Immune cells with inhibited line1 expression

Abstract

The present invention relates to an immune cell stably or transiently affected in the expression of LINE1 (L1) and its use in treating or preventing cancer. The L1 expression may be inhibited or downregulated through an inhibitor, such as a polynucleotide or a small molecule. Further, the invention relates to a method for engineering immune cells.

Description

[0001] DESCRIPTION TITLE IMMUNE CELLS WITH INHIBITED LINE1 EXPRESSION

[0002] Technical field

[0003] The present invention provides an immune cell affected in the expression of Long Interspersed Nuclear Element 1 (LINE1 or L1 ).

[0004] Backg ound art

[0005] Immunotherapies utilizing Adoptive T Cell Transfer (ACT) have marked a groundbreaking advancement in cancer treatment. ACT is a sophisticated form of immunotherapy that harnesses the body's immune system to fight diseases, particularly cancers. This method involves isolating T cells from a patient, genetically modifying them to target and destroy cancer cells, expanding them outside the body, and then reinfusing them into the patient. In particular, Chimeric Antigen Receptors (CARs) are engineered receptors that alter T- cell antigen specificity and reprogram T-cell function against a specific target. CAR-T cells are presently employed in clinical settings to treat several B cell hematologic malignancies, including B lineage leukaemia, non-Hodgkin lymphoma and relapsed and / or refractory multiple myeloma.

[0006] Despite the promise of CAR-T cell therapy, significant challenges remain in its widespread application and long-term efficacy. A substantial fraction of patients experiences relapse or recurrence of diseases after initial treatment. Moreover, a large proportion of tumours, especially solid tumours, are still not eligible for this therapy. The primary limitations in achieving broader success include the poor persistence of CAR-T cells in the tumor microenvironment and their premature exhaustion, which hinders their long-term anti-tumor effectiveness. Additional challenges include low or absent target antigen expression, inadequate tumor penetration, and immune suppression within the tumor microenvironment, particularly in solid tumours.

[0007] Another factor influencing T cell function and identity is the expression of retrotransposable elements (REs), particularly Long Interspersed Nuclear Element 1 (LINE1 ). LINE1 elements are the most abundant superfamily of non-LTR retrotransposons in the human genome, making up approximately 18% of genomic content. Recent research has uncovered that LINE1 transcripts can accumulate at chromatin in tumor-infiltrating lymphocytes (TILs) experiencing a loss of effector functions and undergoing exhaustion. These LINE1 -transcripts have emerged as crucial factors in determining TIL dysfunctional state, serving as epigenetic regulators for an entire class of genes associated with T-cell effector functions. It has been appreciated that a method for enhancing CAR-T cell persistence and antitumor effect is needed that overcomes one or more of these problems.

[0008] Disclosure of the invention

[0009] In a first aspect, the present invention relates to an immune cell wherein said cell is stably or transiently affected in the expression of Long Interspersed Nuclear Element 1 (LINE1 or L1 ). Preferably, the immune cell is an engineered cell, more preferably the cell expresses at least one chimeric antigen receptor (CAR) or at least one artificial T-cell receptor (TCR) subunit.

[0010] In a preferred embodiment, the immune cell is selected from the group consisting of a CAR-T cell, a CAR-NK cell and a CAR-macrophage, preferably, the immune cell is a CAR-T cell.

[0011] The present invention provides a novel approach to enhance the persistence and antitumor effects of immune cells, preferably of cell expressing a CAR. This approach involves the modulation of Long Interspersed Nuclear Element 1 (LINE1 ) expression in immune cell; preferably, the expression of LINE1 (L1 ) is stably or transiently affected in said cell, more preferably is stably or transiently inhibited or downregulated.

[0012] In some embodiments, the expression of L1 is affected through an inhibitor or suppressor, said inhibitor being chosen from a polynucleotide or a small molecule. Preferably, the polynucleotide is selected from the group consisting of an antisense construct, antisense oligonucleotide, RNA interference construct or siRNA or a polynucleotide coding for it.

[0013] In a preferred embodiment, the polynucleotide is an isolated inhibitory nucleic acid targeting LINE1 , preferably, said inhibitory nucleic acid is at least one RNA inhibitor selected from the group consisting of: antisense oligo (ASO), gapmer, mixmer, shRNA, siRNA, stRNA, snRNA, more preferably said inhibitory nucleic acid is modified, and / or comprises one or more modified bonds or bases. One example of modified inhibitory nucleic acid is phosphorothioate bonds and flanking 2’-fluoro-ribonucleotides (2’F-ASO) and 2’-deoxy-2’-fluoro-p-D-arabinonucleid acid (FANA) ASO. Preferably, the polynucleotide comprises a sequence capable of hybridizing or complementary to a sequence comprising or consisting of: SEQ ID NO: 1 , 2 or 3.

[0014] Preferably, the immune cell has its L1 expression affected through a polynucleotide that targets or is complementary to a sequence selected from the group consisting of SEQ ID NO: 4-8 or its corresponding RNA sequence. More preferably, the polynucleotide targets or is complementary to a sequence that has at least 70%, preferably at least 80%, more preferably at least 90% identity with SEQ ID NO: 8. In a preferred embodiment, the polynucleotide is complementary to a sequence having at least 80%, preferably at least 90% identity with a sequence selected from the group consisting of SEQ ID NOs: 9-10.

[0015] Preferably, the polynucleotide sequence that has at least 70%, preferably at least 80%, more preferably at least 90% identity to a sequence selected from the group consisting of SEQ ID NOs: 11 -19.

[0016] In some embodiments, the immune cell is either autologous or allogenic or derived from an iPSC. Preferably, the immune cell expresses a Chimeric Antigen Receptor (CAR) targeting an antigen, more preferably the antigen is selected from: CD19, BCMA, CD22, CD20, CD138, CD33, CD123, PSMA.

[0017] In a second aspect the present invention relates to a composition comprising the immune cell.

[0018] In a third aspect, the present invention relates to the immune cell for use as a medicament.

[0019] In a fourth aspect, the present invention relates to the immune cell for use in the treatment or prevention or in a follow-up of a cancer. Preferably, the cancer is a blood cancer o a solid cancer, more preferably is a B lineage leukaemia, non-Hodgkin lymphoma and multiple myeloma.

[0020] A fifth aspect of the present invention relates to a method for engineering an immune cell comprising at least one step of delivery of an L1 inhibitor or suppressor to said cell. Preferably, said inhibitor is selected from a polynucleotide or a small molecule as above disclosed.

[0021] Brief description of drawings

[0022] Figure 1 shows that chronic over-stimulation with anti-CD3 / CD28 beads drives T cells into an exhausted taste. (A) Schematic representing T cell overstimulation with anti- CD3 / CD28 beads. (B) Expression levels of LINE1 were assessed by RT-qPCR in T cell over-stimulated or not with anti-CD3 / CD28 beads (n = 2 individuals). Data represent mean ± s.e.m. (C) Percentage of PD1 positive cells in T cell over-stimulated or not with antiCD3 / CD28 beads (n = 2 individuals). Data represent mean ± s.e.m. (D) Percentage of IFN-y positive cells in T cell over-stimulated or not with anti-CD3 / CD28 beads (n = 2 individuals). Data represent mean ± s.e.m.

[0023] Figure 2 shows that CAR-T cells do not express LINE1. Expression levels of LINE1 (A), LINE transcripts #1 (B), and LINE1 transcript #2 (C) were assessed by RT-qPCR in CD3+ T cells, CAR T cells, and naive CD4+ T cells isolated from healthy donors (n = 4 individuals). Data represent mean ± s.e.m. LINE1 T cells vs CAR T * P = 0.05, paired Two-tailed t test; LINE1 T cells vs naive CD4+ ** P=0.002, unpaired Two-tailed t test; LINE1 CAR T vs naive CD4+ ** P=0.001 , unpaired Two-tailed t test; Figure 3 shows that CAR T cells die after chronic over-stimulation with anti- CD3 / CD28beads. (A) Schema representing T cell overstimulation with anti-CD3 / CD28 beads. (B) Percentage of live CAR-T and T cells not and overstimulated (n=2 individuals for CART cells; n=3 individuals for T cells). Data represent mean ± s.e.m;

[0024] Figure 4 demonstrates that overstimulation of CAR-T cells by CD19-positive targets increases LINE1 expression accompanied by a reduction of killing abilities without inducing canonical exhaustion markers. (A) Schematic representing CAR T cell overstimulation with the specific antigen CD19 that is expressed by LCL cell line. (B) Expression levels of LINE1 were assessed by RT-qPCR in CAR-T cell over-stimulated or not with LCL (n = 1 individuals). (C) Percentage of PD1 positive cells in CART cell over- stimulated or not with LCL (n = 1 individuals). (D) Percentage of IFN-y positive cells in CART cell over-stimulated or not with LCL (n = 1 individuals). (E) Percentage of dead LCL (CD19+ cells) co-cultured for 12 h with CAR T cells (n = 1 individual). Percentage of LCL self-death is indicated (without CAR T cells);

[0025] Figure 5 shows the inhibition of mTORCI with Rapamycin induces LINE1 expression and reduces the effector functions of CAR-T cells. (A) Schematic representing CAR T cell treatment with the mTORCI inhibitor. Expression levels of LINE1 (B), LINE transcripts #1 (C), and LINE1 transcript #2 (D) were assessed by RT-qPCR in CAR-T cells treated or not with Rapamycin. (n = 2 biological replicates). E) Percentage of dead LCL (CD19+ cells) co-cultured for 12 h with CAR T cells treated or not with Rapamycin (n = 2 biological replicates). Percentage of LCL spontaneous cell death is also indicated (without CAR-T cells, left point. (F) IFN-y-, (G) TNF-a-, (H) GrzB- positive CAR-T cell treated or not with Rapamycin (n = 2 biological replicates). Data represent mean ± s.e.m.; and Figure 6 shows LINE1 expression that modulates the effector functions of CAR-T cells. (A) Schematic representing CAR T cell treatment with the mTORCI inhibitor in presence or absence of LINE1 or control (Scr) ASO. (B) Expression levels of LINE1 by RT-qPCR in CAR-T cells treated or not with Rapamycin in presence or absence of LINE1 or control (Scr) ASO (n = 1 individuals). (C) Percentage of dead LCL (CD19+ cells) co-cultured for 12 h with CAR-T cells treated or not with Rapamycin in presence or absence of LINE1 or control (Scr) ASO (n = 1 individuals). Percentage of LCL spontaneous cell death is indicated (without CAR T cells).

[0026] Detailed description of preferred embodiments of the invention

[0027] In a first aspect, the present invention provides an immune cell in which the expression of Long Interspersed Nuclear Element 1 (LINE1 or L1 ) is affected, preferably the expression of LINE1 is inhibited or downregulated.

[0028] Preferably, the expression of LINE1 is explicitly and actively inhibited or downregulated.

[0029] In a preferred embodiment, the expression of LINE1 is explicitly and actively directly inhibited or down regulated.

[0030] In some embodiments, the immune cell is an immune cell expressing a chimeric antigen receptor (“CAR”) or artificial T-cell receptor (“TCR”) subunit.

[0031] Preferably, the immune cell is selected from the group consisting of: a CAR-T cell, a CAR-NK cell, a CAR-macrophage or a TCR-T cell.

[0032] In a preferred embodiment, the immune cell is a cell expressing a CAR, preferably a CAR-T or a CAR-NK cell, more preferably a CAR-T cell.

[0033] In some embodiments, the expression of L1 is affected through at least one inhibitor preferably selected from: a polynucleotide and a small molecule.

[0034] In some embodiments, the immune cell, preferably the CAR-T or the CAR-NK cell is stably or transiently affected the expression of L1 .

[0035] In some embodiments, the immune cell is stably affected in the L1 expression. This could involve a permanent alteration in the expression of L1 in the immune cell. The stable alteration of L1 expression may result in a long-lasting enhancement of the immune cells' persistence and anti-tumor activity within the tumor microenvironment.

[0036] In other embodiments, the immune cell is transiently affected in the L1 expression. This could involve a temporary alteration in the expression of L1 in the immune cells.

[0037] Preferably, the immune cell has its L1 expression inhibited or downregulated.

[0038] In some embodiments, the immune cell has its L1 expression inhibited or downregulated through various means, such as the use of at least one inhibitor or suppressor.

[0039] In some embodiments, the at least one inhibitor is selected from the group consisting of a polynucleotide, a CRISPR component and a small molecule. For example, the polynucleotides may be RNA interference constructs or small interfering RNAs (siRNAs) that bind to L1 RNA and prevent its translation, thereby down-regulating L1 expression.

[0040] In some embodiments, the polynucleotide is an antisense construct, antisense oligonucleotide, RNA interference construct, or small interfering RNA (siRNA), or a polynucleotide coding for these.

[0041] In some embodiments, the polynucleotide is an isolated inhibitory nucleic acid targeting L1. This inhibitory nucleic acid may be an RNA inhibitor, preferably selected from an antisense oligo (ASO), gapmer, mixmer, shRNA, siRNA, stRNA, and snRNA.

[0042] In other embodiments, the polynucleotide is a gapmer. A gapmer is a type of antisense oligonucleotide that contains a central block of deoxynucleotides flanked by blocks of modified nucleotides, like modified ribonucleotides. The central block of deoxynucleotides is designed to hybridize with the target L1 RNA, while the flanking blocks of modified nucleotides provide stability and resistance to nuclease degradation. When the gapmer binds to the L1 RNA, it may trigger the degradation of the RNA, thereby inhibiting or down-regulating L1 expression. In some embodiments, the gapmer is a gapmer with phosphorothioate bonds and flanking 2’-fluoro-ribonucleotides (2’F-ASO), a 2’-deoxy-2’-fluoro-p-D-arabinonucleid acid (FANA) ASO, and / or it comprises one or more modified bonds or bases. The sequence of the ASO may be capable of hybridizing or complementary to a sequence comprising or consisting of specific sequences. In some cases, the polynucleotide inhibitor comprises or consists of a sequence having a certain level of identity with specific sequences.

[0043] In yet other embodiments, the polynucleotide is a mixmer. A mixmer is a type of antisense oligonucleotide that contains a mixture of deoxynucleotides and modified nucleotides throughout the sequence. The mixmer is designed to hybridize with the target L1 RNA and trigger its degradation, thereby inhibiting or down-regulating L1 expression.

[0044] In yet other embodiments, the polynucleotide is a short hairpin RNA (shRNA). The shRNA is a small RNA molecule that forms a hairpin structure and is processed within the cell to produce siRNA. The siRNA may bind to the L1 RNA and trigger its degradation, thereby inhibiting or down-regulating L1 expression.

[0045] In yet other embodiments, the polynucleotide is a small interfering RNA (siRNA). The siRNA is a short RNA molecule that is complementary to a portion of the L1 RNA. The siRNA may bind to the L1 RNA and trigger its degradation, thereby inhibiting or downregulating L1 expression.

[0046] In yet other embodiments, the polynucleotide is a small nuclear RNA (snRNA). The snRNA is a small RNA molecule that is involved in the splicing of pre-mRNA.

[0047] Alternatively, the CRISPR components may be guide RNAs that direct a CRISPR- associated (Cas) protein to the L1 gene, resulting in the disruption of L1 expression. The small molecules may be drugs that inhibit the activity of the L1 protein. The downregulation of L1 expression in the CAR-T cells may enhance their persistence within the tumor microenvironment and their anti-tumor activity, potentially leading to improved outcomes in the treatment of cancer.

[0048] Preferably, the polynucleotide is complementary to a sequence that has at least 70%, preferably at least 80%, more preferably at least 90% identity to a sequence selected from the group consisting of SEQ ID NOs: 1 -3.

[0049] In some embodiments, the polynucleotide is complementary to a sequence that has 91 , 92, 93, 94, 95, 96, 97, 98, 99 or 100% identity to a sequence selected from the group consisting of SEQ ID NOs: 1 -3.

[0050] In some embodiments, the polynucleotide comprises a sequence of nucleotides that are complementary to 10 to 50 consecutive nucleotides of SEQ ID NO: 1 , 2 or 3, preferably complementary to 15 to 25 consecutive nucleotides of SEQ ID NO: 1 , 2 or 3.

[0051] In some embodiments, the immune cell, preferably a cell expressing a CAR, has its L1 expression affected through a polynucleotide that targets or is complementary to a sequence selected from the group consisting of SEQ ID NO: 4-8 or its corresponding RNA sequence.

[0052] In a preferred embodiment, the immune cell, preferably a cell expressing a CAR, has its L1 expression affected through a polynucleotide that targets or is complementary to a sequence selected from the group consisting of SEQ ID NO: 4-8 or its corresponding RNA sequence.

[0053] In another preferred embodiment, the immune cell, preferably a cell expressing a CAR, has its L1 expression affected through a polynucleotide that targets or is complementary to SEQ ID NO: 8 or its corresponding RNA sequence.

[0054] In a preferred embodiment, the polynucleotide is complementary to at least an Untranslated Region (UTR) of L1 or to at least one Open Reading Frame (ORF), preferably the polynucleotide is complementary and targets at least one portion of the 3’ UTR and / or to ORF2 of L1 .

[0055] Preferably, the polynucleotide is complementary to a sequence that has at least 70%, preferably at least 80%, more preferably at least 90% identity to a sequence selected from the group consisting of SEQ ID NOs: 9-10.

[0056] In some embodiments, the polynucleotide is complementary to a sequence that has 91 , 92, 93, 94, 95, 96, 97, 98, 99 or 100% identity to a sequence selected from the group consisting of SEQ ID NOs: 9-10.

[0057] In some embodiments, the polynucleotide comprises a sequence of nucleotides that are complementary to 10 to 50 consecutive nucleotides of SEQ ID NO: 9 or 10, preferably complementary to 15 to 25 consecutive nucleotides of SEQ ID NO: 9 or 10.

[0058] Preferably, the polynucleotide sequence has at least 70%, preferably at least 80%, more preferably at least 90% identity to a sequence selected from the group consisting of SEQ ID NOs: 11 -19.

[0059] Preferably, the polynucleotide sequence consists of a sequence selected from the group consisting of SEQ ID NOs: 11 -19.

[0060] In a preferred embodiment, the polynucleotide sequence has at least 70%, preferably at least 80%, more preferably at least 90% identity to a sequence selected from the group consisting of SEQ ID NOs: 11 , 16-19.

[0061] Preferably, the polynucleotide sequence consists of a sequence selected from the group consisting of SEQ ID NOs: 11 , 16-19.

[0062] In a preferred embodiment, the polynucleotide is an ASO, preferably a 2’F-ASO or (FANA) ASO.

[0063] In yet other embodiments, the expression of L1 in the immune cell, preferably a cell expressing a CAR, is affected through the use of a small molecule inhibitor.

[0064] In some embodiments, the immune cell with affected L1 expression is autologous. Preferably, autologous immune cell is derived from the patient's own T or NK cells, which are collected, genetically modified to express a CAR, expanded in vitro, and then reinfused back into the patient.

[0065] In other embodiments, the immune cell with affected L1 expression is allogenic. Preferably, allogenic immune cells are derived from the T or NK cells of a healthy donor, which are collected, genetically modified to express a CAR, expanded in vitro, and then infused into the patient.

[0066] In yet other embodiments, the immune cell with affected L1 expression is derived from induced pluripotent stem cells (iPSCs). Preferably, iPSC-derived cells are generated by reprogramming adult cells into a pluripotent state, which are then differentiated into T or NK cells, genetically modified to express a CAR, and expanded in vitro.

[0067] In some embodiments, the immune cell expresses a Chimeric Antigen Receptor (CAR) that targets at least a specific antigen. The antigen targeted by the CAR may vary depending on the type of cancer being treated.

[0068] Preferably, CAR target antigen is selected from CD19, B-cell maturation antigen (BCMA), CD22, CD20, CD138, CD33, CD123 and Prostate-Specific Membrane Antigen (PSMA). This targeted approach may allow for the selective destruction of cancer cells, thereby enhancing the anti-tumor efficacy of the CAR cell therapy.

[0069] In some embodiments, the immune cell with affected L1 expression expresses a CAR that targets CD19. CD19 is a protein expressed on the surface of B cells and is a common target for CAR-T cell therapy in the treatment of B cell malignancies. By targeting CD19, the CAR-T cells may selectively destroy CD19-positive cancer cells, thereby enhancing the anti-tumor efficacy of the therapy.

[0070] In other embodiments, the immune cell with affected L1 expression expresses a CAR that targets B-cell maturation antigen (BCMA). BCMA is a protein expressed on the surface of plasma cells, including malignant plasma cells in multiple myeloma.

[0071] In yet other aspects, embodiments, the immune cell with affected L1 expression expresses a CAR that targets CD22. CD22 is a protein expressed on the surface of B cells and is a target for CAR cell therapy in the treatment of B cell malignancies.

[0072] In other embodiments, the immune cell with affected L1 expression expresses a CAR that targets CD20. CD20 is a protein expressed on the surface of B cells and is a target for CAR cell therapy in the treatment of B cell malignancies.

[0073] In other embodiments, the immune cell with affected L1 expression expresses a CAR that targets CD138. CD138 is a protein expressed on the surface of plasma cells, including malignant plasma cells in multiple myeloma.

[0074] In other embodiments, the immune cell with affected L1 expression expresses a CAR that targets CD33. CD33 is a protein expressed on the surface of myeloid cells and is a target for CAR cell therapy in the treatment of myeloid malignancies.

[0075] In other embodiments, the immune cell with affected L1 expression expresses a CAR that targets CD123. CD123 is a protein expressed on the surface of myeloid cells and is a target for CAR cell therapy in the treatment of myeloid malignancies.

[0076] In other embodiments, the immune cell with affected L1 expression expresses a CAR that targets Prostate-Specific Membrane Antigen (PSMA). PSMA is a protein expressed on the surface of prostate cancer cells and is a target for CAR cell therapy in the treatment of prostate cancer.

[0077] The immune cell with affected L1 expression may be used in the treatment or prevention of various types of cancers, including but not limited to, blood cancers or solid cancers.

[0078] The modulation of L1 expression in the immune cells may impact the function and exhaustion of these cells. L1 has been found to play a role in the regulation of T or NK cell function and identity, with its expression being associated with the transition from T or NK cell quiescence to activation and exhaustion. Therefore, by affecting the expression of L1 in the immune cells, it may be possible to control the activation and exhaustion of these cells, thereby enhancing their anti-tumor activity. For example, the inhibition or down-regulation of L1 expression may prevent the exhaustion of the immune cells, preferably of the immune cells expressing a CAR allowing them to maintain their effector functions for a longer period of time. This could result in a more sustained anti-tumor response, potentially leading to improved outcomes in the treatment of cancer. The targeting of L1 expression may result in the downregulation or inhibition of L1 expression, potentially enhancing the persistence and anti-tumor activity of the CAR-T cells within the tumor microenvironment.

[0079] Further, the invention relates to an immune cell affected in the expression of Long Interspersed Nuclear Element 1 (LINE1 or L1 ). In some embodiments, the immune cell is selected form the group consisting of: a T cell, a B cell, an NK cell or a T cell, B cell, NK cell or Tumor cell, preferably said cell is a CD4+ T cell or a CD8+ T cell, or a dysfunctional T cell, e.g. a Tumor Infiltrating Lymphocyte (TIL).

[0080] In a preferred embodiment, the polynucleotide is complementary to at least an Untranslated Region (UTR) of L1 or to at least one Open Reading Frame (ORF), preferably the polynucleotide is complementary and targets at least one portion of the 3’ UTR and / or to ORF2 of L1 .

[0081] Preferably, the polynucleotide is complementary to a sequence that has at least 70%, preferably at least 80%, more preferably at least 90% identity to a sequence selected from the group consisting of SEQ ID NOs: 9-10.

[0082] In some embodiments, the polynucleotide is complementary to a sequence that has 91 , 92, 93, 94, 95, 96, 97, 98, 99 or 100% identity to a sequence selected from the group consisting of SEQ ID NOs: 9-10.

[0083] Preferably, the polynucleotide sequence has at least 70%, preferably at least 80%, more preferably at least 90% identity to a sequence selected from the group consisting of SEQ ID NOs: 11 -19.

[0084] Preferably, the polynucleotide sequence consists of a sequence selected from the group consisting of SEQ ID NOs: 11 -19.

[0085] In a preferred embodiment, the polynucleotide sequence has at least 70%, preferably at least 80%, more preferably at least 90% identity to a sequence selected from the group consisting of SEQ ID NOs: 11 , 16-19.

[0086] Preferably, the polynucleotide sequence consists of a sequence selected from the group consisting of SEQ ID NOs: 11 , 16-19.

[0087] A second aspect of the present invention relates to a composition comprising the immune cell disclosed above and a pharmaceutically acceptable carrier. The pharmaceutically acceptable carrier may be any suitable carrier known in the art that can facilitate the delivery of the immune cell to a subject's cells or tissues.

[0088] In some embodiments, the composition is formulated as a solution, suspension, emulsion, or other suitable formulation that can facilitate the administration of the immune cell. The composition may be prepared using standard techniques known in the art and may be administered to the subject using various methods, including but not limited to, injection, infusion, inhalation, or other suitable administration methods.

[0089] A third aspect of the present invention relates to the immune cell with affected L1 expression or to the composition comprising said immune cell for use as a medicament. Preferably, the immune cell is as disclosed above in detail. Preferably, the immune cell or a composition comprising said immune cell is administered to an individual in need thereof, more preferably an individual diagnosed with cancer. In some embodiment, the composition comprising the immune cell is formulated for enteral or parenteral administration, preferably for parenteral administration.

[0090] Preferably, the composition is formulated for parenteral administration, more preferably as: a solution a suspension, an injectable preparation, an infusion, a concentrate for injectable preparations, or a powder for injectable preparation.

[0091] In some embodiments, the immune cell or the composition is administered in a single dose or in multiple doses over a period of time, depending on the needs of the patient and the type of cancer being treated.

[0092] In some embodiments, the immune cell is used in immunotherapy, preferably in cancer immunotherapy. Preferably, the immune cell is used for improving the persistence of the T or NK cell, preferably cells expressing a CAR within the tumor microenvironment and its anti-tumor activity. A fourth aspect of the present invention relates to the immune cell with affected L1 expression or a composition comprising said immune cell for use in the treatment or in the prevention or in a follow-up of cancer.

[0093] Preferably, the treatment comprises the administration of the immune cell or of a composition comprising said cell to an individual diagnosed with cancer, with the aim of destroying the cancer cells and reducing the size of the tumor.

[0094] Preferably, the prevention comprises the administration of the immune cells or of a composition comprising said cell to an individual at risk of developing cancer, with the aim of enhancing the patient's immune response and reducing the likelihood of cancer development.

[0095] Preferably, the follow-up comprises the administration of the immune cells to a patient who has undergone treatment for cancer, with the aim of monitoring the patient's response to treatment and detecting any signs of cancer recurrence. The follow-up may be conducted at regular intervals, such as every few months or once a year, depending on the type of cancer and the patient's risk of recurrence. The follow-up may involve various tests and procedures, such as blood tests, imaging studies, and biopsies, in addition to the administration of the immune cells.

[0096] In some embodiments, the immune cells with affected L1 expression is used in the treatment of a solid cancer. Preferably, the solid cancer is selected from the group consisting of lung cancer, breast cancer, colon cancer and prostate cancer.

[0097] The CAR-T cell is administered to a patient diagnosed with a solid cancer, with the aim of destroying the cancer cells and reducing the size of the tumor. The affected L1 expression in the immune cells may enhance their persistence within the tumor microenvironment and their anti-tumor activity, potentially leading to improved outcomes in the treatment of solid cancers.

[0098] In other embodiments, immune cells with affected L1 expression is used in the treatment of blood cancers. Preferably, the blood cancer is selected from the group consisting of a leukaemia, preferably B lineage leukaemia, a lymphoma, preferably Non-Hodgkin lymphoma and a myeloma, preferably multiple myeloma. The immune cells may be administered to a patient diagnosed with a blood cancer, with the aim of destroying the cancer cells and reducing the disease burden. The affected L1 expression in the immune cells, preferably cells expressing a CAR may enhance their persistence within the tumor microenvironment and their anti-tumor activity, potentially leading to improved outcomes in the treatment of blood cancers.

[0099] In some embodiments, the immune cell or the composition comprising it is administered alone.

[0100] In other embodiments, the immune cell or the composition comprising it is administered in combination or in association with at least one therapy for the treatment of cancer. Preferably, the at least one therapy for the treatment of cancer is selected from surgery, radiation therapy, chemotherapy, thermal ablation or other forms of immunotherapy.

[0101] A fifth aspect of the present invention relates to a method for engineering an immune cell, preferably a cell expressing a CAR comprises at least one step of delivery of an L1 inhibitor to said cell.

[0102] In some embodiments, the delivery of the L1 inhibitor is achieved through transfection, electroporation or viral transduction. Preferably, the L1 inhibitor is a polynucleotide, or a small molecule, as previously described. The delivery of the L1 inhibitor or suppressor may result in the stable or transient alteration of L1 expression in the immune cells, potentially enhancing their persistence and anti-tumor activity within the tumor microenvironment.

[0103] In some embodiments, the immune cell is engineered to express at least one LINE1 - interfering molecule. This may involve the introduction of a polynucleotide sequence into the genome of the immune cells that codes for the at least one LINE1 -interfering molecule

[0104] The delivery of the LINE1 inhibitor may be achieved through various means, such as transfection, electroporation, nanoparticles or viral transduction.

[0105] In some embodiments, the delivery of the inhibitor is achieved through direct injection of the inhibitor, preferably of the polynucleotide, more preferably of the ASO in the immune cell.

[0106] In some embodiments, the LINE1 inhibitor, preferably the polynucleotide, more preferably the ASO is delivered by nanoparticles, preferably by lipid nanoparticles in the cell.

[0107] A sixth aspect of the present invention relates to a method of treating a subject in need thereof comprising administering to the subject an immune cell or a composition comprising said cell as above detailed disclosed.

[0108] EXAMPLE

[0109] LINE1 -transcripts originate from CD4+-specific genes upregulated during T-cell activation. In naive CD4+ T-cells, LIN E1 -transcripts are regulated by the transcription factor IRF4 and maintained at chromatin by Nucleolin, acting in cis to hinder H3K36me3 levels and stall gene expression. T-cell activation triggers LINE1 -transcript downregulation by the PTBP1 splicing suppressor and promotes the expression of the corresponding protein-coding genes via the GTF2F1 elongating factor through mTORCI . Here, has been for the first time demonstrated that LINE1 expression fine-tunes the effector response of T cells. Notably, the Applicant has shown that in vitro exhausted T cells and dysfunctional tumor infiltrating lymphocytes (TILs), isolated from various tumors and patients, accumulate LINE1 transcripts at chromatin. Furthermore, pharmacologically targeting LINE1 transcripts with antisense oligonucleotides can reinvigorate the antitumor immune response of exhausted T cells and TILs.

[0110] Material and Methods

[0111] Cell culturing and treatments

[0112] CAR T cells were cultured at a concentration of 0.8 x 106cells / mL in a medium composed of complete RPMI medium (RPMI with GlutaMAX-l (Gibco), 10% (v / v) Fetal Bovine Serum (FBS) (Gibco), 1% (v / v) non-essential amino acids, 1 mM sodium pyruvate, 50 U / mL penicillin, 50 pg / mL streptomycin) supplemented with specific cytokines (5 ng / mL recombinant IL-15 (Miltenyi Biotec), 10 ng / mL recombinant IL-7 (Miltenyi Biotec).

[0113] For CAR T cell chronic stimulation LCL cell lines were irradiated with 3,000 rads. CAR T cells were overstimulated every 3.5 days with 150.000 LCL irradiated cells / 1x106CAR T cells.

[0114] To inhibit tonic mTORCI activity in quiescent neonatal naive CD4+ T cell, 15 x 106cells / mL CAR T cells were cultured in complete RPMI medium with the addition of 250 nM Rapamycin (Merck; cat. num. R8781 ) for 72.

[0115] To downregulate LINE1 , FANA-ASOs (2'-deoxy-2'-fluoro-p-d-arabinonucleic acid) were employed, following the sequences and usage protocol outlined in 39. An unrelated scr ASO was used as a control. ASOs were administered without any transfection reagent, following the manufacturer’s instruction, at a final concentration of 10 pM. CAR T cells were cultured for 48 h in complete RPMI medium supplemented IL-5 and IL-7 and 10 pM of the specific ASOs. RNA knockdown efficiency was assessed through RT-qPCR. T cells were maintained at 37°C in a 5% CO2 humidified incubator.

[0116] T cell staining and flow cytometry analysis

[0117] For intracellular cytokine staining, 5 x 104CAR T cells were stimulated with 50 ng ml-1 phorbol 12-myristate 13-acetate and with 0.5 pg ml-1 ionomycin for 2 h at 37 °C; subsequently, 100 pg ml-1 brefeldin A (Merck) was added for an additional 2 h at 37 °C.

[0118] Cells were washed, fixed and permeabilized for 30 min at 4 °C with the Foxp3

[0119] Transcription Factor Fixation / Permeabilization kit (Invitrogen by Life Technologies) according to the manufacturer’s instructions. Cytokines were stained by incubating 1 pl antibody for every 5 x 104cells diluted in Permeabilization Buffer (Invitrogen by Life Technologies) for 20 min at room temperature. T cells were washed with PBS and then analyzed. For cytokine staining, the following antibodies were used: anti-IFN-y-V450 (clone B27), anti-GrzB-PE (clone GB11 ), anti-TNFa-BUV395 (clone Mab11 ).

[0120] Killing assay

[0121] CAR T inhibited or not for mTORCI were treated with ASOs and co-cultured for 12 h with LCL CD19+ cells at a 1 :2 ratio. After co-culturing, cells were stained with the LIVE / DEAD Fixable Green Dead Cell Stain kit (Invitrogen by Life Technologies, L34969) for 20 min at room temperature, washed with PBS and stained with anti-CD19-PECy7 antibody (clone SJ25C1 ) to recognize LCL cells. LCL were identified as CD19-positive cells, and their viability was assessed as the percentage of dead LCL CD19+ cells. To assess the spontaneous dead of LCL cells, we cultured LCL cells without CAR T cells as a control. An average of 104cells were acquired with the FACSCanto I (BD Biosciences) using BD FACSDiva Software version 8.0.3, and data were analyzed using FlowJo version 10.6.1 software.

[0122] RNA isolation and RT-qPCR

[0123] Total RNA was extracted using the RNeasy Mini kit and the QIAshredder (Qiagen) system following manufacturer’s instruction. DNAse treatment was peformed with the RNase-free DNase Set (Qiagen). The SuperScript III First-Strand Synthesis SuperMix kit (Invitrogen) was used to reverse transcribe total RNA in accordance with the manufacturer's recommendations. Power SYBR Green PCR Master Mix was used to perform real-time quantitative PCR on the StepOnePlus Real-Time PCR System. Gene expression data were normalized on GAPDH as independent housekeeping gene. Normalized Ct values were calculated as 2-ACtor 2-AACt.

[0124] Results

[0125] Experiments comparing CAR-T cells and conventional T cells under overstimulation conditions revealed a fundamental difference in exhaustion response. When conventional T cells were exposed to chronic over stimulation mediated by anti-CD3 / CD28 beads, they exhibited no reduction in viability and a clear induction of exhaustion markers, including PD-1 , along with decreased IFN-y production (Fig. 1 ). CAR-T cells do not express LINE1 , in a manner that is comparable to effector T cells (Fig. 2). However, CAR-T cells subjected to the same overstimulation protocol mentioned before exhibit a marked reduction in viability (Fig. 2). CAR-T cells can be overstimulated by continuous exposure to the CD19 antigen expressed by LCL cell lines while remaining viable. However, in this condition, over-stimulated CAR-T cells expressed LINE-1 but did not display canonical features of T-cell exhaustion. Specifically, CAR-T, did not express PD-1 , preserved IFN-y expression and exhibited only a slight reduction in cytotoxic activity against CD19-positive targets (Fig. 4). Since LINE1 expression is controlled by mTORCI , it has been also investigated whether this mechanism is maintained in CAR-T cells. CAR-T cells have been treated with rapamycin (Figure 5A), an inhibitor of mTORCI activity, and found that LINE1 expression increases (Figure 5B-D), while their killing abilities and effector cytokine production (i.e., IFN-y, TNF-a, GrzB) decrease (Figure 5-F). To directly assess the role of LINE1 in regulating CAR-T cell function, mTORCI -inhibited CAR-T cells have been treated with LINE1 or control ASO (Figure 6A). Targeting LINE1 expression (Figure 6B) enhances the ability of CAR T cells to kill LCL CD19+ cells (Figure 6C).

[0126] These results demonstrate that CAR-T cells do not face comparable functional impairment when exhaustion pathways are triggered by protocols that drive conventional T cells into a dysfunctional state. Furthermore, although CAR-T cells showed increased LINE1 expression upon antigen-driven overstimulation, these data indicate that the exhaustion phenotype observed in conventional T cells was not induced in CAR-T.

[0127] Sequences

[0128] SEQ ID NO:1 (L1 ME4A L1 Homo sapiens)

[0129] CTTGTATCCAGAATATATAAAGAACGCCTACAACTCAACAATAAAAAAACGAATTTCC CAACAAAAAAACGGACAAAGGACACGAANAGACCGTTTACAAAAGAAGAAATGGAAA TAACTANCGAACATGAAAAATGTTCAACCTCACTAATAATCAAAGAAATGCAAATTAA AACAACAATGAGATNCCGTTCTTCNTCGTCTANCAAACTGGCANAGATATAAAAAGAT AATAKCCAGTGTTGGTGAGGATGTGGAGAAACGGGCACTCTCATACACTGCTGGTG GGAGTATAAATTGGTACAACCTTTCTGGAAGGCAATTTGGCAATATNTATCAAAAGCC TTAAAAATGTTCATACCCTTTGACCCAGCAATTCCACTTCTAGGAATCTATCCTAAGG AAATAATCAGAAATGTGNACAAAGATTTACGTACAAAGATGTTCACCGCAGTATTATT TATAATAGCAAAAAATTGGAAACAACCTAAATGTCCAATAATAGGGGANTGGTTAAAT AAATTATGGTACATCCATACAATGGAATATTATGCAGCCATTAAAAATNATGTTTTCGA AGAATATTTAATGACATGGGAAAATGCTCATGATATAATGTTAAGTGAAAAAAGCAGG NTACAAAACTGTATATACAGTATGATCTCAACTTTGTTATAAAATTACATATATAAATG TATACGTATNTACATAGAAAAAAGACTGGAAGGAAATACACCAAAATGTTAACAGTGG TTATCTCTGGGTGGTGGGATTATGGGTGATTTTTATTTTCTTTTTTCTTTGTATTTTCT GTATTTTCCAAATTTTCTACAATGAACATGTATTACTTTTATAATCAGAAAAAAAA

[0130] SEQ ID NO:2 (L1 M4B L1 Homo sapiens)

[0131] AAGGAGTTTCACTTCTGGAATGGCAGCATGAGGAGCTCCGNAGACCCNCTCCCCAG CGAAACAANCATAACTGGTGAAAATTATTTTTAAAAAAACAACCATTTAAAGTCTCTG GAAATTGTCCTAAGGGCATACAGCAAATGAAGAAACATTTATTCAAGAAAATCTACTA AATCTCAGTAAGAACAGTGAGAGTCTGTGGCACTTGAGCCACGACCCGCTCCCACC CTCCCCCCTCCCCAGCTCAGCNTGACAGAAGCTCCACTCCGGGCGGGTGCGGCCA AGAAGACGGGGCTCCCTCTCCCCTCAGCTCCCAGTCAAGGGNTACGGTATCTCNCC GGGAGGGGCAGGCCGCCAGCATTTCTCATCCCCTCCAGCTCCGNGTTGCAGAGGC TAAATTCCAGGTGAGTGTAGCTGAGAGGTCGGGGGCTCCCTTCCTCCACCCAGCCC CCACTCATAGGGCGGAGGCTCTACCCCAGGCGCGGCAGGCCGAGAATACTGGGGC CCTGATTGCCCTCACCCCAGCTCGCTCATAGGGCGGAGGTTCCACGCCGGGAGAG GCAAGCCGAGAAGACCAGAGGCTACCGCCCCCGCCCAGCGCCCTGCTCATAAAGC AGGGGTGTCACTCCGAGAGAAGCGGGCCACTGTCCCCGCCCCCAGCTCCGGAGCA GTGGCTCAGAGATTTTGCCCAGGGGGAGAGGCAGNCCATAAGAACAGAGAGCTCC GAAGCTCTCCCCAAAGGAACTGACTTTATTTGAAACAGAGTGTGGGGAAGTTCAAGC CTAAGGGTACTCTCGAAAACAATGGAGATTTTGGTGGTAAGCAATTAAGAGGAGGCT GGTAGCTCCATGAGAGCAACAAGCTAAACCATAGGCCAGCTAGTTTACCAGAGAGA ACCAGGGAAAGAGACAGCTAAGAAGAGCCCTCCTGGGGTCAGAACAAACCTCAAAG ACTGGCCTCAAAAACTACCCCTRCAAAGGGGCCCGAATTTAATTGGATCAGACTGTG GAG C AATTTATGCCCC AGG GC ATTGTCG AAAAC AATAG AG C AATC AG CCG G CAATTA GTGGAGCCTAACAGCTGGGTGTGATACCAANNGAGGCAGACAGCTTAACAGAGAGA TCAGGGAAAGAGACAGTCAAAGAGAGCCCTGCTAAAACCACTGTCATCCCAGGGTG ACTGTGCGCATGCCCAAGGCTGCGCCCTCTGAGGAGCGACATCAGAGGCTTCACAC TGNGGGGGAAATAGACTTCACTAAAATAGTCCAGCCAAGTCACTAAACAAATAAACA AGCAAAAACAANCACNANGAGCCGGGGGNGGGGAATCAGTATCCAGAGTTGCTACA ATATATTACCTAAAATGTCCAGTTTTCAACAAAAAATTATGAGACATGCAAAGAAACA GGAAAGTGTGACCCATACACAGGAAAAAAAGCAGGCAACAGAAACTGCCTGTGAGA GGGCCCAGATGTCGGATTTAGCAGACAAAGACTTCAAAGCAGCCATTATAAATATGT TCAAAGAACTAAAGGAAACCATGCTTAAAGAAGTAAAGGAAGGTATGATGACAATGT CTCATCAAATAGAGANTATCAATAAAGAGATAGAAATTATAANAAAAAACCAAATGGA AATTCTGGAGTTGAAAAGTACAATAACTGAAATGAAAAATTCACTAGAGGGGCTCAA CAGTAGATTTGANCTGGCAGAAGAAAAGAATCAGTRAACTTGAAGATAGATCAATAG AGATTATGCAATCTGAAGAACAGAAAGAAAAAAAAGAATGAAGAAAAATGAACAGAG CCTCAGAGAAATGTGGGACACCATYAAGCATACCAACATATACATACATGGACAGAC AAACAACATATACATAATGGGAGTACCAGAAGGAGAGGAGAAGAGAGAGAAAGGAG CAGAAAAAATATTTGAAGAAATAATGGCTAAAAACTTCCCAAATTTGATGAAAAACATT AATATTAATCTACACATCCAAGAAGCTCAATAAACTCCAAGTAGGATAAACTCAAAGA GATCCACACCTAGACACATCATAGTCAAAATGTTGAAAGACAAAGACAAAGAGAAAA TCTTGAAAGCAGCAAGAGAAAAATGACTCATCACATACAAGGGAANNNACCTCAATA AGATTAACAGCTGACTTCTCATCAGAAACAATGGAGGCCAGAAGGCAGTGGGATGA CATATTCAAAGTGCTGAAAGAAAAAAAAAAAAAAAAAAAAACAAAAAAAHACANAAAC AAATACAACNYTACCTGTCAACCAAGAATTCTATATCCAGCAAAACTATCTTTCAAAA

[0132] ATGAAGGTGAAATAAAGACATTCCCAGATAAACAAAAACTGAGAGAATTTGTTGCTAG CAGACCTACCTTACAAGAAATACTAAAGGAAGAGTTCTTCAGGCTGAAAGGCAAGTG ACACCAGATAGTAATTCAAATCCACATAAAAAAATAAAGAGACACACACTAAGTAAAG NNCACTAGTAAAGGTAATTATGTAGNAAGACAGTAANTTAATTATNAAAGRCATGTAK GTAATTATAAAAGACAGTATAAATGCATATTTCTTCTTTCTTCTCTTAACTGATTTAAAA AGCAATTGTATAAAACAATATGTATATAATTGTATTGTTGGGCCTATAACATATAGAAA

[0133] TGTAATATATTTGACAATAACAGCACAAAGGAGGTGGGTGGGAGCAAAGCTGTATTG

[0134] GAGTAAGGAAATGACACCAGATGGTAACTTGAATCCACAGGAACAAATGAAGAGAAC

[0135] CAGAAATGGTAAATAAGAAGGTTAATATAACAAACTCTATAAATATATACTTGTTCTCC

[0136] TTTCTTCTCTTCTTTAAAAGACATAAAATTATATAAAGTAATAATTATAACAAATGTATT

[0137] TNTNNNATAATAATGTTGGGTTTGTAACATATATAGATGTATATATATTNNTATTGTAA

[0138] TATGTATAACAATAATAGCACAAAAAAGGAGAAAAAGGAATAGAGCTATATAGGAGTA

[0139] ACATTTCTATATCTCACTGGAATTAAGTTAGTATAAATCTGAAGTAGATTCTGATAANG

[0140] TTAAGATGTATATGGTAAGCCCTAGAGCAACCACTAAGAAAATAACTTAAAAAAATAT

[0141] AGTAAAAAAAATCATTAAAGAAATTAAAATGTTACACTAGAAAATATTCACTTAATGCA

[0142] AAAGAAAGCAGTAAAGGAGGAATAGAGGAACAAAAAAGACATGAGACATATNACATA

[0143] TAGAAAACAAAAAGTAAAATGGCAGATATAAATCCAACTATATCAATATAACATTAAAT

[0144] GTGATTATGGATTAARYAAAATGGCARAAGCTGTCAGNCTNGAGATTTANTNTATATA

[0145] AATCCAANTNNNTNGTTNANATGNTNAGACNGNTAATNCAAATATCAATAATAACATT

[0146] AAATGTGAATGGATTAAACAATCCAATCAAAAGGCAGAGATTGTCAGACTGGATAAA

[0147] AAAAAAAAAACAAGATCCAACTATATGCTGTCTACAGGAGACACACTTTAGATTCAAA

[0148] GATACAAATAGRTTGAAAGTAAAAGGATGGAAAAAGATATATCATGCAAACAGCAAC

[0149] CATAAGAAAGCTGGAGTGGCTATACTAATATCAGACAAAATAGACTTTAAAACAAAAA

[0150] ATGTTACTAGAGATAAAGAGGGACATTTTATTATATAATGATAAAAGGGTCAAAAGGG

[0151] TCAATCCATCAGGAAGATATAACAATTATAAACATATATGCATATANATATATGCACCT

[0152] AACAACAGAGCCCCCAAAATACATGAAGCAAAAACTGACAGAAATGAAGGGAGAAAT

[0153] AGACAATTCAACAATAATAGTTGGAGACTTCAATAYCCCACTTTCAATAATGGATAGA

[0154] ACAACTAGGCAGAAGNNAATANGATCAACAAGGAAATAGAAGACTTGAACAACACTA

[0155] TAAACCAACTAGACCTAACAGACATCTATAGAACATTTATAGAACACTCYATCCAACA

[0156] ACAGCAGAATATACATTCTTCTCAAGTGCACATGGAACATTCTCCAGGATAGACCATA

[0157] TGCTAGGCCATAAAACAAGYCTCAATAAATTTATTTAAAGGATTGAAATAATACAAAG

[0158] TATGTTCTCTGACCACAATGGAATGAAATTAGAAATCAATAACAAAAAATTTGGGAAA

[0159] TTTACAAATATGTGGAAATTAAACAACACACTCCTAAATAACCAATGGGTCAAAGAAG

[0160] AAATCACAAGAGAAATTAGAAAATACTTTGAGATGAATGAAAATGAAGACACAACATA

[0161] CCAAAATTTATGGGATGCAGCTAAAGCAGTGYTTAGAGGAAAATTTATAGCTGTAAAT

[0162] GCCTATATTAAAAAAGAAGAAAGATCTCAAATCAATAACCTAACCTTCTACCTTAAGA

[0163] CACTAAAAAAAGAAGAGCAAACTAAACCTAAAGCAAGCAGAAGGAAGGAAATAATAA

[0164] AGATTAGAGCAGAAATTAATGAAATAGAAGAAAAACAATAGAGAAAATCAATGAAACC

[0165] AAAAGCTGGTTCTTTGAAAAGATCAACAAAATTGACAAACCTTTAGCTAGACTGACCA

[0166] AGAAAAAGAGAAGACTCAAATTACTAAAATCAGAAATGAAAGAGGGAACATTACTACT AACCTTACAGAAATAAAAAGGATTATAAAGGAATACTATGAACAATTGTATGCCAATA AATTNAGATAACTTAGATGAAATGGACAAATTCCTAGAAANYAAGACACACAAACTAC YAAAACTGACTCAAGAAGAAATAGANAATCTGAATAGACCTATAAAANTNAAGAGATT GAATTAGTAATNTAAAAACTNCCYACAAAAAAAGCCCAGNCCCAGATGGCTTCACTG GTGAATTCTCCAAANATTTAAAANAGAATTAATACCAATTATTCACCTNTTCCAAAAAA

[0167] TAGAAGAGGAGGNAAYACTNCCNAACTNATTCTATGAGGCCAGTATTATCCTGATAC CAAAACCAGNCAAAGACATNACAAAAGAAAAGAAAA

[0168] TAAAACAAGYCTCAATAAATTTATTTAAAGGATTGAAATAATACAAAGTATGTTCTCTG

[0169] ACCACAATGGAATGAAATTAGAAATCAATAACAAAAAATTTGGGAAATTTACAAATAT GTGGAAATTAAACAACACACTCCTAAATAACCAATGGGTCAAAGAAGAAATCACAAG

[0170] AGAAATTAGAAAATACTTTGAGATGAATGAAAATGAAGACACAACATACCAAAATTTA TGGGATGCAGCTAAAGCAGTGYTTAGAGGAAAATTTATAGCTGTAAATGCCTATATTA AAAAAGAAGAAAGATCTCAAATCAATAACCTAACCTTCTACCTTAAGACACTAAAAAA AGAAGAGCAAACTAAACCTAAAGCAAGCAGAAGGAAGGAAATAATAAAGATTAGAGC AGAAATTAATGAAATAGAAGAAAAACAATAGAGAAAATCAATGAAACCAAAAGCTGGT TCTTTGAAAAGATCAACAAAATTGACAAACCTTTAGCTAGACTGACCAAGAAAAAGAG AAGACTCAAATTACTAAAATCAGAAATGAAAGAGGGAACATTACTACTAACCTTACAG AAATAAAAAGGATTATAAAGGAATACTATGAACAATTGTATGCCAATAAATTNAGATAA

[0171] CTTAGATGAAATGGACAAATTCCTAGAAANYAAGACACACAAACTACYAAAACTGACT CAAGAAGAAATAGANAATCTGAATAGACCTATAAAANTNAAGAGATTGAATTAGTAAT NTAAAAACTNCCYACAAAAAAAGCCCAGNCCCAGATGGCTTCACTGGTGAATTCTCC AAANATTTAAAANAGAATTAATACCAATTATTCACCTNTTCCAAAAAATAGAAGAGGA GGNAAYACTNCCNAACTNATTCTATGAGGCCAGTATTATCCTGATACCAAAACCAGN CAAAGACATNACAAAAGAAAAGAAAA

[0172] TTAAAGGATTGAAATAATACAAAGTATGTTCTCTGACCACAATGGAATGAAATTAGAA ATCAATAACAAAAAATTTGGGAAATTTACAAATATGTGGAAATTAAACAACACACTCCT

[0173] AAATAACCAATGGGTCAAAGAAGAAATCACAAGAGAAATTAGAAAATACTTTGAGATG AATGAAAATGAAGACACAACATACCAAAATTTATGGGATGCAGCTAAAGCAGTGYTTA G AG G AAAATTTATAG CTGTAAATG CCTATATTAAAAAAG AAG AAAG ATCTC AAATC AA TAACCTAACCTTCTACCTTAAGACACTAAAAAAAGAAGAGCAAACTAAACCTAAAGCA AGCAGAAGGAAGGAAATAATAAAGATTAGAGCAGAAATTAATGAAATAGAAGAAAAA CAATAGAGAAAATCAATGAAACCAAAAGCTGGTTCTTTGAAAAGATCAACAAAATTGA CAAACCTTTAGCTAGACTGACCAAGAAAAAGAGAAGACTCAAATTACTAAAATCAGAA ATGAAAGAGGGAACATTACTACTAACCTTACAGAAATAAAAAGGATTATAAAGGAATA

[0174] CTATGAACAATTGTATGCCAATAAATTNAGATAACTTAGATGAAATGGACAAATTCCT AGAAANYAAGACACACAAACTACYAAAACTGACTCAAGAAGAAATAGANAATCTGAAT

[0175] AGACCTATAAAANTNAAGAGATTGAATTAGTAATNTAAAAACTNCCYACAAAAAAAGC

[0176] CCAGNCCCAGATGGCTTCACTGGTGAATTCTCCAAANATTTAAAANAGAATTAATACC

[0177] AATTATTCACCTNTTCCAAAAAATAGAAGAGGAGGNAAYACTNCCNAACTNATTCTAT

[0178] GAGGCCAGTATTATCCTGATACCAAAACCAGNCAAAGACATNACAAAAGAAAAGAAA A

[0179] GAAAAAGAGAAGACTCAAATTACTAAAATCAGAAATGAAAGAGGGAACATTACTACTA

[0180] ACCTTACAGAAATAAAAAGGATTATAAAGGAATACTATGAACAATTGTATGCCAATAA

[0181] ATTNAGATAACTTAGATGAAATGGACAAATTCCTAGAAANYAAGACACACAAACTACY

[0182] AAAACTGACTCAAGAAGAAATAGANAATCTGAATAGACCTATAAAANTNAAGAGATTG

[0183] AATTAGTAATNTAAAAACTNCCYACAAAAAAAGCCCAGNCCCAGATGGCTTCACTGG

[0184] TGAATTCTCCAAANATTTAAAANAGAATTAATACCAATTATTCACCTNTTCCAAAAAAT

[0185] AGAAGAGGAGGNAAYACTNCCNAACTNATTCTATGAGGCCAGTATTATCCTGATACC

[0186] AAAACCAGNCAAAGACATNACAAAAGAAAAGAAAA

[0187] AAAATCAGAAATGAAAGAGGGAACATTACTACTAACCTTACAGAAATAAAAAGGATTA

[0188] TAAAGGAATACTATGAACAATTGTATGCCAATAAATTNAGATAACTTAGATGAAATGG

[0189] ACAAATTCCTAGAAANYAAGACACACAAACTACYAAAACTGACTCAAGAAGAAATAGA

[0190] NAATCTGAATAGACCTATAAAANTNAAGAGATTGAATTAGTAATNTAAAAACTNCCYA

[0191] CAAAAAAAGCCCAGNCCCAGATGGCTTCACTGGTGAATTCTCCAAANATTTAAAANA

[0192] GAATTAATACCAATTATTCACCTNTTCCAAAAAATAGAAGAGGAGGNAAYACTNCCNA

[0193] ACTNATTCTATGAGGCCAGTATTATCCTGATACCAAAACCAGNCAAAGACATNACAAA

[0194] AGAAAAGAAAA

[0195] SEQ ID NO.3 (L1 MC4 L1 Homo sapiens)

[0196] CTAATATACCTAATATACAAAAAACTCTTAAAATTGAAGGATAAAAAGNCAAAAACCN

[0197] AATANNAAAATGGGNAAAAGACATGAACAGACAATTCACNAAAAATNATAAAATGGC

[0198] CCTTAAGCATATAAAAAGATGTTCANCCTCACNTATAATTAGAGAAACGCAAATTAAA

[0199] ACTACACCGAGATACCATTTCTCACCCANCAGATCGGCAAAAATTAAAAAGTATGGC

[0200] AATATANNCTGTTGGCGAGGCTGTGGGGNAACNGGNACTCTCATACACTGCTGGTG

[0201] GGAGTGCAAATTGGTACAACTNCTTTGGAAGANAATTTGGCAGTNTCTAATAAAACTA

[0202] CACNTGCNTTTACACTTTGACCCATTAGTCCCACTTCTAGAAATTTACCCTANAGAAA

[0203] TACTTCTAACAGNTCAAAAATACACATGTACAGGGATGTTCATAGCAGTNTTATTNNT

[0204] AATNGTAAAANATTGGAAACAATCNAAATGTCCATCAGCAGGAGAATGGNTGAATAA

[0205] ACTATGGTNCATCCACACAATGGAATACTATNCAGCTGTAAAAAAGAATGAGGAAGA

[0206] TCTCTGTAATAATGTGGAGNGATTTCGGAACATNNTNTTNAGTTGAAAAAGCNANGC

[0207] GCAAAAGAGTATATATANTATGCTACCCTTCATATAAGAAAGAAGGGGATATGAGAAA ATATACATATATCTGCTCATTTGTGCAAAAAGAAACACAGAAAAGATAAANCAGANAC

[0208] TAATGAGATTGGTTACCCACAGGGAANNGGTGGGAATGGGGAGGAAAGGACGGAA

[0209] GGAATGGGGGGCAGTGACACTTTTCTGAGTATACCTTTTTGTATAGTTCTAACTTTTG

[0210] NAACCATGTTAATGTTTCACATACTCAAGAAATGAATAANTAAAATCAACAAGGATGG

[0211] GGGANAACTCAAAATGAAATACAAACAGAAACAAATGAACCWAACTGTATTTCAAAT

[0212] GAATAACATAACCACACTGAAGGGGGTNAGGAAGAAAAGAACTAACCCAAGTAACTT

[0213] TTGAACACAGTATTTTGACTATATGCCCTCAGGCTAAAGACAAAAAGAACTNTAAACA

[0214] AATATTGAACTCTAGTTAGTAGGCTTATTTTCCGCAGNGGCATGGGTTAGCAATTCTG

[0215] AAACTACTTTCTGTATATTCTAGGACTGAGCAAATAAGTAAATATATTGNGGATAATG

[0216] GGAGCCAGGTTTCTCACTGTCGGAGAAGGGAGTTACAAATATGGAAAGGGGGAAGA

[0217] CTAGAATGAACCCTGTGGTGTTGGATTGGAATTGGAGGTATCAGTGTGAACTCATGG

[0218] TTTTTAATATANATAGATATACAGACAGACAGATATAGAAATAGATATAGATATATATG

[0219] TGTNTGTGTATATGTGTATGTATATACGTACATATATTTCCTAGCTCTGTCCACTGAG

[0220] AGGGCCTAGAAGCAATGACACCCCAGTAGCAATGAGCACACCTAGCGCCCAGATCT

[0221] TGGTTTCTAAATACCATTCTCCACTAAAAGGAACCAGGGCTCCTTGGAGAAATGGCT

[0222] GATTCCAGGGCTGGGGCAGGGAAAGTACAAGATGAGCCTGGAACATCTTGTTGTGC

[0223] CAGAAAGTAAGGAAGTGCTCAAAGAATGATGGGGACATGTCAAAAGGACACAGGAG

[0224] CCAGCTTGAAGGGGCTCCCACTGGCCAAATCTGGGACAATTTGAGCATCAAAATAAA

[0225] TAATGATAGTAATGGATTATAACCCATTGAATAAAATAAGAATCCATGAGTCCATACT

[0226] GATATAAATAAATAAATAAATAAATGGGGGAGAAGGGAAAGCTCTTCCTTACAGTAGA

[0227] ATGCCAACTAATAAATGTAGAAGGAATGATGGAATTAGAAAATCACCATTTGGCAACC

[0228] ATCATAGTAATAATTGATTCAGGCAAGAATCATCAATGGATGCTAAAACTAGTGGGTG

[0229] AAAGTTTGATGAGNAACAGGATATTTACATAGTCTCAAAGTATCTCCCCACAAAATAC

[0230] TTATTAATTACAAAGGGGAAAATAGTAACTTTACAGTGGAGAAACCTGGCAGACACC

[0231] ACCTTAACCAAGTGATCAAAGTTAACATCACCAGTAATGGGACAAATCGACATCATGT

[0232] GCCTCCTGATATGATGCACTGAGAAGGACACAACATCACTTCTGTGGTATTCCTGCC

[0233] AAAAATGCATAACCTGAATCTAATCATGAGGAAACATCAGACAAACCCAAATTGAGG

[0234] GACATTCTACAAAATAACTGGCCTGTACTCTTCAAAAATGTCAAGGTCATGAAAGACA

[0235] AAGAAAGACTGAGGAACTGTTCCAGATTAAAGGAGACTAAAGAGACATGACAACTAA

[0236] ATGCAACGCGTGATCCTGGATTGGATCCTGGACCAGANTTTTTTTTGCTATAAAGGA

[0237] CATTATTGGGACAACTGGCGAAATTTGAATAAGGTCTGTAGATTAGATAATAGTATTG

[0238] TATCAATGTTAATTTCCTGATTTTGATNATTGTACTGTGGTTATGTAAGAGAA

[0239] TCCACTAAAAGGAACCAGGGCTCCTTGGAGAAATGGCTGATTCCAGGGCTGGGGCA

[0240] GGGAAAGTACAAGATGAGCCTGGAACATCTTGTTGTGCCAGAAAGTAAGGAAGTGC

[0241] TCAAAGAATGATGGGGACATGTCAAAAGGACACAGGAGCCAGCTTGAAGGGGCTCC CACTGGCCAAATCTGGGACAATTTGAGCATCAAAATAAATAATGATAGTAATGGATTA TAACCCATTGAATAAAATAAGAATCCATGAGTCCATACTGATATAAATAAATAAATAAA TAAATGGGGGAGAAGGGAAAGCTCTTCCTTACAGTAGAATGCCAACTAATAAATGTA GAAGGAATGATGGAATTAGAAAATCACCATTTGGCAACCATCATAGTAATAATTGATT CAGGCAAGAATCATCAATGGATGCTAAAACTAGTGGGTGAAAGTTTGATGAGNAACA GGATATTTACATAGTCTCAAAGTATCTCCCCACAAAATACTTATTAATTACAAAGGGG AAAATAGTAACTTTACAGTGGAGAAACCTGGCAGACACCACCTTAACCAAGTGATCA AAGTTAACATCACCAGTAATGGGACAAATCGACATCATGTGCCTCCTGATATGATGC ACTGAGAAGGACACAACATCACTTCTGTGGTATTCCTGCCAAAAATGCATAACCTGA ATCTAATCATGAGGAAACATCAGACAAACCCAAATTGAGGGACATTCTACAAAATAAC TGGCCTGTACTCTTCAAAAATGTCAAGGTCATGAAAGACAAAGAAAGACTGAGGAAC TGTTCCAGATTAAAGGAGACTAAAGAGACATGACAACTAAATGCAACGCGTGATCCT GGATTGGATCCTGGACCAGANTTTTTTTTGCTATAAAGGACATTATTGGGACAACTG GCGAAATTTGAATAAGGTCTGTAGATTAGATAATAGTATTGTATCAATGTTAATTTCCT GATTTTGATNATTGTACTGTGGTTATGTAAGAGAA

[0242] TGGAGAAATGGCTGATTCCAGGGCTGGGGCAGGGAAAGTACAAGATGAGCCTGGA ACATCTTGTTGTGCCAGAAAGTAAGGAAGTGCTCAAAGAATGATGGGGACATGTCAA AAGGACACAGGAGCCAGCTTGAAGGGGCTCCCACTGGCCAAATCTGGGACAATTTG AGCATCAAAATAAATAATGATAGTAATGGATTATAACCCATTGAATAAAATAAGAATCC ATGAGTCCATACTGATATAAATAAATAAATAAATAAATGGGGGAGAAGGGAAAGCTCT TCCTTAC AGTAG AATG CC AACTAATAAATGTAG AAG G AATG ATG G AATTAG AAAATC A CC ATTTG GC AACC ATC ATAGTAATAATTG ATTC AG G C AAG AATCATC AATG G ATG CTA AAACTAGTGGGTGAAAGTTTGATGAGNAACAGGATATTTACATAGTCTCAAAGTATCT CCCCACAAAATACTTATTAATTACAAAGGGGAAAATAGTAACTTTACAGTGGAGAAAC CTGGCAGACACCACCTTAACCAAGTGATCAAAGTTAACATCACCAGTAATGGGACAA ATCGACATCATGTGCCTCCTGATATGATGCACTGAGAAGGACACAACATCACTTCTG TGGTATTCCTGCCAAAAATGCATAACCTGAATCTAATCATGAGGAAACATCAGACAAA CCCAAATTGAGGGACATTCTACAAAATAACTGGCCTGTACTCTTCAAAAATGTCAAG GTCATGAAAGACAAAGAAAGACTGAGGAACTGTTCCAGATTAAAGGAGACTAAAGAG ACATGACAACTAAATGCAACGCGTGATCCTGGATTGGATCCTGGACCAGANTTTTTT TTGCTATAAAGGACATTATTGGGACAACTGGCGAAATTTGAATAAGGTCTGTAGATTA GATAATAGTATTGTATCAATGTTAATTTCCTGATTTTGATNATTGTACTGTGGTTATGT AAGAGAA

[0243] GGGAAAATAGTAACTTTACAGTGGAGAAACCTGGCAGACACCACCTTAACCAAGTGA TCAAAGTTAACATCACCAGTAATGGGACAAATCGACATCATGTGCCTCCTGATATGAT GCACTGAGAAGGACACAACATCACTTCTGTGGTATTCCTGCCAAAAATGCATAACCT

[0244] GAATCTAATCATGAGGAAACATCAGACAAACCCAAATTGAGGGACATTCTACAAAATA

[0245] ACTGGCCTGTACTCTTCAAAAATGTCAAGGTCATGAAAGACAAAGAAAGACTGAGGA

[0246] ACTGTTCCAGATTAAAGGAGACTAAAGAGACATGACAACTAAATGCAACGCGTGATC

[0247] CTGGATTGGATCCTGGACCAGANTTTTTTTTGCTATAAAGGACATTATTGGGACAACT

[0248] GGCGAAATTTGAATAAGGTCTGTAGATTAGATAATAGTATTGTATCAATGTTAATTTC

[0249] CTGATTTTGATNATTGTACTGTGGTTATGTAAGAGAA

[0250] GAAACCTGGCAGACACCACCTTAACCAAGTGATCAAAGTTAACATCACCAGTAATGG

[0251] GACAAATCGACATCATGTGCCTCCTGATATGATGCACTGAGAAGGACACAACATCAC

[0252] TTCTGTGGTATTCCTGCCAAAAATGCATAACCTGAATCTAATCATGAGGAAACATCAG

[0253] ACAAACCCAAATTGAGGGACATTCTACAAAATAACTGGCCTGTACTCTTCAAAAATGT

[0254] CAAGGTCATGAAAGACAAAGAAAGACTGAGGAACTGTTCCAGATTAAAGGAGACTAA

[0255] AGAGACATGACAACTAAATGCAACGCGTGATCCTGGATTGGATCCTGGACCAGANTT

[0256] TTTTTTGCTATAAAGGACATTATTGGGACAACTGGCGAAATTTGAATAAGGTCTGTAG

[0257] ATTAGATAATAGTATTGTATCAATGTTAATTTCCTGATTTTGATNATTGTACTGTGGTT

[0258] ATGTAAGAGAA

[0259] TGTCCTTGTTTTTAGGAAATACACACTGAAGTATTTAGGGGTAANGGGGCATCATGT

[0260] CTGCAACTTACTCTCAAATGGTTCAGAAAAAAAAATATGTATATGNANACAGAGAATG

[0261] ATAAAGCAAATGTGGCAAAATGTTAACATTTGGGGAATCTGGGTGAAGGGTATACGG

[0262] GAATTCTTTGTACTATTCTTGCAACTTTTCTGTAAGTCTGAAATTATTTCAAAATAAAA

[0263] AGTTAAAAAA

[0264] GGTAANGGGGCATCATGTCTGCAACTTACTCTCAAATGGTTCAGAAAAAAAAATATG

[0265] TATATGNANACAGAGAATGATAAAGCAAATGTGGCAAAATGTTAACATTTGGGGAAT

[0266] CTGGGTGAAGGGTATACGGGAATTCTTTGTACTATTCTTGCAACTTTTCTGTAAGTCT

[0267] GAAATTATTTCAAAATAAAAAGTTAAAAAA

[0268] SEQ ID NO: 4

[0269] GCACTAAATGCCTACAAGAGA

[0270] SEQ ID NO: 5

[0271] GATAGACCGCTAGCAAGACTA

[0272] SEQ ID NO: 6

[0273] GAAGTTGAATCTCTGAATAGA

[0274] SEQ ID NO: 7

[0275] GGACCTCTTCAAGGAGAACTA

[0276] SEQ ID NO: 8

[0277] GGAGAGGATGCGGAGAAATAG SEQ ID NO: 9

[0278] TGCAGTGGTGGCATAAAGAGGCCACACACACACNCCACAGCAAAATANACNNAANT

[0279] ANTAATAATAAGATTATAAAANACAAAGNNAANACAAAAGTAAAATGTATGGNTTNAT

[0280] TNCAACAACAATAGAAAANGAAAAANNATTTANTAAATAAACNCTGCAAACATNTGNN

[0281] AAGAAAAATANATCTTATTACATATAAANTTAAATCANTATAAACAATATCCTACAGCC

[0282] AAAGTGAGNNTGTATCTAATCAATACTACCACTTTTTTANTANTTCCTAAAAAAAAACA

[0283] AGAATCNCATACAAACATNANAAAGGTTAGAAAAAAAATANATAACNGAAGNATGAAT

[0284] ANTTAAAAGCTTNCCTTTTCCTACATATTAATGNGTACACCAANCTAAACTAAATTTTT

[0285] CAATTTATTTTATTTTAGATNGCAATATAAGTTTGTTGAAACAAAGGGAGAAAANGTTT

[0286] AANTGNAGGTTTTAACCTTNTACATAAATTTAATTCCAAGAGACTANNGANANTTGCT

[0287] TAATTTATTTAGCNGNAACAATNAGAATATTATANAACAACTATACCATGANATATGAA

[0288] AAATAAAAATGTAATTAACTTNTTTTTATTAATTATANTTAAATTATTATATTATAAATGA

[0289] AAAAAATAAACTAATTTTACAATNAATTTTTTTATTTAAGTAATAAAAAAAAAAATATTG

[0290] AAAG G AAAATTAATNTTACTAATATAAAAATAACTATGTTC AAAAAAAATG ATATTTAAT

[0291] TAAAATGTTAATTTCCAATAAAANAATTTTTGTAAATACTGATTTCATTTTCTTTANNTA

[0292] TTGGGTATTTAAAAGAGTATAATNANAAATGATTNATAAATGAAAAAAATAATTTCAAT

[0293] GTAATGTTCATTTATGCNTTATNAGAATTAAAGTAGATTATTAATAAATTAACTTTATAT

[0294] AANGTTAAAAAAGAATAAATTTTTTATATTNTATNANGANCAATATTTTTAAGATATGTA

[0295] AATAAAAAAATTTTTTTACAAGCAATAACNTATGAAAANAAAATTTTATTAAAATTTTTT

[0296] ATGATTTTTTAAAATTTAATATNTTTTATANATAACTACAATTATTNTTTTATTTATTTNT

[0297] ATAANTATTTTAAATTTAATATCATTTTAAAAATATTATTAATAAGCTNCTTATAAAAATT

[0298] TACTCTAATTATGATTATATTTATACCTTATCATAAATAAAATAAAAAANTTTTAAGTTA

[0299] TANTATCTTATTTAAACAAATNCTTCTTAAGTTTTTTTACTATTTATNACCTATTNGGAN

[0300] ANTATTTAATTTTGTTTNTTTAAACTTAGATNTAGATTNATGAATTTTGAAAATATTTTA

[0301] TTTATTGTTAAATTTAATAAAAAGAANAAAAAAATTACATACTGTATTATTCAATTTATA

[0302] TGTCTATTTTTTAAAAATAANAAAAAAAATCANATCCATGGAGNTCATAATATGCATAA

[0303] TAANNANATAGTTACTTATGTTTTTTATACTGAATAAATTTGTTTGCGAAGGTNATTTA

[0304] ACATAAATATGTNTTGAGATTGGGAAANTGTATATTAAAAAGAATAGNAAGGNAAAAA

[0305] ATATTTGCAAATTTTTGTGANTGATTGAAATTTTTTTATATANATTATATAATTGANNGT

[0306] GTTGGTTGTATAANTTTTTATNAAANTTATATTAAATNTGTTTAAAAAGTCATTAAACTN

[0307] TACAATTTTTAAATAGTAAAAATGNTTAAATTTATTAAAAATTGTTTTTTATATAAATTTT

[0308] ATCTAAGAANAAAANATAATTTNTNAATAAACATATGAAAATTGTCGAGTTNTTCAACA

[0309] TCATTAGNTAATTAAATGAAAATGTAATTTAAAACNATATTGAGANATNTTGTCNCTTT

[0310] TAAATAATAAGAATGGCTAAAATTTAAATTATAAACTATATTTAAATATTTTTTGTTAGT

[0311] ATGTNGAGNANATGGTATTTTTATTGTTCTTTTAATCTTCTNTGNTAGAAANATTAACA

[0312] TTTTNNTTTTAATTTNNGTGTTGGGAATGTAAAATGTTATAACCACTTTGGAAAAACAG

[0313] TTTGGAATTTTCTTATTAAAAGTTAAACATAATTTTGTCATATTAATTACNATATTGATC ATTCTATTTNAATTATTTAGAATTAGATTTTNAATTTAAAANAAAAATTAAATTAACTTN

[0314] TAATTTTTACAAATATAAAAATGTATATCTTTANATANTAAATTTTATATTAGCTTAATTT

[0315] TATAATAGCCAAAAANTAGAAANACAACCAAAATATNTTTCAATNTGTGAATGGATGA

[0316] TNCATTTCTTTNTTAATAAATTGTGGGATATCTCTTCAATAAATTTAATNNTACTATTCA

[0317] TGNAATGAATAGAATTGTTTTATTTTTCAGTTTTGCTNCACTTATTTNGTAAGNAATAA

[0318] AATACTGATAANTTNATTNTAAAACTTNTAGTANATTAAGNATATATATAAAAATGTTT

[0319] GANTNTAATAAATCAATTTACATAAAGAAAAATANTATTTTNATTATTCAANTTATATGA

[0320] AAGTATAACTAGNATATATAAATTCATATAAATTTTAAAAAAAGTCNTTTTTTCTGAAAC

[0321] TATTTAGATTGAATTTAGTANATTTGGCATTGGCNACTTTTAGANGCTTTGTGTAAAAT

[0322] TGATTNACTANTTATTTNATANAAGATGATAGAANTGTTTTTATACCTTCTCTTNNTTG

[0323] GTAATTANAATACATTAAAGTATGTTTCACTTGTTNANNNTNNTAGTATACATNTATTA

[0324] AATANCAANTAAAAANAATATATAATCTAACAAGGATAATGTAAATATTTTTTTTTATAA

[0325] ANAAATTTNAAAGNATNNTAACCNCAATTAACAACAATCATTGNNTATAAAGAAAAAT

[0326] CNTTAATGTGAAATCTGATTTTATTTTTATNTCAGCAAAATAAATAAACTTTTCCTTTNA

[0327] AATTTTTTCTGATACTTNCAAAAAAAGTAAAAGTTATGTTNATGATTTGAAATNAGCAT

[0328] NNGTTGNTTANAATGAGTTATATTTGCATTTAATATGTATANTAGTANCAAGAGNGAA

[0329] AANAGNAGNAAAACACANTCTANAACCTTATGNAGTNAAAATAATNTTTTTATAACNT

[0330] CTGNTNATATAAAATTNAAATTATTTCCAAATCTTTAGTANATATTTTATAATTTCTGGT

[0331] TTNTAAATTAAGAAATTAAAAAAAAAAAAGAAAAAAAAANTAATAAAANNGTATAAATA

[0332] TAAATTAGAAGGAAATATAAANNTAANACAAAAAAGTTATTGAGGTAAAAAATATATAA

[0333] AAATAAAAAAAGANANAACATATAAANTACAAAACTGGTTNTTTTAAAGAATATNAAAA

[0334] ATCTATAAGATTGAATATATAATTCAGTGAGATTAANNTATTTANAAANAAAAACAAAA

[0335] GNAAAAAAAGTTNTG

[0336] TTATATGAAAGTATAACTAGNATATATAAATTCATATAAATTTTAAAAAAAGTCNTTTTT

[0337] TCTGAAACTATTTAGATTGAATTTAGTANATTTGGCATTGGCNACTTTTAGANGCTTT

[0338] GTGTAAAATTGATTNACTANTTATTTNATANAAGATGATAGAANTGTTTTTATACCTTC

[0339] TCTTNNTTGGTAATTANAATACATTAAAGTATGTTTCACTTGTTNANNNTNNTAGTATA

[0340] CATNTATTAAATANCAANTAAAAANAATATATAATCTAACAAGGATAATGTAAATATTT

[0341] TTTTTTATAAANAAATTTNAAAGNATNNTAACCNCAATTAACAACAATCATTGNNTATA

[0342] AAGAAAAATCNTTAATGTGAAATCTGATTTTATTTTTATNTCAGCAAAATAAATAAACT

[0343] TTTCCTTTNAAATTTTTTCTGATACTTNCAAAAAAAGTAAAAGTTATGTTNATGATTTGA

[0344] AATNAGCATNNGTTGNTTANAATGAGTTATATTTGCATTTAATATGTATANTAGTANCA

[0345] AGAGNGAAAANAGNAGNAAAACACANTCTANAACCTTATGNAGTNAAAATAATNTTTT

[0346] TATAACNTCTGNTNATATAAAATTNAAATTATTTCCAAATCTTTAGTANATATTTTATAA

[0347] TTTCTGGTTTNTAAATTAAGAAATTAAAAAAAAAAAAGAAAAAAAAANTAATAAAANNG

[0348] TATAAATATAAATTAGAAGGAAATATAAANNTAANACAAAAAAGTTATTGAGGTAAAAA

[0349] ATATATAAAAATAAAAAAAGANANAACATATAAANTACAAAACTGGTTNTTTTAAAGAA TATNAAAAATCTATAAGATTGAATATATAATTCAGTGAGATTAANNTATTTANAAANAA

[0350] AAAC AAAAG N AAAAAAAGTTNTG

[0351] TAACTAGNATATATAAATTCATATAAATTTTAAAAAAAGTCNTTTTTTCTGAAACTATTT

[0352] AGATTGAATTTAGTANATTTGGCATTGGCNACTTTTAGANGCTTTGTGTAAAATTGAT

[0353] TNACTANTTATTTNATANAAGATGATAGAANTGTTTTTATACCTTCTCTTNNTTGGTAA

[0354] TTANAATACATTAAAGTATGTTTCACTTGTTNANNNTNNTAGTATACATNTATTAAATA

[0355] NCAANTAAAAANAATATATAATCTAACAAGGATAATGTAAATATTTTTTTTTATAAANAA

[0356] ATTTNAAAGNATNNTAACCNCAATTAACAACAATCATTGNNTATAAAGAAAAATCNTTA

[0357] ATGTGAAATCTGATTTTATTTTTATNTCAGCAAAATAAATAAACTTTTCCTTTNAAATTT

[0358] TTTCTGATACTTNCAAAAAAAGTAAAAGTTATGTTNATGATTTGAAATNAGCATNNGTT

[0359] GNTTANAATGAGTTATATTTGCATTTAATATGTATANTAGTANCAAGAGNGAAAANAG

[0360] NAGNAAAACACANTCTANAACCTTATGNAGTNAAAATAATNTTTTTATAACNTCTGNT

[0361] NATATAAAATTNAAATTATTTCCAAATCTTTAGTANATATTTTATAATTTCTGGTTTNTA

[0362] AATTAAGAAATTAAAAAAAAAAAAGAAAAAAAAANTAATAAAANNGTATAAATATAAAT

[0363] TAGAAGGAAATATAAANNTAANACAAAAAAGTTATTGAGGTAAAAAATATATAAAAATA

[0364] AAAAAAGANANAACATATAAANTACAAAACTGGTTNTTTTAAAGAATATNAAAAATCTA

[0365] TAAGATTGAATATATAATTCAGTGAGATTAANNTATTTANAAANAAAAACAAAAGNAAA

[0366] AAAAGTTNTG

[0367] TAATATGTATANTAGTANCAAGAGNGAAAANAGNAGNAAAACACANTCTANAACCTTA

[0368] TGNAGTNAAAATAATNTTTTTATAACNTCTGNTNATATAAAATTNAAATTATTTCCAAAT

[0369] CTTTAGTANATATTTTATAATTTCTGGTTTNTAAATTAAGAAATTAAAAAAAAAAAAGAA

[0370] AAAAAAANTAATAAAANNGTATAAATATAAATTAGAAGGAAATATAAANNTAANACAAA

[0371] AAAGTTATTGAGGTAAAAAATATATAAAAATAAAAAAAGANANAACATATAAANTACAA

[0372] AACTGGTTNTTTTAAAGAATATNAAAAATCTATAAGATTGAATATATAATTCAGTGAGA

[0373] TTAANNTATTTANAAANAAAAACAAAAGNAAAAAAAGTTNTG

[0374] AGTANCAAGAGNGAAAANAGNAGNAAAACACANTCTANAACCTTATGNAGTNAAAAT

[0375] AATNTTTTTATAACNTCTGNTNATATAAAATTNAAATTATTTCCAAATCTTTAGTANATA

[0376] TTTTATAATTTCTGGTTTNTAAATTAAGAAATTAAAAAAAAAAAAGAAAAAAAAANTAAT

[0377] AAAANNGTATAAATATAAATTAGAAGGAAATATAAANNTAANACAAAAAAGTTATTGA

[0378] GGTAAAAAATATATAAAAATAAAAAAAGANANAACATATAAANTACAAAACTGGTTNTT

[0379] TTAAAGAATATNAAAAATCTATAAGATTGAATATATAATTCAGTGAGATTAANNTATTT

[0380] AN AAAN AAAAACAAAAG N AAAAAAAGTTNTG

[0381] CATNACCATCACCACTATAAATNCANNAACAGCAGNCCNTATGATNNTGANTNATGA

[0382] AATCGATNNCAAAAACTTTTTGATCATCCCNAAAAACTGAAACACTGTACCGATTAAA

[0383] AAATNCCATNAACTTCCCATTAAAAGGAT

[0384] CTTCCCATTAAAAGGAT

[0385] GAAGACCTCTCNACCCAGCNCCTGGCAACCCCTGAAAAGAGNAATTNTACTTTTTNT ANCNNNNNNNNANNAATGTAANTNCTGTCTCTATGAATTAGACTATTNTAGCCAATAT

[0386] CTTATATAAATAGAAGTCATACAATATNT

[0387] ATGTATTTTGTATTGNTTGGCTTATTTAACACTTAGCATAATGTNTTCAAGGTTCATAC

[0388] ATGTNGTANCATATATCAGAATTTCATTCCTTTTTACANNGGANNATTTATTCCATTCC

[0389] TTTACAGAAATAAANCAAATTTTGTTTATCCATTCATTTACTAANNAANAAAACAATNA

[0390] NTNNTAAGATTNNCAAGGATATNGNNCANNNANANAACTACTGATNGACATTTGGGT

[0391] TNTTTCCATNTTATAAAANACTTTTTGGCAATTTCAAATAAGGCTGCTATGAACATTCC

[0392] TGTACATATGTCTCTTTGTAGTGATACNTAAGATATTCTNAAGTGAGNNTAAGAAAAA

[0393] TAAANAAATNTNAAAGGACTAATAANATNTTTATTATTTNCGTTAATACCTANNAGNAG

[0394] AAATTCTGANATCANAGAACAGATAAANTAGGGAAATTTATAAATTNTTGGAAANTAA

[0395] ATAACACAGTGTTTTCTAACANTAGGTATACAANATAACTCTCATACNANCAGTGNAT

[0396] AATGTTTTGTATTTAATTTANATGNAGAACTAAAANNAATCAACATTNTGNNAGTCANT

[0397] CANTGNAATGNTCCNAGNGAANATGTATCTATAAATANGTNTNTTAAAAANNAATCAG

[0398] ACATNGTGNTTGTAANNTNNANTNNCNATTAACATGGNCNAAAANNAG

[0399] SEQ ID NO: 10

[0400] GGATCNTGGNTTCCAAGTTTTCTTNTTTTATTTTTTTTTTTTTTACTTATAAANCTTTAT

[0401] TTAAGTTCTAGGGTACATGTGCACACTGCATTACGTGCAGGAAGATTTGTTACATGC

[0402] CAACTTATGTATACATGTGCCATGTTGGGTGTTTGTTTTCCTTTNGCTGCACCCATCG

[0403] GAGGGNAANCCGAAGCANTGCAGGAACTTACAAGAATTAACGTTTNAAGTGAACAN

[0404] GGNANNTCGTCCNCTNCCCAACNNANNCTCCCATTTAGGAAATTAACATAGTACCTT

[0405] TTAGGTATATCTCCTAATGCTATCCCTCCCCACAGCTCCNTCCTCCCCAGCCCCACT

[0406] CCNAAAGACAGTCTNATCCCCGNGGTGTGATGTTCCCCTCTCTGTGTCCATGTGTTC

[0407] TCATTGTNTTCAACTCCCACCTATGAGTGAGAACATGTGGGTGTTGCATTCTNGATTC

[0408] ATGGTTTTCTGTTCTTGTGATAGCTTTGCAAGGNATTGCNTCACTTGCTGAGAATAAT

[0409] GGTTTCCAGCTTCATCCATAGTCCCTGCAAAGGACATGAATTCATGTTTTTTTTGTGA

[0410] GAATAATGGTTTCCAGCTTCATCCATAGTCCCTGCAAAGGACATGAATTCATGTTTTT

[0411] TTTGTGA

[0412] >

[0413] TTTTCCTTTTTTTATGGCTGCATAGTATTCCATGGTGTATATGTGCCACATTTGCGCC

[0414] TTTTCTTTATCCAGTCTATCATTGATGGACATTTGGGTTGGTTCCAAGTCTTTGCTATT

[0415] GTGAATATTGCNGCAATAAACATCCACAAACGTGTGCATGTGTCTTTTTAGTAGCATG

[0416] ATTTCAATATTCCTTTGGGTATATACCCAGTAATGGAACTTGCTGGGTCGAGANGNA

[0417] AATGGTATTTCTAATTCTTGATCNTTGAGGAATCTCCACACTGTCTTCCACAAGATGC

[0418] CAGGAAGGTTGAACTAGTTTACANTCCCACCAGCTCAGTGTAATTTTACCTTGTNTTT

[0419] NCTTCCTCAGTATATAAGTGTTCCTATTTCTCCACATCCTCAGATTAAAAGNTTNATCT

[0420] CCAGTCACTTTAAAGCATCTGTTGTTTCCTTGACCTTTTTAATGATGGCCATTCTAACT

[0421] GGTGTGAGATGGTNGTATTCTCATTGTGGTTTTGATTTGCATTTCTCTGATGACCAGT

[0422] G ATG ATG AG C ATTTG GTTCTTTC ATC ATGG CTTTTG GCTGTTATN NG C ATATGTC AAA TGTCTTAATAGCTGNCAGTCTGTTGCCTAATTTTGAAGAAGTGTCTGTTCATTTCCTT

[0423] TGCCCACTTTTTGATGGGGTGATCTTTNTTTGTGTTNCTTGTAAGTTTGTGTTGAGTT

[0424] CATTGATAGATTCTTGGAGTATTAGCCANTTGTCAGATGAATAGTTTCCAACTATTTT

[0425] CTCTTCTTCTGTAGGGTTGGATCAGGCNACTACAGTGGNTTTTTTNTTTTNNTANTGG

[0426] AATGTTCACTCTGATGATAGTNAATTCTTTTATTGCGCNTGATAATCCTGTGCAGAAA

[0427] AAGCTTTTTAGTTTGATCAGTTCAGATAACTTCAGTAAAACTGTAGCTTTTGTTGCCAT

[0428] TTNTTTTAAGTGAAATTTTAAAAATGAAGTCTTTGACTACCCATACCTAAGTATTGAAT

[0429] GTTTTTNCATATGTTTTAAATCTTCTAGGGTTTTTATGGCTTTAAGTNTTACATTGAAG

[0430] TCTTCCTTAAATACATCTTGAAATTATTTTTTCTATATNTTAAGACCAAAGGATCCTATT

[0431] TCCACNTAGATGATATTGCTAAATAGAATTGATTTTTAAATTTGNNGGGTAAAAAACTT

[0432] TTCCCAGAACCATTTATTGAATAGGGTAAAAANCAGAGTGCCTATCCTTTTCCCAATG

[0433] ATCGCAGCTCCTCTCCAGCAACTTTTCAAAGTTGGGTTGAAAATCAGATTGTTGTTG

[0434] GTATATAGGAATGCTTCTGATGTTTGTANATTAATTTTGTTCCCTGAGACTATGGATC

[0435] AGTTTTTATACCAATGCTAGGAAGCTTAAAACAGAAACTTGAAAAAGATTTTATAAGTA

[0436] TAGCCTTCTAGTATAAGTNANAAGTCAGATAGTATGATNCCTCCAGCTTTCTGGANCT

[0437] GTATACCTTGGATTGANTTCTCTATGATGGCTGCATTNGCNTTCCATGTNANTTTTTT

[0438] TTCTTTATCTCCTCTCAGTAGCATTAAANANACANNTTANAATTAAAAANAGTAATTAT

[0439] TTCCAATTCTGTGAAGAATATCATTGGTGGTTAAAGAGGAAATTGTTGTAATTTATGT

[0440] AAATTGACATGGGCAGTACATAACAGTTTAAAAAGATTTGAACATAACCTTCTAGCAA

[0441] TNTGTGACTATGTGNACATAATTCTAAATCTTTTGGGAGTNTGTTTCCTTGATGTTTN

[0442] NTTATCTAATTCAGTGGTGGTTTATTTCATGGAACAATGTTGTAAAATTTTCTTTAAAN

[0443] AGGACATTTTCCTGCATCACTTCCCCAACATAGTTATATAGTTCTACATTTTTTTATTC

[0444] TTTTTCAGCAAATACGAATGGAATACAACTCATGATNTACCTCTCTGGTTGAATGATC

[0445] ATTGCCTACACACATTATNGTCTGATTCATCAAGGTTGATTATGATATAATGATATGTT

[0446] ACTGGAAGTCAGAGAATAAGGTTTGGTAATCTTTTAAAGGGAAGCCCATCAGACTAA

[0447] TAGGGGNTTTCTCAGTAGAAACTCATCGAAAGCCAGCAAAGAGATTGAGTTTGACTT

[0448] CCTCTCTTCACCATTNAAAACCAATAGACTTACTTTTTCTTTCCTGANTNCTATTAGCC

[0449] AGAATTTCGTAATCCAGCACTATACTAAAAACATCATAAATGAAAGAGCGGAATACAT

[0450] TCCTTTACAGATGTTCT

[0451] TTGANTTCTCTATGATGGCTGCATTNGCNTTCCATGTNANTTTTTTTTCTTTATCTCCT

[0452] CTCAGTAGCATTAAANANACANNTTANAATTAAAAANAGTAATTATTTCCAATTCTGTG

[0453] AAGAATATCATTGGTGGTTAAAGAGGAAATTGTTGTAATTTATGTAAATTGACATGGG

[0454] CAGTACATAACAGTTTAAAAAGATTTGAACATAACCTTCTAGCAATNTGTGACTATGT

[0455] GNACATAATTCTAAATCTTTTGGGAGTNTGTTTCCTTGATGTTTNNTTATCTAATTCAG

[0456] TGGTGGTTTATTTCATGGAACAATGTTGTAAAATTTTCTTTAAANAGGACATTTTCCTG

[0457] CATCACTTCCCCAACATAGTTATATAGTTCTACATTTTTTTATTCTTTTTCAGCAAATAC

[0458] GAATGGAATACAACTCATGATNTACCTCTCTGGTTGAATGATCATTGCCTACACACAT TATNGTCTGATTCATCAAGGTTGATTATGATATAATGATATGTTACTGGAAGTCAGAG

[0459] AATAAGGTTTGGTAATCTTTTAAAGGGAAGCCCATCAGACTAATAGGGGNTTTCTCA

[0460] GTAGAAACTCATCGAAAGCCAGCAAAGAGATTGAGTTTGACTTCCTCTCTTCACCATT

[0461] NAAAACCAATAGACTTACTTTTTCTTTCCTGANTNCTATTAGCCAGAATTTCGTAATCC

[0462] AGCACTATACTAAAAACATCATAAATGAAAGAGCGGAATACATTCCTTTACAGATGTT

[0463] CT

[0464] GCATTAAANANACANNTTANAATTAAAAANAGTAATTATTTCCAATTCTGTGAAGAATA

[0465] TCATTGGTGGTTAAAGAGGAAATTGTTGTAATTTATGTAAATTGACATGGGCAGTACA

[0466] TAACAGTTTAAAAAGATTTGAACATAACCTTCTAGCAATNTGTGACTATGTGNACATA

[0467] ATTCTAAATCTTTTGGGAGTNTGTTTCCTTGATGTTTNNTTATCTAATTCAGTGGTGGT

[0468] TTATTTCATGGAACAATGTTGTAAAATTTTCTTTAAANAGGACATTTTCCTGCATCACT

[0469] TCCCCAACATAGTTATATAGTTCTACATTTTTTTATTCTTTTTCAGCAAATACGAATGG

[0470] AATACAACTCATGATNTACCTCTCTGGTTGAATGATCATTGCCTACACACATTATNGT

[0471] CTGATTCATCAAGGTTGATTATGATATAATGATATGTTACTGGAAGTCAGAGAATAAG

[0472] GTTTGGTAATCTTTTAAAGGGAAGCCCATCAGACTAATAGGGGNTTTCTCAGTAGAA

[0473] ACTCATCGAAAGCCAGCAAAGAGATTGAGTTTGACTTCCTCTCTTCACCATTNAAAAC

[0474] CAATAGACTTACTTTTTCTTTCCTGANTNCTATTAGCCAGAATTTCGTAATCCAGCACT

[0475] ATACTAAAAACATCATAAATGAAAGAGCGGAATACATTCCTTTACAGATGTTCT

[0476] TTACTGGAAGTCAGAGAATAAGGTTTGGTAATCTTTTAAAGGGAAGCCCATCAGACT

[0477] AATAGGGGNTTTCTCAGTAGAAACTCATCGAAAGCCAGCAAAGAGATTGAGTTTGAC

[0478] TTCCTCTCTTCACCATTNAAAACCAATAGACTTACTTTTTCTTTCCTGANTNCTATTAG

[0479] CCAGAATTTCGTAATCCAGCACTATACTAAAAACATCATAAATGAAAGAGCGGAATAC

[0480] ATTCCTTTACAGATGTTCT

[0481] TTTCTCAGTAGAAACTCATCGAAAGCCAGCAAAGAGATTGAGTTTGACTTCCTCTCTT

[0482] CACCATTNAAAACCAATAGACTTACTTTTTCTTTCCTGANTNCTATTAGCCAGAATTTC

[0483] GTAATCCAGCACTATACTAAAAACATCATAAATGAAAGAGCGGAATACATTCCTTTAC

[0484] AGATGTTCT

[0485] TTTGGGGACATAGCTTCCAGCTTTTTTNAATTAAGAATAATGTCTTCAAGGNGCTTCT

[0486] NATATTAAGCTCTAAAATATTTNGAAACATGTACCAGCAATACCTTTTTTTTTAAANGG

[0487] CTGAATAATATTAAATTGTGTTGAATTTTGTCAAAGGCNTTTAAAGAATCTATNAAAAT

[0488] AATCATGTGGTTTTTGTTCTTTGGTTATGAAGAAGCTATGAATTACATTTATTTANTGG

[0489] GCTAAATAAACACCGCATTATCTTNATGAAATGATCAAANCTTGATAGACAAGTATTA

[0490] NCCTTTAATGTTAATGGGCTANATCCACCTTTTGGCTATTATAAATAGAATTGCTAAAT

[0491] GGATAAAGTGTCAAGAAATATTTGTTTTGAATNCTGTATTTTTNTCTGGTTACNCATCT

[0492] CACGGGCTATAGACTCACAAAAGTTCAAATTTAAAGGGATGANATAGGAAAAAAGCT

[0493] CTATTTTTCAATTAATTGGAAAACAAAAAAANAAAAAAAAAGCAGAAGTAGCAGTCCT

[0494] AATTTATATTTCTACTAGAATTAAGCTATGAAAGATCAAAAATCCATCCAGTCCAGGG CATTTTATAATGGGAAAGCTTTTATAAGCTNTTAATTATTGACTCAATTTAACAACAAG

[0495] ATATNGGTCTCTTCAATATTTATATTTCTTCCTTGGTTCAATATAGGAGGGTTCGTATT

[0496] TTTCCAGCAATTTCTCCATTACTNTCTAAATTTTCTAATTTATAAATTGTATAGACTTCT

[0497] TCATAATATTCTCAAATCAAATCTTAGGATACTTTTATATTCCTGTGGCAANATTAGAC

[0498] GTATTATCGAGTTTGAAATTTTATATTATGATAATTTGGATTTGCACTCTTCTTTTTTTA

[0499] TCTATTAGACCTAATATAGCTCTTAAGTGATATCTACANNTTTTTCAACCTCCAAAAAA

[0500] ACAACAGAATGTATTCGTTTACATTCTTCTTATGGAAGCATCACAAAATCTCAATTTCT

[0501] TTAAATTCTTCTATATNATTGGAAATATCAAACTTTTCTTCTTCAGCAATTTTTAAAGAT

[0502] NGTTTGCTCTTGATTTTTTCT

[0503] GCTATTATAAATAGAATTGCTAAATGGATAAAGTGTCAAGAAATATTTGTTTTGAATNC

[0504] TGTATTTTTNTCTGGTTACNCATCTCACGGGCTATAGACTCACAAAAGTTCAAATTTA

[0505] AAGGGATGANATAGGAAAAAAGCTCTATTTTTCAATTAATTGGAAAACAAAAAAANAA

[0506] AAAAAAAGCAGAAGTAGCAGTCCTAATTTATATTTCTACTAGAATTAAGCTATGAAAG

[0507] ATCAAAAATCCATCCAGTCCAGGGCATTTTATAATGGGAAAGCTTTTATAAGCTNTTA

[0508] ATTATTGACTCAATTTAACAACAAGATATNGGTCTCTTCAATATTTATATTTCTTCCTT

[0509] GGTTCAATATAGGAGGGTTCGTATTTTTCCAGCAATTTCTCCATTACTNTCTAAATTTT

[0510] CTAATTTATAAATTGTATAGACTTCTTCATAATATTCTCAAATCAAATCTTAGGATACTT

[0511] TTATATTCCTGTGGCAANATTAGACGTATTATCGAGTTTGAAATTTTATATTATGATAA

[0512] TTTGGATTTGCACTCTTCTTTTTTTATCTATTAGACCTAATATAGCTCTTAAGTGATAT

[0513] CTACANNTTTTTCAACCTCCAAAAAAACAACAGAATGTATTCGTTTACATTCTTCTTAT

[0514] GGAAGCATCACAAAATCTCAATTTCTTTAAATTCTTCTATATNATTGGAAATATCAAAC

[0515] TTTTCTTCTTCAGCAATTTTTAAAGATNGTTTGCTCTTGATTTTTTCT

[0516] GAAGTAGCAGTCCTAATTTATATTTCTACTAGAATTAAGCTATGAAAGATCAAAAATC

[0517] CATCCAGTCCAGGGCATTTTATAATGGGAAAGCTTTTATAAGCTNTTAATTATTGACT

[0518] CAATTTAACAACAAGATATNGGTCTCTTCAATATTTATATTTCTTCCTTGGTTCAATAT

[0519] AGGAGGGTTCGTATTTTTCCAGCAATTTCTCCATTACTNTCTAAATTTTCTAATTTATA

[0520] AATTGTATAGACTTCTTCATAATATTCTCAAATCAAATCTTAGGATACTTTTATATTCCT

[0521] GTGGCAANATTAGACGTATTATCGAGTTTGAAATTTTATATTATGATAATTTGGATTTG

[0522] CACTCTTCTTTTTTTATCTATTAGACCTAATATAGCTCTTAAGTGATATCTACANNTTTT

[0523] TCAACCTCCAAAAAAACAACAGAATGTATTCGTTTACATTCTTCTTATGGAAGCATCA

[0524] CAAAATCTCAATTTCTTTAAATTCTTCTATATNATTGGAAATATCAAACTTTTCTTCTTC

[0525] AGCAATTTTTAAAGATNGTTTGCTCTTGATTTTTTCT

[0526] TTTTATATTATGATAATTTGGATTTGCACTCTTCTTTTTTTATCTATTAGACCTAATATA

[0527] GCTCTTAAGTGATATCTACANNTTTTTCAACCTCCAAAAAAACAACAGAATGTATTCG

[0528] TTTAC ATTCTTCTTATG G AAG C ATC AC AAAATCTC AATTTCTTTAAATTCTTCTATATN A

[0529] TTGGAAATATCAAACTTTTCTTCTTCAGCAATTTTTAAAGATNGTTTGCTCTTGATTTTT

[0530] TCT TATCAAACTTTTCTTCTTCAGCAATTTTTAAAGATNGTTTGCTCTTGATTTTTTCT

[0531] AGTTCTTTTAATTGTGATATTAGAATTTTAATTTAATATCTTTCTAGCTTTTCTATGTAA

[0532] GNAATATAGTGCAATAAAATTATATCTCTTTACAAAGAATTACATGTATCACAAATACA

[0533] ATCAAATANATGGTATCTTTGTTTTGAATAGTTTCAAAAACTAGCTCTTGAATTCTGCC

[0534] TGATTCATTTNGACATTTAAAGCTAAATAAAGTAAATNAAGGACAAAANTAAAGATTG

[0535] NTAGTTATTTAATTTCCATGTATTTGTNTGGTTTTGAAATAGTTTCTTATTAGAAATGAA

[0536] TTCAATTTATAATTGAATTAAGGTATAAAAATATCTTGGATACAATTTAAGCTCTTTTTG

[0537] TTTATAGTGAGAATTGTTTAATCCAGTACCTAAATGAGGTATATTTTGAAAGTAAGAAA

[0538] GATGTGATATAAAATTGAACAGATTCTAAATTATGTTCATACTATTGGGGTGAAAAGA

[0539] ACTATAGAAGTCTGAATAAGATACATTTGCAAAAGGGTAACTAGTAGAAGACAAGAA

[0540] NAAACTAAGATAAATTTGCCTNGAGCCGAAGATCTGAAAAATATTGAAAGTAAAAAAG

[0541] AACTTTNTAAAANAANGTCTCCCAAAAATATTTCAATGAATTCAGAAGATTTATTTTTT

[0542] GAAAAAAATAAAAACTTGGCTTTAAAAATAATTGACCACTAGTACCNCTGNNCTTATA

[0543] CTAGATTACATAAATAAAAAAAAAAAAAGAAAATTTTCAAATTTTATTAAATTTAAAAAT

[0544] GATAAAGGGTATATTACTACATATCCCGACCTTCTTACAGAAATANAAACTNTCATCA

[0545] TAGAAGTGTACTATNTTTTATCACCTATAAGAATAAAAACTAGAAAATTTTTTTTGTTTT

[0546] AAATATAAATTGANAATGGANAAATTTTTTCAAANCATTTNNTTTAGAACCATAAACAA

[0547] TAAACTCTCCCAATATCTNTAATTAGGAAGAAATTGAATCTCTGAAATAGAGCAAAAA

[0548] CAGGCTCTGAAATTGAGTCAATAATCAATAACTTACCAACCAATGTCTGACAAAAAAT

[0549] GTCCATAATACCAGAAGGATTTAAACCATTTNATTAAATNCAAAATTTATTCCAGATAT

[0550] ATAAAGAGAATTTGGTACCATTATTACTAANATTAGTTTTATCTNAACTATTTTTTCCAA

[0551] TTTTAAATAGTAAAG GTAG C AN AATG GTATATCTTCTCCTTATTC AT

[0552] ACTATTGGGGTGAAAAGAACTATAGAAGTCTGAATAAGATACATTTGCAAAAGGGTA

[0553] ACTAGTAGAAGACAAGAANAAACTAAGATAAATTTGCCTNGAGCCGAAGATCTGAAA

[0554] AATATTGAAAGTAAAAAAGAACTTTNTAAAANAANGTCTCCCAAAAATATTTCAATGAA

[0555] TTCAGAAGATTTATTTTTTGAAAAAAATAAAAACTTGGCTTTAAAAATAATTGACCACT

[0556] AGTACCNCTGNNCTTATACTAGATTACATAAATAAAAAAAAAAAAAGAAAATTTTCAAA

[0557] TTTTATTAAATTTAAAAATGATAAAGGGTATATTACTACATATCCCGACCTTCTTACAG

[0558] AAATANAAACTNTCATCATAGAAGTGTACTATNTTTTATCACCTATAAGAATAAAAACT

[0559] AGAAAATTTTTTTTGTTTTAAATATAAATTGANAATGGANAAATTTTTTCAAANCATTTN

[0560] NTTTAGAACCATAAACAATAAACTCTCCCAATATCTNTAATTAGGAAGAAATTGAATCT

[0561] CTGAAATAGAGCAAAAACAGGCTCTGAAATTGAGTCAATAATCAATAACTTACCAACC

[0562] AATGTCTGACAAAAAATGTCCATAATACCAGAAGGATTTAAACCATTTNATTAAATNC

[0563] AAAATTTATTCCAGATATATAAAGAGAATTTGGTACCATTATTACTAANATTAGTTTTAT

[0564] CTN AACTATTTTTTCC AATTTTAAATAGTAAAG GTAG CAN AATG GTATATCTTCTCCTT

[0565] ATTCAT

[0566] GGAAGAAATTGAATCTCTGAAATAGAGCAAAAACAGGCTCTGAAATTGAGTCAATAAT CAATAACTTACCAACCAATGTCTGACAAAAAATGTCCATAATACCAGAAGGATTTAAA

[0567] CCATTTNATTAAATNCAAAATTTATTCCAGATATATAAAGAGAATTTGGTACCATTATT

[0568] ACTAANATTAGTTTTATCTNAACTATTTTTTCCAATTTTAAATAGTAAAGGTAGCANAA

[0569] TGGTATATCTTCTCCTTATTCAT

[0570] TCAATAACTTACCAACCAATGTCTGACAAAAAATGTCCATAATACCAGAAGGATTTAA

[0571] ACCATTTNATTAAATNCAAAATTTATTCCAGATATATAAAGAGAATTTGGTACCATTAT

[0572] TACTAANATTAGTTTTATCTNAACTATTTTTTCCAATTTTAAATAGTAAAGGTAGCANA

[0573] ATGGTATATCTTCTCCTTATTCAT

[0574] TTTATGAGGCCAGCNTCATCCTGATACCAAAACCTGGGCATATTTTATATACAATAGT

[0575] TAAAAACAAAAAAAGAGAANTTTAGACCAATTTGNATATCCTTGATGAAAATGGATGC

[0576] AAACAATCCTTTATTTCNCCATCTTTAAAATTTTCCTTGCAAACCAATTCTAACAGCAT

[0577] ATCAAAAAGCTAATCCACCATGATCAAGTGAGGGATTAAATCCCTAAGATTCAAGGC

[0578] TGTTTGAAAATATGTTCAAATTTATAAATGTAATTCATAAAATAAACAAAAACTAAATTT

[0579] AAACAACCACATGANTATATAATTATTTGAATGAAGAAATTTTCTTCTATAAAATTTAA

[0580] CATTATTCCTTTTCNTTTCATTATTAATACTTTCTTCTAGAAAGATCTATATATTGAAGG

[0581] AACATTTCTATCTGCTCAATATAATAATATCCATTTATGACAAACTNNNACAGAAAATA

[0582] TTTTTCATTNTAACTGAAATGGACAAAGTNATTAACTTGAAAGCATTTTACCTTTATTG

[0583] AAANCTTNTTTGAACAATACAACNATGATGTGTCTNCTTTCTACTCTCTCATCNCTCC

[0584] TATTNAATATATTATTGGAAGTTCTGCTCAATTGCCATGGAACTATCATGNAAGAAAN

[0585] GAATTTTATTTCTTTATNATAAAGAAATAAAAAGTTTTCAAATTTTTTTAAGAAAAGAGA

[0586] AAGTAAAATTTTCTCTGTTCTGCTTGCAGTTTACATAGCTTAATTTTTTCTTTATCTAGA

[0587] AGAAAANCTCATAATCTTTACATATCATCACCATAAACATTTTNCTCATTAAGNATTAT

[0588] AAACNAAAACTTCAGTTAAAGTCTTAACGTATACAAACATNAATTTTTAAAAATTTGTA

[0589] GAATTCCCTATTCTATCTAACANCAGTATTCTTAAAAATCTATGANAGGAGTGTATTCA

[0590] ATACAGAAAAATGTTTTAGTTTTTACTATTNATTTTTAGATTTAAAAAAANATATTTCGA

[0591] AAATTACTGAATTTTTTAATCTAGTAAAATCTTTTATATANNTTTGTGTTTTTCTTATTAT

[0592] TTTNATTTNTATTTCATTAATGAAAAATAAATTTGTAAACATTTTTTCATTCCATATAATT

[0593] TNAATTAACTGANGAGAAATATATTTTCTTATCATTGCANCCTCATTGACAATTGTTAC

[0594] TAAAANTTTCTTCATATAGTAATTAAAATACTTAGGTATTCAGCTATCTAATATTCAANT

[0595] ATATAAAAAATTACTCTAATAATATAAACAAAATTCNTAAAACACTGCTCTAAAATTATA

[0596] TAAATTTAAATATATAAAAATTATATAATATTAGGATAATATTTTATAATACTCAATATG

[0597] GATAAGAAAAATCAATATACATTAAAATTGTCCATATTACTNATAAAANTAGTAAAATT

[0598] TACATTAANTTAATATATTTAAGTACAATAATANTGGTTCAATATATCAATATAATTTTC

[0599] ATTCAACATAATTCAAATATTTTAAAANAGATTAAAATAATNTAAAAAATAATATTTATT

[0600] AATATTTATTATATTAAAGTCTTTAAATTGTAATGAAAAAAAAAAATATTTTAATTTTTTA

[0601] CATATGCATAAAAGTATCTTACAAAANATTNATGATTGAATATAGAATTTCCAAAATTA

[0602] AATTTTAAAAAAAATAACAATNTAAGAACAAATCATGTTAGAAGATAATCATATANTAT TCTCTAA

[0603] AGGATAATATTTTATAATACTCAATATGGATAAGAAAAATCAATATACATTAAAATTGT

[0604] CCATATTACTNATAAAANTAGTAAAATTTACATTAANTTAATATATTTAAGTACAATAAT

[0605] ANTGGTTCAATATATCAATATAATTTTCATTCAACATAATTCAAATATTTTAAAANAGAT

[0606] TAAAATAATNTAAAAAATAATATTTATTAATATTTATTATATTAAAGTCTTTAAATTGTAA

[0607] TGAAAAAAAAAAATATTTTAATTTTTTACATATGCATAAAAGTATCTTACAAAANATTNA

[0608] TGATTGAATATAGAATTTCCAAAATTAAATTTTAAAAAAAATAACAATNTAAGAACAAA

[0609] TCATGTTAGAAGATAATCATATANTATTCTCTAA

[0610] TTTATTAATATTTATTATATTAAAGTCTTTAAATTGTAATGAAAAAAAAAAATATTTTAAT

[0611] TTTTTACATATGCATAAAAGTATCTTACAAAANATTNATGATTGAATATAGAATTTCCA

[0612] AAATTAAATTTTAAAAAAAATAACAATNTAAGAACAAATCATGTTAGAAGATAATCATA

[0613] TANTATTCTCTAA

[0614] TATGATTTTTTAAATTATATATTTACAATACAATGCTATTAGTAAATTAAAATTCTNAAA

[0615] ACAGTATTGTTATTAAATATACNAAAACAGACATTTATAAAATAAGAATTATAGTTGAC

[0616] CAATGGAACAGCACTCACAAAAGAGTCCTTAGAAATTTTAATTGATATCCANCACCAT

[0617] GTACATCTAATTTGATTTTTGACAAACCTGACATGAAAAAATATTNAATAGATAAATGA

[0618] ACACATACGTTTATTGAAATATCTTTTTTATNGGGGAAATAGAGGTTTNTCTATTCAAT

[0619] AAATAAATTTGGTGCTGGGAAAATATAGGATAGGCATTTCCACAATAATAATACATAC

[0620] CACTTTNCTANATGCAGATTTGAAAATTGAAACTTTATTTTGGAACCTTTNCTTNNTAC

[0621] ACCATATACAAAAATTAACTCAAGATGGATTAAAGACTTTAAAATTGTAAGACTTAATA

[0622] TCTNCAAACANCNTGAAAAAAATATGCTCCTTTATGATTTAACTANNTANAANCCAAC

[0623] CATAAAAATCCTAGAATATTTAGAAAACCTAGGCTAAAACCATTCAGGACATAGCAAA

[0624] NAGCATTGGGCAAAGANTTCTCTTGACTAAAAACACCAAAAGCAGNANCAAAAGCAA

[0625] TGGCAACAAACAGGCAAAAATTGACAAATGGGATTTTAAAATTAAACTAAAGAGCTTC

[0626] TGCACAGCAAAAGAAACTATCATAACAGAGTGAANAGACAACCTACAGAATGNCGAG

[0627] AGAAAATTTTTTANAAAATATTTGCAAACTATTCATGCTGACCCANCTAAAGGNCTAAT

[0628] ATCCAGCTTAAGAAAATCTATAATGAACTCAAACAAATTTACAATAAACCACCAAAAAA

[0629] AAACAACCCCATCAAAAAGTGGGCAAAGGATATGAATAGACACTTCTCAAAAGAAGT

[0630] GANATTGTTTCACATTTAGATNGTTATCCAAAAATCTACACATGAAAAAATGCTCATCA

[0631] TCACACTGGTCATCAGAGAAGAATAGANATCCAAAGAAAATGCAAATCAAAACCACA

[0632] ATGAGATACCATTCCTCACACCAGTTAGAATGGCTATAATTAAAGCAAAAAAAATAAC

[0633] CNACTATAAAAAAGTCAAGAAATAACAAATGCTGGAGAGGATGTGGAAGAAATAGGA

[0634] ACTCTCTTATACACTGTTGGTGGGAATTGTCAATGAAAACTAATATATCGTTACAACC

[0635] ATNTTNTGTGGAAAACAGTATGGCNATTCCTCAAAAAAATAAATAGAGAACTAAAAAT

[0636] ATGCGAATTACCATTTGACCCAGCAATCCCATTACTGGGTATNATATATACCCAAAGG

[0637] ATTATAAATCATTCTGCTCATCACTACTAAAAGAATATGAACAAATAAGCACACNTATG

[0638] TCTCTTATTGCAGCACTATTCACAATAGCCCAGNGCAAAGATATGGAATCAACCCAA ATGANTATTTTTATCCATCCAACAATGATAGANTGGTGATATAACGAAAATGTGGCAC

[0639] ACAAAAATACNTAAACACCATGGAATATTATTTCAGCCATAAAAAANAAAGTGAAATC

[0640] ATGCTCCTANTCACTTTGTGCAGGAAGATTACCCCACNNCATGGATTGAACTGGCTG

[0641] GAAAATGGTANGACTCATTAAAAAATTTCTTCAGTAAANTAAAAAACGCAGGAACAAA

[0642] AAAACCAAACACCGCTTCTTATGTTCTCACCAATACTCATATGTGGGAGCTAAACAAT

[0643] NGAGAACACATGGAATAATAAAAAACACANGAAGGGGAACATCACACACCGTGGGG

[0644] ACTGTTGGAAGGGTGGGGGGTGGGCAGGAGGGATAGTTCATTAGGAGAAATATACC

[0645] TAATGGGTATGGCAAAATGCAAAAATAGTTNTGTAAATGAANTGAGTTAGATGGGTG

[0646] CAGCANAACNACAACCTGAATANANTATACTTATNTTTGGCACATGTATACACATATG

[0647] TAACAAACCTGCACATTANGCACATGTACCAAAACCCTAAAACTTGATAAAAGTATAA

[0648] TAAAAAAAAAATATACTTTCCCGTGCTGTTATTATTCAGCCATCNNTTAACNTACCTAT

[0649] ACATNAAATNAACAATAAAAATAAATACAACTNTNATNTATNAATAAAAAT

[0650] SEQ ID NO: 11

[0651] TGTCCATCAACAGATGAATG

[0652] SEQ ID NO: 12

[0653] GGGTTTCTTTTTGGGGTGATG

[0654] SEQ ID NO: 13

[0655] AGTGGTGATGGTTGCACAAC

[0656] SEQ ID NO: 14

[0657] GTGACTGCTAATTGGAACAGGG

[0658] SEQ ID NO: 15

[0659] GAGAAGAAGAATTACTAGGAG

[0660] SEQ ID NO: 16

[0661] TACACTGTTGGTGGGAATTG

[0662] SEQ ID NO: 17

[0663] ATTCATTTTATGAGGCCAGC

[0664] SEQ ID NO: 18

[0665] AACAACCCAAATGTCCATCA

[0666] SEQ ID NO: 19

[0667] CC A ATG G A AT ACTATG C AG C

Claims

CLAIMS1 . An immune cell in which the expression of Long Interspersed Nuclear Element 1 (LINE1 or L1 ) is stably or transiently inhibited or downregulated, wherein said cell expresses at least one chimeric antigen receptor (CAR) or at least one artificial T-cell receptor (TCR) subunit.

2. The immune cell according to claim 1 , wherein the immune cell is selected from the group consisting of: a CAR-T cell, a CAR-NK cell, a CAR-macrophage and TCR-T cell.

3. The immune cell according to claim 1 or 2, wherein the immune cell is a CAR-T cell.

4. The immune cell according to anyone of claims 1 -3, wherein the expression of L1 is affected through an inhibitor or suppressor, said inhibitor being chosen from a polynucleotide and a small molecule.

5. The immune cell according to claim 4, wherein the polynucleotide is selected from the group consisting of: an antisense construct, antisense oligonucleotide, RNA interference construct or siRNA or a polynucleotide coding for it.

6. The immune cell according to claim 4 or 5, wherein the polynucleotide is an isolated inhibitory nucleic acid targeting L1 .

7. The immune cell according to any one of claims 4-6, wherein the polynucleotide is complementary to a sequence having at least 95% identity with a sequence selected from the group consisting of SEQ ID NOs: 9-10.

8. The immune cell according to any one of claims 4-7, wherein the polynucleotide comprises or consists of a sequence having at least 80% of identity, preferably at least 90% identity with a sequence selected from the group consisting of: SEQ ID NO: 1 1 -19.

9. The immune cell according to any one of claims 1 -8, wherein the cell is either autologous or allogenic or derived from an iPSC.

10. The immune cell according to any one of claims 1 -9, wherein the cell expresses at least one CAR targeting an antigen selected from: CD19, BCMA, CD22, CD20, CD138, CD33, CD123, PSMA.1 1 . The immune cell according to any one of claims 1 -10 for use as a medicament.

12. The immune cell according to any one of claims 1 -10 for use in the treatment or prevention or in a follow-up of a cancer.

13. The immune cell for use according to claim 12, wherein the cancer is a solid cancer.

14. The immune cell for use according to claim 12 or 13, wherein the cancer is a blood cancer.

15. The immune cell for use according to claim 14, wherein the blood cancer, is a leukaemia, preferably a B lineage leukaemia, or a lymphoma, preferably a non-Hodgkin lymphoma, or a myeloma, preferably a multiple myeloma.

16. An immune cell in which the expression of Long Interspersed Nuclear Element 1 (LINE1 or L1 ) is stably or transiently inhibited or downregulated, through a polynucleotide, wherein the polynucleotide comprises or consists of a sequence having at least 80% of identity with a sequence selected from the group consisting of: SEQ ID NO: 11 -19.

17. The immune cell according to claim 16, wherein the polynucleotide comprises or consists of a sequence having at least 90% of identity with a sequence selected from the group consisting of: SEQ ID NO: 11 -19.

18. An in vitro method for engineering an immune cell comprising at least one step of delivery of a nucleic acid inhibitor of LINE1 to said immune cell, wherein said immune cell expresses at least one chimeric antigen receptor (CAR) or at least one artificial T-cell receptor (TCR) subunit.

Images