ATG7 inhibitors for treatment of pancreatic cancer

Abstract

The present invention relates to oligonucleotides targeting ATG7(2), and their use as a medicament, in particular for treatment of pancreatic cancer.

Description

[0001] P7341 PC00

[0002] Atg7 inhibitors for treatment of pancreatic cancer

[0003] Technical field

[0004] The present invention relates to oligonucleotides targeting ATG7(2), and their use as a medicament, in particular for treatment of pancreatic cancer.

[0005] Background

[0006] Macro-autophagy, hereafter referred to as autophagy, is a degradation process that enables the degradation of damaged organelles and other cellular components. In turn, autophagy provides the cell with recovered materials that can be used in diverse pathways.

[0007] Autophagy is initiated at the endoplasmic reticulum where a phagophore is formed around the targeted organelles, named cargo, before fusing with a lysosome, leading to the degradation of the cargo in an autolysosome.

[0008] 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.

[0009] Defects in autophagy, and in particular in the function of ATG7, have been implicated in a variety of diseases, including neurodegenerative disorders, cancer, and metabolic conditions. For instance, impaired autophagy due to ATG7 deficiency has been associated with the accumulation of protein aggregates in neurons, contributing to the pathogenesis of diseases such as Alzheimer's and Parkinson's. Conversely, overactivation of autophagy can lead to excessive degradation of essential cellular components, potentially leading to cell death and tissue damage.

[0010] Autophagy has been identified as both tumour suppressive or tumour protective, as the degradation of damaged organelles can prevent tumour formation, but can promote proliferation and drug resistance in established tumours. Both autophagy and ATG7 P7341 PC00 specifically have been identified as prognosis markers in diverse cancer types. Mainly, ATG7 has been linked with pancreatic adenocarcinoma (PAAD), in which high ATG7 expression is linked with poor prognosis and rapid cancer progression. However, ATG7 has also been shown to play an autophagy-independent role in pancreatic adenocarcinoma (PAAD).

[0011] 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.

[0012] Summary

[0013] Interestingly, a shorter isoform of ATG7, designated as ATG7(2), has been identified by the present inventor (M.H. Ogmundsdottir ef al., 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, ATG7(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.

[0014] The present disclosure provides evidence of ATG7(2) role in the progression and metastasis of pancreatic adenocarcinoma (PAAD), and the use of oligonucleotides targeting ATG7(2) as a medicament.

[0015] In an aspect, the present disclosure is directed to an oligonucleotide targeting a polynucleotide encoding ATG7 variant 2 (ATG7(2)).

[0016] In an aspect, the present disclosure is directed to an oligonucleotide targeting a polynucleotide encoding ATG7 variant 2 (ATG7(2)) for use as a medicament.

[0017] In an aspect, the present disclosure is directed to an oligonucleotide targeting a polynucleotide encoding ATG7 variant 2 (ATG7(2)) for use in a method of treating pancreatic cancer. P7341 PC00

[0018] In an aspect, the present disclosure is directed to an oligonucleotide targeting a polynucleotide encoding ATG7(2) for use in a method of decreasing proliferation and / or migration of pancreatic cancer cells.

[0019] In an aspect, the present disclosure is directed to a composition comprising the oligonucleotide, or the oligonucleotide for use as described herein.

[0020] In an aspect the present disclosure is directed to a method of decreasing migration and / or proliferation of a pancreatic cancer cell, said method comprising administration of a therapeutically effective amount of a oligonucleotide targeting a polynucleotide encoding ATG7(2) to an individual in need thereof.

[0021] In an aspect the present disclosure is directed to a method of treating pancreatic cancer, said method comprising administration of a therapeutically effective amount of an oligonucleotide targeting a polynucleotide encoding ATG7(2) to an individual in need thereof.

[0022] In an aspect the present disclosure is directed to a method of treating pancreatic cancer, the method comprising administering the oligonucleotide as described herein.

[0023] In an aspect the present disclosure is directed to the use of the oligonucleotide as described herein, for the manufacture of a medicament.

[0024] 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 pancreatic cancer.

[0025] Description of Drawings

[0026] Figure 1 ATG7(2) is linked with poor prognosis in PAAD

[0027] (A) Alternative splicing of ATG7 exon 17 leads to the transcription of isoforms ATG7(1) and ATG7(2). (B)-(D) Kaplan-Meier survival plots of ATG7 ; ATG7(1) or ATG7(2) in

[0028] PAAD. High and low expression groups were determined by median mRNA expression. (E) Expression plot of ATG7(1) and ATG7(2) in PAAD versus pancreas. (F) Expression plot of ATG7(1) and ATG7(2) in PAAD at different stages (Stage IA to Stage IV). (C-D) One way ANOVA was performed between the different groups, with * : p<0,05 ; ** p<0,01 ; *** p<0,001 ; **** p<0,0001. P7341 PC00

[0029] Figure 2. ATG7(2) is involved in PAAD immunity

[0030] (A) Expression plot of ATG7(1) and ATG7(2) in different immune subtypes of cancer samples. (B) Box plot of survival time or time before new tumour event in the same samples. (C) Correlation of ATG7 expression with M2 macrophages and CAFs infiltration across cancers. Black : positive correlation between ATG7 expression and tumour infiltration ; White : negative correlation between ATG7 expression and tumour infiltration ; * : significant correlation p<0,05. (A-B) One way ANOVA was performed between the different groups, with * : p<0,05 ; ** : p<0,01 ; *** : p<0,001 ; **** : p<0,0001.

[0031] Figure 3. ATG7(2) promotes PAAD cells migration and proliferation in vitro (A) CRISPR / Cas9 gRNAs targeting exon 3 for ATG7 knock-out ; or exon 17 for ATG7(1) knock-out. (B), (C) and (D) mRNA guantification of ATG7, ATG7(1) and ATG7(2), respectively, using gPCR in Capan-1 Control and ATG7(1)- / - cells. (E) Western-Blot analysis of Capan-1 Control and ATG7(1)- / - cells. (F), (G) and (H) mRNA guantification of ATG7, ATG7(1) and ATG7(2), respectively, using gPCR in Capan-1 Control and ATG7- / - cells. (I) Western-Blot analysis of Capan-1 Control and ATG7- / - cells. (J) and (K) Normalized proliferation and migration ratios of Capan-1 Control and ATG7(1)- / - cells. (L) Normalized migration ratios of ATG7- / - cells. (J-L) One way ANOVA was performed between the different groups, with * : p<0,05 ; ** : p<0,01 ; *** : p<0,001 ; **** p<0,0001.

[0032] Figure 4. ATG7(2) knock-down decreases PAAD cells proliferation in vitro

[0033] (A) Arrows indicate the location and direction of the seguences targeted by siRNA. (B), (C), (D) Quantification of ATG7, ATG7(1) or ATG7(2) mRNA expression in Capan-1 WT or (E), (F), (G) Capan-1 ATG7(1)- / - cells transfected with siControl, siATG7, siATG7(1) or siATG7(2). (H) Evaluation of proliferation by guantifying Ki67 mRNA expression in Capan-1 WT or Capan-1 ATG7(1)- / - cells transfected with siControl, siATG7, siATG7(1) or siATG7(2). (B-H) One way ANOVA was performed between the different groups, with * : p<0,05 ; ** : p<0,01 ; *** : p<0,001 ; **** : p<0,0001.

[0034] Figure 5. Specificity of siATG7(2) was confirmed in MEF cells Quantification of ATG7, ATG7(1) or ATG7(2) mRNA expression in MEFs Atg7- / - cells transfected with plasmids expressing ATG7(1) (A), (B), (C) or ATG7(2) (D), (E), (F); and transfected with siControl, siATG7, siATG7(1) or siATG7(2). (A-F) One way P7341 PC00

[0035] ANOVA was performed between the different groups, with * : p<0,05 ; ** : p<0,01 ; *** : p<0,001 ; **** p<0,0001.

[0036] Detailed description

[0037] Definitions

[0038] Unless 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.

[0039] 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 items, the language "comprising" can include analogous embodiments described in terms of “consisting of’ and / or “consisting essentially of”.

[0040] As used in the specification and items, 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".

[0041] As used in the specification and items, 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.

[0042] 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.

[0043] 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, P7341 PC00 chemical synthesis. An isolated nucleic acid is one which is readily manipulable by 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.

[0044] 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.

[0045] Throughout the text any reference to the oligonucleotide, also refers to the oligonucleotide for use, or uses of said oligonucleotide as described herein.

[0046] Oligonucleotide targeting ATG7 variant 2 (ATG7(2)).

[0047] The present disclosure describes oligonucleotides for targeting ATG7 variant 2 (ATG7(2)), such as a polynucleotide encoding ATG7(2), and their use as a P7341 PC00 medicament, such as their use in a method of treating pancreatic cancer; or such as a method of decreasing proliferation and / or migration of pancreatic cancer cells.

[0048] Both autophagy and ATG7 specifically have been identified as prognosis markers in diverse cancer types. Mainly, ATG7 has been linked with pancreatic adenocarcinoma (PAAD), in which high ATG7 expression is linked with poor prognosis and rapid cancer progression. However, ATG7 has also been shown to play an autophagy-independent role in pancreatic adenocarcinoma (PAAD).

[0049] The present disclosure describes that it is specifically ATG7 variant 2 (ATG7(2)) which is associated with cancer development, such as pancreatic cancer, and with promoting proliferation and / or migration of pancreatic cancer cells.

[0050] Thus, in an aspect of the invention, the present disclosure is directed to an oligonucleotide targeting a polynucleotide encoding ATG7(2).

[0051] In an aspect of the invention, the present disclosure is directed to an oligonucleotide targeting a polynucleotide encoding ATG7(2) for use as a medicament.

[0052] In an aspect of the invention, the present disclosure is directed to an oligonucleotide targeting a polynucleotide encoding ATG7(2) for use in a method of treating pancreatic cancer.

[0053] In an aspect of the invention, the present disclosure is directed to an oligonucleotide targeting a polynucleotide encoding ATG7(2) for use in a method of decreasing proliferation and / or migration of pancreatic cancer cells.

[0054] In some embodiments, ATG7(2) is human ATG7(2).

[0055] 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 use of a siRNA specific for ATG7(2) decreases PAAD cells proliferation in vitro. 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. P7341 PC00

[0056] 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).

[0057] 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).

[0058] 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 the RNA 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.

[0059] 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).

[0060] Thus, as described herein: exon 16 refers to the sequence: GGGCTATGCCATTGCCAGCAGCAGTGACGATCGGATGAATGAGCCTCCAACCTCT CTTGGGCTTGTGCCTCACCAG

[0061] Thus, as described herein: exon 17 refers to the sequence: ATCCGGGGATTTCTTTCACGGTTTGATAATGTCCTTCCCGTCAGCCTGGCATTTGA CAAATGTACAGCTTGTTCTTCCAAA

[0062] Thus, as described herein: exon 18 refers to the sequence: P7341 PC00

[0063] GTTCTTGATCAATATGAACGAGAAGGATTTAACTTCCTAGCCAAGGTGTTTAATTCT TCACATTCCTTCTTAGAAGACTTGACTGGTCTTACATTGCTGCATCAAGAAACCCA AGCTGCTGAG

[0064] Antisense oligonucleotides are capable of inhibiting, such as preventing, 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 prevents 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.

[0065] 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.

[0066] 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.

[0067] 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. P7341 PC00

[0068] 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.

[0069] Thus, in summary, in some embodiments, the oligonucleotide is: a siRNA; a shRNA; an antisense oligonucleotide; a guide RNA; or a morpholino oligomer.

[0070] 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, the oligonucleotide is a guide RNA (gRNA). In some embodiments, the oligonucleotide is a morpholino oligomer.

[0071] In some embodiments, the oligonucleotide is a RNA.

[0072] Sequences and Complementarity

[0073] Differential 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.

[0074] 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: 35), 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) P7341 PC00 showed silenced expression of both isoforms upon treatment with the siRNA, revealing a lack in specificity for ATG7(2).

[0075] To overcome this technical difficulty, additional siRNA sequences were designed manually to avoid long GC 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.

[0076] 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).

[0077] In some embodiments, ATG7(2) is as set forth in SEQ ID NO: 24.

[0078] In some embodiments, ATG7(2) is encoded by a nucleotide sequence comprising a nucleotide sequence as set forth in SEQ ID NO: 36

[0079] 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 comprises a sequence complementary to a splice site known and / or predicted to be associated with exon skipping of exon 17.

[0080] Thus, in some embodiments the oligonucleotide comprises a sequence complementary to ATG7(2) mRNA.

[0081] 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).

[0082] In some embodiments, ATG7(1) is as set forth in SEQ ID NO: 26. P7341 PC00

[0083] In some embodiments, ATG7(1) is encoded by a nucleotide sequence comprising a nucleotide sequence as set forth in SEQ ID NO: 37.

[0084] 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 sequences

[0085] In some embodiments, the oligonucleotide prevents and / or inhibits translation of ATG7(2). Such inhibition can be caused, for example, by the selective degradation of the ATG7(2) mRNA.

[0086] In some embodiments, the oligonucleotide does not target ATG7 variant 1 (ATG7(1)).

[0087] In some embodiments, the oligonucleotide comprises a sequence complementary to: a fragment of exon 16 of ATG7 and to a fragment of exon 18 of ATG7.

[0088] In some embodiments, the oligonucleotide does not comprise a sequence complementary to exon 17 of ATG7. In other words, in some embodiments, the oligonucleotide is not complementary to ATG7(1).

[0089] 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 comprises a sequence complementary to ATG7(2) mRNA but not to ATG7(1) mRNA; only ATG7(2) mRNA comprises as junction between exon 16 and exon 18.

[0090] In some embodiments, the oligonucleotide is a small interfering RNA (siRNA).

[0091] 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 P7341 PC00 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.

[0092] 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.

[0093] In some embodiments, said oligonucleotide is complementary to a 3’ fragment of exon 16 of ATG7 and to a 5’ fragment of exon 18 of ATG7.

[0094] 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.

[0095] 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); and b. at least 3 nt, such as at least 4, such as at least 5 nt complementary to exon 18 of ATG7(2).

[0096] 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).

[0097] 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 P7341 PC00 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).

[0098] 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).

[0099] In some embodiments, 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).

[0100] 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).

[0101] 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 P7341 PC00 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).

[0102] 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).

[0103] 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.

[0104] In some embodiments, said oligonucleotide comprises a strand comprising SEQ ID NO: 16 or a functional variant thereof, wherein said functional variant has 1 or 2 individual ribonucleotide substitutions compared to SEQ ID NO: 16.

[0105] 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 individual ribonucleotide substitution compared to SEQ ID NO: 16. In other embodiments, said oligonucleotide comprises a strand comprising SEQ ID NO: 16 or a functional variant thereof, wherein said functional variant has 2 individual ribonucleotide substitutions compared to SEQ ID NO: 16.

[0106] 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 P7341 PC00 nucleotides, such as 16 nucleotides attached at the 3’ of SEQ ID NO: 16 or said functional variant thereof.

[0107] 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: 16 or said functional variant thereof.

[0108] In some embodiments, said oligonucleotide comprises a strand comprising SEQ ID NO: 22 or SEQ ID NO: 23, 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: 22 or SEQ ID NO: 23.

[0109] Thus, in some embodiments, said oligonucleotide comprises a strand comprising SEQ ID NO: 22, 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: 22. In some embodiments, said oligonucleotide comprises a strand comprising SEQ ID NO: 23, 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: 23.

[0110] In some embodiments, said oligonucleotide comprises a strand of SEQ ID NO: 22 or SEQ ID NO: 23, 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: 22 or SEQ ID NO: 23.

[0111] Thus, in some embodiments, said oligonucleotide comprises a strand of SEQ ID NO: 22, or a functional variant thereof, wherein said functional variant has 1 , 2 or 3 individual ribonucleotide substitutions compared to SEQ ID NO: 22. In some embodiments, said oligonucleotide comprises a strand of SEQ ID NO: 22, or a functional variant thereof, wherein said functional variant has 1 individual ribonucleotide substitution compared to SEQ ID NO: 22. In some embodiments, said oligonucleotide comprises a strand of SEQ ID NO: 22, or a functional variant thereof, wherein said P7341 PC00 functional variant has 2 individual ribonucleotide substitutions compared to SEQ ID NO: 22. In some embodiments, said oligonucleotide comprises a strand of SEQ ID NO: 22, or a functional variant thereof, wherein said functional variant has 3 individual ribonucleotide substitutions compared to SEQ ID NO: 22.

[0112] In some embodiments, said oligonucleotide comprises a strand of SEQ ID NO: 23, or a functional variant thereof, wherein said functional variant has 1 , 2 or 3 individual ribonucleotide substitutions compared to SEQ ID NO: 23. In some embodiments, said oligonucleotide comprises a strand of SEQ ID NO: 23, or a functional variant thereof, wherein said functional variant has 1 individual ribonucleotide substitution compared to SEQ ID NO: 23. In some embodiments, said oligonucleotide comprises a strand of SEQ ID NO: 23, or a functional variant thereof, wherein said functional variant has 2 individual ribonucleotide substitutions compared to SEQ ID NO: 23. In some embodiments, said oligonucleotide comprises a strand of SEQ ID NO: 23, or a functional variant thereof, wherein said functional variant has 3 individual ribonucleotide substitutions compared to SEQ ID NO: 23.

[0113] In some embodiments, said oligonucleotide comprises a sense strand of SEQ ID NO: 22, SEQ ID NO: 23, or SEQ ID NO: 16 or functional variant thereof, and a complementary antisense strand.

[0114] Thus, in some embodiments said oligonucleotide comprises a sense strand of SEQ ID NO: 22, or functional variant thereof, and a complementary antisense strand. In other embodiments, said oligonucleotide comprises a sense strand of SEQ ID NO: 23, or functional variant thereof, and a complementary antisense strand. In further embodiments, said oligonucleotide comprises a sense strand of SEQ ID NO: 16, or functional variant thereof, and a complementary antisense strand.

[0115] 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. P7341 PC00

[0116] The oligonucleotides, or oligonucleotides for use, provided herein are useful for use as a medicament; for use in methods related to the treatment of pancreatic cancer; or use in methods related to decreasing the proliferation and / or migration of pancreatic cancer cells. The usefulness of said oligonucleotides, or oligonucleotides for use, for the applications disclosed above derives from their capacity to selectively inhibit the activity of ATG7(2), which is implicated in the development of cancer, without disrupting the activity of ATG7(1), which has a role in autophagy.

[0117] Nucleotide modifications

[0118] Nucleotide 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.

[0119] In 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.

[0120] 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 P7341 PC00 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.

[0121] 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.

[0122] 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.

[0123] In 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.

[0124] 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 P7341 PC00

[0125] 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.

[0126] In some embodiments, the oligonucleotide comprises a modification of the nucleic acid backbone, the nucleobase, the ribose sugar and / or 2'-ribose substitutions.

[0127] 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.

[0128] 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.

[0129] In 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.

[0130] 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, in a method of treating pancreatic cancer, or in a method of decreasing the proliferation and / or migration of pancreatic cancer cells. Furthermore, in some embodiments the above P7341 PC00 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.

[0131] In some embodiments, the oligonucleotide is a double stranded oligonucleotide.

[0132] 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.

[0133] Medical use of the oligonucleotide

[0134] In an aspect, the present disclosure is directed to an oligonucleotide as described herein for use as a medicament.

[0135] The present inventors have found that siRNA specifically and selectively targeting ATG7(2) decreases proliferation and migration rates of pancreatic cancer cells, see Fig. 4.H-I and the relative text in the Examples herein.

[0136] In an aspect the present disclosure is directed to a method of treating cancer, the method comprising administering the oligonucleotide as described herein.

[0137] In an aspect the present disclosure is directed to a method of treating pancreatic cancer, the method comprising administering the oligonucleotide as described herein. Without being bound by theory, the examples disclosed herein support that an oligonucleotide can be used to slow down tumour progression and metastasis.

[0138] In some embodiments, the pancreatic cancer is an exocrine pancreatic cancer or an endocrine pancreatic cancer. The oligonucleotide, in some embodiments, is as described elsewhere herein, for example in the section “Oligonucleotide”.

[0139] In some embodiments, the pancreatic cancer is pancreatic adenocarcinoma (PAAD), pancreatic neuroendocrine tumor, acinar cell carcinoma of the pancreas, cystadenocarcinomas, pancreatoblastoma, adenosquamous carcinomas, signet ring P7341 PC00 cell carcinomas, hepatoid carcinomas, colloid carcinomas, undifferentiated carcinomas, undifferentiated carcinomas with osteoclast-like giant cells and / or pancreatic mucinous cystic neoplasms.

[0140] In some embodiments, the pancreatic cancer is pancreatic adenocarcinoma (PAAD). In some embodiments, the pancreatic cancer is pancreatic neuroendocrine tumor. In some embodiments, the pancreatic cancer is acinar cell carcinoma of the pancreas. In some embodiments, the pancreatic cancer is cystadenocarcinomas. In some embodiments, the pancreatic cancer is pancreatoblastoma. In some embodiments, the pancreatic cancer is adenosquamous carcinomas. In some embodiments, the pancreatic cancer is signet ring cell carcinomas. In some embodiments, the pancreatic cancer is hepatoid carcinomas. In some embodiments, the pancreatic cancer is colloid carcinomas. In some embodiments, the pancreatic cancer is undifferentiated carcinomas. In some embodiments, the pancreatic cancer is undifferentiated carcinomas with osteoclast-like giant cells. In some embodiments, the pancreatic cancer is pancreatic mucinous cystic neoplasms.

[0141] In some embodiments, the pancreatic cancer is PAAD.

[0142] 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%.

[0143] 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%.

[0144] 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%. P7341 PC00

[0145] 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.

[0146] Administration form of the oligonucleotide

[0147] The 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.

[0148] 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.

[0149] In 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.

[0150] In some embodiments, the oligonucleotide, or oligonucleotide for use, is administered as: a. a naked oligonucleotide; P7341 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.

[0151] 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, is 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 P7341 PC00 administered as a prodrug, such as wherein the prodrug is metabolized in an active form in the body.

[0152] 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.

[0153] In some embodiments, the composition further comprises an acceptable carrier.

[0154] In an aspect of the invention, the present disclosure is directed to a method of decreasing migration and / or proliferation of a pancreatic cancer cell, said method comprising administration of a therapeutically effective amount of a oligonucleotide targeting a polynucleotide encoding ATG7(2), as described herein, or the composition described above, to an individual in need thereof.

[0155] 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.

[0156] In an aspect of the invention, the present disclosure is directed to a method of treating pancreatic cancer, said method comprising administration of a therapeutically effective amount of an oligonucleotide targeting a polynucleotide encoding ATG7(2) to an individual in need thereof.

[0157] In an aspect of the invention, the present disclosure is directed to a method of treating pancreatic cancer, the method comprising administering the oligonucleotide as described herein.

[0158] 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.

[0159] 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 cancer. P7341 PC00

[0160] 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 pancreatic cancer.

[0161] Items

[0162] 1. An oligonucleotide targeting ATG7 variant 2 (ATG7(2)).

[0163] 2. An oligonucleotide targeting ATG7 variant 2 (ATG7(2)) for use as a medicament.

[0164] 3. An oligonucleotide targeting ATG7 variant 2 (ATG7(2)) for use in a method of treating pancreatic cancer.

[0165] 4. An oligonucleotide targeting ATG7(2) for use in a method of decreasing proliferation and / or migration of pancreatic cancer cells.

[0166] 5. The oligonucleotide for use according to any one of the preceding items, wherein said ATG7(2) is human ATG7(2).

[0167] 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: 24.

[0168] 7. 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: 36

[0169] 8. The oligonucleotide, or the oligonucleotide for use according to any one of the preceding items, wherein said oligonucleotide is selective for ATG7(2).

[0170] 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. P7341 PC00

[0171] 10. The oligonucleotide, or the oligonucleotide for use according to any one of the preceding items, wherein the oligonucleotide inhibits ATG7(2) translation.

[0172] 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)).

[0173] 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: 26.

[0174] 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: 37.

[0175] 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.

[0176] 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.

[0177] 16. The oligonucleotide, or the oligonucleotide for use according to any one of the preceding items, wherein the oligonucleotide comprises a sequence complementary to an ATG7 mRNA comprising a junction between exon 16 and exon 18 of ATG7.

[0178] 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).

[0179] 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. P7341 PC00

[0180] 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).

[0181] 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).

[0182] 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.

[0183] 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: 16 or a functional variant thereof, wherein said functional variant has 1 or 2 individual ribonucleotide substitutions compared to SEQ ID NO: 16.

[0184] 23. 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: 16 or said functional variant thereof.

[0185] 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: 16 or said functional variant thereof. P7341 PC00

[0186] 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: 22 or SEQ ID NO: 23, 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: 22 or SEQ ID NO: 23.

[0187] 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: 22 or SEQ ID NO: 23, 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: 22 or SEQ ID NO: 23.

[0188] 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: 22, SEQ ID NO: 23, or SEQ ID NO: 16 or functional variant thereof, and a complementary antisense strand.

[0189] 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.

[0190] 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.

[0191] 30. 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.

[0192] 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. P7341 PC00

[0193] 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.

[0194] 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.

[0195] 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.

[0196] 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.

[0197] 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.

[0198] 37. 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.

[0199] 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.

[0200] 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 P7341 PC00 antisense oligonucleotide, a guide RNA, a LNA (Locked Nucleic Acid) antisense oligonucleotide, or a morpholino oligomer.

[0201] 40. The oligonucleotide, or the oligonucleotide for use according to any one of the preceding items, wherein the oligonucleotide is a double stranded oligonucleotide.

[0202] 41 . The oligonucleotide, or the oligonucleotide for use according to any one of the preceding items, wherein the oligonucleotide is a siRNA.

[0203] 42. The oligonucleotide, or the oligonucleotide for use according to any one of the preceding items, wherein the oligonucleotide comprises a overhang.

[0204] 43. The oligonucleotide, or the oligonucleotide for use according to any one of the preceding items, wherein the pancreatic cancer is an exocrine pancreatic cancer or an endocrine pancreatic cancer.

[0205] 44. The oligonucleotide, or the oligonucleotide for use according to any one of the preceding items, wherein the pancreatic cancer is pancreatic adenocarcinoma (PAAD), pancreatic neuroendocrine tumor, acinar cell carcinoma of the pancreas, cystadenocarcinomas, pancreatoblastoma, adenosquamous carcinomas, signet ring cell carcinomas, hepatoid carcinomas, colloid carcinomas, undifferentiated carcinomas, undifferentiated carcinomas with osteoclast-like giant cells, pancreatic mucinous cystic neoplasms.

[0206] 45. The oligonucleotide, or the oligonucleotide for use according to any one of the preceding items, wherein the pancreatic cancer is PAAD.

[0207] 46. 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. P7341 PC00

[0208] 47. 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; 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.

[0209] 48. A composition comprising the oligonucleotide, or the oligonucleotide for use according to any one of the preceding items.

[0210] 49. The composition according to item 47, wherein the composition further comprises an acceptable carrier.

[0211] 50. A method of decreasing migration and / or proliferation of a pancreatic cancer cell, said method comprising administration of a therapeutically effective amount of a oligonucleotide targeting ATG7(2) according to any one of items 1 to 42, or the composition according to any one of items 48 to 49, to an individual in need thereof.

[0212] 51. A method of treating pancreatic cancer, said method comprising administration of a therapeutically effective amount of an oligonucleotide targeting ATG7(2) to an individual in need thereof.

[0213] 52. Method of treating pancreatic cancer, the method comprising administering the oligonucleotide according to any one of items 1 to 42, or the composition according to any one of items 1 to 49. P7341 PC00

[0214] 53. Use of the oligonucleotide according to any one items 1 to 42, or the composition according to any one of items 1 to 49, for the manufacture of a medicament.

[0215] 54. Use of the oligonucleotide according to any one items 1 to 42, or the composition according to any one of items 1 to 49, for the manufacture of a medicament for treating pancreatic cancer.

[0216] Examples

[0217] Example 1.

[0218] Aim: To explore the potential role of ATG7(2) in PAAD progression and metastasis, we decided to use CRISPR / Cas9 technology on human PAAD Capan-1 cells to selectively knock out ATG7(1) or total ATG7. After phenotyping and characterization of these cells, siRNAs were designed to selectively target ATG7(2) in vitro.

[0219] Materials and Methods

[0220] Cell culture

[0221] Capan-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 without Glucose (Gibco A1443001) was used to induce starvation while DMEM high Glucose (Gibco 41965039) with 20% FBS (Gibco 11995073) was used on the control condition. Mouse Embryonic Fibroblasts (MEFs) Atg7' / _were obtained from Dr. Masaaki Komatsu and cultivated in DMEM high Glucose with 10% FBS. Cells were transfected to stably over-express flag-ATG7(1) or flag- ATG7(2) in a Doxycycline inducible manner, as previously described (Ostacolo et al. 2024).

[0222] CRISPR / Cas9 gRNA and Cas9 transfection

[0223] For each reaction, 1 L of 100uM crRNA and 1 L of 100uM tracrRNA (IDT Alt-R CRISPR- Cas9 5’ ATTO 550 #1077024) were mixed in a final volume of 100pL of nuclease free duplex buffer (IDT #1072570) and incubated at 95°C for 5 minutes to generate gRNAs. 1 ,6pL of Cas9 (IDT Alt-R S. p. Cas9 #1081059) was diluted in a final volume of 100pL in OptiMEM (Gibco 11058021). In a well of a 24-well plate coated with collagen, 88pL OptiMEM, 6pL gRNA and 6pL diluted Cas9 were mixed and incubated at room temperature for 5 minutes to generate the RNP complex. 99pL of OptiMEM and 1 L of P7341 PC00

[0224] RNAiMax (Gibco 11058021) were added to the RNP complex and incubated 20 minutes at room temperature. 1x104- 2x104cells were added to the well in a final volume of 500pL IM DM 20% FBS and grown at 37°C with 5% CO2. Control reaction was performed with DEPC water instead of crRNA.

[0225] The following sequences were targeted by the crRNA :

[0226] ATG7(1) : 5’- AGATCCGGGGATTTC -3’ (SEQ ID NO: 1)

[0227] ATG7 : 5’- AACTCCAATGTTAAG -3’ (SEQ ID NO: 2)

[0228] Single cell cloning

[0229] After 48 hours, the cells were suspended and diluted to a concentration of 1 - 2 cells / mL. 7mL of cell suspension was added to 10cm tissue dishes coated with collagen. 24 dishes were used per CRISPR reaction. The cells were grown at 37°C with 5% CO2 for 17 days and 5 tracrRNA positive clones were picked from each dish and transferred to a collagen- coated well on a 24 well plate. The 120 clones from each reaction were grown until confluent.

[0230] DNA extraction

[0231] Cell pellets from confluent clones were collected and resuspended in 30pL of Igepal buffer (52,5mM Tris-HCI ; 52,5mM KCI ; 3,3075mM MgCI2; 0,26125% Igepal ; 0,525% Tween 20 ; 0,025% proteinase K (NEB P8107)) and heated 90 minutes at 60°C, followed by 15 minutes at 95°C. Samples were centrifuged 15 minutes at 16 000g and 20pL of the supernatant were collected. DNA was quantified using NanoDrop One Spectrophotometer (ThermoFisher) and samples were diluted to a finale concentration of 50ng.pL'1in DEPC water.

[0232] PCR reaction

[0233] For each reaction, 0,5pL of 10pM forward primer, 0,5pL of 10pM reverse primer, 1 ,5pL DEPC water, 12,5pL OneTaq 2X Master Mix with standard buffer (NEB #M0482) and 10pL of 50ng.pL'1were mixed and underwent the following program : 94°C 30sec - (94°C 30sec - 45°C 60sec - 68°C 60sec) x30 - 68°C 5 minutes.

[0234] The following primers were used :

[0235] ATG7(1) : 5’- TTGAGTATACCAGTGGACCTGT -3’ (SEQ ID NO: 3)

[0236] 5’- CCCCACAAAAAGCCCTTAGTT -3’ (SEQ ID NO: 4) ATG7 : 5’- TAGCATTGCTGTGGGAAGAGA -3’ (SEQ ID NO: 5)

[0237] 5’- CAGCACTCATCTCTATGGGAGG -3’ (SEQ ID NO: 6) P7341 PC00

[0238] Sanger sequencing

[0239] The PCR products were cleaned-up using ExoSAP-IT™ (ThermoFisher #78200.200. ML) : 0,5pL ExoSAP was added to each 5pL of PCR product and heated at 37°C for 30 minutes and 90°C for 10 minutes. For each sequencing reaction, 3,2pL of BigDye (ref : demander a Decode), 0.8pL DEPC water, 0,5pL sequencing primer and 0,5pL ExoSAP product were mixed and underwent the following program : 95°C 3min - (95°C 20sec - 55°C 7sec - 60°C 4min) x30. Sequencing was read by DeCode Genetics, Inc. and results were compared with WT ATG7 sequence using Benchling Biology Software.

[0240] The following sequencing primers were used :

[0241] ATG7(1) : 5’- AGAAACCAGACGTGCATTTCA -3’ (SEQ ID NO: 7)

[0242] ATG7 : 5’- TGAACGCTGCTATTTCTGACTTG -3’ (SEQ ID NO: 8)

[0243] Quantitative Real-Time PCR

[0244] Total RNA was extracted from cells using the Quick-RNA Miniprep Kit (Zymo Research R1055) following the manufacturer's protocol. RNA samples were diluted to a final concentration of 100ng.pL"1in nuclease-free water. Reverse transcription was performed using the High-Capacity cDNA Reverse Transcription Kit (Applied Biosystems no. 4368814) following the manufacturer's protocol for 10pL reaction volume. cDNA samples were diluted 1 :50 in nuclease-free water. qPCR was performed using Luna Universal qPCR Master Mix (New England Biolabs Inc., #M3003) following the manufacturer's protocol for 10pL reaction volume with 4pL of cDNA. The data were normalized to actin mRNA. The following primers were used at a final concentration of 0,25pM :

[0245] ACTIN : 5’- TTCTACAATGAGCTGCGTGTGG -3’ (SEQ ID NO: 9)

[0246] 5’- GTGTTGAAGGTCTCAAACATGAT -3’ (SEQ ID NO: 10)

[0247] ATG7 : 5’- CGTTGCCCACAGCATCATCTTC -3’ (SEQ ID NO: 11)

[0248] 5’- CACTGAGGTTCACCATCCTTGG -3’ (SEQ ID NO: 12) ATG7(1) : 5’- CAGAAGGGGGCTATGCCATT -3’ (SEQ ID NO: 13) 5’- AACCGTGAAAGAAATCCCCG -3’ (SEQ ID NO: 14) ATG7(2) : 5’- GCATCCAGAAGGGGGCTATG -3’ (SEQ ID NO: 15) 5’- GATCAAGAACCTGGTGAGGCA -3’ (SEQ ID NO: 17) KI67 : 5’- GAAAGAGTGGCAACCTGCCTTC -3’ (SEQ ID NO: 18) 5’- GCACCAAGTTTTACTACATCTGCC -3’ (SEQ ID NO: 19) P7341 PC00

[0249] Western Blotting

[0250] Samples were collected in 1X RIPA buffer (Abeam ab156034) with 1 % protease inhibitor (ThermoFisher 1861281) or through another specified method. Protein loading dye (NEB B7703S) was added to the samples following manufacturer’s instructions and the samples were boiled for 5 min at 95°C. Proteins were separated by SDS-polyacrylamide gel electrophoresis (SDS-PAGE) at 150V on 12% acrylamide gels. Then, the proteins were transferred onto PVDF membranes (Thermo Fisher Scientific, 88520) at 25V overnight or 400mA for 2h on ice. Membranes were blocked in 5% BSA (Sigma, A9647) in TBS with 0,1 % Tween-20 (Sigma, P1379) for 30min at RT. Primary antibodies, such as anti-actin (Millipore, MAB1501), anti-FLAG (Sigma, F3165), anti-LC3B (Cell Signaling Technology, #2775), and anti-p62 (Cell Signaling Technology, #5114), were incubated overnight at 4°C or 3h at room temperature. Finally, DyLight 800 anti-mouse and DyLight 680 anti-rabbit (Cell Signaling Technology) were applied for 45min at RT. All the antibodies were used at the dilutions recommended by manufacturers. Odyssey imaging system and Image Studio version 2.0 (LI-COR Biosciences) were used to scan the blots. Proliferation and migration

[0251] Proliferation assay

[0252] Capan-1 cells were seeded at a density of 2000 cells per well on a 96 well plate coated with collagen Rat tail I and placed in the IncuCyte® Live-Cell imaging system. For knockdown, cells were reverse transfected with siRNA when seeded (see siRNA knock-down) and treatment was repeated with forward transfection every 72 hours. 1 picture was taken of each seeded well with a 4x zoom every 4 hours over 11 days. For each well, cell confluency was normalized to the confluency of the first image (Oh) directly in the IncuCyte® 2022A Rev1 software. For each condition, 5 technical replicates were done, and the experiment was performed 3 times for statistical analysis.

[0253] Migration assay

[0254] Capan-1 cells were seeded at a density of 35000 cells per well on a 96 well plate (Sartorius IncuCyte® ImageLock 4379) coated with collagen Rat tail I and incubated for 24h at 37°C with 5%CO2. For knock-down, cells were reverse transfected with siRNA when seeded (see siRNA knock-down). Wounds were created using a woundmaker (Sartorius 4563). The pins of the woundmaker were washed 5min in sterile water and 5min in ethanol 70%. The plate was placed on the woundmaker and the cover was removed before placing the pins in the wells. The switch of the woundmaker was pressed down and the pins were removed from the wells without releasing the switch. The cells were rinsed once with PBS and treatment was added to the wells if applicable. The pins P7341 PC00 were subsequently washed 5min in Alconox 0,5% ; 5min in Virkon S 1 % ; 5min in sterile water and 5min in ethanol 70%. The plate was placed in the IncuCyte® Live-Cell imaging system. 2 pictures were taken of each seeded well with a 10x zoom every 2 hours over 3 days. For each well, relative wound density was calculated directly in the IncuCyte® 2022A Rev1 software. For each condition, 5 technical replicates were done, and the experiment was performed 3 times for statistical analysis. siRNA knock-down

[0255] Cells were reverse- or forward-transfected with siRNA using OptiMEM™ (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: 20) siATG7(1) : 5’- GGCAUUUGACAAAUGUACAgc -3’ (SEQ ID NO: 21) siATG7(2): 5’- UCACCAGGUUCUUGAUCAAtt -3’ (SEQ ID NO: 22)

[0256] 5’- UGUGCCUCACCAGGUUCUUtt -3’ (SEQ ID NO:23)

[0257] Results

[0258] ATG7(2) is linked with poor prognosis in PAAD

[0259] Alternative splicing of ATG7 pre-mRNA leads to the expression of two main isoforms termed ATG7(1) and ATG7(2) (Fig. 1.A). Survival and RNA expression data of cancer patients were downloaded from TCGA database and analysed on the Gepia webpage. Results showed that the correlation of high ATG7 expression with poor prognosis in PAAD is through ATG7(2) (Fig. 1.B-D). RNA expression data from Gtex were downloaded and expression of ATG7 isoforms was compared between pancreatic tissue and PAAD samples (Fig. 1.E) and showed increased expression of both isoforms in PAAD compared with normal pancreatic tissue. However, subsequent analysis of ATG7 isoforms expression in PAAD stages showed similar levels of A TG7(1) in all stages and increased ATG7(2) expression in Stage IV PAAD (Table 1), suggesting that ATG7(2) could play a role in PAAD progression and metastasis.

[0260] Table 1. Expression plot of ATG7(1) and ATG7(2) in PAAD clinical stages with statistical analysis table. P7341 PC00

[0261] ATG7(2) is involved in PAAD immunity

[0262] PANCAN data of samples from different immune subtypes were downloaded from TCGA and ATG7 isoform specific expression was analysed in these samples and showed that ATG7(1) is more expressed in immunologically quiet cancer samples while ATG7(2) is more expressed in immunologically active cancer samples (Fig. 2. A). This suggests that ATG7(2) could play a role in cytokine secretion and immune infiltration in cancer. Analysis of survival and new tumour formation in these samples showed that immunologically quiet cancers are significantly linked with greater survival time and higher delay to the apparition of a new tumour compared with immunologically active cancers (Fig. 2.B). This suggests that A TG7(2) effect on PAAD survival could be through immune regulation. Data of tumour infiltrating immune cells was downloaded from the TIMER2.0 database (Li etal. 2020) and correlation analysis of ATG7 expression in these samples revealed that ATG7 expression is linked with CAFs and M2 infiltration across cancers, including PAAD (Table 2).

[0263] Table 2. Correlation of ATG7 expression with M2 macrophages and CAFs infiltration across cancers. Black : positive correlation between ATG7 expression and tumour infiltration ; White : negative correlation between ATG7 expression and tumour infiltration ; * : significant correlation p<0,05. P7341 PC00

[0264] Together, these results suggest that the effect of ATG7 on PAAD previously attributed to autophagy could be caused by ATG7(2).

[0265] ATG7(2) promotes PAAD cell migration and proliferation in vitro

[0266] CRISPR / Cas 9 was used in the Capan-1 cells to selectively knock-out either total ATG7 expression by inducing nonsense mutation in Exon 3, or ATG7(1) expression by targeting the splice site of Exon 17 (Fig. 3.A). Knock-out of ATG7(1) was confirmed using qPCR (Fig. 3.B-D). The qPCR results showed that the different clones of ATG7(1)' / _cells expressed different levels of ATG7(2) mRNA, which were reflected by the total ATG7 mRNA expression in these cells (Fig. 3.B-D). Moreover, Western-Blot analysis of these clones showed endogenous ATG7 expression, suggesting that ATG7(2) is expressed endogenously at protein level (Fig. 3.E.). Staining for p62 and LC3B showed impaired autophagy activity in the ATG7(1)' / _cells compared with the control cells (Fig. 3.E). This was evident by the increase in the autophagy receptor cargo protein p62 and a lack of the lower lipidated band of LC3B in the ATG7(1)' / _cells. This confirms that ATG7(2) is unable to carry out autophagy activity and that selective knock-out of ATG7(1) is sufficient to impair autophagy. qPCR analysis of the ATG7_ / _clones showed minimal mRNA expression of ATG7, ATG7(1) and ATG7(2) (Fig. 3.F-H). Additional Western-Blot analysis of these cells confirmed the absence of ATG7 protein expression (Fig. 3.1). Staining of p62 and LC3B revealed that autophagy activity was impaired, exhibiting similar profile to the ATG7(1)' / _cells. Proliferation and migration assays showed that ATG7(1)' / _cells proliferate and migrate at a higher rate than the control cells (Fig. 3.J-K), while the ATG7' / _cells proliferate and migrate at a lower rate than the control cells (Fig. 3.L-M). This suggests that ATG7(1) knock-out facilitates Capan-1 cells proliferation and P7341 PC00 migration while total ATG7 knock-out impairs the cells ability to proliferate and migrate. Additionally, the ATG7(1)' / _clone with the highest A TG7(2) expression level was also the one exhibiting the highest migration and proliferation rates (Fig. 3.B-D and J-K). Together, these results suggest that ATG7(2) is a promoter of PAAD migration and proliferation in vitro.

[0267] ATG7(2) knock-down decreases PAAD cells proliferation in vitro 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). Two different siRNA sequences were tested for ATG7(2) knock-down. Efficiency and specificity of the siRNAs was confirmed in MEF Atg7_ / _cells expressing either ATG7(1) or ATG7(2) (Fig. 5.A-F). Capan-1 WT and ATG7(1)' / _cells were transfected with the siRNAs and qPCR results revealed that A TG7(2) knock-down leads to reduced A TG7(1) expression in WT cells (Fig. 4.B-D). Additionally, transfection of the ATG7(1)' / _clone with the highest ATG7(2) expression revealed that the second siATG7(2) sequence was the most efficient one, with 60% knock-down efficiency (Fig. 4.E-G). Additionally, siATG7(2) lead to decreased proliferation and migration rates of Capan-1 Control and ATG7(1)' / _cells compared with siControl in the same cells (Fig. 4.H-I). siATG7(2) also showed reduced expression of genes positively regulated by YAP (Fig. 4. J). Active YAP has been linked with poor prognosis in PAAD in different studies. When active, it acts as an oncoprotein that promotes the expression of genes involved in cell proliferation and tumor-microenvironment modulation. YAP has been shown to be up-regulated in metastatic PAAD, and has been identified as a therapeutic target. Together, these results suggest that targeting ATG7(2) in pancreatic cancer cells can decrease proliferation and, therefore, become a potential therapeutic approach for the treatment of PAAD.

[0268] Conclusion

[0269] The data indicate that ATG7(2) is unable to carry out autophagy activity, opposite to ATG7(1), and that ATG7(2), but not ATG7(1), is a promoter of PAAD migration and proliferation in vitro. Moreover, that data indicate that a siRNA targeting ATG7(2) and selective for ATG7(2) decreases proliferation of pancreatic cancer cells.

[0270] Example 2. P7341 PC00

[0271] Sequence overview

[0272] SEQ ID NO: 1: ATG7(1) : 5’- AGATCCGGGGATTTC -3’

[0273] SEQ ID NO: 2: ATG7 : 5’- AACTCCAATGTTAAG -3’

[0274] SEQ ID NO: 3: ATG7(1) : 5’- TTGAGTATACCAGTGGACCTGT -3’

[0275] SEQ ID NO: 4: 5’- CCCCACAAAAAGCCCTTAGTT -3’

[0276] SEQ ID NO: 5: ATG7 : 5’- TAGCATTGCTGTGGGAAGAGA -3’

[0277] SEQ ID NO: 6: 5’- CAGCACTCATCTCTATGGGAGG -3’

[0278] SEQ ID NO: 7: ATG7(1) : 5’- AGAAACCAGACGTGCATTTCA -3’

[0279] SEQ ID NO: 8: ATG7 : 5’- TGAACGCTGCTATTTCTGACTTG -3’

[0280] SEQ ID NO: 9: ACTIN : 5’- TTCTACAATGAGCTGCGTGTGG -3’

[0281] SEQ ID NO: 10: 5’- GTGTTGAAGGTCTCAAACATGAT -3’

[0282] SEQ ID NO: 11 : ATG7 : 5’- CGTTGCCCACAGCATCATCTTC -3’

[0283] SEQ ID NO: 12: 5’- CACTGAGGTTCACCATCCTTGG -3’

[0284] SEQ ID NO: 13: A TG7(1) : 5’- CAGAAGGGGGCTATGCCATT -3’

[0285] SEQ ID NO: 14: 5’- AACCGTGAAAGAAATCCCCG -3’

[0286] SEQ ID NO: 15: ATG7(2) : 5’- GCATCCAGAAGGGGGCTATG -3’

[0287] SEQ ID NO: 16: 5’- CAGGTTCTT -3’

[0288] SEQ ID NO: 17: 5’- GATCAAGAACCTGGTGAGGCA -3’

[0289] SEQ ID NO: 18: KI67 : 5’- GAAAGAGTGGCAACCTGCCTTC -3’

[0290] SEQ ID NO: 19: 5’- GCACCAAGTTTTACTACATCTGCC -3’

[0291] SEQ ID NO: 20: siATG7 : 5’- GAAGCUCCCAAGGACAUUAtt -3’

[0292] SEQ ID NO: 21 : siATG7(1) : 5’- GGCAUUUGACAAAUGUACAgc -3’

[0293] SEQ ID NO: 22: siATG7(2): 5’- UCACCAGGUUCUUGAUCAAtt -3’

[0294] SEQ ID NO: 23: 5’- UGUGCCUCACCAGGUUCUUtt -3’

[0295] SEQ ID NO: 24: ATG7(2) polypeptide:

[0296] MAAATGDPGLSKLQFAPFSSALDVGFWHELTQKKLNEYRLDEAPKDIKGYYYNGDSA

[0297] GLPARLTLEFSAFDMSAPTPARCCPAIGTLYNTNTLESFKTADKKLLLEQAANEIWESI

[0298] KSGTALENPVLLNKFLLLTFADLKKYHFYYWFCYPALCLPESLPLIQGPVGLDQRFSLK

[0299] QIEALECAYDNLCQTEGVTALPYFLIKYDENMVLVSLLKHYSDFFQGQRTKITIGVYDP

[0300] CNLAQYPGWPLRNFLVLAAHRWSSSFQSVEVVCFRDRTMQGARDVAHSIIFEVKLPE

[0301] MAFSPDCPKAVGWEKNQKGGMGPRMVNLSECMDPKRLAESSVDLNLKLMCWRLVP

[0302] TLDLDKWSVKCLLLGAGTLGCNVARTLMGWGVRHITFVDNAKISYSNPVRQPLYEFE

[0303] DCLGGGKPKALAAADRLQKIFPGVNARGFNMSIPMPGHPVNFSSVTLEQARRDVEQL

[0304] EQLIESHDWFLLMDTRESRWLPAVIAASKRKLVINAALGFDTFWMRHGLKKPKQQG P7341 PC00

[0305] AGDLCPNHPVASADLLGSSLFANIPGYKLGCYFCNDWAPGDSTRDRTLDQQCTVSR

[0306] PGLAVIAGALAVELMVSVLQHPEGGYAIASSSDDRMNEPPTSLGLVPHQVLDQYERE GFNFLAKVFNSSHSFLEDLTGLTLLHQETQAAEIWDMSDDETI

[0307] SEQ ID NO: 36: ATG7(2) polynucleotide.

[0308] SEQ ID NO: 26: ATG7(1) polypeptide:

[0309] MAAATGDPGLSKLQFAPFSSALDVGFWHELTQKKLNEYRLDEAPKDIKGYYYNGDSA

[0310] GLPARLTLEFSAFDMSAPTPARCCPAIGTLYNTNTLESFKTADKKLLLEQAANEIWESI

[0311] KSGTALENPVLLNKFLLLTFADLKKYHFYYWFCYPALCLPESLPLIQGPVGLDQRFSLK

[0312] QIEALECAYDNLCQTEGVTALPYFLIKYDENMVLVSLLKHYSDFFQGQRTKITIGVYDP

[0313] CNLAQYPGWPLRNFLVLAAHRWSSSFQSVEVVCFRDRTMQGARDVAHSIIFEVKLPE

[0314] MAFSPDCPKAVGWEKNQKGGMGPRMVNLSECMDPKRLAESSVDLNLKLMCWRLVP

[0315] TLDLDKWSVKCLLLGAGTLGCNVARTLMGWGVRHITFVDNAKISYSNPVRQPLYEFE

[0316] DCLGGGKPKALAAADRLQKIFPGVNARGFNMSIPMPGHPVNFSSVTLEQARRDVEQL

[0317] EQLIESHDWFLLMDTRESRWLPAVIAASKRKLVINAALGFDTFVVMRHGLKKPKQQG

[0318] AGDLCPNHPVASADLLGSSLFANIPGYKLGCYFCNDWAPGDSTRDRTLDQQCTVSR

[0319] PGLAVIAGALAVELMVSVLQHPEGGYAIASSSDDRMNEPPTSLGLVPHQIRGFLSRFD NVLPVSLAFDKCTACSSKVLDQYEREGFNFLAKVFNSSHSFLEDLTGLTLLHQETQAA EIWDMSDDETI

[0320] SEQ ID NO: 37: ATG7(1) polynucleotide.

[0321] SEQ ID NO: 28 - exon 16

[0322] GGGCTATGCCATTGCCAGCAGCAGTGACGATCGGATGAATGAGCCTCCAACCTCT

[0323] CTTGGGCTTGTGCCTCACCAG

[0324] SEQ ID NO: 29 - exon 17

[0325] ATCCGGGGATTTCTTTCACGGTTTGATAATGTCCTTCCCGTCAGCCTGGCATTTGA

[0326] CAAATGTACAGCTTGTTCTTCCAAA

[0327] SEQ ID NO: 30 - exon 18

[0328] GTTCTTGATCAATATGAACGAGAAGGATTTAACTTCCTAGCCAAGGTGTTTAATTCT

[0329] TCACATTCCTTCTTAGAAGACTTGACTGGTCTTACATTGCTGCATCAAGAAACCCA AGCTGCTGAG P7341 PC00

[0330] SEQ ID NO: 31 - siATG7(2) no. 1 ; Target RNA sequence ccucaccagguucuugaucaa

[0331] SEQ ID NO: 32 - siATG7(2) no. 1 ; Antisense sequence : UUGAUCAAGAACCUGGUGAGG siATG7(2) no 2 :

[0332] SEQ ID NO: 33 - siATG7(2) no. 2; Target RNA sequence cuugugccucaccagguucuu

[0333] SEQ ID NO: 34 - siATG7(2) no. 2; Antisense sequence AAGAACCUGGUGAGGCACAAG

[0334] SEQ ID NO: 35 5’- GGUUCUUGAUCAAUAUGAAcg -3’

[0335] References

[0336] M. H. Ogmundsdottir et al., ‘A short isoform of ATG7 fails to lipidate LC3 / GABARAP’, Sci. Rep., vol. 8, p. 14391 , Sep. 2018, doi: 10.1038 / s41598-018-32694-7.

[0337] K. Ostacolo et al., ‘ATG7(2) Interacts With Metabolic Proteins and Regulates Central Energy Metabolism’, Traffic, vol. 25, no. 4, p. e12933, 2024, doi: 10.1111 / tra.12933.

[0338] T. Li et al., ‘TIMER2.0 for analysis of tumor-infiltrating immune cells’, Nucleic Acids Res., vol. 48, no. W1 , pp. W509-W514, Jul. 2020, doi: 10.1093 / nar / gkaa407.

Claims

44P7341 PC00Claims1 . An oligonucleotide targeting a polynucleotide encoding human ATG7 variant 2 (ATG7(2)) as set forth in SEQ ID NO: 36, 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: 37.

4. The oligonucleotide according to any one of the preceding claims, wherein said oligonucleotide comprises a sequence complementary to ATG7(2) mRNA, such as 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:45P7341 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 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 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: 16 or a functional variant thereof, wherein said functional variant has 1 or 2 individual ribonucleotide substitutions compared to SEQ ID NO: 16, 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: 16 or said functional variant thereof, and a complementary antisense strand.

14. The oligonucleotide according any one of the preceding claims, wherein said oligonucleotide further comprises 1 to 16 nucleotides attached at the 5’ of SEQ ID NO: 16 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 of SEQ ID NO: 22 or SEQ ID NO: 23, or a functional variant thereof, wherein said functional variant has 1 , 2, 3, 4, 5, or 646P7341 PC00 individual ribonucleotide substitutions compared to any one of SEQ ID NO: 22 or SEQ ID NO: 23, 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: 22, SEQ ID NO: 23, or SEQ ID NO: 16, 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 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-methyl (2'-O-Me), such as 2'-O-methoxyethyl (2' MOE), such as 2'-fluoro, such as 2’-O-P7341 PC00 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).P7341 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 pancreatic cancer.

37. An oligonucleotide targeting human ATG7 variant 2 (ATG7(2)) according to any one of the preceding claims, for use in a method of decreasing proliferation and / or migration of pancreatic cancer cells.

38. The oligonucleotide for use according to any one of claims 35 and 37, wherein the pancreatic cancer is an exocrine pancreatic cancer or an endocrine pancreatic cancer.

39. The oligonucleotide for use according to any one of claims 35 to 38, such as wherein the pancreatic cancer is pancreatic adenocarcinoma (PAAD), pancreatic neuroendocrine tumor, acinar cell carcinoma of the pancreas, cystadenocarcinomas, pancreatoblastoma, adenosquamous carcinomas, signet ring cell carcinomas, hepatoid carcinomas, colloid carcinomas, undifferentiated carcinomas, undifferentiated carcinomas with osteoclast-like giant cells, or pancreatic mucinous cystic neoplasms.

40. The oligonucleotide for use according to any one of claims 35 to 39, 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.49P7341 PC0041. The oligonucleotide for use according to any one of claims 35 to 40, 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; g. a prodrug, such as wherein the prodrug is metabolized in an active form in the body.

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

43. The composition according to claim 42, wherein the composition further comprises an acceptable carrier.

44. A method of decreasing migration and / or proliferation of a pancreatic cancer cell, said method comprising administration of a therapeutically effective amount of a oligonucleotide targeting ATG7(2) according to any one of claims 1 to 34, or the composition according to any one of claims 42 to 43, to an individual in need thereof.

45. A method of treating pancreatic cancer, 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: 36, 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.P7341 PC0046. Method of treating pancreatic cancer, the method comprising administering the oligonucleotide according to any one of claims 1 to 34, or the composition according to any one of claims 42 to 43.

47. Use of the oligonucleotide according to any one claims 1 to 34, or the composition according to any one of claims 42 to 43, for the manufacture of a medicament.

48. Use of the oligonucleotide according to any one claims 1 to 34, or the composition according to any one of claims 42 to 43, for the manufacture of a medicament for treating pancreatic cancer.

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