium, remote zone of the heart, and / or border zone of the heart. The inhibitory nucleic acid molecules of the disclosure, and compositions thereof, may be delivered to an endothelial cell in the subject (e.g., an endothelial cell, a cardiomyocyte, a fibroblast, a vascular smooth muscle cell, and / or a leukocyte in the subject’s coronary endothelium, remote zone, and / or border zone). Such delivery can promote angiogenesis in the subject, e.g., by stimulating endothelial cell proliferation.

[0249] Dosage

[0250] The actual dosage amount of a composition of the present disclosure administered to a subject can be determined by physical and physiological factors such as body weight, severity of condition, previous or concurrent therapeutic interventions, idiopathy of the patient and on the route of administration. Depending upon the dosage (e.g., mg / kg) and the route of administration, the number of administrations of a preferred dosage and / or an effective amount may vary according to the response of the subject. The practitioner responsible for administration will, in any event, determine the concentration of active ingredient(s) in a composition and appropriate dose(s) for the individual subject. Administration may occur any suitable number of times per day, and for as long as necessary. Subjects may be adult or pediatric humans, with or without comorbid diseases.

[0251] Routes of Administration

[0252] The compositions utilized in the methods described herein can be administered to a subject by any suitable route of administration. For example, a composition containing an inhibitory nucleic acid of the disclosure may be administered intramuscularly, intravenously, intradermally, percutaneously, intraarterially, intraperitoneally, intralesionally, intracranially, intraarticularly, intraprostatically, intrapleurally, intratracheally, intranasally, intravitreally, intravaginally, intrarectally, topically, intratumorally, peritoneally, subcutaneously, subconjunctivally, intravesicularlly, mucosally, intrapericardially, intraumbilically, intraocularly, orally, topically, locally, by inhalation, by injection, by infusion, by continuous infusion, by localized perfusion bathing target cells directly, by catheter, by lavage, in cremes, or in lipid compositions.

[0253] In some embodiments, the compositions utilized in the methods described herein can be administered to the subject intravenously. In some embodiments, the compositions utilized in the methods described herein can be administered to the subject subcutaneously. In some embodiments, the compositions utilized in the methods described herein can be administered to the subject intraarticularly. In some embodiments, the compositions utilized in the methods described herein can be administered to the subject intramuscularly.

[0254] EXAMPLE

[0255] The following example is put forth so as to provide those of ordinary skill in the art with a description of how the compositions and methods described herein may be used and evaluated and are intended to be purely exemplary of the invention and is not intended to limit the scope of what the inventors regard as their invention.

[0256] Example 1. MicroRNA-342-3p Rescues Post-Ischemic Cardiac Function in Diabetic Mice via Activation of the HGF-MET Pathway

[0257] This example describes the discovery of miR-342-3p’s role in regulating angiogenesis under physiologic and diabetic conditions. Additionally, this example describes the discovery of decorin (DCN) as a therapeutic target for promoting post-ischemic angiogenesis, which can be used to treat or reduce the likelihood of a medical complication resulting from myocardial infarction (Ml).

[0258] Briefly, microRNAs (miRNAs) were sequenced from cardiac endothelial cells (ECs) obtained from Ldlr / - mice on either chow or high fat sucrose containing (HFSC) diet at 0-, 3-, 7- and 14-days post-MI. Among the top differentially expressed miRNAs at day 14 was miR-342-3p (FIG. 11 A). In vitro, overexpression of miR-342-3p promotes angiogenic sprouting and migration in endothelial cells (ECs) by activating the hepatocyte growth factor (HGF)-mesenchymal-epithelial transition factor (MET) signaling pathway. Delivery of miR-342-3p mimics to the border zone increased capillary density, EC proliferation, and cardiac function post-MI. Ejection fraction (EF) and fractional shortening (FS) of miR-342-3p injected hearts were significantly higher by 145% and 173%, respectively, at day 14 compared to controls (p = 0.0204 and p = 0.0095). Capillary density as well as ki67+ endothelial cells (ECs) were more abundant by 107% and 154%, respectively (p = 0.0371 and p = 0.0242) vs control. DCN was identified as a direct target of miR-342-3p and validated by Western blot (WB) and luciferase reporter assays. RNA sequencing and subsequent WBs implicated HGF-MET signaling as one of the top upregulated pathways in miR-342-3p overexpressing ECs (FIG. 11 B). Hypoxia (2% O2) induced miR-342-3p expression (p = 0.0023) while suppressing DCN levels. However, both high glucose (25 mM) and palmitate (100 pM) treatments had the opposite effect. DCN and MET binding was confirmed by co-immunoprecipitation assay. These findings establish a critical role for miR-342-3p in regulating cardiac angiogenesis post- ischemic injury in diabetic mice. Under diabetic conditions, the hypoxia induced expression of miR-342-3p is blunted, leading to DCN mediated degradation of MET receptors, which impairs angiogenesis (FIG. 7). These findings, as discussed further below, indicate that miR-342-3p plays a protective role in post- ischemic angiogenesis in diabetic hearts following Ml.

[0259] Results

[0260] Identification of commonly dysregulated miRNAs between healthy and diabetic mice post-MI To investigate the role of miRNAs in post-ischemic angiogenesis, a diet induced diabetes model was utilized. Male Ldlr mice on chow or high fat sucrose containing (HFSC) diet for 14 weeks were subjected to permanent left anterior descending (LAD) coronary ligation. Mice were sacrificed at 0-, 3-, 7- or 14-days post-surgery. EC and non-EC fractions were obtained from the hearts using cluster of differentiation (CD)-31 magnetic bead-based cell isolation. RNA from the EC fraction was sequenced and the raw data analyzed for differentially expressed (fold change (FC) > 1 .5, adjusted p value < 0.05) miRNAs between chow and HFSC groups. The resulting list of miRNAs had unique expression kinetics across different time points, indicating the diverse roles played by miRNAs during various phases following ischemic insult, e.g., inflammation, angiogenesis, and vascular remodeling. For this study, day- 14, the peak of the angiogenesis phase, was interrogated. miR-342-3p was among the top dysregulated miRNAs from this list. From the miRNA-seq data, miR-342-3p was higher in the chow group vs the HFSC group at day 14 (FIG. 1 A), hinting at its potential pro-angiogenic role in healthy ECs. As diabetic mice have been shown to have an impaired angiogenic response, lower expression of miR-342 in the coronary endothelium of HFSC diet fed mice could be one of the factors impeding proper post-ischemic angiogenesis.

[0261] In vitro overexpression of miR-342-3p promotes angiogenesis in endothelial cells

[0262] To test whether miR-342 promotes angiogenesis in ECs, human umbilical vein endothelial cells (HUVECs) were transfected with either non-specific (NSm) or miR-342 mimic (rniR-342m), followed by spheroid sprouting assay, scratch wound closure assay, and BrdU incorporation (proliferation) assay. Spheroids overexpressing miR-342 (rniR-342mgroup) exhibited significantly more sprout numbers compared to the control group (NSm) (FIG. 1 B and FIG. 1 C). The cumulative sprout length was also higher in rniR-342mgroup. Concurrently, spheroids composed of miR-342 inhibitor (miR-342i) transfected HUVECs showed much lower number of sprouts as well as cumulative sprout length vs non-specific (NS) control (NSi) (FIG. 1 B and FIG. 1 C). To assess migration and wound healing aspects of miR-342, scratch closure assays were performed. Scratches created in a monolayer of rniR-342mtransfected HUVECs closed significantly faster compared to those in the NSm group (FIG. 1 D and FIG. 1 E). This was confirmed by area under curve (AUC) analysis (FIG. 1 F). In contrast, the miR-342i scratches failed to close completely even after 24 hours (FIG. 1 D and FIG. 1 E). To quantify the proliferation, HUVECs transfected with either control or miR-342 mimic were incubated with the nucleoside analogue, BrdU (5-Bromo-2'- deoxyuridine). Cells in the synthesis (s) phase of cell division incorporated this thymidine analog in their DNA. A colorimetric test was used to quantify the BrdU levels in both groups. rniR-342mHUVECs had significantly higher BrdU incorporation vs control, while inhibition of miR-342 reduced endothelial proliferation dramatically (FIG. 1 G)

[0263] Identification of DCN as a target of miR-342-3p

[0264] Predicted miR-342-3p targets were cross-referenced with the top downregulated mRNAs (day 0 vs day 14) in the chow group (FIG. 2A) and arrived at four potential targets. Of these, only DCN was significantly downregulated at the protein level (FIG. 2B and FIG. 2C). DCN is a small leucine rich protein which stabilizes collagen fibers and is a component of the extracellular matrix. Soluble DCN is secreted and has been shown to bind to various extracellular receptors by competing with their canonical ligands. While miR-342 overexpression repressed DCN protein, its inhibition promoted DCN expression, suggesting that DCN is a novel target of miR-342. DCN mRNA was confirmed as a target of miR-342 using a 3’-UTR luciferase reporter assay (FIG. 2D). Luciferase activity was significantly reduced in wildtype DCN 3’ UTR transfected cells when co-transfected with miR-342 mimic vs scramble control (FIG. 2D). Mutation of the 3’ UTR seed sequence abolished transcriptional repression by miR-342 (FIG. 2D). miR-342 overexpression promotes HGF-MET signaling in ECs

[0265] In diabetes, many signaling pathways are dysregulated, leading to impaired angiogenesis. To determine the signaling pathways downstream of miR-342, RNA from miR-342 overexpressing HUVECs was sequenced and analyzed for differentially expressed (DE) genes (DEGs). RNA-seq results showed that one of the top upregulated genes was MET, which codes for the c-MET receptor. HGF is the natural ligand of the MET receptor. HGF-MET signaling is a well-studied pathway involved in cell proliferation[26- 28]. It has been implicated in angiogenesis in diverse tissue types and in different disease contexts[29- 31]. Concordantly, Ingenuity Pathway Analysis (IPA) revealed HGF-MET signaling as one of the top upregulated pathways (FIG. 2E). MET receptor mRNA and protein was upregulated in ECs transfected with miR-342 mimic vs control at the transcript and protein level (FIG. 2F and FIG. 2G), validating this studies RNA-seq results. miR-342 also induced HGF mRNA expression (FIG. 2H). This result is also mirrored in the RNA-seq data from the initial Ml study, where MET mRNA levels were higher in the chow group at day 14 vs HFSC (data not shown). miR-342-3p expression is responsive to hypoxia, high glucose, and palmitate treatments

[0266] As the cell type continuously exposed to the blood, ECs can sense and respond to diverse stimuli such as oxygen concentration, circulating glucose, fatty acid levels, or paracrine factors. To recapitulate the effects of ischemia in the diabetic environment (hyperglycemia / dyslipidemia), ECs were pre-treated with either 25 mM mannitol or 25 mM glucose for 48 hours under normoxic conditions, followed by 0, 2, 16 or 24 hours of hypoxia (2% O2). In a parallel experiment, ECs were treated with 100 uM Palmitate or 100 uM BSA and 2% O2 for 0, 2, 16 or 24 hours. Hypoxia induced miR-342-3p expression at 2 hours and 16 hours vs control (normoxia), while suppressing DCN levels at the same time points. MET mRNA expression reached significance over control after 16 and 24 hours of hypoxia as well (FIG. 3C). Treatment with 25 mM glucose significantly attenuated the induction of miR-342 and MET transcripts, while boosting DCN mRNA expression at all time points (FIGS. 3A-3C). Similarly, palmitate treatment blocked hypoxia-driven induction of miR-342 and MET mRNA, while significantly promoting DCN expression (FIGS. 3D-3F). This pattern of glucose or palmitate mediated induction of DCN also held true at the protein level (FIG. 8A and FIG. 8B). These data show that not only is miR-342 responsive to hypoxia, glucose and palmitate, but also that the inverse relationship between it and DCN is preserved even under these pathologic conditions. MET expression pattern under hypoxic and hyperglycemic or dyslipidemic conditions also supports its role as the downstream effector of miR-342’s angiogenic program. Under normoxia, high glucose treatment (72 hours) significantly reduced MET protein, while inducing DCN production (FIG. 3B and FIG. 3C). Concurrently, miR-342 expression was suppressed in the high glucose treated ECs (FIG. 3A). In vivo administration of miR-342-3p mimic rescues cardiac function and promotes angiogenesis following ischemia / reperfusion injury

[0267] The observed pro-angiogenic effects of miR-342 in vitro suggests that exogenous administration or overexpression of miR-342 can rescue impaired post-ischemic angiogenesis in diabetic mice. Male Ldlr- / - mice were placed on a HFSC diet for 6 weeks, followed by temporary left anterior descending (LAD) coronary ligation surgery. Mice were subjected to 45 minutes of myocardial ischemia followed by reperfusion. Immediately before reperfusion, either lipid encapsulated non-specific control or miR-342 mimic was injected around the border zone of the heart (FIG. 4A). Echocardiography was used to assess cardiac function at days 3, 7 and 14 post-surgery (FIG. 4B). Mice continued their special diet during this period. Ejection fraction (EF) and fractional shortening (FS) was significantly improved in the miR-342 injected group vs control at day 14 (FIG. 4C and FIG. 4D). Following sacrifice at day 14, miR-342-3p overexpression was confirmed in the border zone by qPCR (FIG. 4D). DCN expression was significantly reduced in the miR-342 group vs control, while MET mRNA was upregulated (FIG. 4E and FIG. 4F). Immunofluorescence on heart sections revealed increased capillary density and EC proliferation in miR- 342 mimic vs control group (FIGS. 4G-I and FIG. 11 C).

[0268] Pro-angiogenic effects of miR-342 are dependent on the HGF-MET pathway

[0269] To determine if HGF-MET signaling is responsible for the pro-angiogenic effects of miR-342, ECs were transfected with scramble or miR-342 mimic and added a selective small molecule MET inhibitor, PHA-665752 or its vehicle (DMSO) and performed a proliferation assay using BrdU pulse-chase approach. While miR-342 overexpression in the vehicle group dramatically increased proliferation, the addition of the MET inhibitor significantly blunted this effect (FIG. 5A). This experiment was repeated under high glucose or mannitol conditions. Results show that the inhibitor reduced proliferation under basal conditions; but in the high glucose group ECs proliferated less, even in the absence of this inhibitor. The addition of PHA-665752 under high glucose conditions did not synergistically attenuate proliferation (FIG. 5E). On the other hand, miR-342 promoted proliferation in the mannitol group while the inhibitor blunted this effect. Although miR-342 significantly induced EC proliferation in the glucose treated group, the maximal proliferation achieved was lower compared to that in the mannitol group (FIG. 5E). Similar to the NSm group, addition of the MET inhibitor did not significantly reduce EC proliferation induced by miR- 342 overexpression. Similar results were obtained when HGF was added to the groups (FIG. 5F).

[0270] DCN exerts anti-proliferative effects by inhibiting HGF-MET signaling in ECs

[0271] In a similar set of experiments, recombinant DCN protein (rDCN) was added to ECs instead of the MET inhibitor under basal and high glucose conditions. ECs were transfected with either scramble control or miR-342 mimic with or without rDCN (10 nM). Under basal conditions, rDCN significantly reduced the proliferative effect of miR-342, while its addition had no effect on the scramble control group (FIG. 5B). Supplementation with HGF significantly increased proliferation in NSm and rniR-342mtreated groups, while addition of MET inhibitor or rDCN reduced it in the rniR-342mtreated group only (FIG. 5C and FIG. 5D). When added to cells treated with mannitol and transfected with control mimic (NSm), rDCN reduced proliferation (FIG. 5G and FIG. 5H). High glucose treated NSm cells were significantly less proliferative vs rnannitol-NSm. miR-342 rescued proliferation under high glucose conditions. Addition of rDCN did not inhibit proliferation in either NSm or rniR-342mgroups in the high glucose treated group (FIG. 5G and FIG. 5H). These results demonstrate the attenuation of miR-342 induced proliferation by DCN. As shown earlier, inhibition of MET signaling abolishes miR-342-induced endothelial proliferation. The effects of the MET inhibitor (PHA-665752) and rDCN on EC proliferation are strikingly similar across the experiments, supporting DCN as an inhibitor of the MET receptor and counters the pro-proliferative effect of miR-342.

[0272] DCN knockdown in ECs phenocopies pro-angiogenic effects of miR-342 overexpression

[0273] Having established DCN as a target of miR-342, the effect of its silencing on endothelial proliferation and angiogenesis was investigated, along with whether siRNA mediated knockdown (KD) of DCN phenocopied the effects of miR-342 overexpression. miR-342 overexpression suppressed DCN protein while boosting MET receptor expression (FIGS. 6A-6C). siDCN treatment knocked down DCN protein effectively while also significantly increasing MET protein (FIGS. 6D-6F). The increase in MET expression in siDCN treated cells phenocopies the effect of miR-342 overexpression. To this end, mannitol or glucose treated ECs were transfected with siDCN (ThermoFisher, USA) or a non- specific / scramble control (siNS) followed by a BrdU incorporation assay. As observed in similar experiments described above, high glucose treatment significantly reduced endothelial proliferation vs control. KD of DCN increased proliferation under both mannitol and high glucose conditions (p = 0.021 vs NSm and 0.0573 vs NSm, respectively) (FIG. 6G). DCN KD significantly upregulated MET protein under basal conditions, mirroring the effect of miR-342 overexpression (FIG. 6D and FIG. 6E). To assess the effect of DCN KD on angiogenesis, spheroid sprouting assays were performed on ECs transfected with either siNS or siDCN and cultured in either mannitol or high glucose containing media. In the mannitol treated group, siDCN spheroids had significantly more sprouts vs siNS spheroids (FIG. 6I and FIG. 6M). DCN KD also significantly increased the number of sprouts under high glucose conditions. The cumulative sprout length was also higher in siDCN group vs siNS under both mannitol and glucose conditions. Parallel experiments tested the effects of miR-342 overexpression (rniR-342mor NSm) on spheroid sprouting under similar conditions. rniR-342mspheroids exhibited enhanced sprouting vs NSm under both conditions (FIG. 6H and FIG. 6L). Further, supplementing the culture medium with rDCN abrogated the effects of miR-342 overexpression (FIG. 6J and FIG. 6N). Similarly, the addition of rDCN also suppressed siDCN mediated sprouting (FIG. 6K and FIG. 60). Taken together, these results demonstrate that DCN deficiency phenocopies the effects of miR-342 overexpression on angiogenesis and that DCN supplementation antagonizes these effects. miR-342-3p acts via the HGF-MET signaling pathway in both mice and humans

[0274] A BrdU proliferation assay demonstrated that EC proliferation was induced by HGF (50 ng / mL) supplementation in HUVECs transfected with a miR-342 mimic, relative to control (FIG. 9A). The addition of PHA-665752 blunted EC proliferation observed by the miR-342 mimic under normal (mannitol) conditions. HUVECs proliferated less under high glucose conditions even in the absence of the inhibitor, which did not synergistically reduce proliferation (FIG. 9B). miR-342 overexpression induced EC proliferation, while supplementation with rDCN attenuated proliferation in a manner similar to that of the MET inhibitor (FIG. 9C). A BrdU proliferation assay in HUVECS transfected with the miR-342 mimic was also performed with the addition of HGF (FIG. 9D). Notably, rDCNn acted similarly as in FIG. 9C, even in the presence of added HGF (FIG. 9E). Taken together, these data demonstrate that miR-342-3p acts via the HGF-MET signaling pathway in both mice and humans

[0275] Dose-response curves of PHA-665752 and DCN using the scratch closure assay.

[0276] Scratch closure rates in response to vehicle (DMSO) or increasing concentrations of PHA- 665752, a MET inhibitor, showed a slower rate of scratch closure in ECs when treated with 100 nM of the inhibitor (FIG. 10A and FIG. 10B). Scratch closure rates in response to increasing concentrations of rDCN protein show an effective level of inhibition of scratch closure at a 25 nM dose. Knocking down DCN, as described above, promotes EC proliferation and, therefore, scratch closure (FIGS. 1 D-1 F).

[0277] Materials and Methods

[0278] In vivo delivery of miR-342 mimics:

[0279] Male Ldlr- / - mice were placed on a high fat sucrose containing (HFSC) diet for 4 weeks, followed by temporary left anterior descending (LAD) coronary artery ligation surgery. Mice were subjected to 45 minutes of myocardial ischemia followed by reperfusion. Immediately before reperfusion, either lipid encapsulated miR-342-3p mimic or non-specific mimic (40 ng per animal) was injected into the border zone of the heart. Mimics were encapsulated with 20% (v / v) Lipofectamine 2000 reagent and delivered as a 20 pL volume (2 injections of 10 pL each) on the left and right margins of the border zone. Echocardiography was used to assess cardiac function at days 3, 7 and 14 post-surgery. Mice continued their special diet during this period. Mimics were encapsulated in Lipofectamine 2000 as described below:

[0280] 1 . 100 pM (7.47 pg / pL) solution of commercially available human miR-342-3p mimic (e.g., 5’-

[0281] UCUCACACAGAAAUCGCACCCGU-3’ - SEQ ID NO: 13), which is same sequence as mouse miR-342-3p mimic, or non-specific control was prepared by dissolving 250 nmol or 1867 ug of powder in 250 pL of nuclease-free water. One pL of this stock solution was added to 99 pL of sterile 1x PBS to get a 74.70 ng / pL working solution.

[0282] 2. 5.35 pL of the working solution from above was mixed with 94.65 pL of sterile 1x PBS (solution A) .

[0283] 3. 80 pL of sterile 1x PBS was mixed with 20 pL of Lipofectamine 2000 transfection reagent (solution B) .

[0284] 4. Solutions A and B were mixed together by gentle pipetting and incubated at room temperature (RT) for 15 minutes.

[0285] 5. The negative control or scramble mimic was prepared in a similar fashion.

[0286] 6. From the above mixture, 20 pL solution / mouse was used for injections. miR-342-3p sequences (same human and mouse):

[0287] Human - 5’-UCUCACACAGAAAUCGCACCCGU-3’ (SEQ ID NO: 13) Mouse - 5’-UCUCACACAGAAAUCGCACCCGU-3’ (SEQ ID NO: 13) siDecorin sequences used:

[0288] They each can target Human and Mouse - 5’-AGACAUCAACCUAGUUCACUGAAGA-3’ (SEQ ID NO: 1 ) 5’-GACAUCAACCUAGUUCACUGAAGAC-3’ (SEQ ID NO: 2) 5’-AGACAUCAACCUAGUUCACUGAAGA-3’ (SEQ ID NO: 3) 5’-AGACAUCAACCUAGUUCACUGAAGA-3’ (SEQ ID NO: 4) 5’-AGACAUCAACCUAGUUCACUGAAGA-3’ (SEQ ID NO: 5) 5’-CUAGUUCACUGAAGACGGGUGGAC-3’ (SEQ ID NO: 6)

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[0324] Numbered Embodiments

[0325] 1 . An inhibitory nucleic acid molecule comprising sufficient complementarity to a target nucleic acid molecule, wherein (i) the inhibitory nucleic acid molecule is at least 15 nucleotides in length, and (ii) the target nucleic acid molecule comprises a nucleotide sequence having at least 80% sequence identity to SEQ ID NO: 14. 2. The inhibitory nucleic acid molecule of embodiment 1 , wherein the target nucleic acid molecule comprises a nucleotide sequence having at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to SEQ ID NO: 14.

[0326] 3. The inhibitory nucleic acid molecule of embodiment 1 or 2, wherein the target nucleic acid molecule comprises the nucleotide sequence of SEQ ID NO: 14.

[0327] 4. The inhibitory nucleic acid molecule of any one of embodiments 1 -3, wherein the inhibitory nucleic acid molecule is 15 to 6,850 nucleotides in length.

[0328] 5. The inhibitory nucleic acid molecule of embodiment 4, wherein the inhibitory nucleic acid molecule is 15 to 49 nucleotides in length, 50 to 99 nucleotides in length, or 100 to 6,850 nucleotides in length.

[0329] 6. The inhibitory nucleic acid molecule of embodiment 5, wherein the inhibitory nucleic acid molecule is 20 to 28 nucleotides in length.

[0330] 7. The inhibitory nucleic acid molecule of embodiment 6, wherein the inhibitory nucleic acid molecule is 23 or 25 nucleotides in length.

[0331] 8. The inhibitory nucleic acid molecule of any one of embodiments 1 -7, wherein the inhibitory nucleic acid molecule comprises at least 85% complementarity to the target nucleic acid molecule.

[0332] 9. The inhibitory nucleic acid molecule of embodiment 8, wherein the inhibitory nucleic acid molecule comprises at least 90% complementarity to the target nucleic acid molecule.

[0333] 10. The inhibitory nucleic acid molecule of embodiment 9, wherein the inhibitory nucleic acid molecule comprises at least 95% complementarity to the target nucleic acid molecule.

[0334] 11 . The inhibitory nucleic acid molecule of embodiment 10, wherein the inhibitory nucleic acid molecule is complementary to the target nucleic acid molecule.

[0335] 12. The inhibitory nucleic acid molecule of any one of embodiments 1 -11 , further comprising a modification.

[0336] 13. The inhibitory nucleic acid molecule of embodiment 12, wherein the modification comprises:

[0337] (a) a non-natural or modified nucleoside or nucleotide; and / or

[0338] (b) a covalently or non-covalently conjugated moiety.

[0339] 14. The inhibitory nucleic acid molecule of embodiment 13, wherein:

[0340] (a) the non-natural or modified nucleoside or nucleotide is selected from the group consisting of: a locked nucleic acid (LN A), a 2'-O-methyl (2'-O-Me) modified nucleoside, a phosphorothioate (PS) bond between nucleosides, and a 2'-fluoro (2’-F) modified nucleoside; and / or

[0341] (b) the covalently or non-covalently conjugated moiety is selected from the group consisting of: a targeting moiety, a hydrophobic moiety, a cell penetrating peptide, or a polymer.

[0342] 15. The inhibitory nucleic acid molecule of any one of embodiments 1 -14, wherein the inhibitory nucleic acid molecule is selected from the group consisting of: a small interfering RNA (siRNA), a doublestranded RNA (dsRNA), a microRNA (miRNA), a short hairpin RNA (shRNA), an anti-sense oligonucleotide (ASO), and a gapmeR.

[0343] 16. The inhibitory nucleic acid molecule of embodiment 15, wherein the inhibitory nucleic acid molecule is an siRNA.

[0344] 17. The inhibitory nucleic acid molecule of embodiment 16, wherein the siRNA comprises an antisense strand comprising at least 88% sequence identity to any one of SEQ ID NOs: 1 -6. 18. The inhibitory nucleic acid molecule of embodiment 17, wherein the antisense strand comprises at least 92% sequence identity to any one of SEQ ID NOs: 1 -6.

[0345] 19. The inhibitory nucleic acid molecule of embodiment 18, wherein the antisense strand comprises at least 96% sequence identity to any one of SEQ ID NOs: 1 -6.

[0346] 20. The inhibitory nucleic acid molecule of embodiment 19, wherein the antisense strand comprises the nucleotide sequence of any one of SEQ ID NOs: 1 -6.

[0347] 21 . The inhibitory nucleic acid molecule of any one of embodiments 17-20, wherein the siRNA further comprises a sense strand comprising at least 88%, at least 92%, or at least 96% sequence identity to any one of SEQ ID NOs: 7-12.

[0348] 22. The inhibitory nucleic acid molecule of embodiment 21 , wherein the siRNA comprises

[0349] (a) the antisense strand of SEQ ID NO: 1 and the sense strand of SEQ ID NO: 7;

[0350] (b) the antisense strand of SEQ ID NO: 2 and the sense strand of SEQ ID NO: 8;

[0351] (c) the antisense strand of SEQ ID NO: 3 and the sense strand of SEQ ID NO: 9;

[0352] (d) the antisense strand of SEQ ID NO: 4 and the sense strand of SEQ ID NO: 10;

[0353] (e) the antisense strand of SEQ ID NO: 5 and the sense strand of SEQ ID NO: 11 ; or

[0354] (f) the antisense strand of SEQ ID NO: 6 and the sense strand of SEQ ID NO: 12.

[0355] 23. The inhibitory nucleic acid molecule of any one of embodiments 15-22, wherein the siRNA contains 3’ overhangs selected from the group consisting of:

[0356] (i) a single uracil overhang at one or more 3’ ends of the siRNA;

[0357] (ii) a double uracil overhang at one or more 3’ ends of the siRNA;

[0358] (iii) a single thymine overhang at one or more 3’ ends of the siRNA;

[0359] (iv) a double thymine overhang at one or more 3’ ends of the siRNA; or

[0360] (v) a single cytosine and single thymine overhang at one or more 3’ ends of the siRNA.

[0361] 24. The inhibitory nucleic acid molecule of any one of embodiments 15-23, wherein the siRNA targets the nucleotide sequence of any one of SEQ ID NOs: 15-20.

[0362] 25. The inhibitory nucleic acid molecule of embodiment 15, wherein the inhibitory nucleic acid molecule is a miRNA.

[0363] 26. The inhibitory nucleic acid molecule of embodiment 25, wherein the miRNA comprises a modification selected from:

[0364] (a) a non-natural or modified nucleoside or nucleotide; and / or

[0365] (b) a covalently or non-covalently conjugated moiety.

[0366] 27. The inhibitory nucleic acid molecule of embodiment 26, wherein:

[0367] (a) the non-natural or modified nucleoside or nucleotide is selected from the group consisting of: a locked nucleic acid (LNA), a 2'-O-methyl (2'-O-Me) modified nucleoside, a phosphorothioate (PS) bond between nucleosides, and a 2’-fluoro (2'-F) modified nucleoside; and / or

[0368] (b) the covalently or non-covalently conjugated moiety is selected from the group consisting of: a targeting moiety, a hydrophobic moiety, a cell penetrating peptide, or a polymer.

[0369] 28. The inhibitory nucleic acid molecule of embodiment 26 or 27, wherein the miRNA comprises a nucleotide sequence comprising at least 86% sequence identity to SEQ ID NO: 13.

[0370] 29. The inhibitory nucleic acid molecule of embodiment 28, wherein the miRNA comprises a nucleotide sequence comprising at least 91% sequence identity to SEQ ID NO: 13. 30. The inhibitory nucleic acid molecule of embodiment 29, wherein the miRNA comprises a nucleotide sequence comprising at least 95% sequence identity to SEQ ID NO: 13.

[0371] 31 . The inhibitory nucleic acid molecule of embodiment 30, wherein the miRNA comprises the nucleotide sequence of SEQ ID NO: 13.

[0372] 32. The inhibitory nucleic acid molecule of embodiment 31 , wherein the miRNA is miR-342-3p.

[0373] 33. The inhibitory nucleic acid molecule of any one of embodiments 1 -32, wherein the inhibitory nucleic acid molecule is formulated in a delivery vehicle.

[0374] 34. The inhibitory nucleic acid molecule of embodiment 33, wherein the delivery vehicle is selected from the group consisting of: a vector, a plasmid, a micelle, a liposome, an exosome, and a lipid nano particle (LNP).

[0375] 35. The inhibitory nucleic acid molecule of embodiment 34, wherein the vector is a viral vector.

[0376] 36. The inhibitory nucleic acid molecule of any one of embodiments 1 -35, wherein the inhibitory nucleic acid molecule is formulated as a pharmaceutical composition.

[0377] 37. The inhibitory nucleic acid molecule of embodiment 36, wherein the pharmaceutical composition comprises a pharmaceutically acceptable excipient, diluent, and / or carrier.

[0378] 38. A method of treating or reducing the likelihood of a medical complication resulting from a myocardial infarction (Ml) in a subject, the method comprising administering the inhibitory nucleic acid molecule of any one of embodiments 1 -37.

[0379] 39. The method of embodiment 38, wherein the medical complication is an arrhythmic-related complication, an ischemic-related complication, a mechanical-related complication, an inflammatory- related complication, and / or a systemic complication.

[0380] 40. The method of embodiment 39, wherein:

[0381] (a) the arrhythmic-related complication is a heart block, an atrial arrhythmia, and / or a ventricular arrhythmia;

[0382] (b) the ischemic-related complication is reinfarction, peri-infarct ischemia, and / or an infarct extension;

[0383] (c) the mechanical-related complication is a mitral valve rupture or tear, a chordae rupture or tear, a ventricular septal defect (VSD), a ventricular free wall rupture, a cardiac tamponade, and / or an aneurysm;

[0384] (d) the inflammatory-related complication is pericarditis and / or Dressier syndrome; and / or

[0385] (e) the systemic complication is cardiogenic shock, cardiomyopathy, heart failure, embolic stroke, systemic embolism, and / or a lower extremity embolism.

[0386] 41 . A method of promoting angiogenesis in a subject, the method comprising administering the inhibitory nucleic acid molecule of any one of embodiments 1 -37.

[0387] 42. The method of embodiment 41 , wherein the subject has previously experienced a myocardial infarction.

[0388] 43. The method of embodiment 41 or 42, wherein the subject has an ischemic injury.

[0389] 44. The method of any one of embodiments 38-43, wherein the subject has a cardiovascular disease.

[0390] 45. The method of embodiment 44, wherein the cardiovascular disease is coronary artery disease, peripheral artery disease, or stroke. 46. The method of any one of embodiments 38-45, wherein the subject has a metabolic disorder, or the subject is at risk of developing the metabolic disorder.

[0391] 47. The method of embodiment 46, wherein the metabolic disorder is diabetes.

[0392] 48. The method of embodiment 47, wherein the subject at risk of developing diabetes is prediabetic and / or has one or more of the following:

[0393] (a) hyperglycemia;

[0394] (b) glucose resistance;

[0395] (c) insulin resistance;

[0396] (d) hyperlipidemia; and

[0397] (e) has a family history of diabetes.

[0398] 49. The method of any one of embodiments 38-48, wherein the inhibitory nucleic acid molecule is administered to the subject intravenously, intraperitoneally, subcutaneously, intraarticularly, or intramuscularly.

[0399] 50. The method of any one of embodiments 38-49, wherein the inhibitory nucleic acid molecule is delivered to the coronary endothelium, remote zone of the heart, and / or border zone of the heart.

[0400] 51 . The method of any one of embodiments 38-50, wherein the inhibitory nucleic acid molecule is delivered to an endothelial cell, a cardiomyocyte, a fibroblast, a vascular smooth muscle cell, and / or a leukocyte.

[0401] 52. The method of any one of embodiments 38-51 , further comprising administering an additional therapeutic agent.

[0402] 53. The method of embodiment 52, wherein the additional therapeutic agent is a statin or hepatocyte growth factor (HGF).

[0403] Other Embodiments

[0404] All publications, patents, and patent applications mentioned in this specification are incorporated herein by reference to the same extent as if each independent publication or patent application was specifically and individually indicated to be incorporated by reference.

[0405] While the invention has been described in connection with specific embodiments thereof, it will be understood that it is capable of further modifications and this application is intended to cover any variations, uses, or adaptations following, in general, the principles and including such departures from the invention that come within known or customary practice within the art to which the invention pertains and may be applied to the essential features hereinbefore set forth, and follows in the scope of the claims.

[0406] Other embodiments are within the claims.

Claims

CLAIMS1 . An inhibitory nucleic acid molecule comprising sufficient complementarity to a target nucleic acid molecule, wherein (i) the inhibitory nucleic acid molecule is at least 15 nucleotides in length, and (ii) the target nucleic acid molecule comprises a nucleotide sequence having at least 80% sequence identity to SEQ ID NO: 14.

2. The inhibitory nucleic acid molecule of claim 1 , wherein the target nucleic acid molecule comprises a nucleotide sequence having at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to SEQ ID NO: 14.

3. The inhibitory nucleic acid molecule of claim 1 or 2, wherein the target nucleic acid molecule comprises the nucleotide sequence of SEQ ID NO: 14.

4. The inhibitory nucleic acid molecule of claim 1 , wherein the inhibitory nucleic acid molecule is 15 to 6,850 nucleotides in length.

5. The inhibitory nucleic acid molecule of claim 4, wherein the inhibitory nucleic acid molecule is 15 to 49 nucleotides in length, 50 to 99 nucleotides in length, or 100 to 6,850 nucleotides in length.

6. The inhibitory nucleic acid molecule of claim 5, wherein the inhibitory nucleic acid molecule is 20 to 28 nucleotides in length.

7. The inhibitory nucleic acid molecule of claim 6, wherein the inhibitory nucleic acid molecule is 23 or 25 nucleotides in length.

8. The inhibitory nucleic acid molecule of claim 1 , wherein the inhibitory nucleic acid molecule comprises at least 85% complementarity to the target nucleic acid molecule.

9. The inhibitory nucleic acid molecule of claim 8, wherein the inhibitory nucleic acid molecule comprises at least 90% complementarity to the target nucleic acid molecule.

10. The inhibitory nucleic acid molecule of claim 9, wherein the inhibitory nucleic acid molecule comprises at least 95% complementarity to the target nucleic acid molecule.11 . The inhibitory nucleic acid molecule of claim 10, wherein the inhibitory nucleic acid molecule is complementary to the target nucleic acid molecule.

12. The inhibitory nucleic acid molecule of claim 1 , further comprising a modification.

13. The inhibitory nucleic acid molecule of claim 12, wherein the modification comprises:(a) a non-natural or modified nucleoside or nucleotide; and / or(b) a covalently or non-covalently conjugated moiety.

14. The inhibitory nucleic acid molecule of claim 13, wherein:(a) the non-natural or modified nucleoside or nucleotide is selected from the group consisting of: a locked nucleic acid (LN A), a 2’-O-methyl (2’-0-Me) modified nucleoside, a phosphorothioate (PS) bond between nucleosides, and a 2'-fluoro (2’-F) modified nucleoside; and / or(b) the covalently or non-covalently conjugated moiety is selected from the group consisting of: a targeting moiety, a hydrophobic moiety, a cell penetrating peptide, or a polymer.

15. The inhibitory nucleic acid molecule of claim 1 , wherein the inhibitory nucleic acid molecule is selected from the group consisting of: a small interfering RNA (siRNA), a double-stranded RNA (dsRNA), a microRNA (miRNA), a short hairpin RNA (shRNA), an anti-sense oligonucleotide (ASO), and a gapmeR.

16. The inhibitory nucleic acid molecule of claim 15, wherein the inhibitory nucleic acid molecule is an siRNA.

17. The inhibitory nucleic acid molecule of claim 16, wherein the siRNA comprises an antisense strand comprising at least 88% sequence identity to any one of SEQ ID NOs: 1 -6.

18. The inhibitory nucleic acid molecule of claim 17, wherein the antisense strand comprises at least 92% sequence identity to any one of SEQ ID NOs: 1 -6.

19. The inhibitory nucleic acid molecule of claim 18, wherein the antisense strand comprises at least 96% sequence identity to any one of SEQ ID NOs: 1 -6.

20. The inhibitory nucleic acid molecule of claim 19, wherein the antisense strand comprises the nucleotide sequence of any one of SEQ ID NOs: 1 -6.21 . The inhibitory nucleic acid molecule of claim 17, wherein the siRNA further comprises a sense strand comprising at least 88%, at least 92%, or at least 96% sequence identity to any one of SEQ ID NOs: 7- 12.

22. The inhibitory nucleic acid molecule of claim 21 , wherein the siRNA comprises(a) the antisense strand of SEQ ID NO: 1 and the sense strand of SEQ ID NO: 7;(b) the antisense strand of SEQ ID NO: 2 and the sense strand of SEQ ID NO: 8;(c) the antisense strand of SEQ ID NO: 3 and the sense strand of SEQ ID NO: 9;(d) the antisense strand of SEQ ID NO: 4 and the sense strand of SEQ ID NO: 10;(e) the antisense strand of SEQ ID NO: 5 and the sense strand of SEQ ID NO: 11 ; or(f) the antisense strand of SEQ ID NO: 6 and the sense strand of SEQ ID NO: 12.

23. The inhibitory nucleic acid molecule of claim 15, wherein the siRNA contains 3’ overhangs selected from the group consisting of:(i) a single uracil overhang at one or more 3’ ends of the siRNA;(ii) a double uracil overhang at one or more 3’ ends of the siRNA;(iii) a single thymine overhang at one or more 3’ ends of the siRNA;(iv) a double thymine overhang at one or more 3’ ends of the siRNA; or(v) a single cytosine and single thymine overhang at one or more 3’ ends of the siRNA.

24. The inhibitory nucleic acid molecule of claim 15, wherein the siRNA targets the nucleotide sequence of any one of SEQ ID NOs: 15-20.

25. The inhibitory nucleic acid molecule of claim 15, wherein the inhibitory nucleic acid molecule is a miRNA.

26. The inhibitory nucleic acid molecule of claim 25, wherein the miRNA comprises a modification selected from:(a) a non-natural or modified nucleoside or nucleotide; and / or(b) a covalently or non-covalently conjugated moiety.

27. The inhibitory nucleic acid molecule of claim 26, wherein:(a) the non-natural or modified nucleoside or nucleotide is selected from the group consisting of: a locked nucleic acid (LNA), a 2'-O-methyl (2'-O-Me) modified nucleoside, a phosphorothioate (PS) bond between nucleosides, and a 2'-fluoro (2’-F) modified nucleoside; and / or(b) the covalently or non-covalently conjugated moiety is selected from the group consisting of: a targeting moiety, a hydrophobic moiety, a cell penetrating peptide, or a polymer.

28. The inhibitory nucleic acid molecule of claim 26 or 27, wherein the miRNA comprises a nucleotide sequence comprising at least 86% sequence identity to SEQ ID NO: 13.

29. The inhibitory nucleic acid molecule of claim 28, wherein the miRNA comprises a nucleotide sequence comprising at least 91 % sequence identity to SEQ ID NO: 13.

30. The inhibitory nucleic acid molecule of claim 29, wherein the miRNA comprises a nucleotide sequence comprising at least 95% sequence identity to SEQ ID NO: 13.31 . The inhibitory nucleic acid molecule of claim 30, wherein the miRNA comprises the nucleotide sequence of SEQ ID NO: 13.

32. The inhibitory nucleic acid molecule of claim 31 , wherein the miRNA is miR-342-3p.

33. The inhibitory nucleic acid molecule of claim 1 , wherein the inhibitory nucleic acid molecule is formulated in a delivery vehicle.

34. The inhibitory nucleic acid molecule of claim 33, wherein the delivery vehicle is selected from the group consisting of: a vector, a plasmid, a micelle, a liposome, an exosome, and a lipid nano particle (LNP).

35. The inhibitory nucleic acid molecule of claim 34, wherein the vector is a viral vector.

36. The inhibitory nucleic acid molecule of claim 1 , wherein the inhibitory nucleic acid molecule is formulated as a pharmaceutical composition.

37. The inhibitory nucleic acid molecule of claim 36, wherein the pharmaceutical composition comprises a pharmaceutically acceptable excipient, diluent, and / or carrier.

38. A method of treating or reducing the likelihood of a medical complication resulting from a myocardial infarction (Ml) in a subject, the method comprising administering the inhibitory nucleic acid molecule of claim 1 .

39. The method of claim 38, wherein the medical complication is an arrhythmic-related complication, an ischemic-related complication, a mechanical-related complication, an inflammatory-related complication, and / or a systemic complication.

40. The method of claim 39, wherein:(a) the arrhythmic-related complication is a heart block, an atrial arrhythmia, and / or a ventricular arrhythmia;(b) the ischemic-related complication is reinfarction, peri-infarct ischemia, and / or an infarct extension;(c) the mechanical-related complication is a mitral valve rupture or tear, a chordae rupture or tear, a ventricular septal defect (VSD), a ventricular free wall rupture, a cardiac tamponade, and / or an aneurysm;(d) the inflammatory-related complication is pericarditis and / or Dressier syndrome; and / or(e) the systemic complication is cardiogenic shock, cardiomyopathy, heart failure, embolic stroke, systemic embolism, and / or a lower extremity embolism.41 . A method of promoting angiogenesis in a subject, the method comprising administering the inhibitory nucleic acid molecule of claim 1 .

42. The method of claim 41 , wherein the subject has previously experienced a myocardial infarction.

43. The method of claim 41 or 42, wherein the subject has an ischemic injury.

44. The method of claim 38, wherein the subject has a cardiovascular disease.

45. The method of claim 44, wherein the cardiovascular disease is coronary artery disease, peripheral artery disease, or stroke.

46. The method of claim 38, wherein the subject has a metabolic disorder, or the subject is at risk of developing the metabolic disorder.

47. The method of claim 46, wherein the metabolic disorder is diabetes.

48. The method of claim 47, wherein the subject at risk of developing diabetes is prediabetic and / or has one or more of the following:(a) hyperglycemia;(b) glucose resistance;(c) insulin resistance;(d) hyperlipidemia; and(e) has a family history of diabetes.

49. The method of claim 38, wherein the inhibitory nucleic acid molecule is administered to the subject intravenously, intraperitoneally, subcutaneously, intraarticularly, or intramuscularly.

50. The method of claim 38, wherein the inhibitory nucleic acid molecule is delivered to the coronary endothelium, remote zone of the heart, and / or border zone of the heart.51 . The method of claim 38, wherein the inhibitory nucleic acid molecule is delivered to an endothelial cell, a cardiomyocyte, a fibroblast, a vascular smooth muscle cell, and / or a leukocyte.

52. The method of claim 38, further comprising administering an additional therapeutic agent.

53. The method of claim 52, wherein the additional therapeutic agent is a statin or hepatocyte growth factor (HGF).