ATG7 inhibitors for treatment of obesity and type 2 diabetes

Abstract

The present invention relates to oligonucleotides targeting ATG7(2), and their use as a medicament for metabolic diseases, such as obesity and type 2 diabetes.

Description

P7411 PC00ATG7 inhibitors for treatment of obesity and type 2 diabetesTechnical fieldThe present invention relates to oligonucleotides targeting ATG7(2), and their use as a medicament for metabolic diseases, such as obesity and type 2 diabetes.BackgroundCells are continuously engaged in the dynamic processes of synthesizing and degrading various biomolecules to maintain homeostasis. Macro-autophagy, hereafter referred to as autophagy, is a highly conserved degradation pathway that plays a pivotal role in this balance by degrading and recycling potentially harmful cellular components. Through autophagy, cells can repurpose these components for anabolic reactions, contributing to overall cellular health and survival, especially under stress conditions.ATG7 is a key enzyme in the autophagy pathway, functioning as an E1 -like activating enzyme essential for the conjugation systems that mediate autophagosome formation. ATG7 activates two ubiquitin-like proteins: ATG12 and ATG8 (also known as LC3). The ATG7-ATG12 conjugation system is critical for the elongation of the isolation membrane and subsequent autophagosome formation, while ATG7 also plays a role in the lipidation of ATG8, facilitating its association with the autophagosomal membrane.In addition to its well-characterized role in autophagy, emerging evidence indicates that ATG7 also participates in non-autophagy-related functions. For example, ATG7 has been implicated in the regulation of lipid metabolism, where it influences the balance between lipid storage and mobilization. Furthermore, ATG7 has been shown to play a role in immune responses, including the regulation of interferon production and the modulation of inflammation.SummaryInterestingly, a shorter isoform of ATG7, designated as ATG7(2), has been identified by the present inventor (M.H. Ogmundsdottir ef a / ., 2018), which is incapable of performing the canonical function of ATG7 in initiating autophagy. Unlike ATG7(1), which includes all exons and is responsible for the standard autophagic activity,P7411 PC00ATG7(2) lacks exon 17, resulting in a 27-amino acid truncation at the C-terminal region. This truncation prevents ATG7(2) from lipidating ATG8 proteins, a key step in autophagosome formation.More recently, the inventors of the present disclosure have described the role of ATG7(2) in mouse embryonic fibroblasts (MEFs) and human hepatic cancer cell lines. It was observed that while the canonical ATG7(1) binds autophagy proteins, ATG7(2) binds glycolysis enzymes and mitochondrial proteins in cancer cells and may suppress both glycolysis and mitochondrial respiration (Ostacolo et al., 2024).In the present disclosure, the inventors describe that an oligonucleotide targeting specifically ATG7(2) can be employed to treat metabolic diseases, such as diabetes or such as obesity. Surprisingly, the present inventors have found that maintaining the expression level of ATG7(2) low, contributes to increased tolerance to stress, in particular oxidative stress, and at the same time a mild repression of glycolysis, which results in a non-obese, non-diabetic phenotype.Thus, an aspect of the present disclosure is directed to an oligonucleotide targeting ATG7 variant 2 (ATG7(2)).In an aspect, the present disclosure is directed to an oligonucleotide targeting a polynucleotide encoding human ATG7 variant 2 (ATG7(2)).In an aspect, the present disclosure is directed to an oligonucleotide targeting a polynucleotide encoding human ATG7 variant 2 (ATG7(2)) for use as a medicament.In an aspect, the present disclosure is directed to an oligonucleotide targeting a polynucleotide encoding human ATG7 variant 2 (ATG7(2)) for use in a method of treating a metabolic disease.In an aspect, the present disclosure is directed to a composition comprising the oligonucleotide, or the oligonucleotide for use as described herein.In an aspect the present disclosure is directed to a method of treating a metabolic disease, said method comprising administration of a therapeutically effective amount ofP7411 PC00 an oligonucleotide targeting a polynucleotide encoding human ATG7(2) to an individual in need thereof.In an aspect the present disclosure is directed to a method of treating a metabolic disease, the method comprising administering the oligonucleotide as described herein.In an aspect the present disclosure is directed to the use of the oligonucleotide as described herein, for the manufacture of a medicament.In an aspect the present disclosure is directed to the use of the oligonucleotide as described herein, for the manufacture of a medicament for treating a metabolic disease.Description of DrawingsFigure 1 : Expression of ATG7. (A-C) Expression of fly Atg7 on mRNA level analysed by qPCR (A-B) or RNA sequencing (C) in the different CRISPR strains, indicated on the X-axis. While the Atg7- / - and hATG7(1) strains lack the fly gene, both hATG7(2) strains express it. The expression of the CRISPR strains was normalized to that of WT. (D) mRNA expression of human ATG7 in the different strains indicated on the X-axis. The expression was normalized to that of hATG7(1), which had the highest expression. The hATG7(2) strains were given names based on their expression levels, i.e. hATG7(2)highhas higher expression of hATG7(2) and hATG7(2)l0Whas lower expression. (E) Confocal imaging of fat bodies stained for human ATG7 and DAPI. The protein expression was mainly cytosolic as expected and the expression levels are comparable to the mRNA expression, i.e. hATG7(1) has the highest expression, then hATG7(2)highand hATG7(2)l0W.Figure 2: Lifespan and stress tolerance. All data was analysed by the Kaplan-Meier method and p-values were calculated with log-rank (Mantel-Cox) test. Symbols for each strain are indicated at the bottom. (A) Lifespan of the different strains. WT (n = 170), Atg7' / _(n = 107), hATG7(1) (n =175), hATG7(2)high(n = 103), hATG7(2)l0W(n = 108). All strains varied significantly from WT with p < 0,0001. Atg7_ / _had a 44% median lifespan reduction, ATG7(1) even worse with 51 %, hATG7(2)highhad a slight decrease of 9%. However, hATG7(2)l0Whad a lifespan extension of 17%. (B) Survival under oxidative stress (10 mM Paraquat). WT (n = 111), Atg7_ / _(n = 115) p = 0,0307,P7411 PC00 hATG7(1) (n = 101) p < 0,0001 , hATG7(2)high(n = 123) p = 0,0018, hATG7(2)l0W(n = 109) p = 0,0176. All strains survived for about 3 days, while hATG7(2)l0Wsurvived for up to 20 days with a longer survival of 56% on average (although the median was not significant). (C) Survival under complete starvation. WT (n = 139), Atg7_ / _(n = 107) p < 0,0001 , hATG7(1) (n = 124) p < 0,0001 , hATG7(2)high(n = 136) p < 0,0001 , hATG7(2)l0W(n = 132) p < 0,0001. Surprisingly, WT had the worst survival under starvation. While most strains survived for about 3 days, hATG7(2)highhad a median survival of 4. Collectively this shows a lifespan increase and increased tolerance to oxidative stress with low hATG7(2) expression and increased tolerance to starvation with hATG7(2) expression.Figure 3: hATG7(2)highlarvae are double in size. (A) Third instar larvae from the strain indicated above. The hATG7(2)high strain produces larvae double as large as the other strains. (B) hATG7(2)highlarvae weigh almost twice as much as the other strains indicated on the X-axis. The larvae were weighed in batches of 10 larvae. This indicates that higher expression of hATG7(2) leads to a size increase.Figure 4: hATG7(2) leads to fat accumulation. (A) Confocal images of fat bodies from third instar larvae. The tissues were stained for lipids (with BODIPY) and nuclei (DAPI). The strain is indicated on the left. (B) Quantification of the confocal images, where the area of lipid droplets were measured. The guantification shows a significant increase in lipids per cell in the hATG7(2)highstrain. This suggests that the larger size of the hATG7highis at least partly explained by obesity.Figure 5: Increased fat accumulation in hATG7(2) on high sugar diet. Fat accumulation on high sugar diet (HSD). (A) Confocal images of fat bodies from third instar larvae cultured on HSD. The tissues were stained for lipids (with BODIPY) and nuclei (DAPI). The strain is indicated on the left. (B) Quantification of the confocal images, where the area of lipid droplets were measured. The guantification shows a significant increase in lipids per cell in both hATG7(2) strains. This indicates that the fat accumulation is affected by diet and expression levels of hATG7(2) in the hATG7(2) strains.Figure 6: hATG7(2) increases blood sugar. Trehalose measurements in adult flies. In fruit flies, circulating sugars are primarily trehalose rather than glucose. Trehalose levels were normalized to total protein content of the flies. (A) hATG7(2)highhave increased trehalose levels under normal conditions. (B) Both hATG7(2) strains exhibitP7411 PC00 an increase in trehalose levels on HSD. This indicates that hATG7(2) increases blood sugar in a dose dependent manner (referring to the expression levels of ATG7(2)).Figure 7 hATG7(2) strains exhibit delayed larval development. Larval development.Days from when the cultures were started until the larvae crawled up from the food. (A) Normal conditions. Atg7- / - develop faster than WT (p < 0,0001), while hATG7(1) is NS. hATG7(2)highand hATG7(2)l0Wdevelop slower (p < 0,001 and p = 0,0023, respectively). (B) Flies cultured on HSD. Atg7- / - and hATG7(1) develop faster (p < 0,0001 and p = 0,0355, respectively). Both hATG7(2) strains develop slower (p < 0,0001 ib both cases). Slower larval development is a phenotype observed in diabetic fruit flies. This further suggests diabetes in the hATG7(2) strains.Figure 8 hATG7(2) strains have decreased motor function hATG7(2) exhibit decreased motor function and die on pupal stage. (A) hATG7(2)highfrequently fail to fully emerge from the pupal case. (B) All strains exhibit decreased climbing in comparison to WT. The y-axis shows the percentage of flies who climbed 2 / 3rds of the vial height in 10 seconds. Considering that the hATG7(2) strains have the fly gene (WT background), this indicates detrimental effects of hATG7(2) on motor function. (C) A large portion of the hATG7(2)highflies died on the pupal stage, or 57% while only 2,7% of WT died. A frequent cause of death was failure to completely emerge from the pupal case as shown in (A). Collectively, this suggests muscle atrophy in the hATG7(2) strains, which is more pronounced in hATG7(2)high(and thus seems to be reliant on expression levels of ATG7(2)). Muscle atrophy is a common phenotype in obesity.Figure 9 mRNA expression of ATG7 is significantly elevated in obese humans, compared to lean. The increased is explained by ATG7(2), but not ATG7(1) or ATG7(3) (also known as isoform c). (A) expression of total ATG7 mRNA in obese and lean humans. (B) The mRNA expression of ATG7(1) is not significantly increased. (C) The mRNA expression of ATG7(2) is significantly increased in obese humans compared to lean (p = 0.0028). Expression levels were compared by a Welch’s t-test. The GSE data accession number: GSE152991.Figure 10 siRNAs that are specific and selective for ATG7(2) (A) Arrows indicate the location and direction of the sequences targeted by siRNA. (B), (C), (D) Quantification of ATG7, ATG7(1) or ATG7(2) mRNA expression in Capan-1 WT or (E), (F), (G)P7411 PC00Capan-1 ATG7(1)- / - cells transfected with siControl, siATG7, siATG7(1) or siATG7(2).Detailed descriptionDefinitionsUnless defined otherwise, all technical and scientific terms used herein have the same meaning as is commonly understood by one of skill in art to which the subject matter herein belongs. As used herein, the following definitions are supplied to facilitate the understanding of the present invention.The term “comprise” is generally used in the sense of include, that is to say permitting the presence of one or more features or components. In addition, as used in the specification and claims, the language "comprising" can include analogous embodiments described in terms of “consisting of’ and / or “consisting essentially of”.As used in the specification and claims, the term "and / or" used in a phrase such as "A and / or B" herein is intended to include "A and B", "A or B", "A", and "B".As used in the specification and claims, the singular forms "a", "an" and "the" include plural references unless the context clearly dictates otherwise. Similarly, terms such as “one or more” or “at least one” include both the singular and plural form of the respective feature.The term “nucleic acid” as used herein refers to polynucleotides such as deoxyribonucleic acid (DNA) and ribonucleic acid (RNA) or a combination of the two and any chemical or enzymatic modification thereof (e.g. methylated DNA, DNA of modified nucleotides). The term should also be understood to include, as equivalents, derivatives, variants and analogs of either RNA or DNA made from nucleotide analogs, single (sense or antisense) and double-stranded polynucleotides.The term “isolated nucleic acid” as used herein refers to a nucleic acid that is separated from its native environment and present in sufficient quantity to permit its identification or use. An isolated nucleic acid may be one that is (i) amplified in vitro by, for example, polymerase chain reaction (PCR); (ii) recombinantly produced or cloned; (iii) purified, as by cleavage and gel separation; or (iv) synthesized by, for example, chemical synthesis. An isolated nucleic acid is one which is readily manipulable byP7411 PC00 recombinant DNA techniques well known in the art. Thus, a nucleotide sequence contained in a vector in which 5' and 3' restriction sites are known or for which polymerase chain reaction (PCR) primer sequences have been disclosed is considered isolated but a nucleic acid sequence existing in its native state in its natural host is not. An isolated nucleic acid may be substantially purified, but need not be. For example, a nucleic acid that is isolated within a cloning or expression vector is not pure in that it may comprise a small percentage of the material of the cell in which it resides. Such a nucleic acid is isolated, however, as the term is used herein because it is readily manipulable by standard techniques known to those of ordinary skill in the art. Any of the nucleic acids provided herein may be isolated.The terms homology, identity and similarity, with respect to a polynucleotide (or polypeptide), as defined herein are used interchangeably and refer to the percentage of nucleic acids (or amino acids) in the candidate sequence that are, homolog, identical or similar, respectively, to the residues of a corresponding native nucleic acids (or amino acids), after aligning the sequences and introducing gaps, if necessary, to achieve the maximum percent identity I similarity, and considering any conservative substitutions according to the NCIIIB rules (http: / / www.chem. qmul.ac.uk / iubmb / misc / naseq.html; NC-llIB, Eur J Biochem (1985)) as part of the sequence identity. In particular, the percentage of similarity refers to the percentage of residues conserved with similar physiochemical properties. Neither 5' or 3' extensions nor insertions (for nucleic acids) or N’ or C’ extensions nor insertions (for polypeptides) result in a reduction of identity or similarity. Methods and computer programs for the alignments are well known in the art. Generally, a given similarity between two sequences implies that the identity between these sequences is at least equal to the similarity; for example, if two sequences are 70% similar to one another, they cannot be less than 70% identical to one another - but could be sharing 80% identity.Throughout the text any reference to the oligonucleotide, also refers to the oligonucleotide for use, or uses of said oligonucleotide as described herein.Oligonucleotide targeting ATG7 variant 2 (ATG7(2)).The present disclosure describes oligonucleotides for targeting ATG7 variant 2 (ATG7(2)), and their use as a medicament, such as their use in a method of treating a metabolic disease, such as diabetes or obesity.P7411 PC00The present disclosure describes that it is specifically ATG7 variant 2 (ATG7(2)) which is associated with metabolic diseases, such as diabetes or obesity.Thus, in an aspect of the invention, the present disclosure is directed to an oligonucleotide targeting ATG7(2).In an aspect of the invention, the present disclosure is directed to an oligonucleotide targeting ATG7(2) for use as a medicament.In an aspect of the invention, the present disclosure is directed to an oligonucleotide targeting ATG7(2) for use in a method of treating a metabolic diseases.In some embodiments, ATG7(2) is human ATG7(2).The present disclosure is directed to means to inhibit specifically ATG7(2), without targeting other isoforms of ATG7, such as ATG7(1). In particular, in the Examples, the inventors have substantiated that ATG7(2) is associated with obesity and diabetes, and that decreasing the level of the ATG7(2) is beneficial to address these metabolic diseases. It will be evident to the skilled person that any equivalent method capable of inhibiting specifically an isoform of a gene might be suitable for carrying out the teaching of the present disclosure.For example, in some embodiments the oligonucleotide inhibits gene expression or translation by neutralizing ATG7(2) mRNA molecules, effectively silencing specifically ATG7(2). Thus, in some embodiments ATG7(1) is not silenced by said oligonucleotide. Thus, in some embodiments, the oligonucleotide is an oligonucleotide suitable for RNA interference (RNAi).Thus, in some embodiments, the oligonucleotide is a small Interfering RNA (siRNA), or short Hairpin RNA (shRNA). In some embodiments, the oligonucleotide is a small Interfering RNA (siRNA). In some embodiments, the oligonucleotide is a short Hairpin RNA (shRNA).In some embodiments, the oligonucleotide binds to the target mRNA and forms a DNA- RNA hybrid. The hybrid is then recognized by the enzyme RNase H, which cleaves theP7411 PC00RNA strand, leading to the degradation of the mRNA, thereby preventing it from being translated into protein. Alternatively and / or concomitantly, in some embodiments the oligonucleotide physically blocks the binding of proteins or other molecules to the mRNA, interfering with processes such as translation, mRNA transport, or polyadenylation. Thus, in some embodiments, the oligonucleotide is an antisense oligonucleotide (ASO). In some embodiments the antisense oligonucleotide is complementary to the junction of exon 16 and exon 18.The terms “exon 16”, “exon 17” and "exon 18” as used herein refer to the corresponding sequences in ATG7(1). Thus “exon 18” is the seventeenth exon in the mature mRNA of ATG(2).Thus, as described herein: exon 16 refers to the sequence: GGGCTATGCCATTGCCAGCAGCAGTGACGATCGGATGAATGAGCCTCCAACCTCT CTTGGGCTTGTGCCTCACCAGThus, as described herein: exon 17 refers to the sequence: ATCCGGGGATTTCTTTCACGGTTTGATAATGTCCTTCCCGTCAGCCTGGCATTTGA CAAATGTACAGCTTGTTCTTCCAAAThus, as described herein: exon 18 refers to the sequence: GTTCTTGATCAATATGAACGAGAAGGATTTAACTTCCTAGCCAAGGTGTTTAATTCT TCACATTCCTTCTTAGAAGACTTGACTGGTCTTACATTGCTGCATCAAGAAACCCA AGCTGCTGAGAntisense oligonucleotides are capable of inhibiting translation of a protein also by other means. For example, in some embodiments, the oligonucleotide binds to specific sequences in pre-mRNA (the precursor to mRNA) and promotes the inclusion of exon 17 of ATG7. This type of antisense-mediated splicing modulation uses antisense oligonucleotides to manipulate the splicing, for example by sterically blocking the binding of splicing factors to pre-mRNA transcripts (also referred to as precursor mRNA). Thus, in some embodiments, the oligonucleotide promotes the inclusion of exon 17 of ATG7. By promoting inclusion of ATG7, in some embodiments, the oligonucleotide decreases the amount of ATG7(2) mRNA. In some embodiments of the present disclosure, upon binding to a target site, the antisense oligonucleotide preventsP7411 PC00 splicing factors from binding. Thus, in some embodiments the oligonucleotide is complementary to a splice site known and / or predicted to be associated with exon skipping of exon 17. The skilled person will understand that tools available to predict sequences in a pre-mRNA which affect splicing can be used to design said antisense oligonucleotides.Locked Nucleic Acid (LNA) antisense oligonucleotides are a specific type of ASO, characterized by its modified nucleotide structure that confers high binding affinity, specificity, and stability. Thus, in some embodiments, the oligonucleotide is a Locked Nucleic Acid (LNA) antisense oligonucleotide.A guide RNA (gRNA) can selectively inhibit a specific isoform of a gene by targeting a unique sequence that is present only in that isoform and not in others. This selectivity can be achieved through careful design of the gRNA, based on principles commonly known in the art.Thus, in some embodiments, a gRNA is designed to be complementary to a sequence comprising the junction of exon 16 and exon 18 as described herein. Thus, in some embodiments the oligonucleotide is a gRNA.Morpholino oligomers (often simply called "morpholinos") are synthetic molecules used to block the translation of specific mRNA sequences or to modify pre-mRNA splicing. They are designed to be highly specific to their target sequences, and they can be tailored to target specific isoforms of a gene by leveraging unique features of those isoforms. Similarly to antisense oligonucleotides, morpholino oligomers can specifically target ATG7(2) by binding the junction of exon 16 and 18, thereby inhibiting translation in a protein; or they can be designed to promote inclusion of exon 17 in the mature mRNA. The skilled person will understand that any known technique to design morpholino oligomers is suitable for carrying out the teachings of the present disclosure.Thus, in summary, in some embodiments, the oligonucleotide is: a siRNA; a shRNA; an antisense oligonucleotide; a guide RNA; or a morpholino oligomer.In some embodiments, the oligonucleotide is a siRNA. In some embodiments, the oligonucleotide is a short hairpin RNA (shRNA). In some embodiments, the oligonucleotide is an antisense oligonucleotide. In some embodiments, theP7411 PC00 oligonucleotide is a guide RNA (gRNA). In some embodiments, the oligonucleotide is a morpholino oligomer.In some embodiments, the oligonucleotide is a RNA.Sequences and ComplementarityDifferential splicing of ATG7 produces two isoforms of ATG7, herein referred to as ATG7(1) and ATG7(2). This differential splicing results in ATG7(1) and ATG7(2) having distinct exons composition. ATG7(1) comprises exons 1 to 19, while ATG7(2) comprises exons 1 to 16 and 18 to 19, i.e. , it does not comprise exon 17.Thus, ATG7(2) comprises a junction between exon 16 and exon 18, which ATG7(1) does not comprise, as said exons are separated by exon 17 in ATG7(1). siRNA targeting ATG7(2) by complementarity with nucleotides in the 3’ region of exon 16 and the 5’ region of exon 18 was designed using the online tool siDirect v2.0. The obtained sequence was 5’- GGUUCUUGAUCAAUAUGAAcg -3’ (SEQ ID NO: 29), with 1 nucleotide only targeting the 3’ region of exon 16, and 20 nucleotides targeting the 5’ region of exon 18. Use of this siRNA in Huh7 cells overexpressing ATG7(1) or ATG7(2) showed silenced expression of both isoforms upon treatment with the siRNA, revealing a lack in specificity for ATG7(2).To overcome this technical difficulty, additional siRNA sequences were designed manually to avoid long GO stretches, ensure compliant melting temperature (35-55°C) of the oligonucleotide and presence of a T / A nucleotide at the 5’ end of the antisense strand. The designed oligonucleotides were aligned on the whole human genome using the NCBI BLAST online tool and the two siRNA sequences without off target binding sites were selected.Provided herein is an oligonucleotide targeting ATG7 variant 2 (ATG7(2)), which can be used as a medicament as described herein. In some embodiments, the oligonucleotide targets human ATG7(2). Thus, in some embodiments, said ATG7(2) is human ATG7(2).In some embodiments, ATG7(2) is as set forth in SEQ ID NO: 18.P7411 PC00In some embodiments, ATG7(2) is encoded by a nucleotide sequence comprising a nucleotide sequence as set forth in SEQ ID NO: 30.Specific targeting of this isoform is achieved, in some embodiments, by designing the oligonucleotide to target a sequence which is found in this variant and not in ATG7(1), such as the junction between exon 16 and exon 18. Alternatively, in some embodiments, the oligonucleotide is complementary to a splice site known and / or predicted to be associated with exon skipping of exon 17.Thus, in some embodiments the oligonucleotide is complementary to ATG7(2) mRNA.Thus, in some embodiments, said oligonucleotide is selective for ATG7(2). In other words, in some embodiments, the oligonucleotide targets ATG7(2), but does not target ATG7(1).In some embodiments, ATG7(1) is as set forth in SEQ ID NO: 20.In some embodiments, ATG7(1) is encoded by a nucleotide sequence comprising a nucleotide sequence as set forth in SEQ ID NO: 31.Selective targeting of ATG7(2) can arise through mechanisms, such as complementary base-pairing between nucleotides in the oligonucleotide and nucleotides in the ATG7(2), wherein the nucleotide sequences of each the oligonucleotide and the ATG7(2) mRNA comprise complementary sense and anti-sense nucleic acid sequencesIn some embodiments, the oligonucleotide prevents and / or inhibits ATG7(2) translation. Such inhibition can be caused, for example, by the selective degradation of the ATG7(2) mRNA.In some embodiments, the oligonucleotide does not target ATG7 variant 1 (ATG7(1)).In some embodiments, the oligonucleotide is complementary to: a fragment of exon 16 of ATG7 and to a fragment of exon 18 of ATG7.In some embodiments, the oligonucleotide is not complementary to exon 17 of ATG7.P7411 PC00In other words, in some embodiments, the oligonucleotide is not complementary to ATG7(1).In some embodiments, the oligonucleotide comprises a sequence complementary to an ATG7 mRNA comprising a junction between exon 16 and exon 18 of ATG7. Thus, in some embodiments the oligonucleotide is complementary to ATG7(2) mRNA but not ATG7(1) mRNA; only ATG7(2) mRNA comprises as junction between exon 16 and exon 18.In some embodiments, the oligonucleotide is a small interfering RNA (siRNA).In some embodiments, said siRNA is complementary to the junction between exon 16 and exon 18 of ATG7(2). Thus, in some embodiments, said siRNA specifically binds to an ATG7(2) mRNA but does not bind to an ATG7(1) mRNA, as only ATG7(2) mRNA comprises the junction between exon 16 and exon 18 of ATG7. It will be evident that to the skilled person that, in some embodiments, the oligonucleotide may be complementary to either ATG7(1) exon 16 mRNA or ATG7(1) exon 18 mRNA, but cannot bind to ATG7(1) mRNA and affect ATG7(1) translation because ATG(1) mRNA lacks the junction between exon 16 and exon 18 of ATG7.In some embodiments, the oligonucleotide hybridizes specifically to the target site under high stringency solution hybridization conditions to target site. As described herein, the target site is the junction between exon 16 and exon 18 in ATG7(2), or a splice site known and / or predicted to be associated with exon skipping of exon 17.In some embodiments, said oligonucleotide binds to a 3’ fragment of exon 16 of ATG7 and to a 5’ fragment of exon 18 of ATG7.The terms nucleotide and nucleotides may be abbreviated to “nt”, said terms are used interchangeably herein. Furthermore, wherein the nucleic acid comprising said nucleotides is a ribonucleic acid, said terms may be used to refer to a ribonucleotide or ribonucleotides.In some embodiments, the oligonucleotide comprises: a. at least 3 nt, such as at least 4, such as at least 5 nt complementary to exon 16 of ATG7(2); andP7411 PC00 b. at least 3 nt, such as at least 4, such as at least 5 nt complementary to exon 18 of ATG7(2).In some embodiments, the oligonucleotide comprises at least 3 nt, such as at least 4, such as at least 5 nt complementary to exon 16 of ATG7(2). In some embodiments, the oligonucleotide comprises at least 3 nt, such as at least 4, such as at least 5 nt complementary to exon 18 of ATG7(2).In some embodiments, the oligonucleotide comprises at least 3 nt complementary to exon 16 of ATG7(2). In some embodiments, the oligonucleotide comprises at least 4 nt complementary to exon 16 of ATG7(2). In some embodiments, the oligonucleotide comprises at least 5 nt complementary to exon 16 of ATG7(2). In some embodiments, the oligonucleotide comprises at least 6 nt complementary to exon 16 of ATG7(2). In some embodiments, the oligonucleotide comprises at least 7 nt complementary to exon 16 of ATG7(2). In some embodiments, the oligonucleotide comprises at least 8 nt complementary to exon 16 of ATG7(2). In some embodiments, the oligonucleotide comprises at least 9 nt complementary to exon 16 of ATG7(2). In some embodiments, the oligonucleotide comprises at least 10 nt complementary to exon 16 of ATG7(2). In some embodiments, the oligonucleotide comprises at least 11 nt complementary to exon 16 of ATG7(2).In some embodiments, the oligonucleotide comprises at least 3 nt complementary to exon 18 of ATG7(2). In some embodiments, the oligonucleotide comprises at least 4 nt complementary to exon 18 of ATG7(2). In some embodiments, the oligonucleotide comprises at least 5 nt complementary to exon 18 of ATG7(2). In some embodiments, the oligonucleotide comprises at least 6 nt complementary to exon 18 of ATG7(2). In some embodiments, the oligonucleotide comprises at least 7 nt complementary to exon 18 of ATG7(2). In some embodiments, the oligonucleotide comprises at least 8 nt complementary to exon 18 of ATG7(2). In some embodiments, the oligonucleotide comprises at least 9 nt complementary to exon 18 of ATG7(2). In some embodiments, the oligonucleotide comprises at least 10 nt complementary to exon 18 of ATG7(2). In some embodiments, the oligonucleotide comprises at least 11 nt complementary to exon 18 of ATG7(2).In some embodiments, the oligonucleotide comprises:P7411 PC00 a. at least the 3 nt, such as at least the 4, such as at least the 5 nt complementary to the 3’ end of exon 16 of ATG7(2); and b. at least 3 nt, such as at least 4, such as at least 5 nt complementary to the 5’ end of exon 18 of ATG7(2).In some embodiments, the oligonucleotide comprises at least 3 nt, such as at least the 4, such as at least the 5 nt complementary to the 3’ end of exon 16 of ATG7(2). In some embodiments, the oligonucleotide comprises at least 3 nt, such as at least 4, such as at least 5 nt complementary to the 5’ end of exon 18 of ATG7(2).In some embodiments, the oligonucleotide comprises at least 3 nt complementary to the 3’ end of exon 16 of ATG7(2). In some embodiments, the oligonucleotide comprises at least 4 nt complementary to the 3’ end of exon 16 of ATG7(2). In some embodiments, the oligonucleotide comprises at least 4 nt complementary to the 3’ end of exon 16 of ATG7(2). In some embodiments, the oligonucleotide comprises at least 5 nt complementary to the 3’ end of exon 16 of ATG7(2). In some embodiments, the oligonucleotide comprises at least 6 nt complementary to the 3’ end of exon 16 of ATG7(2). In some embodiments, the oligonucleotide comprises at least 7 nt complementary to the 3’ end of exon 16 of ATG7(2).In some embodiments, the oligonucleotide comprises at least 3 nt complementary to the 5’ end of exon 18 of ATG7(2). In some embodiments, the oligonucleotide comprises at least 4 nt complementary to the 5’ end of exon 18 of ATG7(2). In some embodiments, the oligonucleotide comprises at least 5 nt complementary to the 5’ end of exon 18 of ATG7(2). In some embodiments, the oligonucleotide comprises at least 6 nt complementary to the 5’ end of exon 18 of ATG7(2). In some embodiments, the oligonucleotide comprises at least 7 nt complementary to the 5’ end of exon 18 of ATG7(2).In some embodiments, said oligonucleotide comprises a double-stranded region of 10 to 30 nt, such as 11 to 29 nt, such as 12 to 28 nt, such as 13 to 27 nt, such as 14 to 26 nt, such as 15 to 25 nt.P7411 PC00In some embodiments, said oligonucleotide comprises a strand comprising SEQ ID NO: 13 or a functional variant thereof, wherein said functional variant has 1 or 2 individual ribonucleotide substitutions compared to SEQ ID NO: 13.Thus, in some embodiments said oligonucleotide comprises a strand comprising SEQ ID NO: 13 or a functional variant thereof, wherein said functional variant has 1 individual ribonucleotide substitution compared to SEQ ID NO: 13. In other embodiments, said oligonucleotide comprises a strand comprising SEQ ID NO: 13 or a functional variant thereof, wherein said functional variant has 2 individual ribonucleotide substitutions compared to SEQ ID NO: 13.In some embodiments, said oligonucleotide further comprises 1 to 16 nucleotides, such as 1 nucleotide, such as 2 nucleotides, such as 3 nucleotides, such as 4 nucleotides, such as 5 nucleotides, such as 6 nucleotides, such as 7 nucleotides, such as 8 nucleotides, such as 9 nucleotides, such as 10 nucleotides, such as 11 nucleotides, such as 12 nucleotides, such as 13 nucleotides, such as 14 nucleotides, such as 15 nucleotides, such as 16 nucleotides attached at the 3’ of SEQ ID NO: 13 or said functional variant thereof.In some embodiments said oligonucleotide further comprises 1 to 16 nucleotides, such as 1 nucleotide, such as 2 nucleotides, such as 3 nucleotides, such as 4 nucleotides, such as 5 nucleotides, such as 6 nucleotides, such as 7 nucleotides, such as 8 nucleotides, such as 9 nucleotides, such as 10 nucleotides, such as 11 nucleotides, such as 12 nucleotides, such as 13 nucleotides, such as 14 nucleotides, such as 15 nucleotides, such as 16 nucleotides attached at the 5’ of SEQ ID NO: 13 or said functional variant thereof.In some embodiments, said oligonucleotide comprises a strand comprising SEQ ID NO: 16 or SEQ ID NO: 17, or a functional variant thereof, wherein said functional variant has 1 , 2, 3, 4, 5, or 6 individual ribonucleotide substitutions compared to any one of SEQ ID NO: 16 or SEQ ID NO: 17.Thus, in some embodiments, said oligonucleotide comprises a strand comprising SEQ ID NO: 16, or a functional variant thereof, wherein said functional variant has 1, 2, 3, 4, 5, or 6 individual ribonucleotide substitutions compared to SEQ ID NO: 16. In someP7411 PC00 embodiments, said oligonucleotide comprises a strand comprising SEQ ID NO: 17, or a functional variant thereof, wherein said functional variant has 1 , 2, 3, 4, 5, or 6 individual ribonucleotide substitutions compared to SEQ ID NO: 17.In some embodiments, said oligonucleotide comprises a strand of SEQ ID NO: 16 or SEQ ID NO: 17, or a functional variant thereof, wherein said functional variant has 1, 2 or 3 individual ribonucleotide substitutions compared to any one of SEQ ID NO: 16 or SEQ ID NO: 17.Thus, in some embodiments, said oligonucleotide comprises a strand of SEQ ID NO: 16, or a functional variant thereof, wherein said functional variant has 1, 2 or 3 individual ribonucleotide substitutions compared to SEQ ID NO: 16. In some embodiments, said oligonucleotide comprises a strand of SEQ ID NO: 16, or a functional variant thereof, wherein said functional variant has 1 individual ribonucleotide substitution compared to SEQ ID NO: 16. In some embodiments, said oligonucleotide comprises a strand of SEQ ID NO: 16, or a functional variant thereof, wherein said functional variant has 2 individual ribonucleotide substitutions compared to SEQ ID NO: 16. In some embodiments, said oligonucleotide comprises a strand of SEQ ID NO: 16, or a functional variant thereof, wherein said functional variant has 3 individual ribonucleotide substitutions compared to SEQ ID NO: 16.In some embodiments, said oligonucleotide comprises a strand of SEQ ID NO: 17, or a functional variant thereof, wherein said functional variant has 1 , 2 or 3 individual ribonucleotide substitutions compared to SEQ ID NO: 17. In some embodiments, said oligonucleotide comprises a strand of SEQ ID NO: 17, or a functional variant thereof, wherein said functional variant has 1 individual ribonucleotide substitution compared to SEQ ID NO: 17. In some embodiments, said oligonucleotide comprises a strand of SEQ ID NO: 17, or a functional variant thereof, wherein said functional variant has 2 individual ribonucleotide substitutions compared to SEQ ID NO: 17. In some embodiments, said oligonucleotide comprises a strand of SEQ ID NO: 17, or a functional variant thereof, wherein said functional variant has 3 individual ribonucleotide substitutions compared to SEQ ID NO: 17.P7411 PC00In some embodiments, said oligonucleotide comprises a sense strand of SEQ ID NO: 16, SEQ ID NO: 17, or SEQ ID NO: 13 or functional variant thereof, and a complementary antisense strand.Thus, in some embodiments said oligonucleotide comprises a sense strand of SEQ ID NO: 16, or functional variant thereof, and a complementary antisense strand. In other embodiments, said oligonucleotide comprises a sense strand of SEQ ID NO: 17, or functional variant thereof, and a complementary antisense strand. In further embodiments, said oligonucleotide comprises a sense strand of SEQ ID NO: 13, or functional variant thereof, and a complementary antisense strand.Thus disclosed herein are oligonucleotides, or oligonucleotides for use, which are useful in selectively targeting ATG7(2) transcripts, for example through complementary base-pairing interactions. In some embodiments, said oligonucleotides, or oligonucleotides for use, do not specifically target ATG7(1), for example, by targeting a section of said ATG7 transcripts comprising an exon junction present in ATG7(2), but not present in ATG7(2), which can result from differential splicing.The oligonucleotides, or oligonucleotides for use, provided herein are useful for use as a medicament; ADD.Nucleotide modificationsNucleotide modifications can influence the properties of the nucleic acids they are modifying. For example, the presence or absence of phosphate groups on the 3’ and / or 5’ terminal nucleotide of an oligonucleotide affects said oligonucleotide’s reactivity, for instance, by determining whether said oligonucleotide can be ligated to another nucleic acid molecule. Other types of modifications have other effects on the properties of the oligonucleotide that modify. For example, some modifications inhibit degradation by nucleases, or otherwise affect the oligonucleotide’s interaction with enzymes, which in turn, affect the stability or morphology of said oligonucleotides, or even their affinity for complementary nucleic acids. The skilled person will understand that any modification known in the art to improve the ability of said nucleotide to silence a specific target is suitable for carrying out the method of the present disclosure.P7411 PC00In some embodiments the ribonucleotide at the 3’ terminus and / or at the 5’ terminus of the sense strand are phosphorylated. In some embodiments the ribonucleotide at the 3’ terminus and at the 5’ terminus of the sense strand are phosphorylated In some embodiments, the ribonucleotide at the 3’ terminus of the sense strand is phosphorylated. In some embodiments, the ribonucleotide at the 5’ terminus of the sense strand is phosphorylated. In some embodiments, the ribonucleotide at the 3’ terminus and / or at the 5’ terminus of the antisense strand are phosphorylated. In some embodiments, the ribonucleotide at the 3’ terminus and at the 5’ terminus of the antisense strand are phosphorylated. In some embodiments, the ribonucleotide at the 3’ terminus of the antisense strand is phosphorylated. In some embodiments, the ribonucleotide at the 5’ terminus of the antisense strand is phosphorylated.In some embodiments, the ribonucleotide at the 3’ terminus and / or at the 5’ terminus of the sense strand are non-phosphorylated. In some embodiments, the ribonucleotide at the 3’ terminus of the sense strand is non-phosphorylated. In some embodiments, the ribonucleotide at the 5’ terminus of the sense strand is non-phosphorylated. In other embodiments, the ribonucleotide at the 3’ terminus and at the 5’ terminus of the antisense strand are non-phosphorylated. In some embodiments, the ribonucleotide at the 3’ terminus of the antisense strand is non-phosphorylated. In some embodiments, the ribonucleotide at the 5’ terminus of the antisense strand is non-phosphorylated.In some embodiments, one or more ribonucleotides of the sense and / or antisense strands of said oligonucleotide are modified. Thus, in some embodiments, one or more ribonucleotides of the sense stand of the oligonucleotide are modified. In some embodiments, one or more ribonucleotides of the antisense strand of said oligonucleotide are modified.In some embodiments a conjugated moiety is attached to one or more ribonucleotides of the sense and / or antisense strands of said oligonucleotide. Said conjugated moiety in some embodiments, affects the properties of said oligonucleotide, such as by altering its stability, binding affinities, or other parameters. In some embodiments, said conjugated moiety improves, increases, or enhances said oligonucleotide’s potency, wherein said oligonucleotide is used as a medicament.P7411 PC00In some embodiments, the oligonucleotide comprises a phosphorothioate (PS) backbone modification; a 5’ capping; a 5'-vinylphosphonate; a 2' ribose modification, such as 2'-O-methyl (2'-O-Me), such as 2'-O-methoxyethyl (2' MOE), such as 2'-fluoro, such as 2’-O-benzyl, such as 2’-O-methyl-4-pyridine (2’-O-CH2Py(4)); a methylene bridge between the 2' and 4' position of a ribose; a 5-methylcytosine (5mC); and / or a non-standard nucleotide.In some embodiments, the oligonucleotide comprises a phosphorothioate (PS) backbone modification. In some embodiments, the oligonucleotide comprises a 5’ capping. In some embodiments, the oligonucleotide comprises a 5'-vinylphosphonate. In some embodiments, the oligonucleotide comprises a 2' ribose modification. In some embodiments, said 2' ribose modification is 2'-O-methyl (2'-O-Me). In some embodiments, said 2' ribose modification is 2'-O-methoxyethyl (2' MOE). In some embodiments, said 2' ribose modification is 2'-fluoro. In some embodiments, said 2' ribose modification is 2’-O-benzyl, In some embodiments, said 2' ribose modification is 2’-O-methyl-4-pyridine (2’-O-CH2Py(4)). In some embodiments, the oligonucleotide comprises a methylene bridge between the 2' and 4' position of a ribose. In some embodiments, the oligonucleotide comprises a 5-methylcytosine (5mC). In some embodiments, the oligonucleotide comprises a non-standard nucleotide.In some embodiments, the oligonucleotide comprises a modification of the nucleic acid backbone, the nucleobase, the ribose sugar and / or 2'-ribose substitutions.In some embodiments, the oligonucleotide comprises a modification of the nucleic acid backbone. In some embodiments, the oligonucleotide comprises a modification of the nucleobase. In some embodiments, the oligonucleotide comprises a modification of the ribose sugar. In some embodiments, the oligonucleotide comprises 2’-ribose substitutions.In some embodiments, the oligonucleotide is conjugated to a moiety or to a nanoparticle formulation. In some embodiments the oligonucleotide is conjugated to a moiety. In some embodiments the oligonucleotide is conjugated to a nanoparticle formulation.P7411 PC00In some embodiments, said moiety is a cell-targeting moiety, and / or a cell-penetrating moiety. In some embodiments, said moiety is a cell-targeting moiety. In some embodiments, said moiety is a cell-penetrating moiety.In some embodiments, the oligonucleotide is conjugated to a Triantennary N- acetylgalactosamine (GalNAc) moiety; cholesterol; and / or a peptide. In some embodiments, the oligonucleotide is conjugated to a Triantennary N- acetylgalactosamine (GalNAc) moiety. In some embodiments, the oligonucleotide is conjugated to cholesterol. In some embodiments, the oligonucleotide is conjugated to a peptide. In some embodiments, said moieties and / or modifications increase the adsorption onto a cell and / or the absorption, uptake or endocytosis of the oligonucleotide into the cell. In some embodiments, said modifications to the oligonucleotide act to enhance, increase or improve the potency of said oligonucleotide, when said oligonucleotide is used a medicament, such as a medicament for use in a method of treating a metabolic disease, such as diabetes or obesity. Furthermore, in some embodiments the above described nucleotide modifications, serve to increase the absorption of the oligonucleotide into the cell, increase the selectivity of said oligonucleotide, and / or increase its stability, potentially thereby increasing its potency.In some embodiments, the oligonucleotide is a double stranded oligonucleotide.In some embodiments, the oligonucleotide comprises a overhang. A person skilled in the art will be able to identify an overhang. For example, said overhang might comprise or consist of one or more single stranded nucleotides at the 3’ terminus or 5’ terminus of said oligonucleotide, wherein the section of the oligonucleotide that is not the overhang is double-stranded.Medical use of the oligonucleotideIn an aspect, the present disclosure is directed to an oligonucleotide as described herein for use as a medicament.In an aspect, the present disclosure is directed to an oligonucleotide targeting ATG7 variant 2 (ATG7(2)) for use in a method of treating a metabolic disease.P7411 PC00In some embodiments, the metabolic disease is obesity, over-weight, an obesity- related disorder, pre-diabetes, impaired fasting glucose, Type 1 Diabetes, Type 2 Diabetes, a diabetes-related disorder, insulin resistance, elevated fasting glucose, hyperglycemia, elevated fasting serum triglyceride level (VLDL triglyceride), low high- density lipoprotein (HDL) levels, a fatty acid metabolism disorder, a cardiovascular disease, elevated blood pressure and atherosclerosis.In some embodiments, the metabolic disease is diabetes or obesity.In some embodiments, the metabolic disease is diabetes. In some embodiments, the metabolic disease is Type 1 Diabetes, Type 2 Diabetes, Gestational Diabetes, Maturity-Onset Diabetes of the Young (MODY), Latent Autoimmune Diabetes in Adults (LADA), Neonatal Diabetes Mellitus, Secondary Diabetes, Wolfram Syndrome (DIDMOAD), Cystic Fibrosis-Related Diabetes (CFRD), Steroid-Induced Diabetes, Type 3c Diabetes (Pancreatogenic Diabetes), Mitochondrial Diabetes.In some embodiments, the metabolic disease is Type 1 Diabetes. In some embodiments, the metabolic disease is Type 2 Diabetes. In some embodiments, the metabolic disease is Gestational Diabetes. In some embodiments, the metabolic disease is Maturity-Onset Diabetes of the Young (MODY). In some embodiments, the metabolic disease is Latent Autoimmune Diabetes in Adults (LADA). In some embodiments, the metabolic disease is Neonatal Diabetes Mellitus. In some embodiments, the metabolic disease is Secondary Diabetes. In some embodiments, the metabolic disease is Wolfram Syndrome (DIDMOAD). In some embodiments, the metabolic disease is Cystic Fibrosis-Related Diabetes (CFRD). In some embodiments, the metabolic disease is Steroid-Induced Diabetes. In some embodiments, the metabolic disease is Type 3c Diabetes (Pancreatogenic Diabetes). In some embodiments, the metabolic disease is Mitochondrial Diabetes.In some embodiments, the metabolic disease is obesity. In some embodiments, the obesity is Class 1 (Mild) Obesity, Class 2 (Moderate) Obesity, Class 3 (Severe or Morbid) Obesity, Monogenic Obesity, Syndromic Obesity, Polygenic or Common Obesity, or Secondary Obesity.In some embodiments, the obesity is Class 1 (Mild) Obesity. In some embodiments, the obesity is Class 2 (Moderate) Obesity. In some embodiments, the obesity is Class 3P7411 PC00(Severe or Morbid) Obesity. In some embodiments, the obesity is Monogenic Obesity.In some embodiments, the obesity is Syndromic Obesity. In some embodiments, the obesity is Polygenic or Common Obesity. In some embodiments, the obesity is Secondary Obesity.In some embodiments, it is beneficial to decrease the amount of ATG7(2) in a cell of at least 10%, such as at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95%, such as at least 99%.In some embodiments, the oligonucleotide for use decreases the amount of ATG7(2) in a cell of at least 10%, such as at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95%, such as at least 99%.In some embodiments, the oligonucleotide for use decreases the amount of ATG7(2) in a cell of 10% to 90%, such as 20% to 90%, 30% to 90%, 40% to 90%, 50% to 90%, 60% to 90%, 70% to 90%, 80% to 90%, such as 10% to 80%, 10% to 70%, 10% to 60%, 10% to 50%, 10% to 40%, 10% to 30%, 10% to 20%, such as 20% to 80%, 30% to 70%, such as 40% to 60%.It is possible to test in a controlled environment whether the oligonucleotide described herein decreases the amount of ATG7(2) of the desired amount. Such test comprises comparing cells that have been contacted with the oligonucleotide to cells derived from the same cell line that have not been contacted with the oligonucleotide.Administration form of the oligonucleotideThe present disclosure describes that the oligonucleotide describe herein can be administered in different forms. The skilled person will understand that any suitable method to deliver an oligonucleotide as described herein to a target cell might be employed.In some embodiments, the administration form of said oligonucleotide is: intravenous administration (IV); subcutaneous administration; intramuscular administration; intrathecal administration; intraventricular administration; oral administration; topical administration; inhalation administration; intraperitoneal administration; and / or intranasal administration.P7411 PC00In some embodiments, the administration form of said oligonucleotide is intravenous administration (IV). In some embodiments, the administration form of said oligonucleotide is subcutaneous administration. In some embodiments, the administration form of said oligonucleotide is intramuscular administration. In some embodiments, the administration form of said oligonucleotide is intrathecal administration. In some embodiments, the administration form of said oligonucleotide is intraventricular administration. In some embodiments, the administration form of said oligonucleotide is oral administration. In some embodiments, the administration form of said oligonucleotide is topical administration. In some embodiments, the administration form of said oligonucleotide is inhalation administration. In some embodiments, the administration form of said oligonucleotide is intraperitoneal administration. In some embodiments, the administration form of said oligonucleotide is intranasal administration.In some embodiments, the oligonucleotide, or oligonucleotide for use, is administered as: a. a naked oligonucleotide; b. an oligonucleotide encapsulated into a nanoparticle, such as a Lipid- based nanoparticle (LNPs), such as a Virus-like nanoparticle (VLP); c. an oligonucleotide encapsulated into a microsphere or a microcapsule; d. conjugated to a targeting ligand, such as a peptide, such as an aptamer; e. incorporated into a vesicle, such as a liposome; f. an oligonucleotide adsorbed onto or incorporated into a hydrogel; g. a prodrug, such as wherein the prodrug is metabolized in an active form in the body.In some embodiments, the oligonucleotide, or oligonucleotide for use, is administered as a naked oligonucleotide. In some embodiments, the oligonucleotide, or oligonucleotide for use, is administered as an oligonucleotide encapsulated into a nanoparticle, such as a Lipid-based nanoparticle (LNPs), such as a Virus-like nanoparticle (VLP). In some embodiments, the oligonucleotide, or oligonucleotide for use, is administered as a Lipid-based nanoparticle (LNPs). In some embodiments, the oligonucleotide, or oligonucleotide for use, is administered as a Virus-like nanoparticle (VLP). In some embodiments, the oligonucleotide, or oligonucleotide for use, isP7411 PC00 administered as an oligonucleotide encapsulated into a microsphere. In some embodiments, the oligonucleotide, or oligonucleotide for use, is administered as an oligonucleotide encapsulated into a microcapsule. In some embodiments, the oligonucleotide, or oligonucleotide for use, is administered as conjugated to a targeting ligand, such as a peptide, such as an aptamer. In some embodiments, the oligonucleotide, or oligonucleotide for use, is administered as conjugated to a targeting peptide. In some embodiments, the oligonucleotide, or oligonucleotide for use, is administered as conjugated to a targeting aptamer. In some embodiments, the oligonucleotide, or oligonucleotide for use, is administered as incorporated into a vesicle, such as a liposome. In some embodiments, the oligonucleotide, or oligonucleotide for use, is administered as incorporated into a liposome. In some embodiments, the oligonucleotide, or oligonucleotide for use, is administered as an oligonucleotide adsorbed onto or incorporated into a hydrogel. In some embodiments, the oligonucleotide, or oligonucleotide for use, is administered as an oligonucleotide adsorbed onto a hydrogel. In some embodiments, the oligonucleotide, or oligonucleotide for use, is administered as an oligonucleotide incorporated into a hydrogel. In some embodiments, the oligonucleotide, or oligonucleotide for use, is administered as a prodrug, such as wherein the prodrug is metabolized in an active form in the body.In an aspect of the invention, the present disclosure is directed to a composition comprising the oligonucleotide, or the oligonucleotide for use as described herein.In some embodiments, the composition further comprises an acceptable carrier.The present disclosure describes that the oligonucleotide, or oligonucleotide for use, can be used in the for the manufacture of medicaments, as well as in methods of treating disease.In an aspect of the invention, the present disclosure is directed to a method of treating a metabolic diseases, wherein the metabolic disease is as described herein, said method comprising administration of a therapeutically effective amount of an oligonucleotide targeting ATG7(2) to an individual in need thereof.P7411 PC00In an aspect of the invention, the present disclosure is directed to a method of treating diabetes, the method comprising administering the oligonucleotide as described herein.In an aspect of the invention, the present disclosure is directed to a method of treating obesity, the method comprising administering the oligonucleotide as described herein.In an aspect of the invention, the present disclosure is directed to the use of the oligonucleotide as described herein, for the manufacture of a medicament.In an aspect of the invention, the present disclosure is directed to the use of the oligonucleotide as described herein, for the manufacture of a medicament for treating a metabolic diseases, wherein the metabolic disease is as described herein.Items1 . An oligonucleotide targeting ATG7 variant 2 (ATG7(2)).2. An oligonucleotide targeting ATG7 variant 2 (ATG7(2)) for use as a medicament.3. An oligonucleotide targeting ATG7 variant 2 (ATG7(2)) for use in a metabolic disease.4. The oligonucleotide targeting ATG7(2) for use according to item 3, wherein the metabolic disease is diabetes or obesity.5. The oligonucleotide, or the oligonucleotide for use according to any one of the preceding items, wherein said ATG7(2) is human ATG7(2).6. The oligonucleotide, or the oligonucleotide for use according to any one of the preceding items, wherein ATG7(2) is as set forth in SEQ ID NO: 18.P7411 PC007. The oligonucleotide, or the oligonucleotide for use according to any one of the preceding items, wherein ATG7(2) is encoded by a nucleotide sequence comprising a nucleotide sequence as set forth in SEQ ID NO: 30.8. The oligonucleotide, or the oligonucleotide for use according to any one of the preceding items, wherein said oligonucleotide is selective for ATG7(2).9. The oligonucleotide, or the oligonucleotide for use according to any one of the preceding items, wherein said oligonucleotide is complementary to ATG7(2) mRNA.10. The oligonucleotide, or the oligonucleotide for use according to any one of the preceding items, wherein the oligonucleotide inhibits ATG7(2) translation.11. The oligonucleotide, or the oligonucleotide for use according to any one of the preceding items, wherein said oligonucleotide does not target ATG7 variant 1 (ATG7(1)).12. The oligonucleotide, or the oligonucleotide for use according to any one of the preceding items, wherein ATG7(1) is as set forth in SEQ ID NO: 20.13. The oligonucleotide, or the oligonucleotide for use according to any one of the preceding items, wherein ATG7(1) is encoded by a nucleotide sequence comprising a nucleotide sequence as set forth in SEQ ID NO: 31.14. The oligonucleotide, or the oligonucleotide for use according to any one of the preceding items, wherein said oligonucleotide is complementary to: a fragment of exon 16 of ATG7 and to a fragment of exon 18 of ATG7.15. The oligonucleotide, or the oligonucleotide for use according to any one of the preceding items, wherein said oligonucleotide is not complementary to exon 17 of ATG7.16. The oligonucleotide, or the oligonucleotide for use according to any one of the preceding items, wherein the oligonucleotide comprises a sequenceP7411 PC00 complementary to an ATG7 mRNA comprising a junction between exon 16 and exon 18 of ATG7.17. The oligonucleotide, or the oligonucleotide for use according to any one of the preceding items, wherein said oligonucleotide binds to the junction between exon 16 and exon 18 of ATG7(2).18. The oligonucleotide, or the oligonucleotide for use according to any one of the preceding items, wherein said oligonucleotide binds to a 3’ fragment of exon 16 of ATG7 and to a 5’ fragment of exon 18 of ATG7.19. The oligonucleotide, or the oligonucleotide for use according to any one of the preceding items, wherein the oligonucleotide comprises: a. at least 3 nt, such as at least 4, such as at least 5 nt complementary to exon 16 of ATG7(2); and b. at least 3 nt, such as at least 4, such as at least 5 nt complementary to exon 18 of ATG7(2).20. The oligonucleotide, or the oligonucleotide for use according to any one of the preceding items, wherein the oligonucleotide comprises: a. at least the 3 nt, such as at least the 4, such as at least the 5 nt complementary to the 3’ end of exon 16 of ATG7(2); and b. at least 3 nt, such as at least 4, such as at least 5 nt complementary to the 5’ end of exon 18 of ATG7(2).21 . The oligonucleotide, or the oligonucleotide for use according to any one of the preceding items, wherein said oligonucleotide comprises a double-stranded region of 10 to 30 nt, such as 15 to 25 nt.22. The oligonucleotide, or the oligonucleotide for use according to any one of the preceding items, wherein said oligonucleotide comprises a strand comprising SEQ ID NO: 13 or a functional variant thereof, wherein said functional variant has 1 or 2 individual ribonucleotide substitutions compared to SEQ ID NO: 13.P7411 PC0023. The oligonucleotide, or the oligonucleotide for use according to item 20, wherein said oligonucleotide further comprises 1 to 16 nucleotides attached at the 3’ of SEQ ID NO: 13 or said functional variant thereof.24. The oligonucleotide, or the oligonucleotide for use according to any one of items 20 and 21 , wherein said oligonucleotide further comprises 1 to 16 nucleotides attached at the 5’ of SEQ ID NO: 13 or said functional variant thereof.25. The oligonucleotide, or the oligonucleotide for use according to any one of the preceding items, wherein said oligonucleotide comprises a strand comprising SEQ ID NO: 16 or SEQ ID NO: 17, or a functional variant thereof, wherein said functional variant has 1 , 2, 3, 4, 5, or 6 individual ribonucleotide substitutions compared to any one of SEQ ID NO: 16 or SEQ ID NO: 17.26. The oligonucleotide, or the oligonucleotide for use according to any one of the preceding items, wherein said oligonucleotide comprises a strand of SEQ ID NO: 16 or SEQ ID NO: 17, or a functional variant thereof, wherein said functional variant has 1 , 2 or 3 individual ribonucleotide substitutions compared to any one of SEQ ID NO: 16 or SEQ ID NO: 17.27. The oligonucleotide, or the oligonucleotide for use according to any one of the preceding items, wherein said oligonucleotide comprises a sense strand of SEQ ID NO: 16, SEQ ID NO: 17, or SEQ ID NO: 13 or functional variant thereof, and a complementary antisense strand.28. The oligonucleotide, or the oligonucleotide for use according to any one of the preceding items, wherein the ribonucleotide at the 3’ terminus and at the 5’ terminus of the sense strand are phosphorylated.29. The oligonucleotide, or the oligonucleotide for use according to any one of the preceding items, wherein the ribonucleotide at the 3’ terminus and at the 5’ terminus of the antisense strand are phosphorylated.P7411 PC0030. The oligonucleotide, or the oligonucleotide for use according to any one of the preceding items, wherein the ribonucleotide at the 3’ terminus and at the 5’ terminus of the sense strand are non-phosphorylated.31 . The oligonucleotide, or the oligonucleotide for use according to any one of the preceding items, wherein the ribonucleotide at the 3’ terminus and at the 5’ terminus of the antisense strand are non-phosphorylated.32. The oligonucleotide, or the oligonucleotide for use according to any one of the preceding items, wherein one or more ribonucleotides of the sense and / or antisense strands of said oligonucleotide are modified.33. The oligonucleotide, or the oligonucleotide for use according to any one of the preceding items, wherein a conjugated moiety is attached to one or more ribonucleotides of the sense and / or antisense strands of said oligonucleotide.34. The oligonucleotide, or the oligonucleotide for use according to any one of the preceding items wherein the oligonucleotide comprises a phosphorothioate (PS) backbone modification; a 5’ capping; a 5'-vinylphosphonate; a 2' ribose modification, such as 2'-O-methyl (2'-O-Me), such as 2'-O-methoxyethyl (2' MOE), such as 2'-fluoro, such as 2’-O-benzyl, such as 2’-O-methyl-4-pyridine (2’-O-CH2Py(4)); a methylene bridge between the 2' and 4' position of a ribose; a 5-methylcytosine (5mC); a non-standard nucleotide.35. The oligonucleotide, or the oligonucleotide for use according to any one of the preceding items wherein the oligonucleotide comprises a modification of the nucleic acid backbone, the nucleobase, the ribose sugar and / or 2'-ribose substitutions.36. The oligonucleotide, or the oligonucleotide for use according to any one of the preceding items, wherein the oligonucleotide is conjugated to a moiety or to a nanoparticle formulation.P7411 PC0037. The oligonucleotide, or the oligonucleotide for use according to any one of the preceding items, wherein the moiety is a cell-targeting moiety, and / or a cellpenetrating moiety.38. The oligonucleotide, or the oligonucleotide for use according to any one of the preceding items, wherein the oligonucleotide is conjugated to Triantennary N- acetylgalactosamine (GalNAc) moiety; cholesterol; and / or a peptide.39. The oligonucleotide, or the oligonucleotide for use according to any one of the preceding items, wherein the oligonucleotide is: a siRNA, a shRNA, an antisense oligonucleotide, a guide RNA, a LNA (Locked Nucleic Acid) antisense oligonucleotide, or a morpholino oligomer.40. The oligonucleotide, or the oligonucleotide for use according to any one of the preceding items, wherein the oligonucleotide is a double stranded oligonucleotide.41 . The oligonucleotide, or the oligonucleotide for use according to any one of the preceding items, wherein the oligonucleotide is a siRNA.42. The oligonucleotide, or the oligonucleotide for use according to any one of the preceding items, wherein the oligonucleotide comprises a overhang.43. The oligonucleotide, or the oligonucleotide for use according to any one of the preceding items, wherein the form of administration of said oligonucleotide is: intravenous administration (IV); subcutaneous administration; intramuscular administration; intrathecal administration; intraventricular administration; oral administration; topical administration; inhalation administration; intraperitoneal administration; and / or intranasal administration.44. The oligonucleotide, or the oligonucleotide for use according to any one of the preceding items, wherein the oligonucleotide is administered as: a. a naked oligonucleotide;P7411 PC00 b. an oligonucleotide encapsulated into a nanoparticle, such as a Lipid- based nanoparticle (LNPs), such as a Virus-like nanoparticle (VLP); c. an oligonucleotide encapsulated into a microsphere or a microcapsule; d. conjugated to a targeting ligand, such as a peptide, such as an aptamer; e. incorporated into a vesicle, such as a liposome; f. an oligonucleotide adsorbed onto or incorporated into a hydrogel; g. a prodrug, such as wherein the prodrug is metabolized in an active form in the body.45. The oligonucleotide, or the oligonucleotide for use according to any one of the preceding items, wherein the oligonucleotide decreases the amount of ATG7(2) in a cell of 10% to 90%.46. A composition comprising the oligonucleotide, or the oligonucleotide for use according to any one of the preceding claims.47. The composition according to claim 46, wherein the composition further comprises an acceptable carrier.48. A method of treating a metabolic disease, said method comprising administration of a therapeutically effective amount of an oligonucleotide targeting ATG7(2) to an individual in need thereof.49. A method of treating a metabolic diseases, the method comprising administering the oligonucleotide according to any one of items 1 to 45, or the composition according to items 46 to 47.50. The method according to any one of items 48 to 49, wherein the metabolic disease is diabetes or obesity.51. Use of the oligonucleotide according to any one items 1 to 45, or of a composition according to any one of items 46 to 47, for the manufacture of a medicament.P7411 PC0052. Use of the oligonucleotide according to any one claims to 45, or of a composition according to any one of items 46 to 47, for the manufacture of a medicament for treating a metabolic disease.53. The use according to item 52, wherein the metabolic disease is diabetes or obesity.54. The use according to item 53, wherein the metabolic disease is diabetes55. The use according to item 54, wherein the metabolic disease is obesity.Items 21. An oligonucleotide targeting human ATG7 variant 2 (ATG7(2)), wherein ATG7(2) comprises a junction between exon 16 and exon 18 of ATG7, for use in a metabolic disease.2. The oligonucleotide for use according to claim 1, wherein the metabolic disease is diabetes or obesity.3. The oligonucleotide for use according to any one of the preceding claims, wherein said oligonucleotide is selective for ATG7(2), and wherein said oligonucleotide does not target ATG7 variant 1 (ATG7(1)).4. The oligonucleotide for use according to any one of the preceding claims, wherein ATG7(2) is as set forth in SEQ ID NO: 18 and wherein ATG7(1) is as set forth in SEQ ID NO: 205. The oligonucleotide for use according to any one of the preceding claims, wherein said oligonucleotide binds to the junction between exon 16 and exon 18 of ATG7(2).6. The oligonucleotide for use according to any one of the preceding claims, wherein said oligonucleotide binds to a 3’ fragment of exon 16 of ATG7 and to a 5’ fragment of exon 18 of ATG7.P7411 PC007. The oligonucleotide for use according to any one of the preceding claims, wherein the oligonucleotide comprises: a. at least the 3 nt, such as at least the 4, such as at least the 5 nt complementary to the 3’ end of exon 16 of ATG7(2); and b. at least 3 nt, such as at least 4, such as at least 5 nt complementary to the 5’ end of exon 18 of ATG7(2).8. The oligonucleotide for use according to any one of the preceding claims, wherein said oligonucleotide comprises a double-stranded region of 10 to 30 nt, such as 15 to 25 nt.9. The oligonucleotide for use according to any one of the preceding claims, wherein said oligonucleotide comprises a strand comprising SEQ ID NO: 13 or a functional variant thereof, wherein said functional variant has 1 or 2 individual ribonucleotide substitutions compared to SEQ ID NO: 13.10. The oligonucleotide for use according to claim 8, wherein said oligonucleotide further comprises 1 to 16 nucleotides attached at the 3’ of SEQ ID NO: 13 or said functional variant thereof, and / or wherein said oligonucleotide further comprises 1 to 16 nucleotides attached at the 5’ of SEQ ID NO: 13 or said functional variant thereof.11 . The oligonucleotide for use according to any one of the preceding claims, wherein said oligonucleotide comprises a strand of SEQ ID NO: 16 or SEQ ID NO: 17, or a functional variant thereof, wherein said functional variant has 1 , 2 or 3 individual ribonucleotide substitutions compared to any one of SEQ ID NO: 16 or SEQ ID NO: 17.12. The oligonucleotide for use according to any one of the preceding claims, wherein said oligonucleotide comprises a sense strand of SEQ ID NO: 16, SEQ ID NO: 17, or SEQ ID NO: 13 or functional variant thereof, and a complementary antisense strand.13. The oligonucleotide for use according to any one of the preceding claims, wherein the oligonucleotide is a siRNA.P7411 PC0014. The oligonucleotide for use according to any one of the preceding claims, wherein the oligonucleotide comprises a overhang.15. The oligonucleotide for use according to any one of the preceding claims, wherein the oligonucleotide decreases the amount of ATG7(2) in a cell of 10% to 90%.ExamplesExample 1.Materials and MethodsGeneration of CRISPR strainsWe used the CRISPR / Cas9 approach to generate Drosophila melanogaster (fruit flies) with knockout (KO) of ATG7 and rescue with either isoform of the human protein, hATG7(1) or hATG7(2). Two gRNAs were used to remove the fly gene, targeting each end of the gene. The gRNA sequences were as follows:5’ CTTGGCTGCTACTTC 3’5’ ACCATGTCTCATGACAAC 3’These were inserted into the pCFD4-U6:1_U6:3tandemgRNAs plasmid (Addgene plasmid #49411). A donor plasmid containing the protein coding sequence of either isoform of the human protein was included for the insertion of the respective isoform. The constructs also contained a dsRed marker for easy screening of positive flies, which exhibit a rosy eye colour. The marker was expressed as a hATG7-dsRed fusion protein. The construct also contained a 1 kb homology sequence both upstream and downstream of the hATG7-dsRed sequence, to facilitate the incorporation of the gene to the target site in the fly genome. These strains were injected by BestGENE Inc. The injection strain was yw;;nos-Cas9(lll-attP2) which expresses Cas9 under the nanos promoter on W

[1118] background. W

[1118] (BDSC #5905) was used as wild type (WT) in all experiments. Correct genotypes were confirmed by Sanger sequencing, carried out by deCODE Genetics.Validation of strainsP7411 PC00Expression of fly Atg7 and human ATG7 were assessed at mRNA level by qPCR and RNA sequencing. Third instar larvae were snap frozen in liquid nitrogen and RNA isolated by TRIzol. cDNA was generated from the RNA using MultiScribe Reverse Transcriptase and random primers (Applied Biosystems, #4368814). The qPCR primers are listed in table 1 below. Expression of fly Atg7 (dAtg7) and human ATG7 (hATG7) were normalized to the expression of two housekeeping genes.Table 1 : Primer sequences used in qPCR.Expression on protein level was assessed for hATG7 by confocal imaging. It was not possible to stain for fly Atg7 due to a lack of commercially available antibody. For confocal imaging, fat bodies (the fat bodies are the primary fat storage site in fruit flies and also serve a similar role as the mammalian liver), from third instar larvae were dissected in 4% formaldehyde. The tissues were incubated for about 30 minutes. Then, 1 mL of blocking solution (TBS-T with 10% goat serum) was added and incubated for 1 hour at room temperature (RT). The samples were then stained with ATG7 primary antibody (Cell Signaling #8558) overnight at 4°C. The antibody was added directly into the blocking buffer (in a dilution of 1 :100). Then the tissues were moved into a new tube containing secondary antibody diluted in TBS-T (Alexa 647 anti-rabbit, Invitrogen #A21244), diluted to 1 :200) and incubated for 1-2 hours at RT in the dark. The tissues were then moved to a new tube containing washing buffer (TBS-T). Lastly, the tissuesP7411 PC00 were moved into a drop of TBS on a dissection plate and the dissection finished before the samples were moved onto a microscope slide and spread out into a single layer. The samples were mounted with VectaShield containing DAPI. The samples were imaged on a Olympus Confocal system.Lifespan, development, motor function, and tolerance to stressTo measure the lifespan of the different fruit fly strains under normal conditions, large crosses were set up and allowed to cross for 3 days at 25°C with 12-hour sunlight. The vials were flipped every three days. The first vials were not used in experiments. Newly eclosed males were placed in vials and kept at 25°C with 12-hour sunlight. About 35 males were placed in each vial. Every three days the vials were flipped and dead flies counted. At least three replicates were made for each strain, resulting in at least 100 individuals from each strain in total.In stress tolerance experiments, males were collected in the same manner as described above and placed on the appropriate medium. For starvation, Kimwipes soaked in water were placed at the bottom of vials. Water was added as needed and dead flies counted every 2 days. For Paraquat experiments, paraquat was stirred into standard cornmeal food to a final concentration of 10 mM. Dead flies were counted every 1-2 days. The flies were kept at 25°C with 12-hour sunlight. The survival curves were compared by the Kaplan-Meier method and p-values calculated by the Mantel- Cox log-rank test.A climbing assay was carried out to evaluate general locomotion in the flies. Between 1-3-day old flies were used. Males from each strain were collected and allowed to recover overnight, to avoid effects of anesthesia. Twenty males were then placed in a long vial. The vials were simultaneously dropped to bring the flies down to the bottom. The flies were recorded for 10 seconds and flies who had reached a mark at approximately 2 / 3rdof the vial height at that time were counted. At least three replicates were made with the same group of flies each time. At least three independent experiments were carried out for each strain.Larval developmental time was measured as the time from when the parents were placed in the vials until the third instar larva crawled up from the food. Prepupa were counted approximately every 12 hours, or at least once in the am and once in the pm. For metamorphosis times, prepupa were counted and moved to new vials using a wet brush. The vials were then monitored and eclosed flies counted every 12 hours. DeadP7411 PC00 pupae were counted by the end of the experiment. The vials were kept at 25°C with 12- hour sunlight.Metabolic experimentsWhen measuring lipid contents of the larvae, a few flies from each strain were allowed to cross for 2 days. The number of parents depended on the fertility of each strain with the goal of obtaining a comparable number of offspring. This was done to avoid population density effects, which would affect the size of the offspring. Commonly, in comparable experiments, an equal number of embryos are placed in vials (Shingleton et al., 2009). However, as these strains had such different survival rates this led to vastly different numbers of offspring between the strains. Therefore, it turned out to be more feasible to optimize the number of parents to generally obtain a similar number of offspring. Flies were either cultured on standard cornmeal food or a high sugar diet (HSD) which consisted of 20% sucrose in their standard cornmeal food.For confocal imaging of lipid droplets, third instar larvae were dissected, stained, and imaged as described above for staining with ATG7. This was excluding the overnight step in primary antibodies as the stain used for lipids (BODIPY 493 / 503, Invitrogen #D3922) is a fluorescent stain with affinity for neutral lipids, but not an antibody. Therefore the tissues were stained in BODIPY, diluted to 1 :1000 in blocking solution, for 1-2 hours at RT in the dark, before they were mounted in VectaShield containing DAPI. The number and area of lipid droplets in fat bodies and midgut were measured using Fiji.Trehalose was measured using the Megazyme kit (Megazyme, Trehalose assay kit #K- TREH). The samples were prepared as described before (Han et al., 2021). In short, 20 flies were collected for each sample and kept on ice. The flies were washed 2 times with 500 pL of cold PBS and the wash buffer removed each time. The flies were homogenized in 200 pL of cold PBS. Then the samples were centrifuged at 5000 RPM for 1 minute. The supernatant was transferred to a new tube, heated at 70°C for 5 minutes, and the samples centrifuged again at max speed for 3 minutes, at 4°C. The supernatant was transferred to a new tube and stored at -20°C until analyzed.A portion of the samples was used to measure the total protein concentration using BCA protein assay (Thermo Fisher #23225). The procedure was carried out as described in the kit user manual. The trehalose levels were normalized to the totalP7411 PC00 protein concentration. Trehalose was measured exactly as described in the kit user manual.Analysis of human RNA sequencing dataA publicly available human RNA sequencing dataset, from subcutaneous adipose tissue, was obtained from the NCBI’s Gene Expression Omnibus database (GEO GSE152991). The study included 45 adult individuals, male and female. The subjects were split into lean (n = 11) and obese (n = 34) groups based on adiposity and insulin sensitivity. The raw FASTQ data was obtained and processed with Kallisto (version 0.51.1). First the reads were cleaned with Trimmomatic and then pseudoalignment was done using human transcriptome GRCh38, release 108, available from Ensembl. org. Reads were quantified in pair-end mode with 100 bootstrap samples. Isoform specific expression estimates for ATG7 were then extracted using R.ResultsTwo strains expressing hATG7(2) were obtained, with different expression levels of the protein (figure 1). Therefore they were referred to as hATG7(2)high(higher expression) and hATG7(2)l0W(lower expression). While knocking out the fly gene was successful in the Atg7~'~ strain and hATG7(1) strain, both the hATG7(2) strains also retained the fly gene despite also incorporating the human gene into their genome. Therefore, the hATG7(2) strains express the human ATG7(2) protein on a wild type (WT) background, rather than KO background and should be compared to WT rather than Atg7' / '.Lower expression ofhATG7(2) leads to a lifespan increase and increased tolerance to stressIt is known that Atg7 KO leads to a lifespan reduction in fruit flies and this was the case for our Atg7' strain which had a 44% shorter median lifespan than that of WT. This phenotype was not rescued by introducing hATG7(1), which even performed worse than the KO with 51% shorter median lifespan than WT. hATG7(2)highexhibited a slight lifespan reduction, with a median of 9%. Surprisingly, hATG7(2)l0Woutlived WT by 17% (figure 2A). Note that no genetical differences were observed between the two hATG7(2) strains apart from their expression levels.Autophagy is important for survival under stress, such as nutrient or oxygen depletion. We tested the tolerance of the different strains under starvation and oxidative stress.P7411 PC00To test oxidative stress, the flies received 10 mM Paraquat in their standard food (figure 2B). The treatment was not well tolerated, and most flies died within 3 days. The median lifespan of WT, Atg7_ / _and hATG7(2)l0Wwere 3 days while hATG7(1) and hATG7(2)highhad a slightly worse median lifespan of 2 days (hATG7(1) performed worse than Atg7' / _, p > 0.0001). However, the hATG7(2)l0Wstrain performed significantly better than WT and lived 58% longer on average, with some flies surviving up to 20 days. On complete starvation, however, this turned around for the hATG7(2) strains (figure 2C). While all strains survived for about 3 days with nothing but water, hATG7(2)highsurvived for about 4 days on average. Again, hATG7(1) performed significantly worse than Atg7_ / _(p = 0.0012). hATG7(2) expressing fruit flies exhibit a diabetic phenotypeI ntriguingly, the two hATG7(2) strains had not compared in health and viability experiments, as discussed above. Most surprising of all, however, was the size difference between the two strains (figure 3). The strain with the higher expression, hATG7(2)highproduced considerably larger larvae and flies with a large abdomen. To address whether the substantial size increase in hATG7(2)highis due to increase in fat reserves, we stained neutral lipids in fat bodies of third instar larvae with BODIPY. The fat bodies are the primary site of metabolism in Drosophila and serve the role of which both white adipose tissue and liver do in mammals (Chatterjee and Perrimon, 2021). Lipid staining in fat bodies revealed that the hATG7(2)highhave significant accumulation of lipids in their fat bodies (figure 4A-B). We wondered whether the phenotype would be affected by diet and repeated the experiment on larvae which had been fed a high sugar diet (HSD) which consisted of 20% sucrose in their standard cornmeal food. The lipid accumulation became more pronounced on the HSD, in which case both hATG7(2) strains exhibited significant lipid accumulation (figure 5A-B).The main circulating sugars in fruit flies is trehalose, a disaccharide consisting of two glucose molecules. It has been shown that long-term effects of HSD are increased levels of both glucose and trehalose. However, glucose spikes after feeding while trehalose remains steady. Therefore, trehalose is a more suitable measurement for insulin resistance. Under normal conditions, hATG7(2)highhad a significant increase in circulating trehalose in comparison to WT. Although not significant, again a slight increase was seen in Atg7' / _, but not in hATG7(1) (figure 6A). Although differences for the hATG7(2) strains were more pronounced on HSD they were not significant due toP7411 PC00 great variance in the samples (figure 6B). Notably, both hATG7(2) strains show a comparable trend towards higher trehalose levels, compared to the other strains.Diabetic fruit fly models have been reported to spend more time in the food (on the larval stage), whether it is due to slower development or dysregulation in pathways mediating their trip from the food to start metamorphosis. Both hATG7(2) strains exhibited delayed larval development (figure 7A) and this became more pronounced on HSD (figure 7B). Lastly, we observed a phenotype in hATG7(2)highwhere they fail to fully emerge from their pupal case (figure 8A). Failure to emerge from the pupal case can indicate muscle weakness or muscle atrophy. We further assessed the motor function of the different strains by a climbing assay (figure 8B). The Atg7' / _and hATG7(1) strains were barely able to climb at all (1 ,2% of the dAtg7_ / _and 0% of hATG7(1), 11 ,5% of hATG7(2)highclimbed and 21 ,5% of hATG7(2)l0W, compared to 64,1% of WT. As the hATG7(2) strains do not lack autophagy, this indicates detrimental effect of ATG7(2) on motor function, independently of autophagy (as these strains have functional autophagy). Moreover, while 2,7% of the WT flies died on pupal stage, 29% of Atg7-Z- did not survive, 28% of hATG7(1), 57% of hATG7(2)highand 12% of hATG7(2)l0W(figure 8C). Despite these differences, only hATG7(2)highwas significantly different from WT. Of the hATG7(2)highwhich died during metamorphosis, roughly half or 55% died due to failure to fully eclose or emerge from the pupal case. This was observed in 0-2% of the other strains. This phenotype along with decreased climbing ability can indicate muscle atrophy, which is commonly observed with obesity. Collectively, these results indicate a diabetic phenotype in the hATG7(2) flies.Expression ofATG7(2) is elevated in obese humansTo assess whether the obesity phenotypes we have observed in our fruit fly models translate to humans we analysed a publicly available RNA sequencing dataset from human subcutaneous adipose tissue. This revealed that expression of ATG7 is indeed elevated in obese humans, compared to the lean control group (figure 9), and this increase is specifically due to isoform ATG7(2), but not ATG7(1). In line with our findings in fruit flies, this suggests that increased expression of ATG7(2) is linked to obesity in humans.P7411 PC00ConclusionAlthough the seemingly contradicting phenotypes of the two hATG7(2) expressing Drosophila strains were puzzling to begin with, the ability to produce a similar phenotypic profile in both strains using diet indicates a metabolic root. Previous in vitro studies in MEFs and human hepatic cancer cell lines, found that ATG7(2) may suppress both glycolysis and mitochondrial respiration. Under normal conditions, the strain with the higher hATG7(2) expression, hATG7(2)high, exhibited diabetic phenotypes. These included fat accumulation, increased trehalose levels, delayed larval development, and decreased motor function. The phenotypes were exaggerated on HSD, in which case the strain with the lower hATG7(2) expression, hATG7(2)l0W, also exhibited fat accumulation and a trend towards increased trehalose levels. This strain also displayed decreased motor function and delayed larval development under normal conditions, the latter of which was more severe on HSD (motor function was not assessed on HSD). Interestingly, hATG7(2)l0Whad a longer lifespan and increased tolerance to stress, most notably oxidative stress. These positive phenotypes could result from a mild repression of glycolysis and in turn more active beta-oxidation. Collectively, the study points towards a metabolic function of ATG7(2), primarily through suppression of glycolysis, indicating that modulation of ATG7(2) expression and maintaining its expression low may result in prevention and / or treatment of metabolic diseases such as obesity and diabetes. Additional functions of the isoform might be at play, such as p53 binding which has been reported previously.Example 2Further characterization of these Drosophila strains will provide valuable insight into the effects of this isoform and implications in diseases. A Drosophila strain expressing a tandem GFP-mCherry fusion protein tagged to the outer mitochondrial membrane is used for assessing mitochondrial integrity and mitochondrial turnover through mitophagy in the hATG7(2) strains. This system relies on the pH sensitivity of GFP, which does not tolerate the acidic environment of lysosomes. The hATG7(2) strains could show defects in mitochondrial turnover using this method in comparison with the control strain.Example 3A UAS-Gal4 expression system is set up for hATG7(2) expression, enabling the assessment of phenotypes in a tissue-specific manner. This will allow us to addressP7411 PC00 whether the phenotypes can be induced and alleviated by turning the expression of hATG7(2) on-and-off. Moreover, we will introduce siATG7(2) using the UAS-Gal4 system, generating a strain that will be crossed with the hATG7(2) CRISPR high and low strains and it is expected that this will revert the metabolic phenotypes that we have observed in these strains.Example 4Cell cultureCapan-1 cells were cultivated on collagen rat tail I (Sigma C3867) coated vessels in IMDM (Gibco 12440053) supplemented with 20% FBS (Gibco A5256701) in an incubator at 37°C with 5% CO2. DMEM high Glucose (Gibco 41965039) with 20% FBS (Gibco 11995073) was to grow the cells. siRNA knock-downCells were reverse- or forward-transfected with siRNA using OptiMEMTM (Gibco 11058021) and Lipofectamine™ RNAiMax transfection reagent (Invitrogen 13778) following manufacturer’s recommendation. The following siRNAs were used : siATG7 : 5’- GAAGCUCCCAAGGACAUUAtt -3’ (SEQ ID NO: 14) siATG7(1) : 5’- GGCAUUUGACAAAUGUACAgc -3’ (SEQ ID NO: 15) siATG7(2): 5’- TCACCAGGTTCTTGATCaa -3’ (SEQ ID NO: 16) 5’- UGUGCCUCACCAGGUUCtt -3’ (SEQ ID NO: 17) siRNAs were designed to target either total ATG7 by binding to exon 2, ATG7(1) by binding to exon 17, and ATG7(2) by binding to the junction of exons 16 and 18 (Fig. 4. A and Fig. 10). Two different siRNA sequences were tested for A TG7(2) knock-down. Efficiency and specificity of the siRNAs was confirmed in MEF Atg7_ / _cells expressing either A TG7(1) or ATG7(2) (Fig. 5.A-F). Capan-1 WT and ATG7(1)-Z- cells were transfected with the siRNAs and qPCR results revealed that A TG7(2) knock-down leads to reduced ATG7(1) expression in WT cells (Fig. 4.B-D). Additionally, transfection of the ATG7(1)' / _clone with the highest A TG7(2) expression revealed that the second siATG7(2) sequence was the most efficient one, with 60% knock-down efficiency (Fig.4.E-G).Sequence overviewP7411 PC00SEQ ID NO: 13: 5’- CAGGTTCTT -3’SEQ ID NO: 14: siATG7 : 5’- GAAGCUCCCAAGGACAUUAtt -3’SEQ ID NO: 15: siATG7(1) : 5’- GGCAUUUGACAAAUGUACAgc -3’SEQ ID NO: 16: siATG7(2): 5’- UCACCAGGUUCUUGAUCAAtt -3’SEQ ID NO: 17: 5’- UGUGCCUCACCAGGUUCUUtt -3’SEQ ID NO: 18: ATG7(2) polypeptide:MAAATGDPGLSKLQFAPFSSALDVGFWHELTQKKLNEYRLDEAPKDIKGYYYNGDSAGLPARLTLEFSAFDMSAPTPARCCPAIGTLYNTNTLESFKTADKKLLLEQAANEIWESIKSGTALENPVLLNKFLLLTFADLKKYHFYYWFCYPALCLPESLPLIQGPVGLDQRFSLKQIEALECAYDNLCQTEGVTALPYFLIKYDENMVLVSLLKHYSDFFQGQRTKITIGVYDPCNLAQYPGWPLRNFLVLAAHRWSSSFQSVEVVCFRDRTMQGARDVAHSIIFEVKLPEMAFSPDCPKAVGWEKNQKGGMGPRMVNLSECMDPKRLAESSVDLNLKLMCWRLVPTLDLDKWSVKCLLLGAGTLGCNVARTLMGWGVRHITFVDNAKISYSNPVRQPLYEFEDCLGGGKPKALAAADRLQKIFPGVNARGFNMSIPMPGHPVNFSSVTLEQARRDVEQLEQLIESHDWFLLMDTRESRWLPAVIAASKRKLVINAALGFDTFVVMRHGLKKPKQQGAGDLCPNHPVASADLLGSSLFANIPGYKLGCYFCNDVVAPGDSTRDRTLDQQCTVSRPGLAVIAGALAVELMVSVLQHPEGGYAIASSSDDRMNEPPTSLGLVPHQVLDQYEREGFNFLAKVFNSSHSFLEDLTGLTLLHQETQAAEIWDMSDDETISEQ ID NO: 30: ATG7(2) polynucleotideP7411 PC00SEQ ID NO: 20: ATG7(1) polypeptide:MAAATGDPGLSKLQFAPFSSALDVGFWHELTQKKLNEYRLDEAPKDIKGYYYNGDSAGLPARLTLEFSAFDMSAPTPARCCPAIGTLYNTNTLESFKTADKKLLLEQAANEIWESIKSGTALENPVLLNKFLLLTFADLKKYHFYYWFCYPALCLPESLPLIQGPVGLDQRFSLKQIEALECAYDNLCQTEGVTALPYFLIKYDENMVLVSLLKHYSDFFQGQRTKITIGVYDPCNLAQYPGWPLRNFLVLAAHRWSSSFQSVEVVCFRDRTMQGARDVAHSIIFEVKLPEMAFSPDCPKAVGWEKNQKGGMGPRMVNLSECMDPKRLAESSVDLNLKLMCWRLVPTLDLDKWSVKCLLLGAGTLGCNVARTLMGWGVRHITFVDNAKISYSNPVRQPLYEFEDCLGGGKPKALAAADRLQKIFPGVNARGFNMSIPMPGHPVNFSSVTLEQARRDVEQLEQLIESHDWFLLMDTRESRWLPAVIAASKRKLVINAALGFDTFWMRHGLKKPKQQGAGDLCPNHPVASADLLGSSLFANIPGYKLGCYFCNDWAPGDSTRDRTLDQQCTVSRPGLAVIAGALAVELMVSVLQHPEGGYAIASSSDDRMNEPPTSLGLVPHQIRGFLSRFD NVLPVSLAFDKCTACSSKVLDQYEREGFNFLAKVFNSSHSFLEDLTGLTLLHQETQAA EIWDMSDDETISEQ ID NO: 31 : ATG7(1) polynucleotideSEQ ID NO: 22 - exon 16GGGCTATGCCATTGCCAGCAGCAGTGACGATCGGATGAATGAGCCTCCAACCTCTCTTGGGCTTGTGCCTCACCAGSEQ ID NO: 23 - exon 17ATCCGGGGATTTCTTTCACGGTTTGATAATGTCCTTCCCGTCAGCCTGGCATTTGACAAATGTACAGCTTGTTCTTCCAAASEQ ID NO: 24 - exon 18GTTCTTGATCAATATGAACGAGAAGGATTTAACTTCCTAGCCAAGGTGTTTAATTCTTCACATTCCTTCTTAGAAGACTTGACTGGTCTTACATTGCTGCATCAAGAAACCCA AGCTGCTGAGSEQ ID NO: 25 - siATG7(2) no. 1 ; Target RNA sequence ccucaccagguucuugaucaaSEQ ID NO: 26 - siATG7(2) no. 1 ; Antisense sequence : UUGAUCAAGAACCUGGUGAGGP7411 PC00 siATG7(2) no 2 :SEQ ID NO: 27 - siATG7(2) no. 2; Target RNA sequence cuugugccucaccagguucuuSEQ ID NO: 28 - siATG7(2) no. 2; Antisense sequence AAGAACCUGGUGAGGCACAAGSEQ ID NO: 29 - 5’- GGUUCUUGAUCAAUAUGAAcg -3’ReferencesOgmundsdottir, M. H., Fock, V., Sooman, L., Pogenberg, V., Dilshat, R., Bindesboll, C., Ogmundsdottir, H. M., Simonsen, A., Wilmanns, M., & Steingrimsson, E. (2018). A short isoform of ATG7 fails to lipidate LC3 / GABARAP. Scientific Reports, 8(1), 14391. https: / / doi.Org / 10.1038 / S41598-018-32694-7Ostacolo, K., Lomana, A. L. G., Larat, C., Hjaltalin, V., Holm, K. Y., ...Ogmundsdottir, M. H. (2024). ATG7(2) interacts with metabolic proteins and regulates central energy metabolism. Traffic, 25(4), e12933. https: / / doi.org / 10.1111 / tra.12933.

Claims

47P7411 PC00Claims1. An oligonucleotide targeting a polynucleotide encoding human ATG7 variant 2 (ATG7(2)) as set forth in SEQ ID NO: 30, wherein ATG7(2) comprises a junction between exon 16 and exon 18 of ATG7 and wherein said oligonucleotide comprises a sequence complementary to the junction between exon 16 and exon 18 of ATG7(2).

2. The oligonucleotide according to claim 1, wherein said oligonucleotide does not target a polynucleotide encoding ATG7 variant 1 (ATG7(1)).

3. The oligonucleotide according to any one of the preceding claims, wherein said oligonucleotide is selective for ATG7(2), and wherein said oligonucleotide does not target ATG7(1) as set forth in SEQ ID NO: 31.

4. The oligonucleotide according to any one of the preceding claims, wherein said oligonucleotide is complementary to ATG7(2) mRNA.

5. The oligonucleotide according to any one of the preceding claims, wherein the oligonucleotide comprises a sequence complementary to an ATG7(2) mRNA comprising a junction between exon 16 and exon 18 of ATG7.

6. The oligonucleotide according to any one of the preceding claims, wherein the oligonucleotide prevents translation of ATG7(2).

7. The oligonucleotide according to any one of the preceding claims, wherein said oligonucleotide comprises a sequence complementary to: a 3’ fragment of exon 16 of ATG7 and to a 5’ fragment of exon 18 of ATG7.

8. The oligonucleotide according to any one of the preceding claims, wherein said oligonucleotide does not comprise a sequence complementary to exon 17 of a polynucleotide encoding ATG7.

9. The oligonucleotide according to any one of the preceding claims, wherein the oligonucleotide comprises:48P7411 PC00 a. at least 3 nucleotides, such as at least 4, such as at least 5 nucleotides complementary to exon 16 of a polynucleotide encoding ATG7(2); and b. at least 3 nucleotides , such as at least 4, such as at least 5 nucleotides complementary to exon 18 of a polynucleotide encoding ATG7(2).

10. The oligonucleotide according to any one of the preceding claims, wherein the oligonucleotide comprises: a. at least the 3 nucleotides, such as at least the 4, such as at least the 5 nucleotides complementary to the 3’ end of exon 16 of a polynucleotide encoding ATG7(2); and b. at least 3 nucleotides , such as at least 4, such as at least 5 nucleotides complementary to the 5’ end of exon 18 of a polynucleotide encoding ATG7(2).11 . The oligonucleotide according to any one of the preceding claims, wherein said oligonucleotide comprises a double-stranded region of 10 to 30 nucleotides , such as 15 to 25 nucleotides.

12. The oligonucleotide according to any one of the preceding claims, wherein said oligonucleotide comprises a strand comprising SEQ ID NO: 13 or a functional variant thereof, wherein said functional variant has 1 or 2 individual ribonucleotide substitutions compared to SEQ ID NO: 13, and a complementary antisense strand.

13. The oligonucleotide according to any one of the preceding claims , wherein said oligonucleotide further comprises 1 to 16 nucleotides attached at the 3’ of SEQ ID NO: 13 or said functional variant thereof, and a complementary antisense strand.

14. The oligonucleotide according to any one of the preceding claims, wherein said oligonucleotide further comprises 1 to 16 nucleotides attached at the 5’ of SEQ ID NO: 13 or said functional variant thereof, and a complementary antisense strand.

15. The oligonucleotide according to any one of the preceding claims, wherein said oligonucleotide comprises a strand comprising SEQ ID NO: 16 or SEQ ID NO:49P7411 PC0017, or a functional variant thereof, wherein said functional variant has 1 , 2, 3, 4, 5, or 6 individual ribonucleotide substitutions compared to any one of SEQ ID NO: 16 or SEQ ID NO: 17, and a complementary antisense strand.

16. The oligonucleotide according to any one of the preceding claims, wherein said oligonucleotide comprises a sense strand of SEQ ID NO: 16, SEQ ID NO: 17, or SEQ ID NO: 13, and a complementary antisense strand.

17. The oligonucleotide according to any one of the preceding claims, wherein the ribonucleotide at the 3’ terminus and at the 5’ terminus of the sense strand are phosphorylated.

18. The oligonucleotide, or the oligonucleotide for use according to any one of the preceding claims, wherein the ribonucleotide at the 3’ terminus and at the 5’ terminus of the antisense strand are phosphorylated.

19. The oligonucleotide according to any one of the preceding claims, wherein the ribonucleotide at the 3’ terminus and at the 5’ terminus of the sense strand are non-phosphorylated.

20. The oligonucleotide according to any one of the preceding claims, wherein the ribonucleotide at the 3’ terminus and at the 5’ terminus of the antisense strand are non-phosphorylated.

21. The oligonucleotide according to any one of the preceding claims, wherein one or more ribonucleotides of the sense and / or antisense strands of said oligonucleotide are modified.

22. The oligonucleotide according to any one of the preceding claims, wherein a conjugated moiety is attached to one or more ribonucleotides of the sense and / or antisense strands of said oligonucleotide.

23. The oligonucleotide according to any one of the preceding claims wherein the oligonucleotide comprises a phosphorothioate (PS) backbone modification; a 5’ capping; a 5'-vinylphosphonate; a 2' ribose modification, such as 2'-O-methyl50P7411 PC00(2'-O-Me), such as 2'-O-methoxyethyl (2' MOE), such as 2'-fluoro, such as 2’-O- benzyl, such as 2’-O-methyl-4-pyridine (2’-O-CH2Py(4)); a methylene bridge between the 2' and 4' position of a ribose; a 5-methylcytosine (5mC); a nonstandard nucleotide.

24. The oligonucleotide according to any one of the preceding claims wherein the oligonucleotide comprises a modification of the nucleic acid backbone, the nucleobase, the ribose sugar and / or 2'-ribose substitutions.

25. The oligonucleotide according to any one of the preceding claims, wherein the oligonucleotide is conjugated to a moiety or to a nanoparticle formulation.

26. The oligonucleotide according to any one of the preceding claims, wherein the moiety is a cell-targeting moiety, and / or a cell-penetrating moiety.

27. The oligonucleotide according to any one of the preceding claims, wherein the oligonucleotide is conjugated to Triantennary N-acetylgalactosamine (GalNAc) moiety; cholesterol; and / or a peptide.

28. The oligonucleotide according to any one of the preceding claims, wherein the oligonucleotide is: a siRNA, a shRNA, an antisense oligonucleotide, a guide RNA, a LNA (Locked Nucleic Acid) antisense oligonucleotide, or a morpholino oligomer.

29. The oligonucleotide according to any one of the preceding claims, wherein the oligonucleotide is a double stranded oligonucleotide.

30. The oligonucleotide according to any one of the preceding claims, wherein the oligonucleotide is a siRNA.

31. The oligonucleotide according to any one of the preceding claims, wherein the oligonucleotide comprises a overhang.

32. The oligonucleotide according to any one of the preceding claims, wherein the oligonucleotide inhibits ATG7(2).P7411 PC0033. The oligonucleotide according to any one of the preceding claims, wherein the oligonucleotide decreases the amount of ATG7(2) in a cell of 10% to 90%.

34. The oligonucleotide according to any one of the preceding claims, wherein the oligonucleotide does not inhibit ATG7(1).

35. An oligonucleotide targeting human ATG7 variant 2 (ATG7(2)) according to any one of the preceding claims, for use as a medicament.

36. An oligonucleotide targeting human ATG7 variant 2 (ATG7(2)) according to any one of the preceding claims, for use in a method of treatment of a metabolic disease.

37. An oligonucleotide targeting human ATG7 variant 2 (ATG7(2)) according to any one of the preceding claims, for use in a method of treatment of diabetes or obesity.

38. The oligonucleotide according to any one of the preceding claims, wherein the form of administration of said oligonucleotide is: intravenous administration (IV); subcutaneous administration; intramuscular administration; intrathecal administration; intraventricular administration; oral administration; topical administration; inhalation administration; intraperitoneal administration; and / or intranasal administration.

39. The oligonucleotide according to any one of the preceding claims, wherein the oligonucleotide is administered as: a. a naked oligonucleotide; b. an oligonucleotide encapsulated into a nanoparticle, such as a Lipid- based nanoparticle (LNPs), such as a Virus-like nanoparticle (VLP); c. an oligonucleotide encapsulated into a microsphere or a microcapsule; d. conjugated to a targeting ligand, such as a peptide, such as an aptamer; e. incorporated into a vesicle, such as a liposome; f. an oligonucleotide adsorbed onto or incorporated into a hydrogel;P7411 PC00 g. a prodrug, such as wherein the prodrug is metabolized in an active form in the body.

40. A composition comprising the oligonucleotide, or the oligonucleotide for use according to any one of the preceding claims.

41. The composition according to claim 40, wherein the composition further comprises an acceptable carrier.

42. A method of treating a metabolic disease, said method comprising administration of a therapeutically effective amount of an oligonucleotide targeting a polynucleotide encoding human ATG7 variant 2 (ATG7(2)) as set forth in SEQ ID NO: 25, wherein ATG7(2) comprises a junction between exon 16 and exon 18 of ATG7, and wherein said oligonucleotide comprises a sequence complementary to the junction between exon 16 and exon 18 of ATG7(2).to an individual in need thereof.

43. A method of treating a metabolic diseases, the method comprising administering the oligonucleotide according to any one of claims 1 to 39, or the composition according to claims 40 to 41.

44. The method according to any one of claims 42 to 43, wherein the metabolic disease is diabetes or obesity.

45. Use of the oligonucleotide according to any one claims 1 to 39, or of a composition according to any one of claims 40 to 41 , for the manufacture of a medicament.

46. Use of the oligonucleotide according to any one claims to 1 to 39, or of a composition according to any one of claims 40 to 41 , for the manufacture of a medicament for treating a metabolic disease.

47. The use according to claim 46, wherein the metabolic disease is diabetes or obesity.

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