A method for the treatment of a tumor with a CXCL12 antagonist and an interferon

Abstract

The present invention is related to a CXCL12 antagonist and an interferon for use in a method for treating a tumor in a subject, wherein the method comprises administering to the subject the CXCL12 antagonist and the interferon, wherein the tumor is an immunologically cold tumor or a tumor refractory to an immunotherapy, wherein preferably the tumor refractory to an immunotherapy is refractory to a checkpoint inhibitor.

Description

[0001] TME Pharma AG

[0002] A method for the treatment of a tumor

[0003] The present invention is related to a CXCL12 antagonist for use in a method for treating a tumor in a subject, wherein the tumor is an immunologically cold tumor; an interferon for use in a method for treating a tumor in a subject, wherein the tumor is an immunologically cold tumor; a CXCL12 antagonist for use in a method for treating a tumor in a subject, wherein the tumor is refractory to immunotherapy; an interferon for use in a method for treating a tumor in a subject, wherein the tumor is refractory to immunotherapy; a CXCL12 antagonist for use in a method for treating a tumor in a subject, wherein the tumor is a solid tumor; an interferon for use in a method for treating a tumor in a subject, wherein the tumor is a solid tumor; a CXCL12 antagonist for use in a method for treating a tumor in a subject, wherein the method comprises administering to the subject the CXCL12 antagonist, and an adjuvant therapy, wherein the tumor is an immunologically cold tumor; an interferon for use in a method for treating a tumor in a subject, wherein the method comprises administering to the subject the interferon, and an adjuvant therapy, wherein the tumor is an immunologically cold tumor; a CXCL12 antagonist for use in a method for treating a tumor in a subject, wherein the method comprises administering to the subject the CXCL12 antagonist, and an adjuvant therapy, wherein the tumor is refractory to immunotherapy; an interferon for use in a method for treating a tumor in a subject, wherein the method comprises administering to the subject the interferon, and an adjuvant therapy, wherein the tumor is refractory to immunotherapy; a CXCL12 antagonist for use in a method for treating a tumor in a subject, wherein the method comprises administering to the subject the CXCL12 antagonist, and an adjuvant therapy, wherein the tumor is a solid tumor; an interferon for use in a method for treating a tumor in a subject, wherein the method comprises administering to the subject the interferon, and an adjuvant therapy, wherein the tumor is a solid tumor; a CXCL12 antagonist for use in a method for treating a tumor in a subject, wherein the method comprises administering to the subject a primary therapy and an adjunct therapy, wherein the adjunct therapy comprises administering to the subject the CXCL12 antagonist, wherein the tumor is an immunologically cold tumor; an interferon for use in a method for treating a tumor in a subject, wherein the method comprises administering to the subject a primary therapy and an adjunct therapy, wherein the adjunct therapy comprises administering to the subject the interferon, wherein the tumor is an immunologically cold tumor; a CXCL12 antagonist for use in a method for treating a tumor in a subject, wherein the method comprises administering to the subject a primary therapy and an adjuncttherapy, wherein the adjunct therapy comprises administering to the subject the CXCL12 antagonist, wherein the tumor is refractory to immunotherapy; an interferon for use in a method for treating a tumor in a subject, wherein the method comprises administering to the subject a primary therapy and an adjunct therapy, wherein the adjunct therapy comprises administering to the subject the interferon, wherein the tumor is refractory to immunotherapy; a CXCL12 antagonist for use in a method for treating a tumor in a subject, wherein the method comprises administering to the subject a primary therapy and an adjunct therapy, wherein the adjunct therapy comprises administering to the subject the CXCL12 antagonist, wherein the tumor is a solid tumor; an interferon for use in a method for treating a tumor in a subject, wherein the method comprises administering to the subject a primary therapy and an adjunct therapy, wherein the adjunct therapy comprises administering to the subject the interferon, wherein the tumor is a solid tumor; a CXCL12 antagonist for use in a method for converting an immunologically cold tumor in a subject into an immunologically hot tumor; an interferon for use in a method for converting an immunologically cold tumor in a subject into an immunologically hot tumor in the subject; a CXCL12 antagonist for use in a method for converting an immunotherapy refractory tumor in a subject into an immunotherapy non-refractory tumor; an interferon for use in a method for converting an immunotherapy refractory tumor in a subject into an immunotherapy non-refractory tumor; and to such methods.

[0004] Cancer morbidity and mortality have continued to rise significantly over the past few years. In 2020, more than 19 million new malignant tumor patients were diagnosed worldwide, with 10 million cancer-related deaths. The cancer incidence rate increased significantly compared with 2018. In many countries, malignant tumors have become the leading cause of death (Sung, Ferlay et al. 2021). With the development of society and technological advancements, precision medicine has gradually replaced conventional radiotherapy (RT) and surgical treatment of malignant tumors (Mimezami, Nicholson et al. 2012, Garraway, Verweij et al. 2013, Collins and Varmus 2015, Rubin 2015, Tsimberidou, Fountzilas et al. 2020). Precision cancer therapy is currently the most promising cancer treatment, including targeted therapy and immunotherapy (Janiaud, Serghiou et al. 2019).

[0005] Immunotherapy is a therapeutic modality that aims to regulate and eradicate tumors by reactivating and sustaining the antitumor immune cycle, as well as reinstating the body’s innate antitumor immune response. It has become the fifth pillar of cancer treatment and mainly includes immune checkpoint inhibitors (ICIs) (Duan, Cui et al. 2020), adoptive cell transfer therapy (Newick, O'Brien et al. 2017, Ma, Li et al. 2019, Lin, Cheng et al. 2021), tumor-specific vaccines, cytokines (CKs), and smallmolecule immunological drugs. Of these, ICIs are currently the most widely used drugs in cancerimmunotherapy. Immunotherapy for tumors has been used for over 100 years, but the efficacy of immunotherapy varies even for patients with the same cancer type (Balkwill and Mantovani 2001).

[0006] CXCL12 expressed by healthy endothelial cells or stromal fibroblasts has long been understood to be a chemokine that potently attracts CXCR4-expressing lymphocytes in the settings of infection and thymic development (Bleul, Farzan et al. 1996, Janas, Varano et al. 2010, Trampont, Tosello-Trampont et al. 2010). In cancer, CXCL12 predominantly expressed by many tumors as well as associated fibroblasts, may play a variety of important roles in cancer progression as well as in the tumor microenvironment (Guo, Wang et al. 2016). Its cognate receptor, CXCR4, is broadly expressed by lymphocytes as well as other immune cells, stromal cells, myeloid cells and even neoplastic cells in some cases (Guyon 2014, Yang, Kumar et al. 2018). In some cancers where CXCL12 and CXCR4 are abnormally expressed, they have been seen directly to mediate tumor proliferation and inhibit apoptosis (Machelon, Gaudin et al. 2011). In others, CXCL12 / CXCR4 signaling has been implicated in metastasis. In models where immune check point inhibitors are applied to activate tumor infdtrating lymphocytes (TIL), high expression of CXCL12 by fibroblasts can thwart these therapies by shielding nests of cancers cells from the TIL without excluding TIL from the TME broadly. Inhibition of CXCL12 / CXCR4 signaling prompted deep infiltration of T cells in the TME in these settings, which was interpreted to indicate that the inhibitors were more efficacious against secreted CXCL12 than against CXCL12 on the cell surface (Feig, Jones et al. 2013, Fearon 2014, Zboralski, Hoehlig et al.

[0007] 2017). CXCR4 has been reported to be expressed in SS cells and examined as a potential target to slow tumor growth (Pazzaglia, Pollino et al. 2019).

[0008] A problem underlying the present invention is the provision of means and a method for treating a tumor in a subject, wherein the tumor is an immunologically cold tumor.

[0009] A further problem underlying the present invention is the provision of means and a method for treating a tumor in a subject, wherein the tumor is refractory to immunotherapy.

[0010] Another problem underlying the present invention is the provision of means and a method for treating a tumor in a subject, wherein the tumor is a solid tumor.

[0011] A problem underlying the present invention is the provision of means and a method for treating a tumor in a subject, wherein the method for treating a tumor comprises an adjuvant therapy and the tumor is an immunologically cold tumor.A further problem underlying the present invention is the provision of means and a method for treating a tumor in a subject, wherein the method for treating a tumor comprises an adjuvant therapy and the tumor is refractory to immunotherapy.

[0012] Another problem underlying the present invention is the provision of means and a method for treating a tumor in a subject, wherein the method for treating a tumor comprises an adjuvant therapy and the tumor is a solid tumor.

[0013] A problem underlying the present invention is the provision of means and a method for treating a tumor in a subject, wherein the method for treating a tumor comprises administering to the subject a primary therapy and an adjunct therapy, wherein the tumor is an immunologically cold tumor.

[0014] A further problem underlying the present invention is the provision of means and a method for treating a tumor in a subject, wherein the method for treating a tumor comprises administering to the subject a primary therapy and an adjunct therapy, wherein the tumor is refractory to immunotherapy.

[0015] Another problem underlying the present invention is the provision of means and a method for treating a tumor in a subject, wherein the method for treating a tumor comprises administering to the subject a primary therapy and an adjunct therapy, wherein the tumor is a solid tumor.

[0016] A problem underlying the present invention is the provision of means and a method for converting an immunologically cold tumor in a subject into an immunologically hot tumor.

[0017] Another problem underlying the present invention is the provision of means and a method for converting an immunotherapy refractory tumor in a subject into an immunotherapy non-refractory tumor.

[0018] These and other problems underlying the present invention are solved by the subject matter of the attached independent claims. Preferred embodiments may be taken from the attached dependent claims.

[0019] Similarly, these and other problems underlying the present invention are solved by the subject matter of the Embodiments disclosed in the following. Additionally, these and other problems underlying thepresent invention are solved by the subject matter of the various aspects of the present invention, including any embodiment thereof, as disclosed herein.

[0020] Embodiment 1. A CXCL12 antagonist for use in a method for treating a tumor in a subject, wherein the method comprises administering to the subject the CXCL12 antagonist and an interferon, wherein the tumor is an immunologically cold tumor.

[0021] Embodiment 2. An interferon for use in a method for treating a tumor in a subject, wherein the method comprises administering to the subject the interferon and a CXCL12 antagonist, wherein the tumor is an immunologically cold tumor.

[0022] Embodiment 3. The CXCL12 antagonist for use of Embodiment 1 and the interferon for use of Embodiment 2, wherein the tumor is refractory to an immunotherapy, preferably the immunotherapy is selected from the group comprising a checkpoint inhibitor, an adoptive cell therapy and a tumor vaccine.

[0023] Embodiment 4. The CXCL12 antagonist for use of Embodiment 3 and the interferon for use of Embodiment 3, wherein the checkpoint inhibitor is a PD-1 inhibitor, a PD-L1 inhibitor, a CTLA-4 inhibitor, a LAG-3 inhibitor, a TIM-3 inhibitor or a TIGIT inhibitor.

[0024] Embodiment 5. The CXCL12 antagonist for use of Embodiment 4 and the interferon for use of Embodiment 4, wherein the PD-1 inhibitor is selected from the group comprising pembrolizumab, nivolumab, cemiplimab, toripalimab, tislelizumab, dostarlimab, retfanlimab, sintilimab, camrelizumab, and balstilimab; wherein the PD-L1 inhibitor is selected from the group comprising atezolizumab, avelumab, durvalumab, cosibelimab, envafolimab, CS1001, PF-06801591; wherein the CTLA-4 inhibitor is selected from the group comprising ipilimumab, tremelimumab, BMS-986218, and zalifrelimab; wherein the LAG-3 inhibitor is selected from the group comprising relatlimab and GSK2831781; wherein the TIGIT inhibitor is selected from the group comprising vibostolimab, domvanalimab, tiragolumab and etigilimab; and wherein the TIM-3 inhibitor is selected from the group comprising sabatolimab, BMS-986258, and cobolimab.

[0025] Embodiment 6. The CXCL12 antagonist for use of any one of Embodiments 1 and 3 to 5 and the interferon for use of any one of Embodiments 2 to 5, wherein the tumor has at least one characteristic selected from the group comprising reduced or lacking expression of PD-L1, reducednumber of tumor-infiltrating lymphocytes, genomic stability and reduced or lacking preexisting antitumor immune response.

[0026] Embodiment 7. The CXCL12 antagonist for use of Embodiment 6 and the interferon for use of Embodiment 6, wherein the tumor-infiltrating lymphocytes are CD3+lymphocytes.

[0027] Embodiment 8. The CXCL12 antagonist for use of any one of Embodiments 1 and 3 to 7 and the interferon for use of any one of Embodiments 2 to 7, wherein the tumor has an immune evasion mechanism.

[0028] Embodiment 9. The CXCL12 antagonist for use of Embodiment 8 and the interferon for use of Embodiment 8, wherein the tumor is an immune-excluded tumor.

[0029] Embodiment 10. The CXCL12 antagonist for use of Embodiment 9 and the interferon for use of Embodiment 9, wherein immune exclusion can be reverted or mitigated by a checkpoint inhibitor.

[0030] Embodiment 11. The CXCL12 antagonist for use of Embodiment 10 and the interferon for use of Embodiment 10, wherein the checkpoint inhibitor is a PD-1 inhibitor, a PD-L1 inhibitor, a CTLA-4 inhibitor, a LAG-3 inhibitor, a TIM-3 inhibitor or a TIGIT inhibitor.

[0031] Embodiment 12. The CXCL12 antagonist for use of Embodiment 11 and the interferon for use of Embodiment 11, wherein the PD-1 inhibitor is selected from the group comprising pembrolizumab, nivolumab, cemiplimab, toripalimab, tislelizumab, dostarlimab, retfanlimab, sintilimab, camrelizumab, and balstilimab; wherein the PD-L1 inhibitor is selected from the group comprising atezolizumab, avelumab, durvalumab, cosibelimab, envafolimab, CS1001, PF-06801591; wherein the CTLA-4 inhibitor is selected from the group comprising ipilimumab, tremelimumab, BMS-986218, and zalifrelimab; wherein the LAG-3 inhibitor is selected from the group comprising relatlimab and GSK2831781; wherein the TIGIT inhibitor is selected from the group comprising vibostolimab, domvanalimab, tiragolumab and etigilimab; and wherein the TIM-3 inhibitor is selected from the group comprising sabatolimab, BMS-986258, and cobolimab.

[0032] Embodiment 13. The CXCL12 antagonist for use of Embodiment 8 and the interferon for use of Embodiment 8, wherein the tumor is an immune-desert tumor.Embodiment 14. The CXCL12 antagonist for use of any one of Embodiments 8 to 13 and the interferon for use of any one of Embodiments 8 to 13, wherein the immune evasion mechanism is one selected from the group comprising loss of MHC class I expression of malignant cells of the tumor and elevated expression of immune checkpoints by malignant and non-malignant cells of the tumor.

[0033] Embodiment 15. The CXCL12 antagonist for use of any one of Embodiments 8 to 14 and the interferon for use of any one of Embodiments 8 to 14, wherein the immune evasion mechanism results from, is caused by and / or is associated with a changed immune cell composition of the tumor microenvironment (TME).

[0034] Embodiment 16. The CXCL12 antagonist for use of Embodiment 15 and the interferon for use of Embodiment 15, wherein the tumor is evading from the subject’s adaptive immune system.

[0035] Embodiment 17. The CXCL12 antagonist for use of Embodiment 16 and the interferon for use of Embodiment 16, wherein the tumor is evading from the subject’s adaptive immune system by inhibiting infiltration of cytotoxic lymphocytes.

[0036] Embodiment 18. The CXCL12 antagonist for use of any one of Embodiments 16 and 17 and the interferon for use of any one of Embodiments 16 and 17, wherein the tumor is evading from the subject’s adaptive immune system by suppressing activity of cytotoxic tumor-infiltrating lymphocytes or inhibiting activation of cytotoxic tumor-infiltrating lymphocytes.

[0037] Embodiment 19. The CXCL12 antagonist for use of Embodiment 15 and the interferon for use of Embodiment 15, wherein the tumor is evading from the subject’s innate immune system.

[0038] Embodiment 20. The CXCL12 antagonist for use of Embodiment 19 and the interferon for use of Embodiment 19, wherein the tumor is characterized by an immunosuppressive tumor microenvironment.

[0039] Embodiment 21. The CXCL12 antagonist for use of Embodiment 20 and the interferon for use of Embodiment 20, wherein the immunosuppressive tumor microenvironment results from, is caused by and / or is associated with presence of a cell selected from the group comprising a myeloid-derived suppressor cell (MDSC), a tumor-associated macrophage (TAM) a tumor-associated neutrophil (TAN), and / or a regulatory T cell (Treg).Embodiment 22. The CXCL12 antagonist for use of Embodiment 21 and the interferon for use of Embodiment 21, wherein the cell is immunomodulatorily active.

[0040] Embodiment 23. The CXCL12 antagonist for use of Embodiment 22 and the interferon for use of Embodiment 22, wherein the immunomodulatory active cell is active by means of a secreted immunomodulator, metabolic reprogramming and / or regulatory cell-to-cell contact.

[0041] Embodiment 24. The CXCL12 antagonist for use of any one of Embodiments 1 and 3 to 23 and the interferon for use of any one of Embodiments 1 to 23, wherein the tumor is a solid tumor.

[0042] Embodiment 25. The CXCL12 antagonist for use of any one of Embodiments 1 and 3 to 24 and the interferon for use of any one of Embodiments 2 to 24, wherein the tumor is selected from the group comprising pancreatic cancer, preferably pancreatic ductal adenocarcinoma (PDAC); triplenegative breast cancer; prostate cancer; ovarian cancer; soft tissue sarcoma and osteosarcoma.

[0043] Embodiment 26. The CXCL12 antagonist for use of Embodiment 25 and the interferon for use of Embodiment 25, wherein the tumor is high-grade serous ovarian cancer.

[0044] Embodiment 27. The CXCL12 antagonist for use of Embodiment 25 and the interferon for use of Embodiment 25, wherein the tumor is advanced castration-resistant prostate cancer.

[0045] Embodiment 28. The CXCL12 antagonist for use of Embodiment 25 and the interferon for use of Embodiment 25, wherein the tumor is synovial sarcoma.

[0046] Embodiment 29. A CXCL12 antagonist for use in a method for treating a tumor in a subject, wherein the method comprises administering to the subject the CXCL12 antagonist and an interferon, wherein the tumor is refractory to immunotherapy.

[0047] Embodiment 30. An interferon for use in a method for treating a tumor in a subject, wherein the method comprises administering to the subject the interferon and a CXCL12 antagonist, wherein the tumor is refractory to immunotherapy.

[0048] Embodiment 31. The CXCL12 antagonist for use of Embodiment 29 and the interferon for use of Embodiment 30, wherein the tumor is refractory to an immunotherapy, preferably the immunotherapyis selected from the group comprising a checkpoint inhibitor, an adoptive cell therapy and a tumor vaccine.

[0049] Embodiment 32. The CXCL12 antagonist for use of Embodiment 31 and the interferon for use of Embodiment 31, wherein the checkpoint inhibitor is a PD-1 inhibitor, a PD-L1 inhibitor, a CTLA-4 inhibitor, a LAG-3 inhibitor, a TIM-3 inhibitor or a TIGIT inhibitor.

[0050] Embodiment 33. The CXCL12 antagonist for use of Embodiment 32 and the interferon for use of Embodiment 32, wherein the PD-1 inhibitor is selected from the group comprising pembrolizumab, nivolumab, cemiplimab, toripalimab, tislelizumab, dostarlimab, retfanlimab, sintilimab, camrelizumab, and balstilimab; wherein the PD-L1 inhibitor is selected from the group comprising atezolizumab, avelumab, durvalumab, cosibelimab, envafolimab, CS1001, PF-06801591; wherein the CTLA-4 inhibitor is selected from the group comprising ipilimumab, tremelimumab, BMS-986218, and zalifrelimab; wherein the LAG-3 inhibitor is selected from the group comprising relatlimab and GSK2831781; wherein the TIGIT inhibitor is selected from the group comprising vibostolimab, domvanalimab, tiragolumab and etigilimab; and wherein the TIM-3 inhibitor is selected from the group comprising sabatolimab, BMS-986258, and cobolimab..

[0051] Embodiment 34. The CXCL12 antagonist for use of any one of Embodiments 29 and 31 to 33 and the interferon for use of any one of Embodiments 30 to 33, wherein the tumor has at least one characteristic selected from the group comprising reduced or lacking expression of PD-L1, reduced number of tumor-infiltrating lymphocytes, genomic stability and reduced or lacking preexisting antitumor immune response.

[0052] Embodiment 35. The CXCL12 antagonist for use of Embodiment 34 and the interferon for use of Embodiment 34, wherein the tumor-infiltrating lymphocytes are CD3+lymphocytes.

[0053] Embodiment 36. The CXCL12 antagonist for use of any one of Embodiments 29 and 31 to 35 and the interferon for use of any one of Embodiments 30 to 35, wherein the tumor has an immune evasion mechanism.

[0054] Embodiment 37. The CXCL12 antagonist for use of Embodiment 36 and the interferon for use of Embodiment 35, wherein the tumor is an immune-excluded tumor.Embodiment 38. The CXCL12 antagonist for use of Embodiment 37 and the interferon for use of Embodiment 37, wherein immune exclusion can be reverted or mitigated by a checkpoint inhibitor.

[0055] Embodiment 39. The CXCL12 antagonist for use of Embodiment 38 and the interferon for use of Embodiment 38, wherein the checkpoint inhibitor is a PD-1 inhibitor, a PD-L1 inhibitor, a CTLA-4 inhibitor, a LAG-3 inhibitor, a TIM-3 inhibitor or a TIGIT inhibitor.

[0056] Embodiment 40. The CXCL12 antagonist for use of Embodiment 39 and the interferon for use of Embodiment 39, wherein the PD-1 inhibitor is selected from the group comprising pembrolizumab, nivolumab, cemiplimab, toripalimab, tislelizumab, dostarlimab, retfanlimab, sintilimab, camrelizumab, and balstilimab; wherein the PD-L1 inhibitor is selected from the group comprising atezolizumab, avelumab, durvalumab, cosibelimab, envafolimab, CS1001, PF-06801591; wherein the CTLA-4 inhibitor is selected from the group comprising ipilimumab, tremelimumab, BMS-986218, and zalifrelimab; wherein the LAG-3 inhibitor is selected from the group comprising relatlimab and GSK2831781; wherein the TIGIT inhibitor is selected from the group comprising vibostolimab, domvanalimab, tiragolumab and etigilimab; and wherein the TIM-3 inhibitor is selected from the group comprising sabatolimab, BMS-986258, and cobolimab.

[0057] Embodiment 41. The CXCL12 antagonist for use of Embodiment 36 and the interferon for use of Embodiment 36, wherein the tumor is an immune-desert tumor.

[0058] Embodiment 42. The CXCL12 antagonist for use of any one of Embodiments 36 to 41 and the interferon for use of any one of Embodiments 36 to 41, wherein the immune evasion mechanism is one selected from the group comprising loss of MHC class I expression of malignant cells of the tumor and elevated expression of immune checkpoints by malignant and non-malignant cells of the tumor.

[0059] Embodiment 43. The CXCL12 antagonist for use of any one of Embodiments 36 to 42 and the interferon for use of any one of Embodiments 36 to 42, wherein the immune evasion mechanism results from, is caused by and / or is associated with a changed immune cell composition of the tumor microenvironment (TME).

[0060] Embodiment 44. The CXCL12 antagonist for use of Embodiment 43 and the interferon for use of Embodiment 43, wherein the tumor evades from the subject’s adaptive immune system.Embodiment 45. The CXCL12 antagonist for use of Embodiment 44 and the interferon for use of Embodiment 44, wherein the tumor evades from the subject’s adaptive immune system by inhibiting infiltration of cytotoxic lymphocytes.

[0061] Embodiment 46. The CXCL12 antagonist for use of any one of Embodiments 44 and 45 and the interferon for use of any one of Embodiments 44 and 45, wherein the tumor is evading from the subject’s adaptive immune system by suppressing activity of cytotoxic tumor-infiltrating lymphocytes or inhibiting activation of cytotoxic tumor-infiltrating lymphocytes.

[0062] Embodiment 47. The CXCL12 antagonist for use of Embodiment 43 and the interferon for use of Embodiment 43, wherein the tumor is evading from the subject’s innate immune system.

[0063] Embodiment 48. The CXCL12 antagonist for use of Embodiment 47 and the interferon for use of Embodiment 47, wherein the tumor is characterized by an immunosuppressive tumor microenvironment.

[0064] Embodiment 49. The CXCL12 antagonist for use of Embodiment 48 and the interferon for use of Embodiment 48, wherein the immunosuppressive tumor microenvironment results from, is caused by and / or is associated with presence of a cell selected from the group comprising a myeloid-derived suppressor cell (MDSC), a tumor-associated macrophage (TAM), a tumor-associated neutrophil (TAN) and / or a regulatory T cell (Tregs).

[0065] Embodiment 50. The CXCL12 antagonist for use of Embodiment 49 and the interferon for use of Embodiment 49, wherein the cell is immunomodulatorily active.

[0066] Embodiment 51. The CXCL12 antagonist for use of Embodiment 50 and the interferon for use of Embodiment 50, wherein the immunomodulatorily active cell is active by means of a secreted immunomodulator, metabolic reprogramming and / or regulatory cell-to-cell contact.

[0067] Embodiment 52. The CXCL12 antagonist for use of any one of Embodiments 29 and 31 to 51 and the interferon for use of any one of Embodiments 30 to 51, wherein the tumor is an immunologically cold tumor.Embodiment 53. The CXCL12 antagonist for use of any one of Embodiments 29 and 31 to 52 and the interferon for use of any one of Embodiments 30 to 52, wherein the tumor is a solid tumor.

[0068] Embodiment 54. The CXCL12 antagonist for use of any one of Embodiments 29 and 31 to 53 and the interferon for use of any one of Embodiments 30 to 53, wherein the tumor is selected from the group comprising pancreatic cancer, preferably pancreatic ductal adenocarcinoma (PDAC); triplenegative breast cancer; prostate cancer; ovarian cancer; soft tissue sarcomas and osteosarcoma.

[0069] Embodiment 55. The CXCL12 antagonist for use of Embodiment 54 and the interferon for use of Embodiment 54, wherein the tumor is high-grade serous ovarian cancer.

[0070] Embodiment 56. The CXCL12 antagonist for use of Embodiment 54 and the interferon for use of Embodiment 54, wherein the tumor is advanced castration-resistant prostate cancer.

[0071] Embodiment 57. The CXCL12 antagonist for use of Embodiment 54 and the interferon for use of Embodiment 54, wherein the tumor is synovial sarcoma.

[0072] Embodiment 58. A CXCL12 antagonist for use in a method for treating a tumor in a subject, wherein the method comprises administering to the subject the CXCL12 antagonist and an interferon, wherein the tumor is a solid tumor.

[0073] Embodiment 59. An interferon for use in a method for treating a tumor in a subject, wherein the method comprises administering to the subject the interferon and a CXCL12 antagonist, wherein the tumor is a solid tumor.

[0074] Embodiment 60. The CXCL12 antagonist for use of Embodiment 58 and the interferon for use of Embodiment 59, wherein the tumor is selected from the group comprising pancreatic cancer, preferably pancreatic ductal adenocarcinoma (PDAC); triple-negative breast cancer; prostate cancer; ovarian cancer; soft tissue sarcomas and osteosarcoma.

[0075] Embodiment 61. The CXCL12 antagonist for use of Embodiment 60 and the interferon for use of Embodiment 60, wherein the tumor is high-grade serous ovarian cancer.Embodiment 62. The CXCL12 antagonist for use of Embodiment 60 and the interferon for use of Embodiment 60, wherein the tumor is advanced castration-resistant prostate cancer.

[0076] Embodiment 63. The CXCL12 antagonist for use of Embodiment 60 and the interferon for use of Embodiment 60, wherein the tumor is synovial sarcoma.

[0077] Embodiment 64. The CXCL12 antagonist for use of Embodiment 58 and the interferon for use of Embodiment 59, wherein the tumor is refractory to an immunotherapy, preferably the immunotherapy is selected from the group comprising a checkpoint inhibitor, an adoptive cell therapy and a tumor vaccine.

[0078] Embodiment 65. The CXCL12 antagonist for use of any one of Embodiments 58 and 60 to 64 and the interferon for use of any one of Embodiments 59 to 64, wherein the checkpoint inhibitor is is a PD-1 inhibitor, a PD-L1 inhibitor, a CTLA-4 inhibitor, a LAG-3 inhibitor, a TIM-3 inhibitor or a TIGIT inhibitor.

[0079] Embodiment 66. The CXCL12 antagonist for use of Embodiment 65 and the interferon for use of Embodiment 65, wherein the PD-1 inhibitor is selected from the group comprising pembrolizumab, nivolumab, cemiplimab, toripalimab, tislelizumab, dostarlimab, retfanlimab, sintilimab, camrelizumab, and balstilimab; wherein the PD-L1 inhibitor is selected from the group comprising atezolizumab, avelumab, durvalumab, cosibelimab, envafolimab, CS1001, PF-06801591; wherein the CTLA-4 inhibitor is selected from the group comprising ipilimumab, tremelimumab, BMS-986218, and zalifrelimab; wherein the LAG-3 inhibitor is selected from the group comprising relatlimab and GSK2831781; wherein the TIGIT inhibitor is selected from the group comprising vibostolimab, domvanalimab, tiragolumab and etigilimab; and wherein the TIM-3 inhibitor is selected from the group comprising sabatolimab, BMS-986258, and cobolimab.

[0080] Embodiment 67. The CXCL12 antagonist for use of any one of Embodiments 58 and 60 to 66 and the interferon for use of any one of Embodiments 59 to 66, wherein the tumor has at least one characteristic selected from the group comprising reduced or lacking expression of PD-L1, reduced number of tumor-infiltrating lymphocytes, genomic stability and reduced or lacking preexisting antitumor immune response.Embodiment 68. The CXCL12 antagonist for use of Embodiment 67 and the interferon for use of Embodiment 67, wherein the tumor-infiltrating lymphocytes are CD3+lymphocytes.

[0081] Embodiment 69. The CXCL12 antagonist for use of any one of Embodiments 58 and 60 to 68 and the interferon for use of any one of Embodiments 59 to 68, wherein the tumor has an immune evasion mechanism.

[0082] Embodiment 70. The CXCL12 antagonist for use of Embodiment 69 and the interferon for use of Embodiment 69, wherein the tumor is an immune-excluded tumor.

[0083] Embodiment 71. The CXCL12 antagonist for use of Embodiment 70 and the interferon for use of Embodiment 69, wherein immune exclusion can be reverted or mitigated by an immune checkpoint inhibitor.

[0084] Embodiment 72. The CXCL12 antagonist for use of Embodiment 71 and the interferon for use of Embodiment 71, wherein the checkpoint inhibitor is a PD-1 inhibitor, a PD-L1 inhibitor, a CTLA-4 inhibitor, a LAG-3 inhibitor, a TIM-3 inhibitor or a TIGIT inhibitor.

[0085] Embodiment 73. The CXCL12 antagonist for use of Embodiment 72 and the interferon for use of Embodiment 72, wherein the PD-1 inhibitor is selected from the group comprising pembrolizumab, nivolumab, cemiplimab, toripalimab, tislelizumab, dostarlimab, retfanlimab, sintilimab, camrelizumab, and balstilimab; wherein the PD-L1 inhibitor is selected from the group comprising atezolizumab, avelumab, durvalumab, cosibelimab, envafolimab, CS1001, PF-06801591; wherein the CTLA-4 inhibitor is selected from the group comprising ipilimumab, tremelimumab, BMS-986218, and zalifrelimab; wherein the LAG-3 inhibitor is selected from the group comprising relatlimab and GSK2831781; wherein the TIGIT inhibitor is selected from the group comprising vibostolimab, domvanalimab, tiragolumab and etigilimab; and wherein the TIM-3 inhibitor is selected from the group comprising sabatolimab, BMS-986258, and cobolimab.

[0086] Embodiment 74. The CXCL12 antagonist for use of Embodiment 69 and the interferon for use of Embodiment 69, wherein the tumor is an immune-desert tumor.

[0087] Embodiment 75. The CXCL12 antagonist for use of any one of Embodiments 69 to 74 and the interferon for use of any one of Embodiments 69 to 74, wherein the immune evasion mechanism isone selected from the group comprising loss of MHC class I expression of malignant cells of the tumor and elevated expression of immune checkpoints by malignant and non-malignant cells of the tumor.

[0088] Embodiment 76. The CXCL12 antagonist for use of any one of Embodiments 69 to 75 and the interferon for use of any one of Embodiments 69 to 75, wherein the immune evasion mechanism results from, is caused by and / or is associated with a changed immune cell composition of the tumor microenvironment (TME).

[0089] Embodiment 77. The CXCL12 antagonist for use of Embodiment 76 and the interferon for use of Embodiment 76, wherein the tumor is evading from the subject’s adaptive immune system.

[0090] Embodiment 78. The CXCL12 antagonist for use of Embodiment 77 and the interferon for use of Embodiment 77, wherein the tumor is evading from the subject’s adaptive immune system by inhibiting infiltration of cytotoxic lymphocytes.

[0091] Embodiment 79. The CXCL12 antagonist for use of any one of Embodiments 77 and 78 and the interferon for use of any one of Embodiments 77 and 78, wherein the tumor is evading from the subject’s adaptive immune system by suppressing activity of cytotoxic tumor-infiltrating lymphocytes or inhibiting activation of cytotoxic tumor-infiltrating lymphocytes.

[0092] Embodiment 80. The CXCL12 antagonist for use of Embodiment 76 and the interferon for use of Embodiment 76, wherein the tumor is evading from the subject’s innate immune system.

[0093] Embodiment 81. The CXCL12 antagonist for use of Embodiment 80 and the interferon for use of Embodiment 80, wherein the tumor is characterized by an immunosuppressive tumor microenvironment.

[0094] Embodiment 82. The CXCL12 antagonist for use of Embodiment 81 and the interferon for use of Embodiment 81, wherein the immunosuppressive tumor microenvironment results from, is caused by and / or is associated with presence of a cell selected from the group comprising a myeloid-derived suppressor cell (MDSC), a tumor-associated macrophage (TAM), a tumor-associated neutrophil (TAN) and / or a regulatory T cell (Treg).Embodiment 83. The CXCL12 antagonist for use of Embodiment 82 and the interferon for use of Embodiment 82, wherein the cell is immunomodulatorily active.

[0095] Embodiment 84. The CXCL12 antagonist for use of Embodiment 83 and the interferon for use of Embodiment 83, wherein the immunomodulatory active cell is active by means of a secreted immunomodulator, metabolic reprogramming and / or regulatory cell-to-cell contact.

[0096] Embodiment 85. A CXCL12 antagonist for use in a method for treating a tumor in a subject, wherein the method comprises administering to the subject the CXCL12 antagonist, an interferon and an adjuvant therapy, wherein the tumor is an immunologically cold tumor.

[0097] Embodiment 86. An interferon for use in a method for treating a tumor in a subject, wherein the method comprises administering to the subject the interferon, a CXCL12 antagonist and an adjuvant therapy, wherein the tumor is an immunologically cold tumor.

[0098] Embodiment 87. The CXCL12 antagonist for use of Embodiment 85 and the interferon for use of Embodiment 86, wherein the tumor is refractory to an immunotherapy, preferably the immunotherapy is selected from the group comprising a checkpoint inhibitor, an adoptive cell therapy and a tumor vaccine.

[0099] Embodiment 88. The CXCL12 antagonist for use of Embodiment 87 and the interferon for use of Embodiment 87, wherein the checkpoint inhibitor is a PD-1 inhibitor, a PD-L1 inhibitor, a CTLA-4 inhibitor, a LAG-3 inhibitor, a TIM-3 inhibitor or a TIGIT inhibitor.

[0100] Embodiment 89. The CXCL12 antagonist for use of Embodiment 88 and the interferon for use of Embodiment 88, wherein wherein the PD-1 inhibitor is selected from the group comprising pembrolizumab, nivolumab, cemiplimab, toripalimab, tislelizumab, dostarlimab, retfanlimab, sintilimab, camrelizumab, and balstilimab; wherein the PD-L1 inhibitor is selected from the group comprising atezolizumab, avelumab, durvalumab, cosibelimab, envafolimab, CS1001, PF-06801591; wherein the CTLA-4 inhibitor is selected from the group comprising ipilimumab, tremelimumab, BMS-986218, and zalifrelimab; wherein the LAG-3 inhibitor is selected from the group comprising relatlimab and GSK2831781; wherein the TIGIT inhibitor is selected from the group comprising vibostolimab, domvanalimab, tiragolumab and etigilimab; and wherein the TIM-3 inhibitor is selected from the group comprising sabatolimab, BMS-986258, and cobolimab.Embodiment 90. The CXCL12 antagonist for use of any one of Embodiments 85 and 87 to 89 and the interferon for use of any one of Embodiments 86 to 89, wherein the tumor has at least one characteristic selected from the group comprising reduced or lacking expression of PD-L1, reduced number of tumor-infiltrating lymphocytes, genomic stability and reduced or lacking preexisting antitumor immune response.

[0101] Embodiment 91. The CXCL12 antagonist for use of Embodiment 90 and the interferon for use of Embodiment 90 wherein the tumor-infiltrating lymphocytes are CD3+lymphocytes.

[0102] Embodiment 92. The CXCL12 antagonist for use of any one of Embodiments 85 and 87 to 91 and the interferon for use of any one of Embodiments 86 to 91, wherein the tumor has an immune evasion mechanism.

[0103] Embodiment 93. The CXCL12 antagonist for use of Embodiment 93 and the interferon for use of Embodiment 92, wherein the tumor is an immune-excluded tumor.

[0104] Embodiment 94. The CXCL12 antagonist for use of Embodiment 93 and the interferon for use of Embodiment 913 wherein immune exclusion can be reverted or mitigated by an immune checkpoint inhibitor.

[0105] Embodiment 95. The CXCL12 antagonist for use of Embodiment 94 and the interferon for use of Embodiment 94, wherein the checkpoint inhibitor is a PD-1 inhibitor, a PD-L1 inhibitor, a CTLA-4 inhibitor, a LAG-3 inhibitor, a TIM-3 inhibitor or a TIGIT inhibitor.

[0106] Embodiment 96. The CXCL12 antagonist for use of Embodiment 95 and the interferon for use of Embodiment 95, wherein the PD-1 inhibitor is selected from the group comprising pembrolizumab, nivolumab, cemiplimab, toripalimab, tislelizumab, dostarlimab, retfanlimab, sintilimab, camrelizumab, and balstilimab; wherein the PD-L1 inhibitor is selected from the group comprising atezolizumab, avelumab, durvalumab, cosibelimab, envafolimab, CS1001, PF-06801591; wherein the CTLA-4 inhibitor is selected from the group comprising ipilimumab, tremelimumab, BMS-986218, and zalifrelimab; wherein the LAG-3 inhibitor is selected from the group comprising relatlimab and GSK2831781; wherein the TIGIT inhibitor is selected from the group comprising vibostolimab, domvanalimab, tiragolumab and etigilimab; and wherein the TIM-3 inhibitor is selected from the group comprising sabatolimab, BMS-986258, and cobolimab.Embodiment 97. The CXCL12 antagonist for use of Embodiment 92 and the interferon for use of Embodiment 92, wherein the tumor is an immune-desert tumor.

[0107] Embodiment 98. The CXCL12 antagonist for use of any one of Embodiments 92 to 97 and the interferon for use of any one of Embodiments 92 to 97, wherein the immune evasion mechanism is one selected from the group comprising loss of MHC class I expression of malignant cells of the tumor and elevated expression of immune checkpoints by malignant and non-malignant cells of the tumor.

[0108] Embodiment 99. The CXCL12 antagonist for use of any one of Embodiments 92 to 98 and the interferon for use of any one of Embodiments 92 to 98, wherein the immune evasion mechanism results from, is caused by and / or is associated with a changed immune cell composition of the tumor microenvironment (TME).

[0109] Embodiment 100. The CXCL12 antagonist for use of Embodiment 99 and the interferon for use of Embodiment 99, wherein the tumor evades from the subject’s adaptive immune system.

[0110] Embodiment 101. The CXCL12 antagonist for use of Embodiment 100 and the interferon for use of Embodiment 100, wherein the tumor evades from the subject’s adaptive immune system by inhibiting infiltration of cytotoxic lymphocytes.

[0111] Embodiment 102. The CXCL12 antagonist for use of any one of Embodiments 100 and 101 and the interferon for use of any one of Embodiments 100 and 101, wherein the tumor evades from the subject’s adaptive immune system by suppressing activity of cytotoxic tumor-infiltrating lymphocytes or inhibiting activation of cytotoxic tumor-infiltrating lymphocytes.

[0112] Embodiment 103. The CXCL12 antagonist for use of Embodiment 99 and the interferon for use of Embodiment 95, wherein the tumor evades from the subject’s innate immune system.

[0113] Embodiment 104. The CXCL12 antagonist for use of Embodiment 103 and the interferon for use of Embodiment 103, wherein the tumor is characterized by an immunosuppressive tumor microenvironment.Embodiment 105. The CXCL12 antagonist for use of Embodiment 104 and the interferon for use of Embodiment 104, wherein the immunosuppressive tumor microenvironment results from, is caused by and / or is associated with presence of a cell selected from the group comprising a myeloid-derived suppressor cell (MDSC), a tumor-associated macrophage (TAM), a tumor-associated neutrophil (TAN) and / or a regulatory T cell (Treg).

[0114] Embodiment 106. The CXCL12 antagonist for use of Embodiment 105 and the interferon for use of Embodiment 105, wherein the cell is immunomodulatorily active.

[0115] Embodiment 107. The CXCL12 antagonist for use of Embodiment 106 and the interferon for use of Embodiment 106, wherein the immunomodulatorily active cell is active by means of a secreted immunomodulator, metabolic reprogramming and / or regulatory cell-to-cell contact.

[0116] Embodiment 108. The CXCL12 antagonist for use of any one of Embodiments 85 and 87 to 107 and the interferon for use of any one of Embodiments 86 to 107, wherein the tumor is a solid tumor.

[0117] Embodiment 109. The CXCL12 antagonist for use of any one of Embodiments 85 and 87 to 108 and the interferon for use of any one of Embodiments 86 to 108, wherein the tumor is selected from the group comprising pancreatic cancer preferably pancreatic ductal adenocarcinoma (PDAC); triplenegative breast cancer; prostate cancer; ovarian cancer; soft tissue sarcoma and osteosarcoma.

[0118] Embodiment 110. The CXCL12 antagonist for use of Embodiment 109 and the interferon for use of Embodiment 109, wherein the tumor is high-grade serous ovarian cancer.

[0119] Embodiment 111. The CXCL12 antagonist for use of Embodiment 110 and the interferon for use of Embodiment 110, wherein the tumor is advanced castration-resistant prostate cancer.

[0120] Embodiment 112. The CXCL12 antagonist for use of Embodiment 110 and the interferon for use of Embodiment 110, wherein the tumor is synovial sarcoma.

[0121] Embodiment 113. A CXCL12 antagonist for use in a method for treating a tumor in a subject, wherein the method comprises administering to the subject the CXCL12 antagonist, an interferon and adjuvant therapy, wherein the tumor is refractory to immunotherapy.Embodiment 114. An interferon for use in a method for treating a tumor in a subject, wherein the method comprises administering to the subject the interferon, a CXCL12 antagonist and an adjuvant therapy, wherein the tumor is refractory to immunotherapy.

[0122] Embodiment 115. The CXCL12 antagonist for use of Embodiment 113 and the interferon for use of Embodiment 114, wherein the tumor is refractory to an immunotherapy, preferably the immunotherapy is selected from the group comprising a checkpoint inhibitor, an adoptive cell therapy and a tumor vaccine.

[0123] Embodiment 116. The CXCL12 antagonist for use of Embodiment 115 and the interferon for use of Embodiment 115, wherein the checkpoint inhibitor is a PD-1 inhibitor, a PD-L1 inhibitor, a CTLA-4 inhibitor, a LAG-3 inhibitor, a TIM-3 inhibitor or a TIGIT inhibitor.

[0124] Embodiment 117. The CXCL12 antagonist for use of Embodiment 116 and the interferon for use of Embodiment 116, wherein the PD-1 inhibitor is selected from the group comprising pembrolizumab, nivolumab, cemiplimab, toripalimab, tislelizumab, dostarlimab, retfanlimab, sintilimab, camrelizumab, and balstilimab; wherein the PD-L1 inhibitor is selected from the group comprising atezolizumab, avelumab, durvalumab, cosibelimab, envafolimab, CS1001, PF-06801591; wherein the CTLA-4 inhibitor is selected from the group comprising ipilimumab, tremelimumab, BMS-986218, and zalifrelimab; wherein the LAG-3 inhibitor is selected from the group comprising relatlimab and GSK2831781; wherein the TIGIT inhibitor is selected from the group comprising vibostolimab, domvanalimab, tiragolumab and etigilimab; and wherein the TIM-3 inhibitor is selected from the group comprising sabatolimab, BMS-986258, and cobolimab.

[0125] Embodiment 118. The CXCL12 antagonist for use of any one of Embodiments 113 and 115 to 117 and the interferon for use of any one of Embodiments 114 to 117, wherein the tumor has at least one characteristic selected from the group comprising reduced or lacking expression of PD-L1, reduced number of tumor-infiltrating lymphocytes, genomic stability and reduced or lacking preexisting antitumor immune response.

[0126] Embodiment 119. The CXCL12 antagonist for use of Embodiment 118 and the interferon for use of Embodiment 118, wherein the tumor-infiltrating lymphocytes are CD3+lymphocytes.Embodiment 120. The CXCL12 antagonist for use of any one of Embodiments 113 and 115 to 119 and the interferon for use of any one of Embodiments 114 to 119, wherein the tumor has an immune evasion mechanism.

[0127] Embodiment 121. The CXCL12 antagonist for use of Embodiment 120 and the interferon for use of Embodiment 120, wherein the tumor is an immune-excluded tumor.

[0128] Embodiment 122. The CXCL12 antagonist for use of Embodiment 121 and the interferon for use of Embodiment 121, wherein immune exclusion can be reverted or mitigated by an immune checkpoint inhibitor.

[0129] Embodiment 123. The CXCL12 antagonist for use of Embodiment 122 and the interferon for use of Embodiment 122, wherein the checkpoint inhibitor is a PD-1 inhibitor, a PD-L1 inhibitor, a CTLA-4 inhibitor, a LAG-3 inhibitor, a TIM-3 inhibitor or a TIGIT inhibitor.

[0130] Embodiment 124. The CXCL12 antagonist for use of Embodiment 123 and the interferon for use of Embodiment 123, wherein the PD-1 inhibitor is selected from the group comprising pembrolizumab, nivolumab, cemiplimab, toripalimab, tislelizumab, dostarlimab, retfanlimab, sintilimab, camrelizumab, and balstilimab; wherein the PD-L1 inhibitor is selected from the group comprising atezolizumab, avelumab, durvalumab, cosibelimab, envafolimab, CS1001, PF-06801591; wherein the CTLA-4 inhibitor is selected from the group comprising ipilimumab, tremelimumab, BMS-986218, and zalifrelimab; wherein the LAG-3 inhibitor is selected from the group comprising relatlimab and GSK2831781; wherein the TIGIT inhibitor is selected from the group comprising vibostolimab, domvanalimab, tiragolumab and etigilimab; and wherein the TIM-3 inhibitor is selected from the group comprising sabatolimab, BMS-986258, and cobolimab.

[0131] Embodiment 125. The CXCL12 antagonist for use of Embodiment 120 and the interferon for use of Embodiment 120, wherein the tumor is an immune-desert tumor.

[0132] Embodiment 126. The CXCL12 antagonist for use of any one of Embodiments 120 to 125 and the interferon for use of any one of Embodiments 120 to 125, wherein the immune evasion mechanism is one selected from the group comprising loss of MHC class I expression of malignant cells of the tumor and elevated expression of immune checkpoints by malignant and non-malignant cells of the tumor.Embodiment 127. The CXCL12 antagonist for use of any one of Embodiments 120 to 126 and the interferon for use of any one of Embodiments 120 to 126, wherein the immune evasion mechanism results from, is caused by and / or is associated with a changed immune cell composition of the tumor microenvironment (TME).

[0133] Embodiment 128. The CXCL12 antagonist for use of Embodiment 127 and the interferon for use of Embodiment 127, wherein the tumor evades from the subject’s adaptive immune system.

[0134] Embodiment 129. The CXCL12 antagonist for use of Embodiment 128 and the interferon for use of Embodiment 128, wherein the tumor evades from the subject’s adaptive immune system by inhibiting infiltration of cytotoxic lymphocytes.

[0135] Embodiment 130. The CXCL12 antagonist for use of any one of Embodiments 128 and 129 and the interferon for use of any one of Embodiments 128 and 129, wherein the tumor is evading from the subject’s adaptive immune system by suppressing activity of cytotoxic tumor-infiltrating lymphocytes or inhibiting activation of cytotoxic tumor-infiltrating lymphocytes.

[0136] Embodiment 131. The CXCL12 antagonist for use of Embodiment 127 and the interferon for use of Embodiment 127, wherein the tumor is evading from the subject’s innate immune system.

[0137] Embodiment 132. The CXCL12 antagonist for use of Embodiment 131 and the interferon for use of Embodiment 131, wherein the tumor is characterized by an immunosuppressive tumor microenvironment.

[0138] Embodiment 133. The CXCL12 antagonist for use of Embodiment 132 and the interferon for use of Embodiment 132, wherein the immunosuppressive tumor microenvironment results from, is caused by and / or is associated with presence of a cell selected from the group comprising a myeloid-derived suppressor cell (MDSC), a tumor-associated macrophage (TAM), a tumor-associated neutrophil (TAN), and / or a regulatory T cell (Treg).

[0139] Embodiment 134. The CXCL12 antagonist for use of Embodiment 133 and the interferon for use of Embodiment 133, wherein the cell is immunomodulatorily active.Embodiment 135. The CXCL12 antagonist for use of Embodiment 134 and the interferon for use of Embodiment 134, wherein the immunomodulatorily active cell is active by means of a secreted immunomodulator, metabolic reprogramming and / or regulatory cell-to-cell contact.

[0140] Embodiment 136. The CXCL12 antagonist for use of any one of Embodiments 113 and 115 to 135 and the interferon for use of any one of Embodiments 114 to 135, wherein the tumor is an immunologically cold tumor.

[0141] Embodiment 137. The CXCL12 antagonist for use of any one of Embodiments 113 and 115 to 136 and the interferon for use of any one of Embodiments 114 to 136, wherein the tumor is a solid tumor.

[0142] Embodiment 138. The CXCL12 antagonist for use of any one of Embodiments 113 and 115 to 137 and the interferon for use of any one of Embodiments 114 to 137, wherein the tumor is selected from the group comprising pancreatic cancer, preferably pancreatic ductal adenocarcinoma (PDAC); triplenegative breast cancer; prostate cancer; ovarian cancer; soft tissue sarcoma and osteosarcoma.

[0143] Embodiment 139. The CXCL12 antagonist for use of Embodiment 138 and the interferon for use of Embodiment 138, wherein the tumor is high-grade serous ovarian cancer.

[0144] Embodiment 140. The CXCL12 antagonist for use of Embodiment 138 and the interferon for use of Embodiment 138, wherein the tumor is advanced castration-resistant prostate cancer.

[0145] Embodiment 141. The CXCL12 antagonist for use of Embodiment 138 and the interferon for use of Embodiment 138, wherein the tumor is synovial sarcoma.

[0146] Embodiment 142. A CXCL12 antagonist for use in a method for treating a tumor in a subject, wherein the method comprises administering to the subject the CXCL12 antagonist, an interferon and an adjuvant therapy, wherein the tumor is a solid tumor.

[0147] Embodiment 143. An interferon for use in a method for treating a tumor in a subject, wherein the method comprises administering to the subject the interferon, a CXCL12 antagonist and an adjuvant therapy, wherein the tumor is a solid tumor.Embodiment 144. The CXCL12 antagonist for use of Embodiment 142 and the interferon for use of Embodiment 143, wherein the tumor is selected from the group comprising pancreatic cancer preferably pancreatic ductal adenocarcinoma (PDAC); triple-negative breast cancer; prostate cancer; ovarian cancer; soft tissue sarcoma and osteosarcoma.

[0148] Embodiment 145. The CXCL12 antagonist for use of Embodiment 144 and the interferon for use of Embodiment 144, wherein the tumor is high-grade serous ovarian cancer.

[0149] Embodiment 146. The CXCL12 antagonist for use of Embodiment 144 and the interferon for use of Embodiment 144, wherein the tumor is advanced castration-resistant prostate cancer.

[0150] Embodiment 147. The CXCL12 antagonist for use of Embodiment 144 and the interferon for use of Embodiment 144, wherein the tumor is synovial sarcoma.

[0151] Embodiment 148. The CXCL12 antagonist for use of any one of Embodiments 142 and 144 to 147 and the interferon for use of any one of Embodiments 142 to 147, wherein the tumor is refractory to an immunotherapy, preferably the immunotherapy is selected from the group comprising a checkpoint inhibitor, an adoptive cell therapy and a tumor vaccine.

[0152] Embodiment 149. The CXCL12 antagonist for use of any one of Embodiments 142 and 144 to 148 and the interferon for use of any one of Embodiments 143 to 148, wherein the checkpoint inhibitor is a PD-1 inhibitor, a PD-L1 inhibitor, a CTLA-4 inhibitor, a LAG-3 inhibitor, a TIM-3 inhibitor or a TIGIT inhibitor.

[0153] Embodiment 150. The CXCL12 antagonist for use of Embodiment 149 and the interferon for use of Embodiment 149, wherein the PD-1 inhibitor is selected from the group comprising pembrolizumab, nivolumab, cemiplimab, toripalimab, tislelizumab, dostarlimab, retfanlimab, sintilimab, camrelizumab, and balstilimab; wherein the PD-L1 inhibitor is selected from the group comprising atezolizumab, avelumab, durvalumab, cosibelimab, envafolimab, CS1001, PF-06801591; wherein the CTLA-4 inhibitor is selected from the group comprising ipilimumab, tremelimumab, BMS-986218, and zalifrelimab; wherein the LAG-3 inhibitor is selected from the group comprising relatlimab and GSK2831781; wherein the TIGIT inhibitor is selected from the group comprising vibostolimab, domvanalimab, tiragolumab and etigilimab; and wherein the TIM-3 inhibitor is selected from the group comprising sabatolimab, BMS-986258, and cobolimab.Embodiment 151. The CXCL12 antagonist for use of any one of Embodiments 142 and 144 to 150 and the interferon for use of any one of Embodiments 143 to 150, wherein the tumor has at least one characteristic selected from the group comprising reduced or lacking expression of PD-L1, reduced number of tumor-infiltrating lymphocytes, genomic stability and reduced or lacking preexisting antitumor immune response.

[0154] Embodiment 152. The CXCL12 antagonist for use of Embodiment 151 and the interferon for use of Embodiment 151, wherein the tumor-infiltrating lymphocytes are CD3+lymphocytes.Embodiment 153. The CXCL12 antagonist for use of any one of Embodiments 142 and 144 to 152 and the interferon for use of any one of Embodiments 143 to 152, wherein the tumor has an immune evasion mechanism.

[0155] Embodiment 154. The CXCL12 antagonist for use of Embodiment 153 and the interferon for use of Embodiment 153, wherein the tumor is an immune-excluded tumor.

[0156] Embodiment 155. The CXCL12 antagonist for use of Embodiment 154 and the interferon for use of Embodiment 154, wherein immune exclusion can be reverted or mitigated by an immune checkpoint inhibitor.

[0157] Embodiment 156. The CXCL12 antagonist for use of Embodiment 155 and the interferon for use of Embodiment 155, wherein the checkpoint inhibitor is a PD-1 inhibitor, a PD-L1 inhibitor, a CTLA-4 inhibitor, a LAG-3 inhibitor, a TIM-3 inhibitor or a TIGIT inhibitor.

[0158] Embodiment 157. The CXCL12 antagonist for use of Embodiment 156 and the interferon for use of Embodiment 156, wherein the PD-1 inhibitor is selected from the group comprising pembrolizumab, nivolumab, cemiplimab, toripalimab, tislelizumab, dostarlimab, retfanlimab, sintilimab, camrelizumab, and balstilimab; wherein the PD-L1 inhibitor is selected from the group comprising atezolizumab, avelumab, durvalumab, cosibelimab, envafolimab, CS1001, PF-06801591; wherein the CTLA-4 inhibitor is selected from the group comprising ipilimumab, tremelimumab, BMS-986218, and zalifrelimab; wherein the LAG-3 inhibitor is selected from the group comprising relatlimab and GSK2831781; wherein the TIGIT inhibitor is selected from the group comprising vibostolimab, domvanalimab, tiragolumab and etigilimab; and wherein the TIM-3 inhibitor is selected from the group comprising sabatolimab, BMS-986258, and cobolimab.Embodiment 158. The CXCL12 antagonist for use of Embodiment 153 and the interferon for use of Embodiment 153, wherein the tumor is an immune-desert tumor.

[0159] Embodiment 159. The CXCL12 antagonist for use of any one of Embodiments 153 to 158 and the interferon for use of any one of Embodiments 153 to 158, wherein the immune evasion mechanism is one selected from the group comprising loss of MHC class I expression of malignant cells of the tumor and elevated expression of immune checkpoints by malignant and non-malignant cells of the tumor.

[0160] Embodiment 160. The CXCL12 antagonist for use of any one of Embodiments 153 to 159 and the interferon for use of any one of Embodiments 153 to 159, wherein the immune evasion mechanism results from, is caused by and / or is associated with a changed immune cell composition of the tumor microenvironment (TME).

[0161] Embodiment 161. The CXCL12 antagonist for use of Embodiment 160 and the interferon for use of Embodiment 160, wherein the tumor is evading from the subject’s adaptive immune system.

[0162] Embodiment 162. The CXCL12 antagonist for use of Embodiment 161 and the interferon for use of Embodiment 161, wherein the tumor is evading from the subject’s adaptive immune system by inhibiting infiltration of cytotoxic lymphocytes.

[0163] Embodiment 163. The CXCL12 antagonist for use of any one of Embodiments 161 and 162 and the interferon for use of any one of Embodiments 161 and 162, wherein the tumor is evading from the subject’s adaptive immune system by suppressing activity of cytotoxic tumor-infiltrating lymphocytes or inhibiting activation of cytotoxic tumor-infiltrating lymphocytes.

[0164] Embodiment 164. The CXCL12 antagonist for use of Embodiment 160 and the interferon for use of Embodiment 160, wherein the tumor is evading from the subject’s innate immune system.

[0165] Embodiment 165. The CXCL12 antagonist for use of Embodiment 164 and the interferon for use of Embodiment 164, wherein the tumor is characterized by an immunosuppressive tumor microenvironment.Embodiment 166. The CXCL12 antagonist for use of Embodiment 165 and the interferon for use of Embodiment 165, wherein the immunosuppressive tumor microenvironment results from, is caused by and / or is associated with presence of a cell selected from the group comprising a myeloid-derived suppressor cell (MDSC), a tumor-associated macrophage (TAM), a tumor-associated neutrophil (TAN), and / or a regulatory T cell (Treg).

[0166] Embodiment 167. The CXCL12 antagonist for use of Embodiment 166 and the interferon for use of Embodiment 166, wherein the cell is immunomodulatorily active.

[0167] Embodiment 168. The CXCL12 antagonist for use of Embodiment 167 and the interferon for use of Embodiment 167, wherein the immunomodulatory active cell is active by means of a secreted immunomodulator, metabolic reprogramming and / or regulatory cell-to-cell contact.

[0168] Embodiment 169. A CXCL12 antagonist for use in a method for treating a tumor in a subject, wherein the method comprises administering to the subject a primary therapy and an adjunct therapy, wherein the adjunct therapy comprises administering to the subject the CXCL12 antagonist and an interferon, wherein the tumor is an immunologically cold tumor.

[0169] Embodiment 170. An interferon for use in a method for treating a tumor in a subject, wherein the method comprises administering to the subject a primary therapy and an adjunct therapy, wherein the adjunct therapy comprises administering to the subject the interferon and a CXCL12 antagonist, wherein the tumor is an immunologically cold tumor.

[0170] Embodiment 171. The CXCL12 antagonist for use of Embodiment 169 and the interferon for use of Embodiment 170, wherein the tumor is refractory to an immunotherapy, preferably the immunotherapy is selected from the group comprising a checkpoint inhibitor, an adoptive cell therapy and a tumor vaccine.

[0171] Embodiment 172. The CXCL12 antagonist for use of Embodiment 171 and the interferon for use of Embodiment 171, wherein the checkpoint inhibitor is a PD-1 inhibitor, a PD-L1 inhibitor, a CTLA-4 inhibitor, a LAG-3 inhibitor, a TIM-3 inhibitor or a TIGIT inhibitor.

[0172] Embodiment 173. The CXCL12 antagonist for use of Embodiment 172 and the interferon for use of Embodiment 172, wherein the PD-1 inhibitor is selected from the group comprisingpembrolizumab, nivolumab, cemiplimab, toripalimab, tislelizumab, dostarlimab, retfanlimab, sintilimab, camrelizumab, and balstilimab; wherein the PD-L1 inhibitor is selected from the group comprising atezolizumab, avelumab, durvalumab, cosibelimab, envafolimab, CS1001, PF-06801591; wherein the CTLA-4 inhibitor is selected from the group comprising ipilimumab, tremelimumab, BMS-986218, and zalifrelimab; wherein the LAG-3 inhibitor is selected from the group comprising relatlimab and GSK2831781; wherein the TIGIT inhibitor is selected from the group comprising vibostolimab, domvanalimab, tiragolumab and etigilimab; and wherein the TIM-3 inhibitor is selected from the group comprising sabatolimab, BMS-986258, and cobolimab.

[0173] Embodiment 174. The CXCL12 antagonist for use of any one of Embodiments 169 and 171 to 173 and the interferon for use of any one of Embodiments 170 to 173, wherein the tumor has at least one characteristic selected from the group comprising reduced or lacking expression of PD-L1, reduced number of tumor-infiltrating lymphocytes, genomic stability and reduced or lacking preexisting antitumor immune response.

[0174] Embodiment 175. The CXCL12 antagonist for use of Embodiment 174 and the interferon for use of Embodiment 174, wherein the tumor-infiltrating lymphocytes are CD3+lymphocytes.

[0175] Embodiment 176. The CXCL12 antagonist for use of any one of Embodiments 169 and 171 to 175 and the interferon for use of any one of Embodiments 170 to 175, wherein the tumor has an immune evasion mechanism.

[0176] Embodiment 177. The CXCL12 antagonist for use of Embodiment 176 and the interferon for use of Embodiment 176, wherein the tumor is an immune-excluded tumor.

[0177] Embodiment 178. The CXCL12 antagonist for use of Embodiment 177 and the interferon for use of Embodiment 177, wherein immune exclusion can be reverted or mitigated by an immune checkpoint inhibitor.

[0178] Embodiment 179. The CXCL12 antagonist for use of Embodiment 178 and the interferon for use of Embodiment 178, wherein the checkpoint inhibitor is a PD-1 inhibitor, a PD-L1 inhibitor, a CTLA-4 inhibitor, a LAG-3 inhibitor, a TIM-3 inhibitor or a TIGIT inhibitor.Embodiment 180. The CXCL12 antagonist for use of Embodiment 179 and the interferon for use of Embodiment 179, wherein the PD-1 inhibitor is selected from the group comprising pembrolizumab, nivolumab, cemiplimab, toripalimab, tislelizumab, dostarlimab, retfanlimab, sintilimab, camrelizumab, and balstilimab; wherein the PD-L1 inhibitor is selected from the group comprising atezolizumab, avelumab, durvalumab, cosibelimab, envafolimab, CS1001, PF-06801591; wherein the CTLA-4 inhibitor is selected from the group comprising ipilimumab, tremelimumab, BMS-986218, and zalifrelimab; wherein the LAG-3 inhibitor is selected from the group comprising relatlimab and GSK2831781; wherein the TIGIT inhibitor is selected from the group comprising vibostolimab, domvanalimab, tiragolumab and etigilimab; and wherein the TIM-3 inhibitor is selected from the group comprising sabatolimab, BMS-986258, and cobolimab.

[0179] Embodiment 181. The CXCL12 antagonist for use of Embodiment 176 and the interferon for use of Embodiment 176, wherein the tumor is an immune-desert tumor.

[0180] Embodiment 182. The CXCL12 antagonist for use of any one of Embodiments 176 to 181 and the interferon for use of any one of Embodiments 176 to 181, wherein the immune evasion mechanism is one selected from the group comprising loss of MHC class I expression of malignant cells of the tumor and elevated expression of immune checkpoints by malignant and non-malignant cells of the tumor.

[0181] Embodiment 183. The CXCL12 antagonist for use of any one of Embodiments 176 to 182 and the interferon for use of any one of Embodiments 176 to 182, wherein the immune evasion mechanism results from, is caused by and / or is associated with a changed immune cell composition of the tumor microenvironment (TME).

[0182] Embodiment 184. The CXCL12 antagonist for use of Embodiment 184 and the interferon for use of Embodiment 183, wherein the tumor evades from the subject’s adaptive immune system.

[0183] Embodiment 185. The CXCL12 antagonist for use of Embodiment 184 and the interferon for use of Embodiment 184, wherein the tumor evades from the subject’s adaptive immune system by inhibiting infiltration of cytotoxic lymphocytes.

[0184] Embodiment 186. The CXCL12 antagonist for use of any one of Embodiments 184 and 185 and the interferon for use of any one of Embodiments 184 and 185, wherein the tumor evades from thesubject’s adaptive immune system by suppressing activity of cytotoxic tumor-infiltrating lymphocytes or inhibiting activation of cytotoxic tumor-infiltrating lymphocytes.

[0185] Embodiment 187. The CXCL12 antagonist for use of Embodiment 183 and the interferon for use of Embodiment 183, wherein the tumor evades from the subject’s innate immune system.

[0186] Embodiment 188. The CXCL12 antagonist for use of Embodiment 187 and the interferon for use of Embodiment 187, wherein the tumor is characterized by an immunosuppressive tumor microenvironment.

[0187] Embodiment 189. The CXCL12 antagonist for use of Embodiment 181 and the interferon for use of Embodiment 188, wherein the immunosuppressive tumor microenvironment results from, is caused by and / or is associated with presence of a cell selected from the group comprising a myeloid-derived suppressor cell (MDSC), a tumor-associated macrophage (TAM), a tumor-associated neutrophil (TAN) and / or a regulatory T cell (Treg).

[0188] Embodiment 190. The CXCL12 antagonist for use of Embodiment 189 and the interferon for use of Embodiment 189, wherein the cell is immunomodulatory active.

[0189] Embodiment 191. The CXCL12 antagonist for use of Embodiment 189 and the interferon for use of Embodiment 189, wherein the immunomodulatory active cell by means of a secreted immunomodulator, metabolic reprogramming and / or regulatory cell-to-cell contact.

[0190] Embodiment 192. The CXCL12 antagonist for use of any one of Embodiments 169 and 171 to 191 and the interferon for use of any one of Embodiments 170 to 191, wherein the tumor is a solid tumor.

[0191] Embodiment 193. The CXCL12 antagonist for use of any one of Embodiments 169 and 171 to 192 and the interferon for use of any one of Embodiments 170 to 192, wherein the tumor is selected from the group comprising pancreatic cancer, preferably pancreatic ductal adenocarcinoma (PDAC); triplenegative breast cancer; prostate cancer; ovarian cancer; soft tissue sarcoma and osteosarcoma.

[0192] Embodiment 194. The CXCL12 antagonist for use of Embodiment 193 and the interferon for use of Embodiment 193, wherein the tumor is high-grade serous ovarian cancer.Embodiment 195. The CXCL12 antagonist for use of Embodiment 193 and the interferon for use of Embodiment 193, wherein the tumor is advanced castration-resistant prostate cancer.

[0193] Embodiment 196. The CXCL12 antagonist for use of Embodiment 193 and the interferon for use of Embodiment 193, wherein the tumor is synovial sarcoma.

[0194] Embodiment 197. A CXCL12 antagonist for use in a method for treating a tumor in a subject, wherein the method comprises administering to the subject a primary therapy and an adjunct therapy, wherein the adjunct therapy comprises administering to the subject the CXCL12 antagonist and an interferon, wherein the tumor is refractory to immunotherapy.

[0195] Embodiment 198. An interferon for use in a method for treating a tumor in a subject, wherein the method comprises administering to the subject a primary therapy and an adjunct therapy, wherein the adjunct therapy comprises administering to the subject the interferon and a CXCL12 antagonist, wherein the tumor is refractory to immunotherapy.

[0196] Embodiment 199. The CXCL12 antagonist for use of Embodiment 197 and the interferon for use of Embodiment 198, wherein the tumor is refractory to an immunotherapy, preferably the immunotherapy is selected from the group comprising a checkpoint inhibitor, an adoptive cell therapy and a tumor vaccine.

[0197] Embodiment 200. The CXCL12 antagonist for use of Embodiment 199 and the interferon for use of Embodiment 199, wherein the checkpoint inhibitor is a PD-1 inhibitor, a PD-L1 inhibitor, a CTLA-4 inhibitor, a LAG-3 inhibitor, a TIM-3 inhibitor or a TIGIT inhibitor.

[0198] Embodiment 201. The CXCL12 antagonist for use of Embodiment 200 and the interferon for use of Embodiment 200, wherein the PD-1 inhibitor is selected from the group comprising pembrolizumab, nivolumab, cemiplimab, toripalimab, tislelizumab, dostarlimab, retfanlimab, sintilimab, camrelizumab, and balstilimab; wherein the PD-L1 inhibitor is selected from the group comprising atezolizumab, avelumab, durvalumab, cosibelimab, envafolimab, CS1001, PF-06801591; wherein the CTLA-4 inhibitor is selected from the group comprising ipilimumab, tremelimumab, BMS-986218, and zalifrelimab; wherein the LAG-3 inhibitor is selected from the group comprising relatlimab and GSK2831781; wherein the TIGIT inhibitor is selected from the group comprisingvibostolimab, domvanalimab, tiragolumab and etigilimab; and wherein the TIM-3 inhibitor is selected from the group comprising sabatolimab, BMS-986258, and cobolimab.

[0199] Embodiment 202. The CXCL12 antagonist for use of any one of Embodiments 197 and 199 to 201 and the interferon for use of any one of Embodiments 198 to 201, wherein the tumor has at least one characteristic selected from the group comprising reduced or lacking expression of PD-L1, reduced number of tumor-infiltrating lymphocytes, genomic stability and reduced or lacking preexisting antitumor immune response.

[0200] Embodiment 203. The CXCL12 antagonist for use of Embodiment 202 and the interferon for use of Embodiment 202, wherein the tumor-infiltrating lymphocytes are CD3+lymphocytes.

[0201] Embodiment 204. The CXCL12 antagonist for use of any one of Embodiments 197 and 199 to 203 and the interferon for use of any one of Embodiments 198 to 203, wherein the tumor has an immune evasion mechanism.

[0202] Embodiment 205. The CXCL12 antagonist for use of Embodiment 204 and the interferon for use of Embodiment 204, wherein the tumor is an immune-excluded tumor.

[0203] Embodiment 206. The CXCL12 antagonist for use of Embodiment 205 and the interferon for use of Embodiment 205, wherein immune exclusion can be reverted or mitigated by an immune checkpoint inhibitor.

[0204] Embodiment 207. The CXCL12 antagonist for use of Embodiment 206 and the interferon for use of Embodiment 206, wherein the checkpoint inhibitor is a PD-1 inhibitor, a PD-L1 inhibitor, a CTLA-4 inhibitor, a LAG-3 inhibitor, a TIM-3 inhibitor or a TIGIT inhibitor.

[0205] Embodiment 208. The CXCL12 antagonist for use of Embodiment 207 and the interferon for use of Embodiment 207, wherein the PD-1 inhibitor is selected from the group comprising pembrolizumab, nivolumab, cemiplimab, toripalimab, tislelizumab, dostarlimab, retfanlimab, sintilimab, camrelizumab, and balstilimab; wherein the PD-L1 inhibitor is selected from the group comprising atezolizumab, avelumab, durvalumab, cosibelimab, envafolimab, CS1001, PF-06801591; wherein the CTLA-4 inhibitor is selected from the group comprising ipilimumab, tremelimumab, BMS-986218, and zalifrelimab; wherein the LAG-3 inhibitor is selected from the group comprisingrelatlimab and GSK2831781; wherein the TIGIT inhibitor is selected from the group comprising vibostolimab, domvanalimab, tiragolumab and etigilimab; and wherein the TIM-3 inhibitor is selected from the group comprising sabatolimab, BMS-986258, and cobolimab.

[0206] Embodiment 209. The CXCL12 antagonist for use of Embodiment 204 and the interferon for use of Embodiment 204, wherein the tumor is an immune-desert tumor.

[0207] Embodiment 210. The CXCL12 antagonist for use of any one of Embodiments 204 to 209 and the interferon for use of any one of Embodiments 204 to 209, wherein the immune evasion mechanism is one selected from the group comprising loss of MHC class I expression of malignant cells of the tumor and elevated expression of immune checkpoints by malignant and non-malignant cells of the tumor.

[0208] Embodiment 211. The CXCL12 antagonist for use of any one of Embodiments 204 to 210 and the interferon for use of any one of Embodiments 204 to 210, wherein the immune evasion mechanism results from, is caused by and / or is associated with a changed immune cell composition of the tumor microenvironment (TME).

[0209] Embodiment 212. The CXCL12 antagonist for use of Embodiment 211 and the interferon for use of Embodiment 211, wherein the tumor evades from the subject’s adaptive immune system.

[0210] Embodiment 213. The CXCL12 antagonist for use of Embodiment 212 and the interferon for use of Embodiment 212, wherein the tumor evades from the subject’s adaptive immune system by inhibiting infiltration of cytotoxic lymphocytes.

[0211] Embodiment 214. The CXCL12 antagonist for use of any one of Embodiments 212 and 213 and the interferon for use of any one of Embodiments 212 and 213, wherein the tumor is evading from the subject’s adaptive immune system by suppressing activity of cytotoxic tumor-infiltrating lymphocytes or inhibiting activation of cytotoxic tumor-infiltrating lymphocytes.

[0212] Embodiment 215. The CXCL 12 antagonist for use of Embodiment 211 and the interferon for use of Embodiment 211, wherein the tumor is evading from the subject’s innate immune system.Embodiment 216. The CXCL12 antagonist for use of Embodiment 211 and the interferon for use of Embodiment 211, wherein the tumor is characterized by an immunosuppressive tumor microenvironment.

[0213] Embodiment 217. The CXCL12 antagonist for use of Embodiment 216 and the interferon for use of Embodiment 216, wherein the immunosuppressive tumor microenvironment results from, is caused by and / or is associated with presence of a cell selected from the group comprising a myeloid-derived suppressor cell (MDSC), a tumor-associated macrophage (TAM), a tumor-associated neutrophil (TAN) and / or a regulatory T cell (Treg).

[0214] Embodiment 218. The CXCL12 antagonist for use of Embodiment 217 and the interferon for use of Embodiment 217, wherein the cell is immunomodulatorily active.

[0215] Embodiment 219. The CXCL12 antagonist for use of Embodiment 218 and the interferon for use of Embodiment 218, wherein the immunomodulatorily active cell is active by means of a secreted immunomodulator, metabolic reprogramming and / or regulatory cell-to-cell contact.

[0216] Embodiment 220. The CXCL12 antagonist for use of any one of Embodiments 197 and 199 to 219 and the interferon for use of any one of Embodiments 198 to 219, wherein the tumor is an immunologically cold tumor.

[0217] Embodiment 221. The CXCL12 antagonist for use of any one of Embodiments 197 and 199 to 220 and the interferon for use of any one of Embodiments 198 to 220, wherein the tumor is a solid tumor.

[0218] Embodiment 222. The CXCL12 antagonist for use of any one of Embodiments 197 and 199 to 221 and the interferon for use of any one of Embodiments 198 to 221, wherein the tumor is selected from the group comprising pancreatic cancer, preferably pancreatic ductal adenocarcinoma (PDAC); triplenegative breast cancer; prostate cancer; ovarian cancer; soft tissue sarcoma and osteosarcoma.

[0219] Embodiment 223. The CXCL12 antagonist for use of Embodiment 222 and the interferon for use of Embodiment 222, wherein the tumor is high-grade serous ovarian cancer.

[0220] Embodiment 224. The CXCL12 antagonist for use of Embodiment 222 and the interferon for use of Embodiment 222, wherein the tumor is advanced castration-resistant prostate cancer.Embodiment 225. The CXCL12 antagonist for use of Embodiment 222 and the interferon for use of Embodiment 222, wherein the tumor is synovial sarcoma.

[0221] Embodiment 226. A CXCL12 antagonist for use in a method for treating a tumor in a subject, wherein the method comprises administering to the subject a primary therapy and an adjunct therapy, wherein the adjunct therapy comprises administering to the subject the CXCL12 antagonist and an interferon, wherein the tumor is a solid tumor.

[0222] Embodiment 227. An interferon for use in a method for treating a tumor in a subject, wherein the method comprises administering to the subject a primary therapy and an adjunct therapy, wherein the adjunct therapy comprises administering to the subject the interferon and a CXCL12 antagonist, wherein the tumor is a solid tumor.

[0223] Embodiment 228. The CXCL12 antagonist for use of Embodiment 226 and the interferon for use of Embodiment 227, wherein the tumor is selected from the group comprising pancreatic cancer, preferably pancreatic ductal adenocarcinoma (PDAC); triple-negative breast cancer; prostate cancer; ovarian cancer; soft tissue sarcoma and osteosarcoma.

[0224] Embodiment 229. The CXCL12 antagonist for use of Embodiment 228 and the interferon for use of Embodiment 228, wherein the tumor is high-grade serous ovarian cancer.

[0225] Embodiment 230. The CXCL12 antagonist for use of Embodiment 228 and the interferon for use of Embodiment 228, wherein the tumor is advanced castration-resistant prostate cancer.

[0226] Embodiment 231. The CXCL12 antagonist for use of Embodiment 228 and the interferon for use of Embodiment 228, wherein the tumor is synovial sarcoma.

[0227] Embodiment 232. The CXCL12 antagonist for use of any one of Embodiments 226 and 228 to 231 and the interferon for use of any one of Embodiments 227 to 231, wherein the tumor is refractory to an immunotherapy, preferably the immunotherapy is selected from the group comprising checkpoint inhibitor, an adoptive cell therapy and a tumor vaccine.

[0228] Embodiment 233. The CXCL12 antagonist for use of any one of Embodiments 226 and 228 to 232 and the interferon for use of any one of Embodiments 227 to 232, wherein the checkpoint inhibitor is aPD-1 inhibitor, a PD-L1 inhibitor, a CTLA-4 inhibitor, a LAG-3 inhibitor, a TIM-3 inhibitor or a TIGIT inhibitor.

[0229] Embodiment 234. The CXCL12 antagonist for use of Embodiment 233 and the interferon for use of Embodiment 233, wherein the PD-1 inhibitor is selected from the group comprising pembrolizumab, nivolumab, cemiplimab, toripalimab, tislelizumab, dostarlimab, retfanlimab, sintilimab, camrelizumab, and balstilimab; wherein the PD-L1 inhibitor is selected from the group comprising atezolizumab, avelumab, durvalumab, cosibelimab, envafolimab, CS1001, PF-06801591; wherein the CTLA-4 inhibitor is selected from the group comprising ipilimumab, tremelimumab, BMS-986218, and zalifrelimab; wherein the LAG-3 inhibitor is selected from the group comprising relatlimab and GSK2831781; wherein the TIGIT inhibitor is selected from the group comprising vibostolimab, domvanalimab, tiragolumab and etigilimab; and wherein the TIM-3 inhibitor is selected from the group comprising sabatolimab, BMS-986258, and cobolimab.

[0230] Embodiment 235. The CXCL12 antagonist for use of any one of Embodiments 226 and 228 to 234 and the interferon for use of any one of Embodiments 227 to 224, wherein the tumor has at least one characteristic selected from the group comprising reduced or lacking expression of PD-L1, reduced number of tumor-infiltrating lymphocytes, genomic stability and reduced or lacking preexisting antitumor immune response.

[0231] Embodiment 236. The CXCL12 antagonist for use of Embodiment 235 and the interferon for use of Embodiment 235, wherein the tumor-infiltrating lymphocytes are CD3+lymphocytes.

[0232] Embodiment 237. The CXCL12 antagonist for use of any one of Embodiments 226 and 228 to 236 and the interferon for use of any one of Embodiments 227 to 236, wherein the tumor has an immune evasion mechanism.

[0233] Embodiment 238. The CXCL12 antagonist for use of Embodiment 237 and the interferon for use of Embodiment 237, wherein the tumor is an immune-excluded tumor.

[0234] Embodiment 239. The CXCL12 antagonist for use of Embodiment 238 and the interferon for use of Embodiment 238 wherein immune exclusion can be reverted or mitigated by an immune checkpoint inhibitor.Embodiment 240. The CXCL12 antagonist for use of Embodiment 237 and the interferon for use of Embodiment 237, wherein the tumor is an immune-desert tumor.

[0235] Embodiment 241. The CXCL12 antagonist for use of any one of Embodiments 237 to 240 and the interferon for use of any one of Embodiments 237 to 240, wherein the immune evasion mechanism is one selected from the group comprising loss of MHC class I expression of malignant cells of the tumor and elevated expression of immune checkpoints by malignant and non-malignant cells of the tumor.

[0236] Embodiment 242. The CXCL12 antagonist for use of any one of Embodiments 237 to 241 and the interferon for use of any one of Embodiments 237 to 241, wherein the immune evasion mechanism results from, is caused by and / or is associated with a changed immune cell composition of the tumor microenvironment (TME).

[0237] Embodiment 243. The CXCL12 antagonist for use of Embodiment 242 and the interferon for use of Embodiment 242, wherein the tumor is evading from the subject’s adaptive immune system.

[0238] Embodiment 244. The CXCL12 antagonist for use of Embodiment 243 and the interferon for use of Embodiment 243, wherein the tumor is evading from the subject’s adaptive immune system by inhibiting infiltration of cytotoxic lymphocytes.

[0239] Embodiment 245. The CXCL12 antagonist for use of any one of Embodiments 243 and 244 and the interferon for use of any one of Embodiments 243 and 233, wherein the tumor is evading from the subject’s adaptive immune system by suppressing activity of cytotoxic tumor-infiltrating lymphocytes or inhibiting activation of cytotoxic tumor-infiltrating lymphocytes.

[0240] Embodiment 246. The CXCL12 antagonist for use of Embodiment 242 and the interferon for use of Embodiment 242, wherein the tumor is evading from the subject’s innate immune system.

[0241] Embodiment 247. The CXCL12 antagonist for use of Embodiment 246 and the interferon for use of Embodiment 246, wherein the tumor is characterized by an immunosuppressive tumor microenvironment.Embodiment 248. The CXCL12 antagonist for use of Embodiment 247 and the interferon for use of Embodiment 247, wherein the immunosuppressive tumor microenvironment results from, is caused by and / or is associated with presence of a cell selected from the group comprising a myeloid-derived suppressor cell (MDSC), a tumor-associated macrophage (TAM), a tumor-associated neutrophil (TAN) and / or a regulatory T cell (Treg).

[0242] Embodiment 249. The CXCL12 antagonist for use of Embodiment 248 and the interferon for use of Embodiment 248, wherein the cell is immunomodulatorily active.

[0243] Embodiment 250. The CXCL12 antagonist for use of Embodiment 249 and the interferon for use of Embodiment 249, wherein the immunomodulatorily active cell is active by means of a secreted immunomodulator, metabolic reprogramming and / or regulatory cell-to-cell contact.

[0244] Embodiment 251. A CXCL12 antagonist for use in a method for converting an immunologically cold tumor in a subject into an immunologically hot tumor in the subject, wherein the method comprises administering to the subject the CXCL12 antagonist and an interferon, wherein upon administration of the CXCL12 antagonist and the interferon the immunologically cold tumor is or will be converted into an immunologically hot tumor.

[0245] Embodiment 252. An interferon for use in a method for converting an immunologically cold tumor in a subject into an immunologically hot tumor in the subject, wherein the method comprises administering to the subject the interferon and a CXCL12 antagonist, wherein upon administration of the interferon and the CXCL12 antagonist the immunologically cold tumor is or will be converted into an immunologically hot tumor.

[0246] Embodiment 253. The CXCL12 antagonist for use of Embodiment 251 and the interferon for use of Embodiment 252, wherein the immunologically cold tumor is refractory to an immunotherapy, preferably the immunotherapy is selected from the group comprising a checkpoint inhibitor, an adoptive cell therapy and a tumor vaccine.

[0247] Embodiment 254. The CXCL12 antagonist for use of Embodiment 253 and the interferon for use of Embodiment 253, wherein the checkpoint inhibitor is a PD-1 inhibitor, a PD-L1 inhibitor, a CTLA-4 inhibitor, a LAG-3 inhibitor, a TIM-3 inhibitor or a TIGIT inhibitor.Embodiment 255. The CXCL12 antagonist for use of Embodiment 254 and the interferon for use of Embodiment 254, wherein the PD-1 inhibitor is selected from the group comprising pembrolizumab, nivolumab, cemiplimab, toripalimab, tislelizumab, dostarlimab, retfanlimab, sintilimab, camrelizumab, and balstilimab; wherein the PD-L1 inhibitor is selected from the group comprising atezolizumab, avelumab, durvalumab, cosibelimab, envafolimab, CS1001, PF-06801591; wherein the CTLA-4 inhibitor is selected from the group comprising ipilimumab, tremelimumab, BMS-986218, and zalifrelimab; wherein the LAG-3 inhibitor is selected from the group comprising relatlimab and GSK2831781; wherein the TIGIT inhibitor is selected from the group comprising vibostolimab, domvanalimab, tiragolumab and etigilimab; and wherein the TIM-3 inhibitor is selected from the group comprising sabatolimab, BMS-986258, and cobolimab.

[0248] Embodiment 256. The CXCL12 antagonist for use of any one of Embodiments 251 and 253 to 255 and the interferon for use of any one of Embodiments 252 to 255, wherein the immunologically cold tumor has at least one characteristic selected from the group comprising reduced or lacking expression of PD-L1, reduced number of tumor-infiltrating lymphocytes, genomic stability and reduced or lacking preexisting antitumor immune response.

[0249] Embodiment 257. The CXCL12 antagonist for use of Embodiment 256 and the interferon for use of Embodiment 256, wherein the tumor-infiltrating lymphocytes are CD3+lymphocytes.

[0250] Embodiment 258. The CXCL12 antagonist for use of any one of Embodiments 251 and 253 to 257 and the interferon for use of any one of Embodiments 252 to 257, wherein the immunologically cold tumor has an immune evasion mechanism.

[0251] Embodiment 259. The CXCL12 antagonist for use of Embodiment 258 and the interferon for use of Embodiment 258, wherein the immunologically cold tumor is an immune-excluded tumor.

[0252] Embodiment 260. The CXCL12 antagonist for use of Embodiment 259 and the interferon for use of Embodiment 259, wherein immune exclusion can be reverted or mitigated by an immune checkpoint inhibitor.

[0253] Embodiment 261. The CXCL12 antagonist for use of Embodiment 260 and the interferon for use of Embodiment 260, wherein the checkpoint inhibitor is a PD-1 inhibitor, a PD-L1 inhibitor, a CTLA-4 inhibitor, a LAG-3 inhibitor, a TIM-3 inhibitor or a TIGIT inhibitor.Embodiment 262. The CXCL12 antagonist for use of Embodiment 261 and the interferon for use of Embodiment 261, wherein the PD-1 inhibitor is selected from the group comprising pembrolizumab, nivolumab, cemiplimab, toripalimab, tislelizumab, dostarlimab, retfanlimab, sintilimab, camrelizumab, and balstilimab; wherein the PD-L1 inhibitor is selected from the group comprising atezolizumab, avelumab, durvalumab, cosibelimab, envafolimab, CS1001, PF-06801591; wherein the CTLA-4 inhibitor is selected from the group comprising ipilimumab, tremelimumab, BMS-986218, and zalifrelimab; wherein the LAG-3 inhibitor is selected from the group comprising relatlimab and GSK2831781; wherein the TIGIT inhibitor is selected from the group comprising vibostolimab, domvanalimab, tiragolumab and etigilimab; and wherein the TIM-3 inhibitor is selected from the group comprising sabatolimab, BMS-986258, and cobolimab.

[0254] Embodiment 263. The CXCL12 antagonist for use of Embodiment 258 and the interferon for use of Embodiment 258, wherein the tumor is an immune-desert tumor.

[0255] Embodiment 264. The CXCL12 antagonist for use of any one of Embodiments 258 to 263 and the interferon for use of any one of Embodiments 258 to 263, wherein the immune evasion mechanism is one selected from the group comprising loss of MHC class I expression of malignant cells of the tumor and elevated expression of immune checkpoints by malignant and non-malignant cells of the tumor.

[0256] Embodiment 265. The CXCL12 antagonist for use of any one of Embodiments 258 to 264 and the interferon for use of any one of Embodiments 258 to 264, wherein the immune evasion mechanism results from, is caused by and / or is associated with a changed immune cell composition of the tumor microenvironment (TME).

[0257] Embodiment 266. The CXCL12 antagonist for use of Embodiment 265 and the interferon for use of Embodiment 265, wherein the immunologically cold tumor evades from the subject’s adaptive immune system.

[0258] Embodiment 267. The CXCL12 antagonist for use of Embodiment 266 and the interferon for use of Embodiment 266, wherein the immunologically cold tumor is evading from the subject’s adaptive immune system by inhibiting infiltration of cytotoxic lymphocytes.Embodiment 268. The CXCL12 antagonist for use of any one of Embodiments 266 and 267 and the interferon for use of any one of Embodiments 266 and 267, wherein the immunologically cold tumor is evading from the subject’s adaptive immune system by suppressing activity of cytotoxic tumor-infiltrating lymphocytes or inhibiting activation of cytotoxic tumor-infiltrating lymphocytes.

[0259] Embodiment 269. The CXCL12 antagonist for use of Embodiment 265 and the interferon for use of Embodiment 265, wherein the immunologically cold tumor is evading from the subject’s innate immune system.

[0260] Embodiment 270. The CXCL12 antagonist for use of Embodiment 269 and the interferon for use of Embodiment 269, wherein the immunologically cold tumor is characterized by an immunosuppressive tumor microenvironment.

[0261] Embodiment 271. The CXCL12 antagonist for use of Embodiment 270 and the interferon for use of Embodiment 270, wherein the immunosuppressive tumor microenvironment results from, is caused by and / or is associated with presence of a cell selected from the group comprising a myeloid-derived suppressor cell (MDSC), a tumor-associated macrophage (TAM), a tumor-associated neutrophil (TAN) and / or a regulatory T cell (Treg).

[0262] Embodiment 272. The CXCL12 antagonist for use of Embodiment 271 and the interferon for use of Embodiment 271, wherein the cell is immunomodulatory active.

[0263] Embodiment 273. The CXCL12 antagonist for use of Embodiment 272 and the interferon for use of Embodiment 272, wherein the immunomodulatory active cell by means of a secreted immunomodulator, metabolic reprogramming and / or regulatory cell-to-cell contact.

[0264] Embodiment 274. The CXCL12 antagonist for use of any one of Embodiments 251 and 253 to 273 and the interferon for use of any one of Embodiments 252 to 273, wherein the immunologically hot tumor is non-refractory to an immunotherapy, preferably the immunotherapy is selected from the group comprising a checkpoint inhibitor, an adoptive cell therapy and a tumor vaccine.

[0265] Embodiment 275. The CXCL12 antagonist for use of Embodiment 274 and the interferon for use of Embodiment 274, wherein the checkpoint inhibitor is a PD-1 inhibitor, a PD-L1 inhibitor, a CTLA-4 inhibitor, a LAG-3 inhibitor, a TIM-3 inhibitor or a TIGIT inhibitor.Embodiment 276. The CXCL12 antagonist for use of Embodiment 275 and the interferon for use of Embodiment 275, wherein the PD-1 inhibitor is selected from the group comprising pembrolizumab, nivolumab, cemiplimab, toripalimab, tislelizumab, dostarlimab, retfanlimab, sintilimab, camrelizumab, and balstilimab; wherein the PD-L1 inhibitor is selected from the group comprising atezolizumab, avelumab, durvalumab, cosibelimab, envafolimab, CS1001, PF-06801591; wherein the CTLA-4 inhibitor is selected from the group comprising ipilimumab, tremelimumab, BMS-986218, and zalifrelimab; wherein the LAG-3 inhibitor is selected from the group comprising relatlimab and GSK2831781; wherein the TIGIT inhibitor is selected from the group comprising vibostolimab, domvanalimab, tiragolumab and etigilimab; and wherein the TIM-3 inhibitor is selected from the group comprising sabatolimab, BMS-986258, and cobolimab.

[0266] Embodiment 277. The CXCL12 antagonist for use of any one of Embodiments 251 and 253 to 276 and the interferon for use of any one of Embodiments 252 to 276, wherein the immunologically hot tumor has at least one characteristic selected from the group comprising presence of tumor-infiltrating lymphocytes, genomic instability of malignant cells and preexisting antitumor immune response.

[0267] Embodiment 278. The CXCL12 antagonist for use of Embodiment 277 and the interferon for use of Embodiment 277, wherein the tumor-infiltrating lymphocytes are CD3+lymphocytes.

[0268] Embodiment 279. The CXCL12 antagonist for use of any one of Embodiments 251 and 253 to 278 and the interferon for use of any one of Embodiments 252 to 278, wherein the immunologically hot tumor is subject to the subject’s adaptive immune system.

[0269] Embodiment 280. The CXCL12 antagonist for use of any one of Embodiments 251 and 253 to 278 and the interferon for use of any one of Embodiments 252 to 278, wherein the immunologically hot tumor is subject to the subject’s innate immune system.

[0270] Embodiment 281. The CXCL12 antagonist for use of any one of Embodiments 251 and 253 to 280 and the interferon for use of any one of Embodiments 252 to 280, wherein the immunologically hot tumor is characterized by a non-immunosuppressive tumor microenvironment.

[0271] Embodiment 282. The CXCL12 antagonist for use of Embodiment 281 and the interferon for use of Embodiment 281, wherein an immunosuppressive tumor microenvironment results from, is caused by and / or is associated with presence of a cell selected from the group comprising a myeloid-derivedsuppressor cell (MDSC), a tumor-associated macrophage (TAM), a tumor-associated neutrophil (TAN) and / or a regulatory T cell (Treg).

[0272] Embodiment 283. The CXCL12 antagonist for use of Embodiment 282 and the interferon for use of Embodiment 282, wherein the cell is immunomodulatory active.

[0273] Embodiment 284. The CXCL12 antagonist for use of Embodiment 280 and the interferon for use of Embodiment 280, wherein the immunomodulatory active cell by means of a secreted immunomodulator, metabolic reprogramming and / or regulatory cell-to-cell contact.

[0274] Embodiment 285. The CXCL12 antagonist for use of any one of Embodiments 251 and 253 to 284 and the interferon for use of any one of Embodiments 252 to 281, wherein the tumor is a solid tumor.

[0275] Embodiment 286. The CXCL12 antagonist for use of any one of Embodiments 251 and 253 to 285 and the interferon for use of any one of Embodiments 252 to 285, wherein the tumor is selected from the group comprising pancreatic cancer, preferably pancreatic ductal adenocarcinoma (PDAC); triplenegative breast cancer; prostate cancer; ovarian cancer; soft tissue sarcoma and osteosarcoma.

[0276] Embodiment 287. The CXCL12 antagonist for use of Embodiment 286 and the interferon for use of Embodiment 286, wherein the tumor is high-grade serous ovarian cancer.

[0277] Embodiment 288. The CXCL12 antagonist for use of Embodiment 286 and the interferon for use of Embodiment 286, wherein the tumor is advanced castration-resistant prostate cancer.

[0278] Embodiment 289. The CXCL12 antagonist for use of Embodiment 286 and the interferon for use of Embodiment 286, wherein the tumor is synovial sarcoma.

[0279] Embodiment 290. A CXCL12 antagonist for use in a method for converting an immunotherapy refractory tumor in a subject into an immunotherapy non-refractory tumor in the subject, wherein the method comprises administering to the subject the CXCL12 antagonist and an interferon, wherein upon administration of the CXCL12 antagonist and the interferon the immunotherapy refractory tumor is or will be converted into an immunotherapy non-refractory tumor.Embodiment 291. An interferon for use in a method for converting an immunotherapy refractory tumor in a subject into an immunotherapy non-refractory tumor in the subject, wherein the method comprises administering to the subject the interferon and a CXCL12 antagonist, wherein upon administration of the interferon and the CXCL12 antagonist the immunotherapy refractory tumor is or will be converted into an immunotherapy non-refractory tumor.

[0280] Embodiment 292. The CXCL12 antagonist for use of Embodiment 290 and the interferon for use of Embodiment 291, wherein the immunotherapy refractory tumor is refractory to an immunotherapy, preferably the immunotherapy is selected from the group comprising a checkpoint inhibitor, an adoptive cell therapy and a tumor vaccine.

[0281] Embodiment 293. The CXCL12 antagonist for use of Embodiment 292 and the interferon for use of Embodiment 292, wherein the checkpoint inhibitor is a PD-1 inhibitor, a PD-L1 inhibitor, a CTLA-4 inhibitor, a LAG-3 inhibitor, a TIM-3 inhibitor or a TIGIT inhibitor.

[0282] Embodiment 294. The CXCL12 antagonist for use of Embodiment 293 and the interferon for use of Embodiment 293, wherein the PD-1 inhibitor is selected from the group comprising pembrolizumab, nivolumab, cemiplimab, toripalimab, tislelizumab, dostarlimab, retfanlimab, sintilimab, camrelizumab, and balstilimab; wherein the PD-L1 inhibitor is selected from the group comprising atezolizumab, avelumab, durvalumab, cosibelimab, envafolimab, CS1001, PF-06801591; wherein the CTLA-4 inhibitor is selected from the group comprising ipilimumab, tremelimumab, BMS-986218, and zalifrelimab; wherein the LAG-3 inhibitor is selected from the group comprising relatlimab and GSK2831781; wherein the TIGIT inhibitor is selected from the group comprising vibostolimab, domvanalimab, tiragolumab and etigilimab; and wherein the TIM-3 inhibitor is selected from the group comprising sabatolimab, BMS-986258, and cobolimab.

[0283] Embodiment 295. The CXCL12 antagonist for use of any one of Embodiments 290 and 292 to 294 and the interferon for use of any one of Embodiments 291 to 294, wherein the immunotherapy refractory tumor has at least one characteristic selected from the group comprising reduced or lacking expression of PD-L1, reduced number of tumor-infiltrating lymphocytes, genomic stability and reduced or lacking preexisting antitumor immune response.

[0284] Embodiment 296. The CXCL12 antagonist for use of Embodiment 295 and the interferon for use of Embodiment 292, wherein the tumor-infiltrating lymphocytes are CD3+lymphocytes.Embodiment 297. The CXCL12 antagonist for use of any one of Embodiments 290 and 292 to 296 and the interferon for use of any one of Embodiments 291 to 296, wherein the immunotherapy refractory tumor evades attack of the immune system.

[0285] Embodiment 298. The CXCL12 antagonist for use of Embodiment 297 and the interferon for use of Embodiment 297, wherein the immunotherapy refractory tumor is an immune-excluded tumor.

[0286] Embodiment 299. The CXCL12 antagonist for use of Embodiment 298 and the interferon for use of Embodiment 298, wherein immune exclusion can be reverted or mitigated by an immune checkpoint inhibitor.

[0287] Embodiment 300. The CXCL12 antagonist for use of Embodiment 299 and the interferon for use of Embodiment 299, wherein the checkpoint inhibitor is a PD-1 inhibitor, a PD-L1 inhibitor, a CTLA-4 inhibitor, a LAG-3 inhibitor, a TIM-3 inhibitor or a TIGIT inhibitor.

[0288] Embodiment 301. The CXCL12 antagonist for use of Embodiment 300 and the interferon for use of Embodiment 300, wherein the PD-1 inhibitor is selected from the group comprising pembrolizumab, nivolumab, cemiplimab, toripalimab, tislelizumab, dostarlimab, retfanlimab, sintilimab, camrelizumab, and balstilimab; wherein the PD-L1 inhibitor is selected from the group comprising atezolizumab, avelumab, durvalumab, cosibelimab, envafolimab, CS1001, PF-06801591; wherein the CTLA-4 inhibitor is selected from the group comprising ipilimumab, tremelimumab, BMS-986218, and zalifrelimab; wherein the LAG-3 inhibitor is selected from the group comprising relatlimab and GSK2831781; wherein the TIGIT inhibitor is selected from the group comprising vibostolimab, domvanalimab, tiragolumab and etigilimab; and wherein the TIM-3 inhibitor is selected from the group comprising sabatolimab, BMS-986258, and cobolimab.

[0289] Embodiment 302. The CXCL12 antagonist for use of Embodiment 297 and the interferon for use of Embodiment 297, wherein the immunotherapy refractory tumor is an immune-desert tumor.

[0290] Embodiment 303. The CXCL12 antagonist for use of any one of Embodiments 297 to 302 and the interferon for use of any one of Embodiments 297 to 302, wherein the immune evasion mechanism is one selected from the group comprising loss of MHC class I expression of malignant cells of the tumor and elevated expression of immune checkpoints by malignant and non-malignant cells of the tumor.Embodiment 304. The CXCL12 antagonist for use of any one of Embodiments 297 to 303 and the interferon for use of any one of Embodiments 297 to 303, wherein the immune evasion mechanism results from, is caused by and / or is associated with a changed immune cell composition of the tumor microenvironment (TME).

[0291] Embodiment 305. The CXCL12 antagonist for use of Embodiment 304 and the interferon for use of Embodiment 304, wherein the immunologically cold tumor evades from the subject’s adaptive immune system.

[0292] Embodiment 306. The CXCL12 antagonist for use of Embodiment 305 and the interferon for use of Embodiment 305, wherein the immunotherapy refractory tumor is evading from the subject’s adaptive immune system by inhibiting infiltration of cytotoxic lymphocytes.

[0293] Embodiment 307. The CXCL12 antagonist for use of any one of Embodiments 305 and 306 and the interferon for use of any one of Embodiments 305 and 306, wherein the immunotherapy refractory tumor is evading from the subject’s adaptive immune system by suppressing activity of cytotoxic tumor-infiltrating lymphocytes or inhibiting activation of cytotoxic tumor-infiltrating lymphocytes.

[0294] Embodiment 308. The CXCL12 antagonist for use of Embodiment 304 and the interferon for use of Embodiment 304, wherein the immunotherapy refractory tumor is evading from the subject’s innate immune system.

[0295] Embodiment 309. The CXCL12 antagonist for use of Embodiment 308 and the interferon for use of Embodiment 308, wherein the immunotherapy refractory tumor is characterized by an immunosuppressive tumor microenvironment.

[0296] Embodiment 310. The CXCL12 antagonist for use of Embodiment 309 and the interferon for use of Embodiment 309, wherein the immunosuppressive tumor microenvironment results from, is caused by and / or is associated with presence of a cell selected from the group comprising a myeloid-derived suppressor cell (MDSC), a tumor-associated macrophage (TAM) a tumor-associated neutrophil (TAN) and / or a regulatory T cell (Treg).

[0297] Embodiment 311. The CXCL12 antagonist for use of Embodiment 310 and the interferon for use of Embodiment 310, wherein the cell is immunomodulatory active.Embodiment 312. The CXCL12 antagonist for use of Embodiment 311 and the interferon for use of Embodiment 311, wherein the immunomodulatory active cell by means of a secreted immunomodulator, metabolic reprogramming and / or regulatory cell-to-cell contact.

[0298] Embodiment 313. The CXCL12 antagonist for use of any one of Embodiments 290 and 292 to 312 and the interferon for use of any one of Embodiments 291 to 313, wherein the immunotherapy nonrefractory tumor is non-refractory to an immunotherapy, preferably the immunotherapy is selected from the group comprising a checkpoint inhibitor, an adoptive cell therapy and a tumor vaccine.

[0299] Embodiment 314. The CXCL12 antagonist for use of Embodiment 313 and the interferon for use of Embodiment 313, wherein the checkpoint inhibitor is a PD-1 inhibitor, a PD-L1 inhibitor, a CTLA-4 inhibitor, a LAG-3 inhibitor, a TIM-3 inhibitor or a TIGIT inhibitor.

[0300] Embodiment 315. The CXCL12 antagonist for use of Embodiment 314 and the interferon for use of Embodiment 314, wherein the PD-1 inhibitor is selected from the group comprising pembrolizumab, nivolumab, cemiplimab, toripalimab, tislelizumab, dostarlimab, retfanlimab, sintilimab, camrelizumab, and balstilimab; wherein the PD-L1 inhibitor is selected from the group comprising atezolizumab, avelumab, durvalumab, cosibelimab, envafolimab, CS1001, PF-06801591; wherein the CTLA-4 inhibitor is selected from the group comprising ipilimumab, tremelimumab, BMS-986218, and zalifrelimab; wherein the LAG-3 inhibitor is selected from the group comprising relatlimab and GSK2831781; wherein the TIGIT inhibitor is selected from the group comprising vibostolimab, domvanalimab, tiragolumab and etigilimab; and wherein the TIM-3 inhibitor is selected from the group comprising sabatolimab, BMS-986258, and cobolimab.

[0301] Embodiment 316. The CXCL12 antagonist for use of any one of Embodiments 290 and 292 to 315 and the interferon for use of any one of Embodiments 291 to 315, wherein the immunotherapy nonrefractory tumor has at least one characteristic selected from the group comprising expression of PD-Ll, presence of tumor-infiltrating lymphocytes, genomic instability of malignant cells and preexisting antitumor immune response.

[0302] Embodiment 317. The CXCL12 antagonist for use of Embodiment 316 and the interferon for use of Embodiment 316, wherein the tumor-infdtrating lymphocytes are CD3+ lymphocytes.Embodiment 318. The CXCL12 antagonist for use of any one of Embodiments 288 and 290 to 317 and the interferon for use of any one of Embodiments 289 to 317, wherein the immunotherapy nonrefractory tumor is subject to the subject’s adaptive immune system.

[0303] Embodiment 319. The CXCL12 antagonist for use of any one of Embodiments 290 and 292 to 318 and the interferon for use of any one of Embodiments 291 to 318, wherein the immunotherapy nonrefractory tumor is subject to the subject’s innate immune system.

[0304] Embodiment 320. The CXCL12 antagonist for use of any one of Embodiments 290 and 292 to 319 and the interferon for use of any one of Embodiments 291 to 319, wherein the immunotherapy nonrefractory tumor is characterized by a non-immunosuppressive tumor microenvironment.

[0305] Embodiment 321. The CXCL12 antagonist for use of Embodiment 320 and the interferon for use of Embodiment 320, wherein an immunosuppressive tumor microenvironment results from, is caused by and / or is associated with presence of a cell selected from the group comprising a myeloid-derived suppressor cell (MDSC), a tumor-associated macrophage (TAM) a tumor-associated neutrophil (TAN), and / or a regulatory T cell (Treg).

[0306] Embodiment 322. The CXCL12 antagonist for use of Embodiment 321 and the interferon for use of Embodiment 321, wherein the cell is immunomodulatorily active.

[0307] Embodiment 323. The CXCL12 antagonist for use of Embodiment 322 and the interferon for use of Embodiment 322, wherein the immunomodulatorily active cell is active by means of a secreted immunomodulator, metabolic reprogramming and / or regulatory cell-to-cell contact.

[0308] Embodiment 324. The CXCL12 antagonist for use of any one of Embodiments 290 and 292 to 323 and the interferon for use of any one of Embodiments 291 to 323, wherein the tumor is a solid tumor.

[0309] Embodiment 325. The CXCL12 antagonist for use of any one of Embodiments 290 and 292 to 324 and the interferon for use of any one of Embodiments 291 to 324, wherein the tumor is selected from the group comprising pancreatic cancer, preferably pancreatic ductal adenocarcinoma (PDAC); triplenegative breast cancer; prostate cancer; ovarian cancer; soft tissue sarcoma and osteosarcoma.Embodiment 326. The CXCL12 antagonist for use of Embodiment 325 and the interferon for use of Embodiment 325, wherein the tumor is high-grade serous ovarian cancer.

[0310] Embodiment 327. The CXCL12 antagonist for use of Embodiment 325 and the interferon for use of Embodiment 325, wherein the tumor is advanced castration-resistant prostate cancer.

[0311] Embodiment 328. The CXCL12 antagonist for use of Embodiment 325 and the interferon for use of Embodiment 325, wherein the tumor is synovial sarcoma.

[0312] Embodiment 329. The CXCL12 antagonist for use of any one of Embodiments 1 to 328 and the interferon for use of any one of Embodiments 2 to 328, wherein the method comprises administering to the subject an adoptive cell therapy.

[0313] Embodiment 330. The CXCL12 antagonist for use of Embodiment 329 and the interferon for use of Embodiment 329, wherein the adoptive cell therapy comprises administering a CAR T cell to the subject.

[0314] Embodiment 331. The CXCL12 antagonist for use of Embodiment 329 and the interferon for use of Embodiment 329, wherein the adoptive cell therapy comprises administering a TCR-T cell to the subject.

[0315] Embodiment 332. The CXCL12 antagonist for use of any one of Embodiments 1 to 331 and the interferon for use of any one of Embodiments 2 to 331, wherein the CXCL12 antagonist and the interferon are administered concomitantly to the subject.

[0316] Embodiment 333. The CXCL12 antagonist for use of any one of Embodiments 1 to 331 and the interferon for use of any one of Embodiments 2 to 331, wherein the CXCL12 antagonist is administered to the subject after the interferon.

[0317] Embodiment 334. The CXCL12 antagonist for use of any one of Embodiments 1 to 333 and the interferon for use of any one of Embodiments 2 to 335, wherein the interferon is a type I interferon.

[0318] Embodiment 335. The CXCL12 antagonist for use of Embodiment 334 and the interferon for use of Embodiment 334, wherein the type I interferon increases expression of MHC I on tumor cells.Embodiment 336. The CXCL12 antagonist for use of any one of Embodiments 334 and 335 and the interferon for use of any one of Embodiments 336 and 337, wherein the type I interferon is interferon alpha.

[0319] Embodiment 337. The CXCL12 antagonist for use of Embodiment 336 and the interferon for use of Embodiment 336, wherein the type I interferon is interferon alpha comprising an amino acid sequence of SEQ ID NO: 242 or SEQ ID NO: 243

[0320] Embodiment 338. The CXCL12 antagonist for use of any one of Embodiments 1 to 333 and the interferon for use of any one of Embodiments 2 to 333, wherein the interferon is a type II interferon.

[0321] Embodiment 339. The CXCL12 antagonist for use of Embodiment 338 and the interferon for use of Embodiment 338, wherein the type II interferon increases expression of MHC I on tumor cells.

[0322] Embodiment 340. The CXCL12 antagonist for use of any one of Embodiments 338 and 339 and the interferon for use of any one of Embodiments 336 and 337, wherein the type II interferon is interferon gamma.

[0323] Embodiment 341. The CXCL12 antagonist for use of Embodiment 340 and the interferon for use of Embodiment 340, wherein the type II interferon is interferon gamma comprising an amino acid sequence of SEQ ID NO: 245.

[0324] Embodiment 342. The CXCL12 antagonist for use of any one of Embodiments 1 to 341 and the interferon for use of any one of Embodiments 2 to 341, wherein the CXCL12 antagonist inhibits the binding of CXCL12 to CXCL12 receptor C-X-C chemokine receptor type 4 (CXCR4) and / or to CXCL12 receptor C-X-C chemokine receptor type 7 (CXCR7).

[0325] Embodiment 343. The CXCL12 antagonist for use of Embodiment 342 and the interferon for use of Embodiments 342, wherein the CXCL12 antagonist is binding to CXCL12.

[0326] Embodiment 344. The CXCL12 antagonist for use of Embodiment 343 and the interferon for use of Embodiments 343, wherein the CXCL12 antagonist is a nucleic acid molecule binding to CXCL12, wherein the nucleic acid molecule is preferably selected from the group comprising an SDF-1 bindingnucleic acid molecule of type B, an SDF-1 binding nucleic acid molecule of type C, an SDF-1 binding nucleic acid molecule of type A and an SDF-1 binding nucleic acid molecule of type D.

[0327] Embodiment 345. The CXCL12 antagonist for use of Embodiment 343 and the interferon for use of Embodiments 344, wherein the SDF-1 binding nucleic acid molecule of type B comprises a central stretch of nucleotides, whereby the central stretch of nucleotides comprises the following nucleotide sequence:

[0328] 5’ GUGUGAUCUAGAUGUADWGGCUGWUCCUAGUYAGG 3’ (SEQ ID NO: 57). Embodiment 346. The CXCL12 antagonist for use of Embodiment 345 and the interferon for use of Embodiments 345, wherein the central stretch of nucleotides comprises the following nucleotide sequence:

[0329] 5’ GUGUGAUCUAGAUGUADUGGCUGAUCCUAGUCAGG 3’ (SEQ ID NO: 58).

[0330] Embodiment 347. The CXCL12 antagonist for use of any one of Embodiments 345 and 346 and the interferon for use of any one of Embodiments 345 and 346, wherein the SDF-1 binding nucleic acid molecule of type B comprises in 5 ’->3’ direction a first terminal stretch of nucleotides, the central stretch of nucleotides, and a second terminal stretch of nucleotides.

[0331] Embodiment 348. The CXCL12 antagonist for use of any one of Embodiments 345 and 346 and the interferon for use of any one of Embodiments 345 and 346, wherein the SDF-1 binding nucleic acid molecule of type B comprises in 5 ’->3’ direction a second terminal stretch of nucleotides, the central stretch of nucleotides, and a first terminal stretch of nucleotides.

[0332] Embodiment 349. The CXCL12 antagonist for use of any one of Embodiments 347 and 348 and the interferon for use of any one of Embodiments 347 and 348, wherein the first terminal stretch of nucleotides comprises a nucleotide sequence of 5’ X1X2SVNS 3’ and the second terminal stretch of nucleotides comprises a nucleotide sequence of 5’ BVBSX3X43’, whereby

[0333] XI is either absent or is A, X2 is G, X3 is C and X4 is either absent or is U;

[0334] or

[0335] XI is absent, X2 is either absent or is G, X3 is either absent or is C and X4 is absent.

[0336] Embodiment 350. The CXCL12 antagonist for use of any one of Embodiments 347 to 349 and the interferon for use of any one of Embodiments 347 to 349, wherein the first terminal stretch ofnucleotides comprises a nucleotide sequence of 5’ X1X2CRWG 3’ and the second terminal stretch of nucleotides comprises a nucleotide sequence of 5’ KRYSX3X43’,

[0337] whereby XI is either absent or A, X2 is G, X3 is C and X4 is either absent or U.

[0338] Embodiment 351. The CXCL12 antagonist for use of any one of Embodiments 347 to 350 and the interferon for use of any one of Embodiments 347 to 350, preferably embodiment 349 or 350, wherein the first terminal stretch of nucleotides comprises a nucleotide sequence of 5’ X1X2CGUG 3’ and the second terminal stretch of nucleotides comprises a nucleotide sequence of 5’ UACGX3X4 3’,

[0339] whereby X 1 is either absent or A, X2 is G, X3 is C, and X4 is either absent or U,

[0340] preferably the first terminal stretch of nucleotides comprises a nucleotide sequence of 5’ AGCGUG 3’ and the second terminal stretch of nucleotides comprises a nucleotide sequence of 5’ UACGCU 3’.

[0341] Embodiment 352. The CXCL12 antagonist for use of any one of Embodiments 347 to 350 and the interferon for use of any one of Embodiments 347 to 350, preferably embodiment 349, wherein the first terminal stretch of nucleotides comprises a nucleotide sequence of 5’ X1X2SSBS 3’ and the second terminal stretch of nucleotides comprises a nucleotide sequence of 5’ BVSSX3X4 3’,

[0342] whereby XI is absent, X2 is either absent or G, X3 is either absent or C, and X4 is absent, preferably the first terminal stretch of nucleotides comprises a nucleotide sequence of 5’ GCGUG 3’ and the second terminal stretch of nucleotides comprises a nucleotide sequence of 5’ UACGC 3’.

[0343] Embodiment 353. The CXCL12 antagonist for use of any one of Embodiments 344 to 352 and the interferon for use of any one of Embodiments 344 to 352, wherein the SDF-1 binding nucleic acid molecule of type B comprises a nucleotide sequence according to any one of SEQ ID NO: 46 to SEQ ID NO: 56, SEQ ID NO: 61 to SEQ ID NO:65, SEQ ID NO: 67 to SEQ ID NO: 72 and SEQ ID NO: 132 or a nucleotide sequence at least 80 % identical thereto,

[0344] preferably any one of SEQ ID NO: 46 to SEQ ID NO: 48, SEQ ID NO: 61, SEQ ID NO: 67 and SEQ ID NO: 132 or a nucleotide sequence at least 80 % identical thereto,more preferably any one of SEQ ID NO: 67 and SEQ ID NO: 132 or a nucleotide sequence at least 80 % identical thereto.

[0345] Embodiment 354. The CXCL12 antagonist for use of Embodiment 344 and the interferon for use of Embodiment 344, wherein the SDF-1 binding nucleic acid molecule of type C comprises a central stretch of nucleotides, whereby the central stretch of nucleotides comprises a nucleotide sequence of GGUYAGGGCUHRXAAGUCGG (SEQ ID NO: 90),

[0346] whereby XA is either absent or is A.

[0347] Embodiment 355. The CXCL12 antagonist for use of Embodiment 354 and the interferon for use of Embodiment 354, wherein the central stretch of nucleotides comprises a nucleotide sequence of 5’ GGUYAGGGCUHRAAGUCGG 3’ (SEQ ID NO: 91), 5’ GGUYAGGGCUHRAGUCGG 3’ (SEQ ID NO: 92) or 5’ GGUUAGGGCUHGAAGUCGG 3’ (SEQ ID NO: 93), preferably 5’ GGUUAGGGCUHGAAGUCGG 3’ (SEQ ID NO: 93).

[0348] Embodiment 356. The CXCL12 antagonist for use of any one of Embodiments 344 to 352 and the interferon for use of any one of Embodiments 354 to 355, wherein the SDF-1 binding nucleic acid molecule of type C comprises in 5 ’->3’ direction a first terminal stretch of nucleotides, the central stretch of nucleotides, and a second terminal stretch of nucleotides.

[0349] Embodiment 357. The CXCL12 antagonist for use of any one of Embodiments 344 to 352 and the interferon for use of any one of Embodiments 354 to 355, wherein the SDF-1 binding nucleic acid molecule of type C comprises in 5 ’->3’ direction a second terminal stretch of nucleotides, the central stretch of nucleotides, and a first terminal stretch of nucleotides.

[0350] Embodiment 358. The CXCL12 antagonist for use of any one of Embodiments 356 to 357 and the interferon for use of any one of Embodiments 356 to 357, wherein the first terminal stretch of nucleotides comprises a nucleotide sequence of 5’ RKSBUSNVGR 3’ (SEQ ID NO: 120) and the second stretch of nucleotides comprises a nucleotide sequence of 5’ YYNRCASSMY 3’ (SEQ ID NO: 121),

[0351] preferably the first terminal stretch of nucleotides comprises a nucleotide sequence of 5’ RKSBUGSVGR 3’ (SEQ ID NO: 122) and the second terminal stretch of nucleotides comprises a nucleotide sequence of 5’ YCNRCASSMY 3’ (SEQ ID NO: 123).Embodiment 359. The CXCL12 antagonist for use of any one of Embodiments 356 to 357 and the interferon for use of any one of Embodiments 356 to 357, wherein the first terminal stretch of nucleotides comprises a nucleotide sequence of 5’ XSSSSV 3’ and the second terminal stretch of nucleotides comprises a nucleotide sequence of 5’ BSSSXS 3’, whereby Xs is either absent or is S,

[0352] preferably the first terminal stretch of nucleotides comprises a nucleotide sequence of 5’ SGGSR 3’ and the second terminal stretch of nucleotides comprises a nucleotide sequence of 5’ YSCCS 3’.

[0353] Embodiment 360. The CXCL12 antagonist for use of any one of Embodiments 356 to 357 and the interferon for use of any one of Embodiments 356 to 357, wherein

[0354] a) the first terminal stretch of nucleotides comprises a nucleotide sequence of 5’ GCCGG 3’ and the second terminal stretch of nucleotides comprises a nucleotide sequence of 5’ CCGGC 3’; or

[0355] b) the first terminal stretch of nucleotides comprises a nucleotide sequence of 5’ CGUGCGCUUGAGAUAGG 3’ (SEQ ID NO: 248) and the second terminal stretch of nucleotides comprises a nucleotide sequence of 5’ CUGAUUCUCACG 3’ (SEQ ID NO: 249); or

[0356] c) the first terminal stretch of nucleotides comprises a nucleotide sequence of 5’ UGAGAUAGG 3 ’ and the second terminal stretch of nucleotides comprises a nucleotide sequence of 5’ CUGAUUCUCA 3’ (SEQ ID NO: 250); or

[0357] d) the first terminal stretch of nucleotides comprises a nucleotide sequence of 5’ GAGAUAGG 3’ and the second terminal stretch of nucleotides comprises a nucleotide sequence of 5’ CUGAUUCUC 3’.

[0358] Embodiment 361. The CXCL12 antagonist for use of any one of Embodiments 354 to 360 and the interferon for use of any one of Embodiments 354 to 360, wherein the type C SDF-1 binding nucleic acid molecule comprises a nucleotide sequence according to any one of SEQ ID NO: 79 to SEQ ID NO: 89, SEQ ID NO: 258 and SEQ ID NO: 259, SEQ ID NO: 94 to SEQ ID NO: 119, SEQ ID NO: 134 and SEQ ID NO: 135,

[0359] preferably any one of SEQ ID NO: 102, SEQ ID NO: 110, SEQ ID NO: 111, SEQ ID NO: 116, SEQ ID NO: 117, SEQ ID NO: 134 and SEQ ID NO: 135.Embodiment 362. The CXCL12 antagonist for use of Embodiments 344 and the interferon for use of Embodiment 344, wherein the SDF-1 binding nucleic acid molecule of type A comprises a central stretch of nucleotides, whereby the central stretch of nucleotides comprises a nucleotide sequence of 5’ AAAGYRACAHGUMAAXAUGAAAGGUARC 3’ (SEQ ID NO: 19),

[0360] whereby XA is either absent or is A.

[0361] Embodiment 363. The CXCL12 antagonist for use of Embodiments 362 and the interferon for use of Embodiment 362, wherein the central stretch of nucleotides comprises a nucleotide sequence of

[0362] 5’AAAGYRACAHGUMAAUGAAAGGUARC 3’ (SEQ ID NO: 20), or

[0363] 5’ AAAGYRACAHGUMAAAUGAAAGGUARC 3’ (SEQ ID NO: 21), or

[0364] 5’ AAAGYAACAHGUCAAUGAAAGGUARC 3 ’(SEQ ID NO: 22), preferably the central stretch of nucleotides comprises a nucleotide sequence of 5’ AAAGYAACAHGUCAAUGAAAGGUARC 3’ (SEQ ID NO: 22).

[0365] Embodiment 364. The CXCL12 antagonist for use of any one of Embodiments 354 to 360 and the interferon for use of any one of Embodiments 354 to 360, wherein the SDF-1 binding nucleic acid molecule of type A comprises in 5 ’->3’ direction a first terminal stretch of nucleotides, the central stretch of nucleotides, and a second terminal stretch of nucleotides.

[0366] Embodiment 365. The CXCL12 antagonist for use of any one of Embodiments 362 to 363 and the interferon for use of any one of Embodiments 362 to 363, wherein the SDF-1 binding nucleic acid molecule of type A comprises in 5 ’->3’ direction a second terminal stretch of nucleotides, the central stretch of nucleotides, and a first terminal stretch of nucleotides.

[0367] Embodiment 366. The CXCL12 antagonist for use of any one of Embodiments 364 to 365 and the interferon for use of any one of Embodiments 364 to 365, wherein the first terminal stretch of nucleotides comprises a nucleotide sequence of 5’ X1X2NNBV 3’ and the second terminal stretch of nucleotides comprises a nucleotide sequence of 5 ’ BNBNX3X43 ’

[0368] whereby Xi is either absent or R, X2 is S, X3 is S and X4 is either absent or Y;

[0369] or

[0370] Xi is absent, X2 is either absent or S, X3 is either absent or S and X4 is absent.Embodiment 367. The CXCL12 antagonist for use of any one of Embodiments 364 to 366, preferably Embodiment 366 and the interferon for use of any one of Embodiments 364 to 366, preferably Embodiment 366, wherein the first terminal stretch of nucleotides comprises a nucleotide sequence of 5’ RSHRYR 3’ and the second terminal stretch of nucleotides comprises a nucleotide sequence of 5’ YRYDSY 3’,

[0371] preferably the first terminal stretch of nucleotides comprises a nucleotide sequence of 5’ GCUGUG 3’ and the second terminal stretch of nucleotides comprises a nucleotide sequence of 5’ CGCAGC 3’.

[0372] Embodiment 368. The CXCL12 antagonist for use of any one of Embodiments 364 to 366, preferably Embodiment 366, and the interferon for use of any one of Embodiments 364 to 366, preferably Embodiment 366, wherein the first terminal stretch of nucleotides comprises a nucleotide sequence of 5’ X2BBBS 3’ and the second terminal stretch of nucleotides comprises a nucleotide sequence of 5’ SBBVX3 3’,

[0373] whereby X2 is either absent or is S and X3 is either absent or is S;

[0374] preferably the first terminal stretch of nucleotides comprises a nucleotide sequence of 5 ’ CUGUG 3 ’ and the second terminal stretch of nucleotides comprises a nucleotide sequence of 5’ CGCAG 3’;

[0375] or the first terminal stretch of nucleotides comprises a nucleotide sequence of 5’ GCGUG 3 ’and the second terminal stretch of nucleotides comprises a nucleotide sequence of 5 ’ CGCGC 3 ’.

[0376] Embodiment 369. The CXCL12 antagonist for use of any one of Embodiments 362 to 368 and the interferon for use of any one of Embodiments 362 to 368, wherein the SDF-1 binding nucleic acid molecule of type A comprises a nucleotide sequence according to any one of SEQ ID NO: 5 to SEQ ID NO: 18, SEQ ID NO: 25 to SEQ ID NO: 29, SEQ ID NO: 31 to SEQ ID NO: 34, SEQ ID NO: 36 to SEQ ID NO: 41, and SEQ ID NO: 133 or a nucleotide sequence at least 80 % identical thereto,

[0377] preferably any one of SEQ ID NO: 5, SEQ ID NO: 8, SEQ ID NO: 11, SEQ ID NO: 25, SEQ ID NO: 31, and SEQ ID NO: 133 or a nucleotide sequence at least 80 % identical thereto,

[0378] more preferably any one of SEQ ID NO: 31 and SEQ ID NO: 133 or a nucleotide sequence at least 80 % identical thereto.Embodiment 370. The CXCL12 antagonist for use of Embodiment 344 and the interferon for use of Embodiment 344, wherein the SDF-1 binding nucleic acid molecule of type D comprises a nucleotide sequence according to any one of SEQ ID NO: 142 to SEQ ID NO: 144 or a nucleotide sequence at least 80 % identical thereto.

[0379] Embodiment 371. The CXCL12 antagonist for use of any one of Embodiments 344 to 370 and the interferon for use of any one of Embodiments 344 to 370, wherein the nucleic acid molecule comprises a modification, whereby the modification is preferably a high molecular weight moiety and / or whereby the modification preferably allows to modify the characteristics of the nucleic acid molecule in terms of residence time in the animal or human body, preferably the human body.

[0380] Embodiment 372. The CXCL12 antagonist for use of Embodiment 371 and the interferon for use of Embodiment 371, wherein the modification is selected from the group comprising a HES moiety, a PEG moiety, biodegradable modifications and combinations thereof.

[0381] Embodiment 373. The CXCL12 antagonist for use of Embodiment 372 and the interferon for use of Embodiment 372, wherein the modification is a PEG moiety consisting of a straight PEG or branched PEG, whereby preferably the molecular weight of the straight or branched PEG is from about 20,000 to 120,000 Da, more preferably from about 30,000 to 80,000 Da and most preferably about 40,000 Da.

[0382] Embodiment 374. The CXCL12 antagonist for use of Embodiment 372 and the interferon for use of Embodiment 372, wherein the modification is a HES moiety, whereby preferably the molecular weight of the HES moiety is from about 10,000 to 200,000 Da, more preferably from about 30,000 to 170.000 Da and most preferably about 150,000 Da.

[0383] Embodiment 375. The CXCL12 antagonist for use of any one of Embodiments 371 to 374 and the interferon for use of any one of Embodiments 371 to 374, wherein the modification is attached to the nucleic acid molecule via a linker, wherein preferably the linker is a biostable or biodegradable linker.

[0384] Embodiment 376. The CXCL12 antagonist for use of any one of Embodiments 371 to 375 and the interferon for use of any one of Embodiments 371 to 375, wherein the modification is attached to the nucleic acid molecule at the 5 ’-terminal nucleotide of the nucleic acid molecule and / or the 3 ’-terminal nucleotide of the nucleic acid molecule and / or to a nucleotide of the nucleic acid molecule between the 5 ’-terminal nucleotide of the nucleic acid molecule and the 3 ’-terminal nucleotide of the nucleic acid molecule.Embodiment 377. The CXCL12 antagonist for use of any one of Embodiments 344 to 376 and the interferon for use of any one of Embodiments 344 to 376, wherein the nucleotides of the nucleic acid molecule or the nucleotides forming the nucleic acid molecule are L-nucleotides.

[0385] Embodiment 378. The CXCL12 antagonist for use of any one of Embodiments 344 to 377 and the interferon for use of any one of Embodiments 344 to 377, wherein the nucleic acid molecule is an L-nucleic acid molecule.

[0386] Embodiment 379. The CXCL12 antagonist for use of Embodiment 344 and the interferon for use of Embodiment 344, wherein the nucleic acid molecule is an L-ribonucleic acid molecule comprising a nucleotide sequence of

[0387] GCGUGGUGUGAUCUAGAUGUAUUGGCUGAUCCUAGUCAGGUACGC (SEQ ID NO: 67).

[0388] Embodiment 380. The CXCL12 antagonist for use of Embodiment 379 and the interferon for use of Embodiment 379, wherein the L-ribonucleic acid molecule comprises a 40 kDa polyethylene glycol moiety, wherein the 40 kDa polyethylene glycol moiety is attached to the 5 ’end of nucleotide sequence.

[0389] Embodiment 381. The CXCL12 antagonist for use of Embodiment 380 and the interferon for use of Embodiment 380, wherein N-[co-methoxypoly(oxy-l,2-ethanediyl], N’-[co-methoxypoly(oxy-l,2-ethanediyl]-acetyl-9-amino-8-oxo-7-aza-nonyloxy phosphate is interspersed between the 5’ terminal nucleotide and the 40 kDa polyethylene glycol moiety.

[0390] Embodiment 382. The CXCL12 antagonist for use of Embodiment 381 and the interferon for use of Embodiment 381, wherein the L-ribonucleic acid molecule is present as a sodium salt, preferably the L-nucleic acid molecule is NOX-A12.

[0391] Embodiment 383. The CXCL12 antagonist for use of Embodiment 342 and the interferon for use of Embodiment 342, wherein the CXCL12 antagonist is an aptamer binding to CXCL12.

[0392] Embodiment 384. The CXCL12 antagonist for use of Embodiment 342 and the interferon for use of Embodiment 342, wherein the CXCL12 antagonist is an antibody binding to CXCL12.

[0393] Embodiment 385 The CXCL12 antagonist for use of Embodiment 384 and the interferon for use of Embodiment 384, wherein the antibody binding to CXCL12 is mAB-30D8, 1131-H12 or 1143-H1.Embodiment 386. The CXCL12 antagonist for use of Embodiment 342 and the interferon for use of Embodiment 342, wherein the CXCL12 antagonist is a CXCL12 binding protein.

[0394] Embodiment 387. The CXCL12 antagonist for use of Embodiment 386 and the interferon for use of Embodiment 386, wherein the protein is a TRAP protein or an anticalin.

[0395] Embodiment 388. The CXCL12 antagonist for use of Embodiment 342 and the interferon for use of Embodiment 342, wherein the CXCL12 antagonist targets CXCR4.

[0396] Embodiment 389. The CXCL12 antagonist for use of Embodiment 388 and the interferon for use of Embodiment 388, wherein the CXCL12 antagonist is binding to CXCR4.

[0397] Embodiment 390. The CXCL12 antagonist for use of any one of Embodiments 388 to 389 and the interferon for use of any one of Embodiments 388 to 389, wherein the CXCR4 binding CXCL12 antagonist is selected from the group comprising a small molecule, a peptide, a synthetic peptide, a recombinant protein, a fusion protein and an antibody.

[0398] Embodiment 391. The CXCL12 antagonist for use of Embodiment 390 and the interferon for use of Embodiment 390, wherein the CXCR4 binding CXCL12 antagonist is a small molecule.

[0399] Embodiment 392. The CXCL12 antagonist for use of Embodiment 391 and the interferon for use of Embodiment 391, wherein the CXCR4 binding CXCL12 antagonist is a small molecule selected from the group comprising plerixafor, mavorixafor, Q-122, HPH-112, BKT-300, X4P-002, X4P-003, X4-136, GP-01CR01, GP-01CR11, GP-01CR21, Pentixather, BKT-170, BMS-585248, CS-3955, CTCE-0012, CTCE-9908, GBV-4086, HPH-112, HPH-211, NB-325, SP-10 and USL-311.

[0400] Embodiment 393. The CXCL12 antagonist for use of Embodiment 392 and the interferon for use of Embodiment 392, wherein the small molecule is selected from the group comprising plerixafor, mavorixafor, Q-122, HPH-112, BKT-300, X4P-002, X4P-003, X4-136, GP-01CR01, GP-01CR11, GP-01CR21, and pentixather.

[0401] Embodiment 394. The CXCL12 antagonist for use of Embodiment 390 and the interferon for use of Embodiment 390, wherein the CXCR4 binding CXCL12 antagonist is a peptide.

[0402] Embodiment 395. The CXCL12 antagonist for use of Embodiment 394 and the interferon for use of Embodiment 394, wherein the CXCR4 binding CXCL12 antagonist is motixafortide.Embodiment 396. The CXCL12 antagonist for use of Embodiment 390 and the interferon for use of Embodiment 390, wherein the CXCR4 binding CXCL12 antagonist is a synthetic peptide.

[0403] Embodiment 397. The CXCL12 antagonist for use of Embodiment 396 and the interferon for use of Embodiment 396, wherein the CXCR4 binding CXCL12 antagonist is a synthetic peptide selected from the group comprising balixafortide, LY-2510924, ALB-408 and POL-5551.

[0404] Embodiment 398. The CXCL12 antagonist for use of Embodiment 397 and the interferon for use of Embodiment 397, wherein the CXCR4 binding CXCL12 antagonist is selected from the group comprising balixafortide, and LY-2510924.

[0405] Embodiment 399. The CXCL12 antagonist for use of Embodiment 390 and the interferon for use of Embodiment 390, wherein the CXCR4 binding CXCL12 antagonist is a recombinant protein.

[0406] Embodiment 400. The CXCL12 antagonist for use of Embodiment 399 and the interferon for use of Embodiment 399, wherein the recombinant protein is selected from the group comprising PTX-9098 andNNL-121.

[0407] Embodiment 401. The CXCL12 antagonist for use of Embodiment 390 and the interferon for use of Embodiment 390, wherein the CXCR4 binding CXCL12 antagonist is a fusion protein.

[0408] Embodiment 402. The CXCL12 antagonist for use of Embodiment 401 and the interferon for use of Embodiment 401, wherein the fusion protein is selected from the group comprising AD-214 and AM-3114.

[0409] Embodiment 403. The CXCL12 antagonist for use of Embodiment 390 and the interferon for use of Embodiment 390, wherein the CXCR4 binding CXCL12 antagonist is an antibody or an antigenbinding fragment of the antibody.

[0410] Embodiment 404. The CXCL12 antagonist for use of Embodiment 403 and the interferon for use of Embodiment 403, wherein the antibody is a monoclonal antibody.

[0411] Embodiment 405. The CXCL12 antagonist for use of any one of Embodiments 403 and 404 the interferon for use of any one of Embodiments 403 and 404, wherein the CXCR4 binding CXCL12 antagonist is an antibody.

[0412] Embodiment 406. The CXCL12 antagonist for use of Embodiment 405 and the interferon for use of Embodiment 405, wherein the antibody is a monoclonal antibody selected from the groupcomprising ulocuplumab, hz-515H7, KY-1051, PF-06747143, ALX-0651, AT-009, LY-2624587 and STIA-220X.

[0413] Embodiment 407. The CXCL12 antagonist for use of Embodiment 406 and the interferon for use of Embodiment 406, wherein the antibody is a monoclonal antibody selected from the group comprising ulocuplumab, hz-515H7 and KY-1051.

[0414] Embodiment 408. The CXCL12 antagonist for use of Embodiment 342 and the interferon for use of Embodiment 342, wherein the CXCL12 antagonist is binding to CXCR7.

[0415] Embodiment 409. The CXCL12 antagonist for use of Embodiment 408 and the interferon for use of Embodiment 408, wherein the CXCR7 binding CXCL12 antagonist is selected from the group comprising a small molecule, a synthetic peptide and an antibody.

[0416] Embodiment 410. The CXCL12 antagonist for use of Embodiment 409 and the interferon for use of Embodiment 409, wherein the CXCR7 binding CXCL12 antagonist is a small molecule.

[0417] Embodiment 411. The CXCL12 antagonist for use of Embodiment 410 and the interferon for use of Embodiment 410, wherein the CXCR7 binding CXCL12 antagonist is a small molecule selected from the group comprising ACT-10041239, CCX-650, CCX-662 and CCX-771.

[0418] Embodiment 412. The CXCL12 antagonist for use of Embodiment 411 and the interferon for use of Embodiment 411, wherein the CXCR7 binding CXCL12 antagonist is ACT-10041239.

[0419] Embodiment 413. The CXCL12 antagonist for use of Embodiment 409 and the interferon for use of Embodiment 409, wherein the CXCR7 binding CXCL12 antagonist is a synthetic peptide.

[0420] Embodiment 414. The CXCL12 antagonist for use of Embodiment 413 and the interferon for use of Embodiment 413, wherein the synthetic peptide is selected from the group comprising LIH-383 and POL-6926.

[0421] Embodiment 415. The CXCL12 antagonist for use of Embodiment 414 and the interferon for use of Embodiment 414, wherein the synthetic peptide is LIH-383.

[0422] Embodiment 416. The CXCL12 antagonist for use of Embodiment 409 and the interferon for use of Embodiment 409, wherein the CXCR7 binding CXCL12 antagonist is an antibody or an antigenbinding fragment of the antibody.Embodiment 417. The CXCL12 antagonist for use of Embodiment 416 and the interferon for use of Embodiment 416, wherein the antibody is a monoclonal antibody.

[0423] Embodiment 418. The CXCL12 antagonist for use of any one of Embodiments 416 and 417 the interferon for use of any one of Embodiments 416 and 417, wherein the antibody is a monoclonal antibody selected from the group comprising JT-07, X-7Ab and STIA-230X.

[0424] Embodiment 419. The CXCL12 antagonist for use of Embodiment 418 and the interferon for use of Embodiment 418, wherein the antibody is a monoclonal antibody selected from the group comprising JT-07 and X-7Ab.

[0425] The problem underlying the present invention is solved in a first aspect which is also a first embodiment of the first aspect, by a CXCL12 antagonist for use in a method for treating a tumor in a subject, wherein the method comprises administering to the subject the CXCL12 antagonist and an interferon, wherein the tumor is an immunologically cold tumor.

[0426] The problem underlying the present invention is solved in a second aspect which is also a first embodiment of the second aspect, by an interferon for use in a method for treating a tumor in a subject, wherein the method comprises administering to the subject the interferon and a CXCL12 antagonist, wherein the tumor is an immunologically cold tumor.

[0427] The problem underlying the present invention is solved in a third aspect which is also a first embodiment of the third aspect, by a CXCL12 antagonist for use in a method for treating a tumor in a subject, wherein the method comprises administering to the subject the CXCL12 antagonist and an interferon, wherein the tumor is refractory to immunotherapy.

[0428] The problem underlying the present invention is solved in a fourth aspect which is also a first embodiment of the fourth aspect, by an interferon for use in a method for treating a tumor in a subject, wherein the method comprises administering to the subject the interferon and a CXCL12 antagonist, wherein the tumor is refractory to immunotherapy.

[0429] The problem underlying the present invention is solved in a fifth aspect which is also a first embodiment of the fifth aspect, by a CXCL12 antagonist for use in a method for treating a tumor in asubject, wherein the method comprises administering to the subject the CXCL12 antagonist and an interferon, wherein the tumor is a solid tumor.

[0430] The problem underlying the present invention is solved in a sixth aspect which is also a first embodiment of the sixth aspect, by an interferon for use in a method for treating a tumor in a subject, wherein the method comprises administering to the subject the interferon and a CXCL12 antagonist, wherein the tumor is a solid tumor.

[0431] The problem underlying the present invention is solved in a seventh aspect which is also a first embodiment of the seventh aspect, by a CXCL12 antagonist for use in a method for treating a tumor in a subject, wherein the method comprises administering to the subject the CXCL12 antagonist, an interferon and an adjuvant therapy, wherein the tumor is an immunologically cold tumor.

[0432] The problem underlying the present invention is solved in an eighth aspect which is also a first embodiment of the eighth aspect, by an interferon for use in a method for treating a tumor in a subject, wherein the method comprises administering to the subject the interferon, a CXCL12 antagonist and an adjuvant therapy, wherein the tumor is an immunologically cold tumor.

[0433] The problem underlying the present invention is solved in a ninth aspect which is also a first embodiment of the ninth aspect, by a CXCL12 antagonist for use in a method for treating a tumor in a subject, wherein the method comprises administering to the subject the CXCL12 antagonist, an interferon and adjuvant therapy, wherein the tumor is refractory to immunotherapy.

[0434] The problem underlying the present invention is solved in a tenth aspect which is also a first embodiment of the tenth aspect, by an interferon for use in a method for treating a tumor in a subject, wherein the method comprises administering to the subject the interferon, a CXCL12 antagonist and an adjuvant therapy, wherein the tumor is refractory to immunotherapy.

[0435] The problem underlying the present invention is solved in an eleventh aspect which is also a first embodiment of the eleventh aspect, by a CXCL12 antagonist for use in a method for treating a tumor in a subject, wherein the method comprises administering to the subject the CXCL12 antagonist, an interferon and an adjuvant therapy, wherein the tumor is a solid tumor.The problem underlying the present invention is solved in a twelfth aspect which is also a first embodiment of the twelfth aspect, by an interferon for use in a method for treating a tumor in a subject, wherein the method comprises administering to the subject the interferon, a CXCL12 antagonist and an adjuvant therapy, wherein the tumor is a solid tumor.

[0436] The problem underlying the present invention is solved in a 13thaspect which is also a first embodiment of the 13thaspect, by a CXCL12 antagonist for use in a method for treating a tumor in a subject, wherein the method comprises administering to the subject a primary therapy and an adjunct therapy, wherein the adjunct therapy comprises administering to the subject the CXCL12 antagonist and an interferon, wherein the tumor is an immunologically cold tumor.

[0437] The problem underlying the present invention is solved in a 14thaspect which is also a first embodiment of the 14thaspect, by an interferon for use in a method for treating a tumor in a subject, wherein the method comprises administering to the subject a primary therapy and an adjunct therapy, wherein the adjunct therapy comprises administering to the subject the interferon and a CXCL12 antagonist, wherein the tumor is an immunologically cold tumor.

[0438] The problem underlying the present invention is solved in a 15thaspect which is also a first embodiment of the 15thaspect, by a CXCL12 antagonist for use in a method for treating a tumor in a subject, wherein the method comprises administering to the subject a primary therapy and an adjunct therapy, wherein the adjunct therapy comprises administering to the subject the CXCL12 antagonist and an interferon, wherein the tumor is refractory to immunotherapy.

[0439] The problem underlying the present invention is solved in a 16thaspect which is also a first embodiment of the 16thaspect, by an interferon for use in a method for treating a tumor in a subject, wherein the method comprises administering to the subject a primary therapy and an adjunct therapy, wherein the adjunct therapy comprises administering to the subject the interferon and a CXCL12 antagonist, wherein the tumor is refractory to immunotherapy.

[0440] The problem underlying the present invention is solved in a 17thaspect which is also a first embodiment of the 17thaspect, by a CXCL12 antagonist for use in a method for treating a tumor in a subject, wherein the method comprises administering to the subject a primary therapy and an adjunct therapy, wherein the adjunct therapy comprises administering to the subject the CXCL12 antagonist and an interferon, wherein the tumor is a solid tumor.The problem underlying the present invention is solved in an 18thaspect which is also a first embodiment of the 18thaspect, by an interferon for use in a method for treating a tumor in a subject, wherein the method comprises administering to the subject a primary therapy and an adjunct therapy, wherein the adjunct therapy comprises administering to the subject the interferon and a CXCL12 antagonist, wherein the tumor is a solid tumor.

[0441] The problem underlying the present invention is solved in an 19thaspect which is also a first embodiment of the 19thaspect, by a CXCL12 antagonist for use in a method for converting an immunologically cold tumor in a subject into an immunologically hot tumor in the subject, wherein the method comprises administering to the subject the CXCL12 antagonist and an interferon, wherein upon administration of the CXCL12 antagonist and the interferon the immunologically cold tumor is or will be converted into an immunologically hot tumor.

[0442] The problem underlying the present invention is solved in a 20thaspect which is also a first embodiment of the 20thaspect, by an interferon for use in a method for converting an immunologically cold tumor in a subject into an immunologically hot tumor in the subject, wherein the method comprises administering to the subject the interferon and a CXCL12 antagonist, wherein upon administration of the interferon and the CXCL12 antagonist the immunologically cold tumor is or will be converted into an immunologically hot tumor.

[0443] The problem underlying the present invention is solved in a 21staspect which is also a first embodiment of the 21staspect, by a CXCL12 antagonist for use in a method for converting an immunotherapy refractory tumor in a subject into an immunotherapy non-refractory tumor in the subject, wherein the method comprises administering to the subject the CXCL12 antagonist and an interferon, wherein upon administration of the CXCL12 antagonist and the interferon the immunotherapy refractory tumor is or will be converted into an immunotherapy non-refractory tumor.

[0444] The problem underlying the present invention is solved in a 22ndaspect which is also a first embodiment of the 22ndaspect, by an interferon for use in a method for converting an immunotherapy refractory tumor in a subject into an immunotherapy non-refractory tumor in the subject, wherein the method comprises administering to the subject the interferon and a CXCL12 antagonist, wherein upon administration of the interferon and the CXCL12 antagonist the immunotherapy refractory tumor is or will be converted into an immunotherapy non-refractory tumor.The problem underlying the present invention is solved in a 23rdaspect which is also a first embodiment of the 23rdaspect, by a method for treating a tumor in a subject, wherein the method comprises administering to the subject a CXCL12 antagonist and an interferon, wherein the tumor is an immunologically cold tumor.

[0445] The problem underlying the present invention is solved in a 24thaspect which is also a first embodiment of the 24thaspect, by a method for treating a tumor in a subject, wherein the method comprises administering to the subject a CXCL12 antagonist and an interferon, wherein the tumor is refractory to immunotherapy

[0446] The problem underlying the present invention is solved in a 25thaspect which is also a first embodiment of the 25thaspect, by a method for treating a tumor in a subject, wherein the method comprises administering to the subject a CXCL12 antagonist and an interferon, wherein the tumor is a solid tumor.

[0447] The problem underlying the present invention is solved in a 26thaspect which is also a first embodiment of the 26thaspect, by a method for treating a tumor in a subject, wherein the method comprises administering to the subject the CXCL12 antagonist, an interferon and an adjuvant therapy, wherein the tumor is an immunologically cold tumor.

[0448] The problem underlying the present invention is solved in a 27thaspect which is also a first embodiment of the 27thaspect, by a method for treating a tumor in a subject, wherein the method comprises administering to the subject a CXCL12 antagonist, an interferon and adjuvant therapy, wherein the tumor is refractory to immunotherapy.

[0449] The problem underlying the present invention is solved in a 28thaspect which is also a first embodiment of the 28thaspect, by a method for treating a tumor in a subject, wherein the method comprises administering to the subject a CXCL12 antagonist, an interferon and an adjuvant therapy, wherein the tumor is a solid tumor.

[0450] The problem underlying the present invention is solved in a 29thaspect which is also a first embodiment of the 29thaspect, by a method for treating a tumor in a subject, wherein the method comprises administering to the subject a primary therapy and an adjunct therapy, wherein the adjuncttherapy comprises administering to the subject a CXCL12 antagonist and an interferon, wherein the tumor is an immunologically cold tumor.

[0451] The problem underlying the present invention is solved in a 30thaspect which is also a first embodiment of the 30thaspect, by a method for treating a tumor in a subject, wherein the method comprises administering to the subject a primary therapy and an adjunct therapy, wherein the adjunct therapy comprises administering to the subject a CXCL12 antagonist and an interferon, wherein the tumor is refractory to immunotherapy.

[0452] The problem underlying the present invention is solved in a 31staspect which is also a first embodiment of the 31staspect, by a method for treating a tumor in a subject, wherein the method comprises administering to the subject a primary therapy and an adjunct therapy, wherein the adjunct therapy comprises administering to the subject a CXCL12 antagonist and an interferon, wherein the tumor is a solid tumor.

[0453] The problem underlying the present invention is solved in a 32ndaspect which is also a first embodiment of the 32ndaspect, by a method for converting an immunologically cold tumor in a subject into an immunologically hot tumor in the subject, wherein the method comprises administering to the subject a CXCL12 antagonist and an interferon, wherein upon administration of the CXCL12 antagonist and the interferon the immunologically cold tumor is or will be converted into an immunologically hot tumor.

[0454] The problem underlying the present invention is solved in a 33rdaspect which is also a first embodiment of the 33rdaspect, by a method for converting an immunotherapy refractory tumor in a subject into an immunotherapy non-refractory tumor in the subject, wherein the method comprises administering to the subject a CXCL12 antagonist and an interferon, wherein upon administration of the CXCL12 antagonist and the interferon the immunotherapy refractory tumor is or will be converted into an immunotherapy non-refractory tumor.

[0455] It will be appreciated that the first aspect, the second aspect and the 23rdaspect are related and that each and any embodiment of one of these aspects, including any embodiment of these aspects, will also be an embodiment of the other of these aspects, including any embodiment of the other of these aspects.It will be appreciated that the third aspect, the fourth aspect and the 24thaspect are related and that each and any embodiment of one of these aspects, including any embodiment of these aspects, will also be an embodiment of the other of these aspects, including any embodiment of the other of these aspects.

[0456] It will be appreciated that the fifth aspect, the sixth aspect and the 25thaspect are related and that each and any embodiment of one of these aspects, including any embodiment of these aspects, will also be an embodiment of the other of these aspects, including any embodiment of the other of these aspects.

[0457] It will be appreciated that the seventh aspect, the eighth aspect and the 26thaspect are related and that each and any embodiment of one of these aspects, including any embodiment of these aspects, will also be an embodiment of the other of these aspects, including any embodiment of the other of these aspects.

[0458] It will be appreciated that the ninth aspect, the tenth aspect and the 27thaspect are related and that each and any embodiment of one of these aspects, including any embodiment of these aspects, will also be an embodiment of the other of these aspects, including any embodiment of the other of these aspects.

[0459] It will be appreciated that the eleventh aspect, the twelfth aspect and the 28thaspect are related and that each and any embodiment of one of these aspects, including any embodiment of these aspects, will also be an embodiment of the other of these aspects, including any embodiment of the other of these aspects.

[0460] It will be appreciated that the 13thaspect, the 14thaspect and the 29thaspect are related and that each and any embodiment of one of these aspects, including any embodiment of these aspects, will also be an embodiment of the other of these aspects, including any embodiment of the other of these aspects.

[0461] It will be appreciated that the 15thaspect, the 16thaspect and the 30thaspect are related and that each and any embodiment of one of these aspects, including any embodiment of these aspects, will also be an embodiment of the other of these aspects, including any embodiment of the other of these aspects.

[0462] It will be appreciated that the 17thaspect, the 18thaspect and the 31staspect are related and that each and any embodiment of one of these aspects, including any embodiment of these aspects, will also be an embodiment of the other of these aspects, including any embodiment of the other of these aspects.It will be appreciated that the 19thaspect, the 20thaspect and the 32ndaspect are related and that each and any embodiment of one of these aspects, including any embodiment of these aspects, will also be an embodiment of the other of these aspects, including any embodiment of the other of these aspects.

[0463] It will be appreciated that the 21staspect, the 22ndaspect and the 33rdaspect are related and that each and any embodiment of one of these aspects, including any embodiment of these aspects, will also be an embodiment of the other of these aspects, including any embodiment of the other of these aspects.

[0464] It will be appreciated by a person skilled in the art that each and any embodiment of any aspect disclosed herein is also an embodiment of each and any of the other aspects, including any embodiments thereof, disclosed herein unless explicitly indicated to the contrary.

[0465] Any reference to the invention or to the present invention refers in particular to each and any aspect of the invention, including any embodiment of such each and any aspect. Also, the terms aspect and aspect of the invention or aspect of the present invention are used interchangeably and synonymously herein, unless indicated otherwise.

[0466] Without wishing to be bound by any theory, the present inventors have surprisingly found that administering both a CXCL12 antagonist and an interferon to a subject suffering from an immunologically cold tumor will transform or convert such immunologically cold tumor into an immunologically hot tumor. Such immunologically hot tumor is susceptible to the subject’s own antitumor immune response and / or to immunotherapy applied or administered to the subject.

[0467] Furthermore and also without wishing to be bound by any theory, the present inventors have surprisingly found that administering both a CXCL12 antagonist and an interferon to a subject suffering from a tumor refractory to immunotherapy will transform or convert such tumor refractory to immunotherapy into a tumor non-refractory to immunotherapy and, respectively, susceptible to immunotherapy. Such tumor non-refractory to immunotherapy is susceptible to immunotherapy applied or administered to the subject and / or to the subject’s anti-tumor immune response.

[0468] Finally and again without wishing to be bound by any theory, the present inventors have surprisingly found that administering both a CXCL12 antagonist and an interferon to a subject suffering from a solid tumor, preferably a pancreatic cancer, a synovial sarcoma or an osteosarcoma, allows aneffective treatment of the solid tumor, in particular if such tumor is an immunologically cold tumor and / or a tumor refractory to immunotherapy.

[0469] It will be appreciated by a person skilled in the art that when reference is made to a distinct tumor entity in a subject, this translates in terms of disease from which the subject is suffering into the distinct cancer which is the same as the distinct tumor entity. In other words, if a person is suffering from, e.g., synovial sarcoma, the tumor in the person is also referred to as synovial sarcoma, and vice versa.”

[0470] In an embodiment of each and any aspect, including any embodiment thereof, the tumor is an immunologically cold tumor. Immunologically cold tumors are cancers that lack an active immune response, making them harder to treat with immunotherapy alone. They often have a hostile tumor microenvironment that suppresses immune activation.

[0471] Immune checkpoint inhibitors (ICIs) have been approved to treat various tumors with demonstrated effectiveness (Miller, Nogueira et al. 2019) but many different tumors do not respond to ICI therapy, and even for tumor entities where ICI therapy is part of the standard of care, a large proportion of patients does not respond. A reason for this is the fact that tumors are capable of developing immune evasion mechanisms; therefore, tumors can adapts to acquire ‘cold’ properties (i.e., lack of effector immune cells in the tumor tissue) (Bai, Chen et al. 2020, Hegde and Chen 2020). Cold tumors are categorized into two types: (i) ‘immune-excluded’ tumors, where immune effector cells accumulate in the tumor periphery but are unable to infiltrate due to an immunosuppressive tumor microenvironment (TME); (ii) ‘immune-desert’ tumors, where also the surrounding tissues are devoid of immune effector cells due to low immunogenicity. In these cases, the non-targeted approach of immunostimulants as monotherapy is likely to be therapeutically futile. Combination therapy of immunostimulants together with ICIs could be promising in paving the way to overcome localized immunosuppression entailed by the TME. In particular, immune-excluded tumors could be reverted to ‘hot’ tumors (i.e., inflamed with a high presence of tumor-infiltrating lymphocytes [TILs]). When administered together with ICIs, these aforementioned immunostimulant drugs have shown a degree of success not only in pre-clinical cancer models (Charych, Hoch et al. 2016, Mariathasan, Turley et al. 2018, Sharma, Khong et al.

[0472] 2020) but also in clinical trials.

[0473] In an embodiment of each and any aspect of the present invention, including any embodiment thereof, the tumor has an immune evasion mechanism. In accordance with this embodiment, the tumor evadesthe host immune system’s surveillance instead of being eliminated. The mechanisms behind this are manifold. The major mechanisms underlying this are (1) camouflage, which hides cancer cells from immune recognition, (2) coercion, which directly or indirectly interferes with immune effector cells, and (3) cytoprotection, which shields malignant cells from immune cytotoxicity (see, e.g., (Galassi, Chan et al. 2024)5).

[0474] A key factor that determines whether certain tumors are cold or hot is related to the molecular characteristics of the tumor cells (microsatellite instability / mismatch repair defects, tumor mutational load, neoantigen load, etc.) (Samstein, Lee et al. 2019). “Hot tumors” as subject to various embodiments of the aspects are characterized by a TME rich in TILs, PD-L1 overexpression, genomic instability, and preexisting antitumor immune responses (Galon and Bruni 2019), whereas cold tumors, which usually do not respond to immunotherapy, have opposite characteristics (Sharma and Allison 2015).

[0475] In an embodiment of each and any aspect of the present invention, including any embodiment thereof, the tumor has at least one characteristic selected from the group comprising reduced or lacking expression of PD-L1, reduced number of tumor-infiltrating lymphocytes, genomic stability and reduced or lacking preexisting antitumor immune response. It will be appreciated by a person skilled in the art that the reference level for deciding whether there is a reduced or lacking expression of PD-Ll, a reduced number of tumor-infdtrating lymphocytes, genomic stability and a reduced or lacking preexisting antitumor immune response, is known to a person skilled in the art and outlined in comprehensive reviews such as in Wang 2023 (Wang, Geng et al. 2023). Similarly, a person skilled in the art is able based on such reference level to determine whether at least one of said characteristics has been reverted which is indicative that the tumor is an immunologically hot tumor.

[0476] The immune escape mechanisms that can lead to establishment of cold tumors are signified by cell-autonomous mechanisms, such as loss of MHC class I expression or the elevated expression of inhibitory immune checkpoints (Bai, Chen et al. 2020, Hegde and Chen 2020). Notably, immune escape can be the consequence of changes in the immune cell composition of the TME that affect both the adaptive and the innate immune systems. Escape from the adaptive immune system can be due to the inhibition of cytotoxic tumor-infdtrating lymphocyte (TIL) immunosurveillance and activation, enhanced by the enrichment of regulatory T (Treg) cells in the TME. Treg cells are normally trained to recognize self-antigens and are thus responsible for maintaining self-tolerance. However, since cancer cells commonly express self-antigens beside unique neoantigens, Treg cells could, in principle,counteract the anti-cancer immune response (Ohue and Nishikawa 2019). With respect to the innate immune system, the presence of myeloid-derived suppressor cells (MDSCs), tumor- associated macrophages (TAMs), and tumor-associated neutrophils (TANs) indicates an immunosuppressive TME associated with poor prognosis (Petty, Owen et al. 2021, Ma, Renz et al. 2022). MDSCs and TANs originate from circulating BM-derived myeloid cells (Ma, Renz et al. 2022), while intra-tumoral TAMs either originate from tissue-resident macrophages or differentiate from recruited MDSCs / monocytes (Petty, Owen et al. 2021). All these cell types show similar immunomodulatory functions, mediated by secreted factors, metabolic reprogramming, or regulatory cell-to-cell contact (Ohue and Nishikawa 2019, Petty, Owen et al. 2021, Ma, Renz et al. 2022). Furthermore, TAMs and MDSCs support tumor vascularization and tumor progression by secreting pro-angiogenic factors and degrading the ECM, respectively (Lugano, Ramachandran et al. 2020, Petty, Owen et al. 2021, Ma, Renz et al. 2022). This highlights the complexity of the crosstalk between TME components that work in concert to maintain tumor viability.

[0477] In an embodiment of each and any aspect of the present invention, including any embodiment thereof, the immune evasion mechanism of the tumor is one selected from the group comprising loss of MHC class I expression of malignant cells of the tumor and elevated expression of immune checkpoints by malignant as well as non-malignant cells of the tumor. In connection therewith, a person skilled in the art will acknowledge that many of the checkpoint ligands are not expressed by the malignant cells but by immune-suppressive immune cells in the stroma. These can be: B7 on antigen-presenting cells (ligand of CTLA-4); MHC II on antigen-presenting cells (ligand of LAG-3); galectin (ligand of TIM-3); VISTA on various immune cells, including T cells, and also on some malignant cells; B7-H3 on malignant cells and immune cells in the tumor microenvironment; TIGIT which is expressed on T cells and NK cells and interacts with its ligands, CD 155 and CD 112, which are expressed on malignant cells; or HLA-G which is a non-classical MHC class I molecule that is expressed by some malignant cells.

[0478] In an embodiment of each and any aspect of the present invention, including any embodiment thereof, the tumor is evading from the subject’s adaptive immune system by suppressing activity of cytotoxic tumor-infiltrating lymphocytes (TILs) or inhibiting activation of cytotoxic tumor-infiltrating lymphocytes. It will be appreciated by a person skilled in the art that these TILs establish or provide immunosurveillance meaning that TILs can kill malignant cells in the tumor. If a tumor is cold, the TILs may be there but are not activated which is dueto suppressive factors in the stroma. Tregs are regulatory T cells, and these cells are generally immunosuppressive. They are thus immunosuppressive lymphocytes and are included along with the immunosuppressive myeloid cells which are MDSCs, TAMs, and TANs.

[0479] In an embodiment of each and any aspect of the present invention, including any embodiment thereof, the immunosuppressive tumor microenvironment results from, is caused by and / or is associated with a presence of a cell selected from the group comprising a myeloid-derived suppressor cell (MDSC), a tumor-associated macrophage (TAM) a tumor-associated neutrophils (TAN), and / or a regulatory T cell (Treg). It will be appreciated by a person skilled in the art that such immunosuppressive tumor microenvironment results from, is caused by and / or is associated with a presence of a multitude of said cells.

[0480] Typical examples of immunologically cold tumors where targeted immunotherapy has remained to be challenging include pancreatic cancer (pancreatic ductal adenocarcinoma or PDAC) (Hartupee, Nagalo et al. 2024); ovarian cancer, with high-grade serous ovarian cancer being the most common and aggressive form (Peng, He et al. 2022); prostate cancer, particularly in its advanced castrationresistant form (Stultz and Fong 2021); triple-negative breast cancer (Liu, Hu et al. 2023); and also rare cancers such as soft tissue sarcomas (Rytlewski, Milhem et al. 2021) including the subtype of synovial sarcoma (Mitchell, Pollack et al. 2021), and osteosarcoma, the most common form of primary bone sarcoma (Yu and Yao 2024).

[0481] In an embodiment of each and any aspect, including any embodiment thereof, the tumor is an immunologically cold tumor is refractory to immunotherapy. In an embodiment of each and any aspect, including any embodiment thereof, the tumor is refractory to immunotherapy. In an embodiment of each and any aspect, including any embodiment thereof, the tumor is non-refractory to or sensitive to immunotherapy. In these embodiments in particular, but also whenever is referred herein to immunotherapy, more preferably immunotherapy includes, but is not limited to a checkpoint inhibitor and, respectively, administering a checkpoint inhibitor to a subject; adoptive cell therapy and, respectively, administering an adoptive cell therapy to a subject, and tumor vaccine and, respectively, administering a tumor vaccine to a subject.

[0482] Immune checkpoints are regulators of the immune system. These pathways are crucial for selftolerance, which prevents the immune system from attacking cells indiscriminately. However, somecancers can protect themselves from attack by stimulating immune checkpoint targets. The success of immune checkpoint blockade in cancer therapy, in which antibodies are used to block signaling pathways that inhibit T cell responses to tumors, illustrates the critical roles of these inhibitory checkpoints in obstructing antitumor immunity.

[0483] PD-1 inhibitory receptor PD-L1 regulates T cell activation, effector T cell responses, T cell tolerance, and T cell exhaustion (Francisco, Sage et al. 2010). PD-1 is rapidly induced on naive T cells following TCR engagement, countering T cell activation. When T cells are repetitively stimulated (as in cancer and chronic infection), PD-1 expression is sustained at high levels and T cells enter a dysfunctional state, termed exhaustion (Barber, Wherry et al. 2006). Thus, PD-1 is expressed on both activated and exhausted T cells, and PD-1 expression alone does not signify T cell exhaustion. PD-1 is not only expressed on conventional CD4+ and CD8+ T cells, but also on regulatory T cells, B cells, NK cells, and NKT cells (Schildberg, Klein et al. 2016). PD-1 has two ligands, PD-L1 (also called B7-H1 or CD274) and PD-L2 (also called B7-DC or CD273). PD-L1 is widely expressed on many types of hematopoietic cells (T cells, B cells, macrophages, dendritic cells) and nonhematopoietic cells (epithelial, stromal, and endothelial cells). PD-L2 is expressed mainly on hematopoietic cells (DCs, macrophages, B cells, and Th2 cells). Upregulation of PD-1 ligands by proinflammatory stimuli may serve as a negative feedback mechanism to attenuate effector T cell responses, protecting tumors from immune attack.

[0484] CTLA-4 and CD28 are both receptors expressed on the surface of T cells, CD80 / 86 are receptors on the surface of antigen-presenting cells. The contact of CTLA-4 with CD80 / 86 receptors results in quiescence of the T cell which remains unactivated. In contrast, the interaction of CD28 on the surface of T cells with CD80 / 86 on the antigen-presenting cell results in activation of the T cell. Pharmacological blocking of CTLA-4 on the T cell surface increases the likelihood of CD28-CD80 / 86 binding, resulting in maintenance of T cell activation (Kareva 2017).

[0485] LAG-3 is an immune checkpoint receptor that is highly upregulated on exhausted T cells in the tumor microenvironment. LAG3 transmits inhibitory signals to T cells upon binding to MHC class II and other ligands, rendering T cells dysfunctional. Consequently, LAG3 is a major target for cancer immunotherapy with many anti-LAG3 monoclonal antibodies (mAbs) that block LAG3 inhibitory activity in clinical trials (Mariuzza, Shahid et al. 2024).

[0486] TIM-3 is an immune checkpoint receptor that suppresses antitumor responses by negatively regulating the activity of CD8 T cells and antigen-presenting cells (Anderson, Joller et al. 2016). It is upregulatedon CD8 T cells following anti-PD-1 therapy, suggesting it may be involved in resistance to PD-1 inhibition (Koyama, Akbay et al. 2016). Preclinical studies have found that concurrent inhibition of TIM-3 and PD-1 pathways is more effective in reducing tumor growth and restoring adaptive immunity than blocking TIM-3 or PD-1 alone (Pagliano, Morrison et al. 2022).

[0487] TIGIT is an emerging immune checkpoint that inhibits immune cell responses at multiple steps of the cancer-immunity cycle. TIGIT impairs T cell priming by dendritic cells, prevents tumor cell killing by NK cells and cytotoxic T cells, and enhances the immune suppressive activity of regulatory T cells. Consequently, TIGIT constitutes a major target in cancer immunotherapy (Harjunpaa and Guillerey 2020).

[0488] Each of the above checkpoint inhibitors constitutes an embodiment of each and any aspect of the invention, including each and any embodiment thereof.

[0489] Adoptive cell therapy (ACT) comprises different strategies to enhance the activity of T lymphocytes and other effector cells that orchestrate the antitumor immune response, including chimeric antigen receptor (CAR) T cell therapy, T-cell receptor (TCR) gene-modified T cells, and therapy with tumorinfiltrating lymphocytes (TILs). The outstanding results of CAR-T cells in some hematologic malignancies have launched the investigation of ACT in patients with refractory solid malignancies. However, certain characteristics of solid tumors, such as their antigenic heterogeneity and immunosuppressive microenvironment, hamper the efficacy of antigen-targeted treatments (Albarran, San Roman et al. 2024).

[0490] ACT encompasses several techniques that use the T lymphocytes themselves, after a process of artificial modification or genetic engineering, to improve their antitumor activity (Morotti, Albukhari et al. 2021). This implies the extraction of autologous T lymphocytes from the patient, and their manipulation and amplification in vitro. Meanwhile, the patient undergoes treatment with lymphodepleting chemotherapy - usually fludarabine plus cyclophosphamide - to deplete ineffective and immune-suppressing lymphocytes. After this process, the improved cell product is reinfused into the patient.

[0491] The difference between different ACT modalities lies in the characteristics of T cell modification in vitro: CAR-T and TCR gene-modified T cells are genetically engineered to incorporate modified membrane receptors with a high affinity for selected tumor antigens. However, in contrast to hematologic malignancies, solid tumors are composed of polyclonal cell populations with hugeantigenic heterogeneity, which hampers the efficacy of antigen-targeted therapies. TIL therapy is based on the activation and expansion of infiltrating T cells extracted from the tumor itself, which are intrinsically reactive against tumor antigens, setting an interesting rationale for its use against the changing and heterogeneous cell population of solid cancers.

[0492] CAR-T cells are genetically modified lymphocytes that incorporate a chimeric antigen receptor (CAR) composed by three parts, (i) an extracellular domain with a single-chain fragment variable (scFv) that allows antigen recognition, (ii) a transmembrane domain - linked to the extracellular part through a spacer, and (iii) an intracellular domain. This includes a CD3 complex which activates the downstream signaling pathways, and several costimulatory domains (usually CD28 and / or 4-1BB) that intensify the cytoplasmatic activity of T cells unchained by antigenic recognition (Zhang, Liu et al. 2017). In recent years, innovations in the structure and manufacturing of CAR-T cells have led to significant improvements in their clinical efficacy, especially with the development of fourth-generation CAR-T cells (Huang, Li et al. 2020). Fifth generation CARs equipped with three costimulatory domains and able to secrete anti-PDLl scFv blockade molecules, targeted against B cell maturation antigen (BCMA) have shown heightened antitumor efficacy and decrease of T cell exhaustion in patients with multiple myeloma (Yuti, Sawasdee et al. 2023).

[0493] Due to their molecular structure, CAR-T cells only recognize extracellular antigens, and are particularly efficient when their scFv has a high affinity for the targeted protein and it is homogeneously expressed by tumor cells. This explains the efficacy of CAR-T cells in patients with leukemia and lymphoma, which comprise a clonal population of cells that uniformly express certain antigens such as CD19 on their membrane (Shank, Do et al. 2017). Since 2018, tisagenlecleucel and axicabtagene-citoleucel have EMA approval for the treatment of B cell LLA and refractory nonHodgkin lymphoma, and in 2020, brexucabtagene-autoleucel was approved by the EMA for mantle lymphoma, achieving complete response rates of over 50% in heavily pre-treated patients (Sermer and Brentjens 2019).

[0494] The outstanding results of CAR-T cells in hematologic malignancies have led to their investigation in solid tumors, mainly using overexpressed epithelial antigens as targets. EGFR, HER2, CEA, mesothelin and soluble antigen GD2 have been frequent targets of CAR-T therapies, although many other antigens have been the object of preclinical studies (Marofi, Motavalli et al. 2021).

[0495] Nearly 500 clinical trials evaluating CAR-T cells in solid tumors have been registered before 2023, with many still ongoing (Wang, Gauthier et al. 2023). Most completed studies are phase 1 / 2 trials thathave reported modest results, with only occasional and generally brief clinical responses; a general breakthrough for CAR T cell therapies of solid cancers is still to come.

[0496] The overall process for manufacturing ot CAR-T cells involves isolation of the starting cell population from the leukapheresis product, T-cell activation, genetic modification, ex vivo expansion, final product formulation, and product release testing (Ayala Ceja, Khericha et al. 2024). The manufacturing processes for several FDA-approved CAR-T cell products have been described in literature with broad similarities as well as unique features for tisacel (KYMRIAH), axicel (YESCARTA), brexucel (TECARTUS), lisocel (BREYANZI), idecabtagene vicleucel (ABECMA), and ciltacabtagene autoleucel (CARVYKTI) (Ayala Ceja, Khericha et al. 2024).

[0497] T cells expressing an engineered T-cell receptor (TCR-T cells) represent an alternative modality of ACT, with several advantages compared to CAR T therapy (Baulu, Gardet et al. 2023). In contrast to CAR T cells, TCR-T- cells can recognize not only membrane proteins but also intracellular antigens presented by MHC molecules, covering a wider repertoire of tumor neoantigens and therapeutic targets. Intrinsic features of T cells, such as high antigen sensitivity and the use of physiological signaling pathways, improve TCR-T-cells antitumor functions and reduce the risk of on-target off-tumor toxicity.

[0498] However, TCR-T-cells also have some disadvantages compared to CAR-T cells, such as their weaker avidity for target antigens and their limitation to a certain human leukocyte antigen (HLA) haplotype, which restricts the number of patients that potentially benefit from each modified TCR. In addition, as antigen-targeted modified T lymphocytes, all the limitations of CAR-T therapy in solid tumors -antigenic heterogeneity, difficulty of T cell trafficking, and detrimental effects of immunosuppressive TME- are also applicable to TCR-T-cells (Foy, Jacoby et al. 2023).

[0499] To date, there is only one TCR T cell therapy approved for clinical use, afamicel (TECELRA), to treat metastatic synovial sarcoma (Keam 2024). TECELRA is prepared from the patient’s peripheral blood mononuclear cells (PBMCs), which are obtained via a standard leukapheresis procedure. The PBMCs are enriched for T cells and are then transduced with a replication-incompetent lentiviral vector containing the MAGE-A4 TCR transgene. The transduced T cells are expanded, washed, formulated into a suspension, and cryopreserved. The product must pass a sterility test before release and shipping as a frozen suspension in one or more infusion bag(s). The product is thawed prior to infusion back into the patient (FDA 2024).Both CAR-T and TCR-T cells harbor inherent limitations for the treatment of patients with solid tumors, which may explain the worse clinical outcomes than those obtained with CAR-T therapy in hematologic malignancies.

[0500] In contrast to the clonal nature of lymphomas and leukemias, solid tumors - particularly in the context of metastatic disease - are characterized by the progressive acquisition of somatic mutations that lead to polyclonal expansion of different cellular lineages, giving rise to genomic instability and antigenic heterogeneity.

[0501] Significant intratumoral heterogeneity in neo-epitope expression and clonal expansion of the adaptive immune system in distant regions of the same disease have also been demonstrated in hepatitis B virus (HBV)-related liver cancer (Losie, Craig et al. 2020). In Her2+ breast cancer, spatial transcriptomic studies have shown intra-patient heterogeneity in the expression of gene signatures that determine cellular interactions with T lymphocytes and other immune cells (Andersson, Larsson et al. 2021). In another study, the expression of several antigens in tumor cells from 7 GBM patients was determined before and after a single infusion of EGFR-targeted CAR-T cells, and in all patients a significant decrease in EGFR expression, but an important increase in several other antigens with known immunosuppressor functions was observed (O'Rourke, Nasrallah et al. 2017). These results shed light on how antigen-specific T cell therapies might quickly promote the selection of resistant cellular subclones with immunosuppressive activity.

[0502] T cells with multi-antigenic recognition have been proposed as a potential strategy to overcome this problem. Some preclinical studies have suggested that CAR-T cells with bispecific adapters can facilitate the eradication of antigenically different tumors (Lee, Marks et al. 2019). Moving a step forward, CAR-T cells equipped with synthetic Notch (synNotch) receptors might be able to induce CAR expression only after the recognition of tumor-specific antigens, creating precise prime-and-kill recognition circuits (Yu, Yi et al. 2019) (81). Although these are promising strategies, they have not yet been evaluated in clinical trials, and further research is required to assess their feasibility.

[0503] Ensuring contact between CAR-T cells and tumor cells is not a problem in leukemias and lymphomas, since malignant cells concentrate within the blood and lymph nodes; however, it is a significant obstacle in solid metastatic tumors. Several studies have shown that metastatic lesions from solid cancers have significantly lower lymphocytic infiltration than primary tumors, suggesting that the lossof T cell trafficking to the tumor site is a relevant mechanism for immune escape and may facilitate tumor progression (Ogiya, Niikura et al. 2016).

[0504] In patients with metastatic melanoma it was shown that the expression of certain chemokines is significantly higher in lesions enriched with tumor-infiltrating lymphocytes than in those with poor lymphocytic infiltration, suggesting that these molecules play an important role in T cell recruitment (Harlin, Meng et al. 2009). For instance, high expression of the chemokine CXCL12 in the tumor microenvironment can shield nests of cancers cells from the immune effector cells in experimental models, an observation which might also be a relevant mechanism that limits efficacy of adoptive cell therapies. Inhibition of CXCL12 / CXCR4 signaling prompted deep infiltration of T cells in the tumor microenvironment in these models (Feig, Jones et al. 2013, Fearon 2014, Zboralski, Hoehlig et al.

[0505] 2017).

[0506] Targeting the tumor vasculature and microenvironment to modulate the chemotactic response is an exciting research topic to improve CAR-T cell trafficking to solid tumors (Donnadieu, Dupre et al.

[0507] 2020).

[0508] In addition to hampering T cell trafficking, the tumor microenvironment has inhibitory effects on the lymphocytes that get to infiltrate the tumor site, which is an obstacle for CAR T or TCR T cell function. In addition, sustained exposure to tumor antigens and inflammatory signals is thought to progressively mitigate the function and proliferation of modified T cells, leading to CAR T cell ‘exhaustion’. Targeting T cell intrinsic pathways (PD-1 / PD-L1 axis, TOX / NR4A, TGF-b, CBL-B), using CRISPR technology to modulate the surface expression of CAR, and uncoupling antigen recognition from CAR activation signaling, are possible approaches to overcome exhaustion and improve adoptive cell therapy clinical outcomes in solid tumors (Gumber and Wang 2022).

[0509] Each of the above adoptive cell therapies constitutes an embodiment of each and any aspect of the invention, including each and any embodiment thereof.

[0510] In an embodiment of each and any aspect of the invention, including any embodiment thereof, the tumor is a soft tissue sarcoma.

[0511] Soft tissue sarcomas represent a heterogeneous group of extraskeletal mesenchymal tumors which are subclassified in approx. 70 subtypes, each characterized by a distinct morphology, that often translates into a specific clinical behaviour as well as into specific therapeutic approaches (Sbaraglia, Bellan etal. 2021). The nomenclature of soft tissue sarcomas is derived from the mature tissue each tumor most closely resembles. The incidence of soft tissue sarcomas among all children age younger than 20 years diagnosed with cancer is approximately 7%-8%, while representing only l%-2% of all cancer diagnoses in adults (Ferrari, Sultan et al. 2011). Within the adolescent and young adult population, defined as patients between age 15 and 39 years, the incidence of synovial sarcoma is 2%-4% (Lewis, Seibel et al. 2014).

[0512] Among the types of soft tissue sarcoma are (American Cancer Society, www.cancer.org / cancer / types / soft-tissue-sarcoma / about / soft-tissue-sarcoma.html, accessed Feb 17, 2025):

[0513] • Adult fibrosarcoma, which usually affects fibrous tissue in legs, arms, or trunk; it is most common between the ages of 20 and 60;

[0514] • Alveolar soft-part sarcoma, a rare cancer that most commonly start in the legs and mostly affects young adults;

[0515] • Angiosarcoma which can start in blood vessels (hemangiosarcomas) or lymph vessels (lymphangiosarcomas), sometimes in a part of the body that has been treated with radiation;

[0516] • Clear cell sarcoma, a rare cancer that often starts in tendons of the arms or legs;

[0517] • Desmoplastic small round cell tumor, a rare sarcoma of teens and young adults, most frequently found in the abdomen;

[0518] • Epithelioid sarcoma, which mostly starts in tissues under the skin of the hands, forearms, feet, or lower legs. Teens and young adults are often affected;

[0519] • Fibromyxoid sarcoma, low-grade or Evans’ tumor is a slow-growing cancer that most often starts as a painless growth in the trunk or arms and legs It is more common in young to middle aged adults;

[0520] • Gastrointestinal stromal tumor (GIST), which starts in the digestive tract. See Gastrointestinal Stromal Tumor (GIST) for more details;

[0521] • Kaposi sarcoma, is a type of sarcoma that starts in the cells lining lymph or blood vessels; • Leiomyosarcoma, a type of cancer that starts in smooth muscle tissue, often in the abdomen;

[0522] • Liposarcomas, malignant tumors of fat tissue, which can start anywhere in the body, but most often in the thigh, behind the knee, and inside the back of the abdomen. They occur mostly in adults between 50 and 65 years old;

[0523] • Malignant mesenchymoma, a rare type of sarcoma that shows features of fibrosarcoma and features of at least 2 other types of sarcoma;• Malignant peripheral nerve sheath tumors, which include neurofibrosarcomas, malignant schwannomas, and neurogenic sarcomas. These sarcomas start in the cells that surround a nerve;

[0524] • Myxofibrosarcomas, low-grade which are mostly found in the arms and legs of elderly people.

[0525] • Rhabdomyosarcoma, which is the most common type of soft tissue sarcoma in children;

[0526] • Synovial sarcoma, a malignant tumor of the tissue around joints. The most common locations are the hip, knee, ankle, and shoulder. This tumor is more common in children and young adults, but it can occur in older people;

[0527] • Undifferentiated pleomorphic sarcoma (UPS), previously called malignant fibrous histiocytoma (MFH), which is most often found in the arms or legs. Less often, it can start inside at the retroperitoneum. This sarcoma is most common in older adults. It mostly tends to grow into other tissues around the place it started, but it can spread to distant parts of the body.

[0528] Each of the above forms of soft tissue sarcoma constitutes an embodiment of each and any aspect of the invention, including each and any embodiment thereof.

[0529] In an embodiment of each and any aspect of the invention, including any embodiment thereof, the tumor is a synovial sarcoma.

[0530] Synovial sarcoma is a deadly soft tissue malignancy with a predilection for adolescent and young adult victims (Stacchiotti and Van Tine 2018). It is a model immunologically quiet (also called cold) tumor with respect to many of the most important metrics for immunotherapy success including T cell infiltration, MHC expression, and inflammatory gene signatures (Pollack, He et al. 2017) (Blay, von Mehren et al. 2023). It almost never responds to checkpoint inhibitors and in a study of over 50 synovial sarcoma patients treated with PD-L1 inhibition there was not a single responder (Chawla, Van Tine et al. 2022). It has a low tumor mutational burden and frequently is infiltrated with inhibitory macrophages. Systemic administration of IFNy can increase T cell infiltration and MHC expression in synovial sarcoma patients but this has not led to improved outcomes for synovial sarcoma patients either with IFNy as a single agent or in combination with PD-1 inhibition (Zhang, Kohli et al. 2019). IFNy is an FDA approved cytokine therapy, used as a standard of care treatment for osteopetrosis and chronic granulomatous disease. In the 1990s, IFNy was evaluated as a single agent cancer immunotherapy in several prospective randomized trials (Windbichler, Hausmaninger et al.

[0531] 2000, Giannopoulos, Constantinides et al. 2003, Alberts, Marth et al. 2008). IFNy can enhance expression of surface major histocompatibility complex molecules and increase the processing andpresentation of tumor-specific antigens (Shankaran, Ikeda et al. 2001), thus facilitating T-cell recognition and cytotoxicity (Gao, Shi et al. 2016, Ayers, Lunceford et al. 2017, Manguso, Pope et al.

[0532] 2017, Patel, Sanjana et al. 2017).

[0533] Synovial sarcoma is among the best disease models for targeted immunotherapy because of its consistently strong and homogenous expression of cancer testis antigens (CTAs). These proteins are normally expressed early in development or within immune-privileged compartments, such as developing germ cells, but are abnormally expressed in some cancers and uniquely so in synovial sarcoma. This type of tumor generally expresses NY-ESO-1, PRAME, and MAGE-A4, among others, elevating synovial sarcoma as a prime target for CTA-specific vaccines or adoptive cellular therapies (ACTs). In fact, synovial sarcoma recently became the first cancer ever to have an engineered T cell receptor-based therapy (TCR-T), afamitresgene autoleucel ("afamicel") targeting MAGE-A4, approved by the FDA. However, despite excellent overall response rates (ORRs) in the 50% range, TCR-T responses are durable in <15% of patients (Tan, Gakhar et al. 2021, Zhang, Liu et al. 2022). Reversing the cold TME is key to successful immunotherapy in synovial sarcoma. Although synovial sarcoma is an uncommon cancer, its immunobiology is broadly important for many cancers.

[0534] CXCR4 has been reported to be expressed in synovial sarcoma cells and examined as a potential target to slow tumor growth (Pazzaglia, Pollino et al. 2019).

[0535] Synovial sarcoma is a relatively rare malignancy representing a soft tissue sarcoma (STS) of uncertain differentiation (Gazendam, Popovic et al. 2021). It accounts for 5–10% of all soft tissue sarcomas (Toro, Travis et al. 2006, Mastrangelo, Coindre et al. 2012, WHO 2020). The age-adjusted incidence is 0.81 / 1,000,000 in children and 1.42 / 1,000,000 in adults with approximately 1000 patients diagnosed with synovial sarcoma in the United States each year(Sultan, Rodriguez-Galindo et al. 2009). Synovial sarcoma is unique from other soft tissue sarcomas as it presents at a younger mean age of onset and commonly occurs in adolescents and young adults (mean age of 39 years at diagnosis) and affects both sexes equally (Aytekin, Ozturk et al. 2020). Synovial sarcoma is the most common nonrhabdomyosarcoma soft tissue sarcoma in children, representing 30% of soft tissue sarcomas diagnosed in childhood (McGrory, Pritchard et al. 2000, Sultan, Rodriguez-Galindo et al. 2009).

[0536] The 5-year survival rates range from 59–75% (Lewis, Antonescu et al. 2000, Trassard, Le Doussal et al. 2001, Sultan, Rodriguez-Galindo et al. 2009, Smolle, Parry et al. 2019, Aytekin, Ozturk et al.

[0537] 2020). There has been a trend for improved survival over time with early cohorts from the 1960squoting a 25–51% five-year survival rate. Local and metastatic relapse of soft-tissue sarcomas generally occur in the first two years following treatment and thus surveillance and follow-up is most intensive in this period (Wilson, Gazendam et al. 2020). However, synovial sarcoma is unique in this regard in that it tends to recur much later, with occurrence of local recurrence after a mean of 3.6 years and of metastases after a mean of 5.7 years (Krieg, Hefti et al. 2011).

[0538] There are several well documented key prognostic factors including tumor size, grade and anatomical location, patient age at diagnosis, negative surgical margins and adjuvant radiotherapy (Lewis, Antonescu et al. 2000, Ferrari, Gronchi et al. 2004, Ferguson, Griffin et al. 2006, Guadagnolo, Zagars et al. 2007, Canter, Qin et al. 2008, Vlenterie, Ho et al. 2015). Osseous or neurovascular invasion, adult age, large tumor size and unplanned excision have been linked to worse prognosis (Singer, Baldini et al. 1996, Lewis, Antonescu et al. 2000, Ferrari, Gronchi et al. 2004, Ferguson, Griffin et al.

[0539] 2006, Vlenterie, Ho et al. 2015). Similar to other soft tissue sarcomas, tumor size and grade has repeatedly been shown to have prognostic value in patients with synovial sarcoma (Ferrari, Gronchi et al. 2004, Eilber, Brennan et al. 2007, Naing, Monjazeb et al. 2015). Tumor location has also been demonstrated to be of predictive value, with non-extremity based synovial sarcomas tending to have worse overall survival (Ferrari, Bisogno et al. 2008, Aytekin, Ozturk et al. 2020). However, this may be in part due to the lack of early symptoms and later stage of presentation. Although synovial sarcoma has a similar clinical presentation in children and adults, there is a growing evidence that they have different outcomes, with children having significantly better survival rates (Sultan, Rodriguez-Galindo et al. 2009, Vlenterie, Ho et al. 2015, Ferrari, Chi et al. 2017, Smolle, Parry et al. 2019).

[0540] Metastasis develops in approximately half of patients and for them, the median overall survival (OS) is 2-years for metastatic SS and the 5 year survival is only 10% (Kampe, Rosen et al. 1993, Ladanyi, Antonescu et al. 2002, Sultan, Rodriguez-Galindo et al. 2009, Su, Cheng et al. 2010, Constantinidou, Pollack et al. 2013, Pollack, Somaiah et al. 2020). Because most patients are young, each death takes an outsized number of years of life (Chen, Borker et al. 2014).

[0541] Although many synovial sarcomas originate near articular structures, the name synovial sarcoma is a misnomer in that these lesions do not originate from intra-articular synovium, but from primitive mesenchymal cells (Jo and Fletcher 2014). Synovial sarcomas most commonly present as soft tissue masses but cases of primary synovial sarcoma of bone have been reported (Caracciolo, Henderson-Jackson et al. 2019). These lesions can occur anywhere in the body, with the majority arising in the extremities, particularly in the lower extremity in the anatomic structures adjacent to the knee joint(Sultan, Rodriguez-Galindo et al. 2009, Smolle, Parry et al. 2019). Synovial sarcomas are considered the most common soft tissue sarcoma of the foot (McGrory, Pritchard et al. 2000, Deshmukh, Mankin et al. 2004).

[0542] Synovial sarcomas often do not present with the typical soft tissue sarcoma presentation of a large and quickly growing painless mass (Smolle, Parry et al. 2019). Instead, the majority of synovial sarcomas are slow growing and the mean duration of symptoms before diagnosis is approximately 2 years (Chotel, Unnithan et al. 2008). In comparison to other soft tissue sarcomas, there may be longstanding pain or joint contractures at the tumor site that precede swelling (De Silva, Barrett et al.

[0543] 2003). Only half of patients have clinical findings consistent with soft tissue sarcoma according to the UK National Institute for Health and Care Excellence (NICE) guidelines (De Silva, Barrett et al. 2003, Chotel, Unnithan et al. 2008). Given the insidious onset, younger age at presentation and atypical presenting symptoms, these patients may be initially clinically misdiagnosed with benign processes including myositis, synovitis, bursitis or tendonitis.

[0544] Plain radiographs are not required for diagnosis but are typically performed as part of the initial workup and can identify adjacent bony remodeling, bone invasion or calcification of the soft tissue mass (Bixby, Hettmer et al. 2010, Bakri, Shinagare et al. 2012, Kao 2019). Typically, synovial sarcoma presents as a well-defined or lobulated soft tissue mass on plain radiographs. Punctate calcifications, particularly around the periphery of the lesion, are visualized in one third of patients. Occasionally, more extensive calcification can be visualized and can mimic bone forming tumors including osteosarcoma and myositis ossificans (O'Sullivan, Harris et al. 2008).

[0545] Similar to other soft tissue sarcomas, MRI with and without contrasting agents is the gold standard for diagnostic imaging for synovial sarcoma (O'Sullivan, Harris et al. 2008). MRI defines the local extent of the soft tissue mass and surrounding edema and provides excellent visualization of the mass with respect to the surrounding anatomy, which is critical for preoperative planning. The utilization of gadolinium contrast can differentiate between hemorrhagic or necrotic areas and areas of solid viable tumor. As with most soft tissue sarcomas, synovial sarcomas are typically heterogenous with low intensity on T1 and high intensity on T2 -weighted images with post-gadolinium enhancement (O'Sullivan, Harris et al. 2008).

[0546] Although synovial sarcomas generally present as a non-specific heterogenous mass, there are some unique features, which can aid in differentiation from other soft tissue sarcomas. Synovial sarcomaspredominately present as well-defined, heterogeneously enhancing solid tumors that are multilobulated in nature. A triple signal intensity demonstrating areas of hyperintensity, isointensity and hypointensity indicating the mix of cystic and hemorrhagic areas, cellular elements and fibrotic areas can be characteristic (O'Sullivan, Harris et al. 2008). Smaller tumors, particularly those smaller than 5 cm in diameter, often show homogeneous enhancement which can be mistaken for a benign process (Liang, Mao et al. 2015). Several findings on MRI have been found to be predictive of highgrade lesions including the absence of calcifications and presence of hemorrhage and the triple signal intensity (Tateishi, Hasegawa et al. 2004).

[0547] Computed tomography (CT) with contrasting agents can be utilized when an MRI is contraindicated or unavailable. Synovial sarcoma appears hypointense compared to muscle with heterogeneity in larger lesions (Baheti, Tirumani et al. 2015). CT allows for better visualization of soft-tissue calcifications and local bone reaction (O'Sullivan, Harris et al. 2008).

[0548] A biopsy and pathologic assessment are required to differentiate synovial sarcoma from other soft tissue sarcoma subtypes and define the tumor grade. As with all soft tissue sarcomas, a biopsy should be performed prior to definitive surgery to avoid inadequate resection and misdiagnosis (von Mehren, Randall et al. 2016). Options for biopsy include incisional biopsies, core needle biopsies (CNB) and fine needle aspirations (FNA). Historically, open incisional biopsies (IB) have been considered the gold standard for soft tissue lesions as they provide larger volumes of tissue. When compared to CNB and FNA, IB tend to have higher diagnostic accuracy but this comes with a higher rates of complications when compared to percutaneous techniques (Kasraeian, Allison et al. 2010). Core needle biopsies retrieve more tissue than FNA and have higher diagnostic accuracy (Yang and Damron 2004). Fine needle aspirates are rarely used in soft tissue sarcomas due to the small quantity of sample material obtained and a limited ability to assess lesional architecture (Kasraeian, Allison et al. 2010). Advances in diagnostic imaging has allowed for image guided percutaneous biopsies which has improved the diagnostic accuracy of these techniques (Narvani, Tsiridis et al. 2009). Given the lower morbidity and relatively high diagnostic accuracy of CNB, image guided CNB are the preferred method of biopsy, particularly for deeper tumors. When open biopsies are performed, the biopsy principles must be observed to reduce biopsy-related complications (Mankin, Mankin et al. 1996).

[0549] Staging investigations are imperative and allow for a better understanding of disease prognosis and risk of recurrence or metastases. The tumor stage also helps to formulate a treatment plan by a multidisciplinary sarcoma team. Staging of synovial sarcoma involves cross-sectional imaging of theaffected extremity, systematic staging with a chest CT, and pathological assessment. The two most commonly utilized staging systems are the American Joint Committee on Cancer (AJCC) system and the Enneking staging system (Enneking 1988, Amin 2017). The Enneking staging system relies on tumor grade, local extent of disease and presence of metastases. The AJCC staging system is based on anatomical site of primary tumor and tumor size (pT) tumor grade, nodal involvement (N), and presence of metastases (M).

[0550] Synovial sarcomas are malignant and metastasize, most commonly to the lungs with up to 13% of patients having distant metastases at the time of diagnosis (Aytekin, Ozturk et al. 2020). Historically, it was thought that synovial sarcoma had a predilection for metastases to lymph nodes, necessitating the need for further advanced imaging and possible sentinel node biopsy (Maduekwe, Homicek et al.

[0551] 2009). However, these findings were based on small case series and more recent SEER database findings suggest that the rates of lymph node metastases are in line with other soft tissue sarcomas and do not require additional workup beyond thorough physical examination (Jacobs, Morris et al. 2018).

[0552] The gross pathology appearance of synovial sarcoma is usually tan or grey and may be multinodular or multicystic (Jo and Fletcher 2014). Most synovial sarcomas are 3-10 cm in diameter. Smaller lesions (<1 cm) can occur in hands and feet (Michal, Fanburg-Smith et al. 2006, WHO 2020). Histologically, synovial sarcoma is a monomorphic spindle cell sarcoma with variable epithelial differentiation (WHO 2020). It presents as one of three variants: monophasic, biphasic or poorly differentiated. In the monophasic variant, the tissue is comprised entirely of spindle-cells whereas in biphasic synovial sarcoma, there are epithelial and spindle-cell components present (Jo and Fletcher 2014). In one third of synovial sarcomas, areas of calcifications and / or ossification can be found. In both monophasic and biphasic variants, there may be poorly differentiated areas with increased cellularity, greater nuclear atypia and high mitotic activity (6 mitoses / mm2>10 mitoses per 1.7 mm2) (WHO 2020). Occasionally, the entire tumor shows poorly differentiated morphology. On immunohistochemistry, diffuse expression of bcl-2 is typically seen. In 60% of cases, these tumors stain positive for CD99 (WHO 2020). Immunohistochemistry also demonstrates strong and diffuse nuclear staining for the transcriptional corepressor TLE1 found in the large majority of synovial sarcomas (WHO 2020). NY-ESO-1 is also expressed strongly in most synovial sarcomas and can help differentiate it from other spindle cell neoplasms (Lai, Robbins et al. 2012).

[0553] Synovial sarcoma bears one of the simplest genomes across cancers, with low tumor mutational burden (TMB) and even rare copy number variations identified in neoplastic cells (Luk, ME et al.2024). Synovial sarcoma is defined and driven by the fusion oncoprotein SS18:: SSX from the translocation t(X;18) fusing SS18, which codes for a BAF chromatin remodeling complex component, in-frame to one of the SSX genes, SSX1, 2, or 4, which code for putative transcriptional repressors (Fiore, Sambri et al. 2021). This translocation is present in >95% of cases (Stegmaier, Leuschner et al.

[0554] 2017) and leads to the expression of different SS18: SSX oncogenic fusion proteins, which drive sarcomagenesis. Subtypes include SS18:SSX1 and SS18:SSX2 and less commonly SS18:SSX4 (dos Santos, de Bruijn et al. 2001). Both fluorescence in situ hybridization and reverse transcription polymerase chain reaction testing have been validated in the diagnosis of this translocation (Amary, Berisha et al. 2007). Almost all SS18-SSX2 synovial sarcomas show monophasic morphology and are significantly more common in women. Rare cases are associated with t(X;20) and SS18L1-SSX1 fusion transcript (WHO 2020).

[0555] The mainstay of treatment for synovial sarcoma remains surgical excision with negative margins with the addition of radiotherapy and / or chemotherapy based on patient and tumor characteristics (von Mehren, Randall et al. 2016). Historically, patients were often treated with amputation but advances in adjuvant therapy and cross-sectional imaging have allowed the majority of patients to be treated with limb-salvage surgery (Ghert, Abudu et al. 2005). The goal of limb-salvage surgery in synovial sarcoma is to achieve oncologic control of the tumor while providing the patient with a functional limb postoperatively. Negative surgical margins are of upmost importance as they predict both local recurrence and overall survival (Guadagnolo, Zagars et al. 2007, Biau, Ferguson et al. 2012, Bilgeri, Klein et al. 2020). While no specific guidelines exist regarding ideal negative margins in synovial sarcoma, surgical management is similar to that of other soft tissue sarcomas. For superficial tumors or small (<5 cm) deep tumors not intimately associated with critical structures, a wide excision with negative margins (1–2 cm) alone could be considered sufficient (Ferrari, Chi et al. 2017). In tumors that are closely associated with neurovascular structures or bone, the epineurium, adventitia or periosteum is utilized as the margin to allow for a functional limb postoperatively (Kawaguchi, Ahmed et al. 2004, Guadagnolo, Zagars et al. 2007). In these cases, very close or microscopically positive margins may occur and radiotherapy is essential to decrease local recurrence risk (O'Donnell, Griffin et al. 2014, Gundle, Gupta et al. 2017). Carefully planned microscopically positive margins on a fixed structure (bone, nerve, vessel) have been shown to not increase the risk for local recurrence in setting of neoadjuvant radiotherapy (O'Donnell, Griffin et al. 2014). Given the heterogeneity and lack of standardization in the literature, recommendations regarding margin management must be taken into context of the individual patient. Both tumor and anatomic factors should be taken into consideration when determining acceptable surgical margins (Sambri, Caldari et al. 2021).Due to its atypical presentation (slow-growing, painful mass), synovial sarcomas have a high rate of presentation following an unplanned excision, with up to 50% of patients presenting after unplanned excision (Chandrasekar, Wafa et al. 2008, Chotel, Unnithan et al. 2008). Unplanned excisions of synovial sarcomas result in high rates of residual disease, particularly for larger and deeper tumors and increased risk for local recurrence even after re-excision (Chandrasekar, Wafa et al. 2008, Potter, Adams et al. 2008). In the case of referral following unplanned excision, patients should be re-staged, and the original histology reviewed at a sarcoma referral center. Tumor bed excision should be performed in patients with residual disease with the goal of complete tumor resection and negative margins (Pretell-Mazzini, Barton et al. 2015). Typically, these resections are extensive in nature given that areas of potential contamination must be removed, with may necessitate reconstructive procedures (Potter, Adams et al. 2008). There is a paucity of data to guide the utilization of radiotherapy in this population; however, radiotherapy is recommended as it would be in a primary presentation (Pretell-Mazzini, Barton et al. 2015).

[0556] In select cases, limb-salvage techniques are not recommended, and primary amputation may be required. Amputation is considered in patients with tumor location that necessitates excision of vital structures which would result in poor limb function. Patients who present following an unplanned excision may require amputation if there is extensive contamination of vital structures or major joints. Finally, older patients or those with extensive medical comorbidities may not be able to tolerate a major operation and amputation can be considered (Ghert, Abudu et al. 2005).

[0557] Neoadjuvant or adjuvant radiation therapy is recommended for larger tumors (>5 cm), or in any case where a close margin may be required to preserve a major neurovascular structure or bone (von Mehren, Randall et al. 2016). In large registry database studies, radiotherapy has been shown to improve local control and may have overall survival benefit in patients with synovial sarcoma (Naing, Monjazeb et al. 2015, Song, Park et al. 2017, Gingrich, Marrufo et al. 2020). Radiotherapy can be administered pre or postoperatively with differing protocols. Preoperative radiation is associated with higher wound complication rates whereas postoperative radiation can cause fibrosis and joint stiffness which may lead to worse long term functional outcomes (Davis, O'Sullivan et al. 2005). Regardless of timing, intensity-modulated radiation therapy (IMRT) is becoming the preferred method of radiation delivery in patients with soft tissue sarcoma (von Mehren, Randall et al. 2016, Wang, Song et al.

[0558] 2019). Intensity-modulated radiation therapy allows for a higher dose of radiation to more closely contour the tumor, which reduces the volume of radiation to the surrounding normal tissues. The utilization of preoperative IMRT for soft tissue sarcoma has been shown to reduce woundcomplications and need for reconstructive soft-tissue flaps (O'Sullivan, Griffin et al. 2013). Radiation therapy may be considered in isolation in patients with multiple medical comorbidities or patients with metastatic disease where the risks of surgery outweigh the potential benefits (Ramu, Houdek et al.

[0559] 2017).

[0560] Unlike the majority of soft tissue sarcomas, synovial sarcoma appears to be more chemosensitive, although there is still controversy surrounding which subgroups of patients benefit from systemic therapy (Eilber, Brennan et al. 2007). In general, chemotherapy is reserved for patients with high-risk tumors or advanced disease and is thought to be more effective in younger patients (Canter, Qin et al.

[0561] 2008, Vlenterie, Ho et al. 2015). In children and adolescents with intermediate or high-risk tumors (i.e., >5 cm, nodal involvement, positive margins), adjuvant or neoadjuvant chemotherapy is generally undertaken with the most common agents being ifosfamide and doxorubicin. In the absence of available data from randomized controlled trials, prospective multicentred cohort studies have demonstrated adequate response to chemotherapy (Ferrari, De Salvo et al. 2015). Recent data has demonstrated that pediatric or adolescent patients with low-risk tumors (Grade 2 or Grade 3 < 5 cm) can be successfully treated with surgical intervention without systemic therapy (Ferrari, Chi et al.

[0562] 2017). However, the role of chemotherapy in adult patients with synovial sarcoma is less clear (Eilber, Brennan et al. 2007, Canter, Qin et al. 2008, Italiano, Penel et al. 2009).

[0563] Chemotherapy is also considered in metastatic or unresectable disease (Spurrell, Fisher et al. 2005, Vlenterie, Ho et al. 2015). In general, anthracycline-based chemotherapy is first line for advanced soft tissue sarcoma and the addition of ifosfamide is dependent on the subtype of the disease. Ifosfamide has well documented efficacy in synovial sarcoma in the palliative setting and should be considered in patients who undergo chemotherapy if the toxicities can be tolerated (Spurrell, Fisher et al. 2005, Sleijfer, Ouali et al. 2010). Front-line treatment is therefore often doxorubicin with ifosfamide, but this combination rarely — if ever — achieves a complete response of disseminated disease (Judson, Verweij et al. 2014).

[0564] Trabectedin, a chemotherapy approved for liposarcoma and leiomyosarcoma, has demonstrated activity in translocation-associated sarcomas like synovial sarcoma, but also rarely achieves more than temporarily stable disease (Kawai, Araki et al. 2015). Pazopanib is another option, however it is not specifically labelled for synovial sarcoma, but for advanced non-adipocytic soft tissue sarcomas who have received prior chemotherapy. This is a multi-tyrosine kinase inhibitor whose mechanism ofaction is related to the selective inhibition of tumor angiogenesis and to the inhibition of key kinases involved in oncogenic pathways (Hurwitz, Dowlati et al. 2009, Gril, Palmieri et al. 2011)

[0565] There is currently only one marketed systemic treatment specifically approved for synovial sarcoma, afamitresgene autoleucel. This is an autologous, MAGE-A4 targeted TCR engineered cell therapy, which was launched in 2024 in the US under accelerated approval. The treatment is approved for treatment of adults with unresectable or metastatic synovial sarcoma who have received prior chemotherapy, are HLA-A*02:01P, -A*02:02P, -A*02:03P, or -A*02:06P positive and whose tumor expresses the MAGE-A4 antigen therapy. However, despite excellent overall response rates for synovial sarcoma patients with cancer testis antigen targeted strategies, the responses to such therapies are durable only in <15% of SS patients (Tan, Gakhar et al. 2021, Zhang, Liu et al. 2022). Some lack of durability of response may be related to the TCR-T themselves, and it has been shown that rare persisting cells from the peripheral blood of patients previously treated with TCR-T can be reinvigorated using IL- 15, but significant problems exist within the tumor microenvironment (TME) as well (Kohli, Yao et al. 2021).

[0566] Systemic treatments in clinical development for synovial sarcoma include (i) catequentinib, a multityrosine kinase inhibitor in clinical Phase 3; (ii) letetresgene autoleucel, a TCR T cell therapy, in clinical Phase 2; (iii) mipetresgene autoleucel, a TCR T cell therapy, in clinical Phase 2; (iv) atezolizumab, an anti-PD-Ll monoclonal antibody, in clinical Phase 2; (v) tazemetostat, a histone methyltransferase inhibitor, in clinical Phase 2; (vi) TC-N201, a TCR T cell therapy, in clinical Phase 1 / 2; and (vii) DS-2243 with no disclosed mechanism of action, in clinical Phase 1.

[0567] Each of the above forms and stages of synovial sarcoma constitutes an embodiment of each and any aspect of the invention, including any embodiment thereof. Also, each of the above methods for treating synovial sarcoma may be applied to the subject in accordance with the present invention as defined in the aspects of the invention, including any embodiment thereof. Finally, each of the above methods for diagnosing synovial sarcoma may be applied to the subject in accordance with the present invention as defined in the aspects of the invention, including any embodiment thereof.

[0568] In an embodiment of each and any aspect of the invention, including any embodiment thereof, the tumor is an osteosarcoma.Osteosarcoma is a common primary malignant tumor of the bone, with a peak incidence in adolescents and adults >60 years of age (Mirabello, Troisi et al. 2009). Although osteosarcoma can present in any bone in the body, the most common sites are around the knee and the proximal humerus (Bielack, Kempf-Bielack et al. 2002). It can also arise in individuals with a history of cancer as secondary osteosarcoma. The diagnosis of osteosarcoma is made following the biopsy of a mass located most commonly at the metaphysis of the long bones based on imaging findings of patients who presented with pain, decreased mobility and, oftentimes, a palpable mass. Histologically, conventional osteosarcomas most commonly appear as spindle cell tumors that produce malignant osteoid and are consequently thought to derive from the malignant transformation of cells of the mesenchymal lineage at an undefined stage of differentiation towards becoming osteoblasts (Klein and Siegal 2006). Microscopically, based on the predominant matrix being produced, the tumors can be subdivided into chondroblastic, fibroblastic, osteoblastic and telangiectatic tumors, suggesting that they maintain some of the pluripotency of their early undifferentiated mesenchymal precursors (Piperdi, Austin-Page et al.

[0569] 2012). Osteosarcomas can also be divided into three major groups as being low, intermediate and high grade (Bertoni and Bacchini 1998). The grade of the tumor serves as a relative indicator of the risk of developing metastatic disease. Low-grade, or parosteal, osteosarcomas are typically indolent and are treated by surgical removal alone. High-grade tumors have a high risk of developing metastasis in the lungs, lymph nodes, and other bones (Kager, Zoubek et al. 2003, Isakoff, Bielack et al. 2015) and require surgery and adjuvant chemotherapy as treatment. Unfortunately, the outcomes for patients with osteosarcomas have remained relatively stagnant since the advent and remarkable improvement in tumor survival associated with modern chemotherapy in the 1980s (Smith, Seibel et al. 2010).

[0570] Although rare, osteosarcoma is the most common primary malignancy of bone, with an incidence in children and adolescents of ~3-4.5 cases per million population per year (Mirabello, Troisi et al.

[0571] 2009). In the USA, osteosarcoma accounts for <1% of all new cancer diagnoses with -1,000 new cases diagnosed per year, half of these cases occurring in children and adolescents (Mirabello, Troisi et al. 2009). The global incidence rates in younger age groups (individuals <24 years) are relatively consistent across the USA, Europe and Asia. Data regarding differences between ethnic groups are limited, but higher rates of osteosarcoma have been observed in African American children and young adults in the USA than in white individuals (Mirabello, Troisi et al. 2009). Greater geographic variation in osteosarcoma incidence in individuals >60 years has been observed, but data are insufficient to determine whether these differences are due to varying criteria for disease classification in registries, differences in environmental exposures, such as prior radiotherapy for other cancer types, or genetic predisposition.Approximately 80% of patients with osteosarcoma present with radiographically localized disease. Those patients with radiographically confirmed non-metastatic osteosarcoma have a 5 -year event-free survival of -60%. In patients who present with a primary lesion and an isolated pulmonary nodule, 5-year event-free survival is generally <40%. For individuals with a primary lesion and multiple pulmonary nodules or radiographically detectable metastatic disease at other sites, 5-year event-free survival is <20% (Gill and Gorlick 2021).

[0572] Osteosarcoma incidence has a bimodal age distribution with a primary peak in adolescents and young adults and a second, smaller peak in the seventh and eighth decade of life (Mirabello, Troisi et al.

[0573] 2009). It is particularly uncommon in young children <10 years of age in whom the genetic etiology may be different to that in adolescents (Mirabello, Zhu et al. 2020). Osteosarcoma in adults (age >40 years) and elderly populations (age >60 years) tends to occur secondary to other conditions, such as Paget disease of bone, as transformation of other benign bone conditions or as a late effect of therapeutic irradiation.

[0574] Osteosarcoma is slightly more common in males, with an average male-to-female ratio of 1.4:1. A Surveillance, Epidemiology, and End Results (SEER) analysis in a US population provides additional insight into demographic differences that relate to age, sex and race / ethnicity of patients with osteosarcoma (Cole, Gianferante et al. 2022). The age-adjusted incidence rate was 1.9 per million for those aged 0-9 years, 6.7 per million for those aged 10–24 years, 1.9 per million for those aged 25–59 years and 2 per million for those aged >60 years. In the USA, Hispanic males aged 10-24 had the highest incidence rate compared with any other age group or sex. Data from all age groups combined revealed that the Black population had the highest overall incidence. Notably, the incidence of osteosarcoma in children and adolescents has increased from the 1970s to the 2000s but has declined in adults >60 years of age. Some of the increase in this population may be related to the increased incidence of subsequent osteosarcomas over the past decade, which may be attributable to the increasing number of childhood survivors. In patients >60 years old, the decreased incidence of osteosarcoma may be attributable to a decrease in the rate of Paget-related osteosarcoma. Patient sex does not seem to markedly influence prognosis but reports suggest that males may have a slightly worse outcome than females and older patients have a worse outcome than young patients with osteosarcoma (Cole, Gianferante et al. 2022).

[0575] Osteosarcoma is defined histologically as a tumor of osteoid-producing cells, which often exist within an admixture of adipogenic, muscle, spindle, fibroblastic and chondroblastic cells (Klein and Siegal2006). This microscopic phenotype has long fuelled the assumption that osteosarcoma arises from a multipotent mesenchymal precursor. Epidemiological observations support this interpretation as tumors were found to arise most frequently within the metaphyses of long bones in children, adolescents and young adults during times of peak linear growth, suggesting that the bone-producing and cartilage-producing cells that proliferate rapidly during those growth spurts are those susceptible to the transformation giving rise to osteosarcoma (Jones 2011, Mutsaers and Walkley 2014). Introduction of TP53 mutations into partially differentiated osteogenic stem cells generated osteosarcoma-like cells in vitro (Lin, Jewell et al. 2017). Similarly, genetically engineered mouse models have been most successful when introducing TP53 and other mutations using promoters for genes activated along the path that leads from mesenchymal stem cells to osteoblasts (Jones 2011). Together, these data support the hypothesis that osteosarcomagenesis occurs within a proliferating population of partially differentiated osteoblast precursor cells. While mesenchymal differentiation is not as well characterized as the haematopoietic system, a number of transcription factors have been identified as key regulators of clusters of genes involved in the development of various cell types. Some of these are highly expressed in the context of osteosarcoma and relate to its osteogenic phenotype and include SOX9, RUNX2 and Osterix. Some of these transcription factors themselves are influenced by tumor suppressor genes and oncogenes such as TP53 and MYC.

[0576] The genomic landscape of osteosarcoma tumors is usually dominated by widespread structural rearrangements, suggesting that several different mutational mechanisms, including chromothripsis, chromoplexy, kataegis and other structure-altering mechanisms, are involved (Cortes-Ciriano, Lee et al. 2020). These rearrangements give rise to genome-wide copy number alterations, usually dominated by copy number loss, including of PTEN and CDKN2A / B, but with recurrent amplifications of MYC, VEGFA and CCNE1 (Sayles, Breese et al. 2019). Osteosarcoma tumors often show signs of whole-genome duplication, which probably occurs in response to stresses imposed by pervasive copy number losses (Wu, Beird et al. 2020). This genomic complexity has long been interpreted as a sign of chromosomal instability, but emerging data suggest that the mechanisms triggering complexity are active early in the process of malignant transformation (Sayles, Breese et al. 2019).

[0577] Apart from these characteristic structural alterations, large-scale sequencing has identified only moderate levels of point mutations with few recurrently mutated genes (Chen, Bahrami et al. 2014, Perry, Kiezun et al. 2014, Behjati, Tarpey et al. 2017, Wu, Beird et al. 2020). The single most frequently altered gene is the tumor suppressor TP53, which is lost in >90% of osteosarcoma. TP53 is an extremely well-known tumor suppressor gene that has been referred to as the ‘guardian of thegenome’. Its normal function is to induce apoptosis in cells that acquire mutations. Given the chaotic genome typically present in osteosarcoma, abrogating that guardian function is necessary for cancer cell survival. Deletion of RBI also occurs in up to 30% of osteosarcoma tumors, often through loss of heterozygosity (Chen, Bahrami et al. 2014, Perry, Kiezun et al. 2014). Using these genomic aberrations to infer the evolution of the tumors, loss of TP53 and RBI likely occurs early in the transformation process (with TP53 inactivation required to propagate abnormal genomes (Overholtzer, Rao et al. 2003), followed by rapid accumulation of driver lesions such as MYC amplification, PTEN loss and deletion of ATRX, which seems to activate alternative lengthening of telomeres and is associated with decreased survival (Lawlor, Veronese et al. 2019).

[0578] Many patients later diagnosed with osteosarcoma first seek medical care with concerns for persisting pain in an extremity (Meltzer and Helman 2021). A question frequently asked to assess the severity of pain is whether the pain keeps them up at night; in the context of osteosarcoma, this is typically answered affirmatively. The pain is often accompanied by swelling at the same site and patients might misassociate these symptoms with recent minor injuries. The loss of structural integrity due to tumor-related osteolysis puts patients at risk for pathological fractures, which occur in -10% of patients and can complicate initial management (Zhou, Lu et al. 2017).

[0579] The workup for the presenting symptoms usually includes plain-film radiographs, potentially revealing large lesions that are causing destruction of normal trabecular bone with poorly defined margins (Papagelopoulos, Galanis et al. 2000). Lesions often stimulate periosteal new bone formation, which can give rise to the characteristic Codman triangle. The associated soft tissue mass can exhibit variable patterns of ossification, leading to the characteristic radial sunburst pattern often associated with osteosarcoma. Even if conventional radiographs are highly suggestive of osteosarcoma diagnoses, MRI covering the entire length of the affected bone should still be performed (Strauss, Erezza et al.

[0580] 2021). MRI can better characterize the associated soft tissue masses and facilitates planning for biopsy and eventual surgical resection. MRI also often reveals skip metastases frequently captured by local site imaging or, if more distant, suggested by bone scan, with implications for management and prognosis.

[0581] The diagnosis of osteosarcoma requires the pathological evaluation of a bone tissue biopsy sample, which can be obtained using either a minimally invasive core needle biopsy approach or an open biopsy. For core needle biopsies, adequate sampling of the tissue has to be ensured as osteosarcoma lesions can be quite heterogeneous and diagnostic features (such as malignant osteoid) can vary fromsample to sample. For additional downstream molecular diagnostics of fresh or frozen tissue, adequate sampling is even more important. Open biopsies are often preferred as they provide larger amounts of intact tissue. Fine-needle aspirates are usually inadequate for a definitive diagnosis of osteosarcoma due to the lack of sufficient histological context and the resulting difficulty to assess tumor grade and are therefore not recommended.

[0582] The histological diagnosis of osteosarcoma depends on the identification of malignant cells producing osteoid and irregular woven bone within fields of malignant tumor cells. The tumor cells usually exhibit marked atypia with a high degree of pleiotropism, and multiple morphologies (spindle, epithelioid, small round and giant cell) may exist within the same tumor. Although SATB2 and osteocalcin immunostaining and negative immunostaining to rule out alternative diagnostic entities can help guide a diagnostic work-up, no immunological or molecular marker has yet been identified that confirms a diagnosis of osteosarcoma (Beird, Bielack et al. 2022).

[0583] The post-diagnostic staging work-up aims to identify and characterize established metastatic disease. All patients presenting with newly diagnosed disease should undergo CT imaging of the chest, which has the highest efficiency in identifying lung nodules. Skeletal imaging with PET or technetium bone scans is important to identify covert bony disease. In 2008, guidelines from the Children’s Oncology Group were published, and are still widely accepted, advocating PET imaging with accompanying whole-body CT or whole-body MRI, with isotope bone scans if these modalities are not available (Meyer, Nadel et al. 2008). This work-up recommendation reveals lung metastases in 15-20% of patients, and occasionally identifies tumors within other bones or, very rarely, lesions at other sites. Staging is guided primarily by the Musculoskeletal Tumor Society (MSTS) staging system for sarcomas (Wolf and Enneking 1996), which defines tumors as being either low or high grade, confined to an anatomic compartment or violating anatomic barriers, and localized or metastatic. Most patients present with high-grade lesions that have both bony and soft tissue components, making the presence or absence of metastasis the primary risk-stratifying feature at diagnosis. A small number of patients present with localized, lower-grade parosteal and periosteal lesions (Grimer, Bielack et al. 2005).

[0584] At baseline, children and adolescents who present with localized osteosarcoma have an overall survival of -60% (Gorlick, Janeway et al. 2013). Patients who present with lung metastasis have the worst prognosis, with 3-year survival rates of <30% (Aljubran, Griffin et al. 2009). Fractures may also be an indicator of more aggressive disease. Patients experiencing fractures have higher rates of lung metastasis, both at presentation and subsequent to treatment (Zhou, Lu et al. 2017). The response toneoadjuvant chemotherapy, assessed in the definitive resection specimen, has clear prognostic value and has been used in previous clinical trials to stratify patients into good responders and poor responders; however, intensification of treatment did not improve outcomes in the poor responder group (Marina, Smeland et al. 2016).

[0585] The complex multi-modality management of osteosarcoma requires an expert multidisciplinary team that includes paediatric, medical and radiation oncologists, surgeons, pathologists, radiologists and specialist nurses (Strauss, Frezza et al. 2021). Combination chemotherapy and complete surgical resection are essential for cure. This applies to both patients with localized disease and those with primary metastatic osteosarcoma, provided complete surgical removal of all known metastatic deposits has been achieved. Current treatment paradigms offer patients with newly diagnosed, resectable osteosarcoma long-term survival rates of 60-70% (Smeland, Bielack et al. 2019). However, outcomes have hardly improved in the past decades, and the intensive chemotherapy regimens used are associated with important acute and long-term toxic effects and considerable impact on quality of life. In addition, patients with unresectable primary or metastatic disease at diagnosis and those with disease relapse have extremely poor outcomes (Bielack, Kempf-Bielack et al. 2002, Kempf-Bielack, Bielack et al. 2005).

[0586] Until the 1980s, the extremely high propensity of osteosarcoma to form pulmonary metastases led to an almost universally fatal disease outcome with only local surgical management available. Progress was only made with the introduction of systemic chemotherapy, which was soon administered neoadjuvantly (Jaffe, Puri et al. 2013). Multiple studies using a combined approach of neoadjuvant chemotherapy and surgery showed long-term, disease-free survival rates in the range of 60-70% in young patients with apparently localized disease (Smeland, Bielack et al. 2019). Doxorubicin, high-dose methotrexate with leucovorin rescue, cisplatin, and ifosfamide have since been established as the most active agents in osteosarcoma as both neoadjuvant and adjuvant therapies. The most efficacious regimens employ at least three of these drugs, yet adding a fourth agent may not lead to further benefits (Anninga, Gelderblom et al. 2011).

[0587] Although a minority of international investigators apply other, partially divergent protocols, most experts routinely use the neoadjuvant MAP regimen of high-dose methotrexate, doxorubicin and cisplatin as their treatment standard. This choice of regimen is based on the largest osteosarcoma study ever performed, EURAMOS-1 (Marina, Smeland et al. 2016). This prospective, randomized trial, based on the MAP regimen, unequivocally proved that long-term outcomes could not be furtherimproved by postoperative treatment alterations and augmentations for poor responders. In addition, maintenance therapy with IFNa was not of any benefit in those with a good response (Bielack, Smeland et al. 2015).

[0588] Attempts to further improve disease outcomes have generally not been met with undisputed progress. Immunotherapy with the macrophage-activator muramyl tripeptide-phosphatidyl ethanolamine encapsulated in liposomes (L-MTP-PE) was investigated in a US population (Meyers, Schwartz et al.

[0589] 2008). The results were hotly disputed at the time and left many questions open (Bielack, Marina et al.

[0590] 2008). The use of L-MTP-PE in patients with metastatic osteosarcoma was not found to improve event-free or overall survival and should not be used outside of clinical trials (Chou, Kleinerman et al.

[0591] 2009). Patients with unresectable or widely metastatic osteosarcoma who are deemed incurable are generally managed with the same systemic therapy options, including MAP chemotherapy and local tumor control, and outcomes are very poor, with <30% of patients surviving long term (Kager, Zoubek et al. 2003). Due to the toxic effects of treatment, quality of life must be balanced against potential treatment benefits for those individuals.

[0592] There is no standard-of-care systemic therapy for patients >40 years of age with a poor outcome and few clinical trials to inform practice (Grimer, Cannon et al. 2003). Guidelines suggest that adult patients (defined as older than 40) should be treated similarly to pediatric and young adult patients. However, adult patients may require tailored regimens, especially with regards to high-dose methotrexate. Retrospective analysis of the European Musculoskeletal Oncology Society of patients over 40 years did demonstrate that adult patients may benefit from aggressive treatment with surgery and chemotherapy, with outcomes possibly being related to decreased chemotherapy administered to some of the elderly patients (Grimer, Cannon et al. 2003).

[0593] Data on lower-grade lesion management remain sparse; however, most clinicians agree that grade 1 localized parosteal tumors can be treated surgically. The general principle of treating subvariants in osteosarcoma is based on their grade. Low-grade lesions, including low-grade central lesions, are treated by local surgical control only. Intermediate-grade lesions, including periosteal lesions and most osteosarcomas that affect the jaw, similarly need local control, which is also typically surgery. For intermediate-grade osteosarcomas, the role of chemotherapy is controversial and certainly not associated with the same risk-to-benefit relationship as high-grade osteosarcomas. Osteosarcomas in other craniofacial locations can be high grade and are treated with chemotherapy and local control insuch case. In craniofacial locations, local control can become challenging and is most often approached by multidisciplinary surgical oncologists and reconstructive teams.

[0594] Primary osteosarcoma resection should be conducted by experts in surgical reconstruction to preserve bone function while achieving complete resection. Otherwise, intralesional or marginal resections increase local recurrence rate, which is associated with reduced overall survival (Picci, Sangiorgi et al.

[0595] 1994, Bielack, Kempf-Bielack et al. 2002). Limb salvage is feasible for most patients with extremity tumors via reconstruction using an endoprosthetic implant, or allogeneic or autologous bone graft. Minimally invasive and non-invasive growing implants enable limb-salvage reconstruction as well as future limb-length equality for patients with skeletal immaturity (Ruggieri, Mavrogenis et al. 2013). Amputation remains optimal for some patients with large tumors when limb preservation is not possible or the expected functional differences between limb-sparing surgery and amputation are small and the risks of limb-sparing surgery high. Technologies such as transosseous suture fixation devices and advances in prosthetics offer the potential for improved functionality for these patients (Hebert, Rehani et al. 2017). Local recurrence rates for extremity osteosarcoma are low and generally less than 5%, suggesting that, in most cases, complete resection is achieved. In selected patients with osteosarcoma, radiation is considered postoperatively, particularly in patients with close surgical margins and a poor grade of necrosis in the resection specimen.

[0596] Surgery is also an important local control modality for metastatic sites, with long-term survival improving with the resection of lung metastases. Here, the number of metastases and completeness of excision seem to affect outcomes (Kager, Zoubek et al. 2003).

[0597] Although osteosarcoma is regarded as a radio-resistant disease, radiotherapy as local control may be considered if resection of a primary tumor is not possible or anticipated to lead to unacceptable morbidities such as in pelvic, trunk or craniofacial primary sites of disease (Strauss, Frezza et al.

[0598] 2021). Heavy particles offer a technical advantage to deliver the high doses of 60 Gy or ideally 70 Gy deemed necessary for disease control (Ciemik, Niemierko et al. 2011). Proton beam therapy and carbon ion radiotherapy are, therefore, increasingly used for patients with inoperable disease or disease at challenging primary sites. Five-year local control rates of 62-67% in patients with inoperable pelvic and trunk sarcomas are encouraging (Ciemik, Niemierko et al. 2011, Matsunobu, Imai et al. 2012). The combination of carbon ion radiotherapy and proton beam therapy for inoperable osteosarcoma was found to be feasible (Seidensaal, Mattke et al. 2021). A comprehensive evaluation of particle beam therapy, in this setting, is a priority.Each of the above forms and stages of osteosarcoma constitutes an embodiment of each and any aspect of the invention, including any embodiment thereof. Also, each of the above methods for treating osteosarcoma may be applied to the subject in accordance with the present invention as defined in the aspects of the invention, including any embodiment thereof. Finally, each of the above methods for diagnosing osteosarcoma may be applied to the subject in accordance with the present invention as defined in the aspects of the invention, including any embodiment thereof.

[0599] In an embodiment of each and any aspect of the invention, including any embodiment thereof, the tumor is a pancreatic cancer.

[0600] The normal pancreas consists of digestive enzyme-secreting acinar cells, bicarbonate-secreting ductal cells, centro-acinar cells that are the geographical transition between acinar and ductal cells, hormone-secreting endocrine islets and relatively inactive stellate cells. The majority of malignant neoplasms of the pancreas are adenocarcinomas; rare pancreatic neoplasms include neuroendocrine tumors which can secrete hormones such as insulin or glucagon and acinar carcinomas which can release digestive enzymes into the circulation. Specifically, ductal adenocarcinoma is the most common malignancy of the pancreas; this tumor - which is commonly referred to as pancreatic cancer - presents a substantial health problem (Kleeff, Korc et al. 2016). Pancreatic cancer is associated with an extremely poor prognosis for several reasons (Siegel, Miller et al. 2023). It is usually diagnosed at advanced stages, which is often due to nonspecific and — in some cases — no symptoms, a lack of sensitive and specific tumor markers and difficulties in imaging early-stage tumors. Pancreatic cancer is aggressive, with perineural and vascular local growth and early distant metastases that preclude curative surgical resection in most patients. Pancreatic cancer is characterized by a remarkable resistance (or tolerance) to most conventional treatment options, including chemotherapy, radiotherapy, molecularly targeted therapy and immunotherapy. Finally, pancreatic cancer harbors multiple genetic and epigenetic alterations and have complex and dense tumor microenvironments. All of these factors result in an overall 5-year survival rate of <7%, with almost all survivors at this time point being the 10-20% of patients who undergo surgical resection; for these patients, the 5-year survival rate is -15-25% (He, Ahuja et al. 2014).

[0601] In 2015, an estimated 367,000 new cases of pancreatic cancer were diagnosed worldwide, >50% of which occurred in high-income countries (Kleeff, Korc et al. 2016). In the United States, the highest rates were registered among black individuals (12-15 cases per 100,000 men and 8-10 cases per 100,000 women); these rates are -30-50% higher than in their white counterparts. In Oceania,indigenous populations are most affected, with an >30% excess, although long-term validated statistics are unavailable. The lowest rates, which are probably influenced by under-diagnosis, were recorded in India and in Northern and Central Africa. Mortality rates closely parallel incidence rates. Increases in reported incidence and mortality have taken place since the 1970s, in particular, in high-income countries where these increases can be partially attributed to diagnostic improvements. Although some reports suggest that incidence and mortality have levelled off in Western Europe and in North America, the latest data show the incidence continuing to rise in the United Kingdom and in North America — probably as a result of an ageing population (Kleeff, Korc et al. 2016).

[0602] Age is the major determinant of pancreatic cancer. Most patients are diagnosed at >50 years of age, with peak incidence in the seventh and eighth decades of life. In terms of preventable risk factors, tobacco smoking is the most important and most studied. Smokers have a twofold to threefold higher risk of developing pancreatic cancer than non-smokers; a dose-risk relationship has been noted, as has a favorable effect of smoking cessation. The proportion of cases attributable to tobacco smoking has been estimated to be 15-30% in various populations (Bosetti, Lucenteforte et al. 2012). Obesity and low physical activity are also linked to pancreatic cancer (Genkinger, Kitahara et al. 2015). Some nutritional and dietary factors, including high intake of (saturated) fats, low intake of vegetables and fruits and consumption of red and processed meats, are also associated with risk. A positive association with heavy alcohol consumption has been observed, but no such association has been shown with moderate drinking (Tramacere, Scotti et al. 2010). Diabetes mellitus is both a risk factor for disease and a consequence of early-stage pancreatic cancer; longterm diabetes mellitus approximately doubles the risk of pancreatic cancer (Bosetti, Rosato et al. 2014).

[0603] Pancreatic cancer most frequently arises from pancreatic intraepithelial neoplasia (PanIN), the classic pre-neoplastic lesions, but can also arise from larger precursor lesions, i.e. intraductal papillary mucinous neoplasms (IPMNs) and mucinous cystic neoplasms (Hezel, Kimmelman et al. 2006). Pancreatic cancer exhibits aberrant autocrine and paracrine signaling cascades that promote pancreatic cancer cell proliferation, migration, invasion and metastasis. For example, many signaling molecules — such as TGFa, IGF1, FGFs and HGF — and their respective tyrosine kinase receptors — such as EGFR, ERBB2 (also known as HER2), HER3, IGF1R, FGFRs and HGFR (also known as MET) — activate multiple pathways that enhance pancreatic cancer cell mitogenic self-sufficiency and promote migration and invasion (Preis and Korc 2011). These signaling cascades are activated in conjunction with the activation of anti-apoptotic and pro-survival pathways, such as signal transducer and activator of transcription 3 (STAT3), nuclear factor-κB (NF-κB) and AKT. Genes that are normally activeduring development, such as WNT, SHH and NOTCH, are also reactivated in some pancreatic cancers (Pasca di Magliano, Biankin et al. 2007).

[0604] The signaling in pancreatic cancer is complex, with multiple nodes and aberrant crosstalk pathways. For example, one of the aberrant signaling nodes in pancreatic cancer is characterized by enhanced activity of HGFR and EGFR, increased expression of neuropilin 1, CD44 expression and pi integrin, and is aggravated by the ability of HGFR to form heterodimers with EGFR (Tanizaki, Okamoto et al.

[0605] 2011). These alterations occur in the context of oncogenic KRAS and loss of CDKN2A, which encodes the tumor suppressor pl6 (Hezel, Kimmehnan et al. 2006). Pancreatic cancer also exhibits metabolic abnormalities and insensitivity to growth inhibitory pathways. Loss of negative growth constraints is best exemplified by aberrant TGFp signaling, which occurs as a consequence of increased expression of TGFp isoforms (Gore, Deitz et al. 2014). Although TGFp is a physiological tumor suppressor, it promotes tumor progression in pancreatic cancer and many other solid tumors by exerting paracrine effects within the tumor microenvironment that lead to enhanced growth and metastasis. TGFp can also directly induce pancreatic cancer cell proliferation by activating non-canonical signaling through MAPK phosphorylation, SRC and AKT phosphorylation, and by upregulating WNT7B expression through canonical SMAD4-dependent mechanisms (Gore, Deitz et al. 2014).

[0606] The molecular pathology of pancreatic cancer is dominated by activating mutations in KRAS, which are present in >90% of tumors. Inactivating mutations of TP53, CDKN2A and SMAD4 occur in 50-80% of pancreatic cancers, whereas other genes, including ARID 1 A, MLL3 and TGFBR2, are mutated in approx. 10% of tumors. However, few genes stand out among the myriad of infrequently mutated genes, which mostly occur at a prevalence of less than 2% (Chang, Grimmond et al. 2014). Although point mutations of individual genes reveal some aspects of disease pathophysiology, other genomic events contribute to carcinogenesis and can provide insight into molecular mechanisms. For example, CDKN2A is commonly inactivated by methylation, and CDKN2A and SMAD4 are commonly inactivated through homozygous deletion. Copy number alterations, which can be difficult to interpret due to the large number of genes that are amplified or lost in these regional events, also seem to play an important part. Four subtypes of pancreatic cancer have been proposed based on the number and location of structural variants (Waddell, Pajic et al. 2015). The first three subtypes comprise tumors with stable genomes with few (<50) structural variants, those with scattered structural events (50-200) and those with unstable genomes (>200 structural variants). Those tumors with unstable genomes are suggestive of defects in DNA maintenance and are a potential biomarker for platinum and poly (ADP-ribose) polymerase (PARP) inhibitor responsiveness (Waddell, Pajic et al. 2015). The fourth group, termed locally rearranged, is defined by >50 events localized to 1-3 chromosomes. These events are typically amplifications that encompass oncogenes, which can be the targets of existing therapeutics, or genomic catastrophes such as chromothripsis.

[0607] Pancreatic cancers characteristically have an abundant and dense collagenous stroma (desmoplasia) (Seymour, Hruban et al. 1994), resulting in a considerable hypoxic environment for cancer cells. This stroma is composed of extracellular matrix (ECM) proteins — collagens, fibronectin and laminin — as well as non-collagenous proteins such as glycoproteins, proteoglycans and glycosaminoglycans. Other factors in the stroma that possibly mediate the interaction of cancer cells with the ECM include growth factors, osteopontin, periostin and serine protein acidic and rich in cysteine. Cellular elements of the stroma include pancreatic stellate cells, which produce the collagenous matrix (activated stellate cells have also been referred to as cancer-associated fibroblasts in the literature), infiltrating immune cells, endothelial cells and neuronal cells. The immune cell complement in pancreatic cancer includes T cells (a majority being CD4+ regulatory T cells), myeloid-derived suppressor cells, macrophages and mast cells (Clark, Hingorani et al. 2007). Overall, immune cell infiltration suggests an immunosuppressive phenotype, even at the earliest stages of pancreatic cancer. A growing body of evidence suggests that CD4+ regulatory T cells in the stroma play a crucial part in warding off the host immune system. This feature is clinically important because factors that mediate the suppression of active antitumor immunity — such as the ligand for programmed cell death protein 1 (PD-1), PD-L1, expressed on cancer cells — form the basis of novel immunotherapeutic approaches study in pancreatic cancer.

[0608] The clinical manifestations of pancreatic cancer are nonspecific and include jaundice, unexplained weight loss, epigastric pain that radiates to the back, nausea, new-onset diabetes mellitus and, rarely, migratory thrombophlebitis (Wolfgang, Herman et al. 2013). When a diagnosis of a pancreatic cancer is suspected, imaging must be conducted. Multidetector CT provides excellent resolution of the pancreas and surrounding vasculature and can be used to evaluate other organs for spread of the disease (Wolfgang, Herman et al. 2013). A pancreas-specific protocol with dual-phase or multi-phase dynamic contrast is usually used, including early arterial phase images (to evaluate the involvement of the coeliac trunk, the superior mesenteric artery and other arteries), pancreatic phase images (to evaluate pancreatic lesions) and portal venous phase images (to evaluate the involvement of the portal vein, the superior mesenteric vein and other veins) (Lee and Lee 2014). Most pancreatic cancers form solid hypodense lesions. Cancers of the pancreas head typically obstruct both the pancreatic ducts andthe bile ducts, producing upstream dilatation of both ducts (the so-called double duct sign), whereas carcinomas of the corpus and tail of the gland will only obstruct distal parts of the pancreatic duct. Multidetector CT can define lesions in the pancreas, but is also the current gold standard to evaluate the resectability of any lesion. Involvement of the large vessels adjacent to the pancreas (such as the superior mesenteric artery and vein), might render the tumor borderline resectable, or locally advanced distant organs, such as the liver, would indicate advanced stage disease and that the patient is best treated with palliative therapy only (chemotherapy in most cases). MRI can provide excellent resolution of the pancreatic ducts and of any cysts that might be present. MRI might also be more sensitive than MDCT in detecting and evaluating liver metastases (Motosugi, Ichikawa et al. 2011). Given that pathology is required for establishing a diagnosis of pancreatic cancer, most patients will undergo endoscopic ultrasonography with fine-needle aspiration biopsy (Shin, Topazian et al. 2015). Endoscopic ultrasonography, although highly operator dependent, provides excellent resolution of the pancreas and peripancreatic vessels and lymph nodes, and can be used to obtain tissue for definitive diagnosis. PET can supplement these technologies, particularly when evaluating enlarged lymph nodes and larger masses of uncertain clinical significance, such as a persistent mass present after therapy (Parikh, Marcus et al. 2015). CA19-9 biomarker levels, although not useful for screening asymptomatic individuals, can be useful in monitoring patients with an established pancreatic cancer (Poruk, Gay et al. 2013). CA19-9 levels in the thousands of IU per ml are suggestive of metastatic disease, whereas a significant decline in levels after treatment suggests a good response. Other blood tests that are particularly useful in managing patients with pancreatic cancer include serum bilirubin and fasting blood glucose levels.

[0609] Biomarkers can be used for detection and response evaluation in palliative and neoadjuvant settings, as well as for post-resection follow-up. Several types of diagnostic biomarkers could be useful in pancreatic cancer. The most widely used biomarker is serum CAI 9-9. Unfortunately, CAI 9-9 lacks sufficient sensitivity or specificity to be useful for early pancreatic cancer diagnosis, but is routinely used to monitor disease progression, recurrence and / or therapy response (Poruk, Gay et al. 2013). Similarly, the presence of circulating tumor cells derived from pancreatic cancer could be diagnostic but are present in only some patients with metastatic disease. By contrast, circulating tumor DNA encoding mutant KRAS has been detected at the time of diagnosis in 43% of patients with localized disease, suggesting that circulating tumor DNA consisting of a panel of mutated genes such as mutated KRAS or mutated TP53 could serve as a non-invasive early diagnostic test (Bettegowda, Sausen et al.

[0610] 2014).The management of pancreatic cancer requires a comprehensive approach that integrates various treatment modalities, including surgery, radiotherapy, chemotherapy, and emerging therapies such as immunotherapy and targeted therapy. Given the aggressive nature of pancreatic cancer and the complexity of its treatment, a multimodality strategy is essential to address the disease at different stages and to improve patient outcomes.

[0611] Surgery remains the only potentially curative option for pancreatic cancer. Major pancreatectomy such as pancreaticoduodenectomy (also called the Whipple procedure) is safe when performed at high-volume centers with reported perioperative 30-day mortality rates of less than 3% (Cameron, Riall et al. 2006) (Kotecha, Tree et al. 2024). Based on the severity of the blood vessel involvement, localized pancreatic cancers are categorized into resectable pancreatic cancer, borderline resectable pancreatic cancer (BRPC), and locally advanced unresectable pancreatic cancer (LAPC) (Tempero, Malafa et al.

[0612] 2017). At diagnosis, approx. 10-20% of patients present with resectable tumours, 30-40% present with BRPC or locally advanced / unresectable pancreatic cancer LAPC, and 50-60% present with metastatic or systemic disease. Although upfront surgical resection followed by adjuvant chemotherapy is the standard of care for patients with resectable pancreatic cancer, the role of neoadjuvant therapy in resectable pancreatic cancer remains controversial. Given that surgery alone is not sufficient to achieve long-term survival (Traverse 2006), adjuvant chemotherapy is standard following tumor resection (Neoptolemos, Stocken et al. 2010, Oettle, Neuhaus et al. 2013).

[0613] Radiotherapy is one of the most commonly used local therapy approaches. Radiotherapy can be used as a neoadjuvant or adjuvant therapy to improve tumor resection rate or to reduce recurrence rate or as a definitive therapy in the localized, unresectable setting to improve local control. Many adjuvant studies have included radiotherapy, usually chemoradiotherapy, in addition to a course of systemic chemotherapy (Regine, Winter et al. 2011). Of note, the exact indications for radiation across disease stages remain controversial. In part, this is reflective of variation in outcomes across historical studies that used techniques which are no longer applicable. However, recent data using modem techniques have shown increasing signals for the benefit that radiation therapy may offer across the neoadjuvant, adjuvant, and definitive settings (Wang, Yang et al. 2024).

[0614] For patients with advanced or metastatic disease, chemotherapy is the only systemic therapy that offers a meaningful benefit. Over the past decade, significant advancements have been made in chemotherapy options for pancreatic cancer, including the introduction of FOLFIRINOX (oxaliplatin, irinotecan, leucovorin, and fluorouracil), gemcitabine plus nab-paclitaxel, S-l, liposomal irinotecan(Nal-IRI), gemcitabine plus capecitabine (Gem + Cap), modified FOLFIRINOX (mFOLFIRINOX), and NALIRIFOX (liposomal irinotecan, oxaliplatin, leucovorin, and fluorouracil) (Wang, Yang et al.

[0615] 2024). Immunotherapy has also made strides, with pembrolizumab, which targets the PD-1 / PD-L1 immune checkpoint pathway, showing improved outcomes in MSI-H / dMMR and TMB-H solid tumors. Erlotinib, an EGFR-targeting agent, showed slight improvement when combined with gemcitabine. Recent clinical investigations have highlighted the efficacy of PARP inhibitors such as olaparib in significantly prolonging survival for patients with germline BRCA-mutated pancreatic cancer, leading to FDA approval for maintenance treatment in cases with non-progressing disease following at least 16 weeks of first-line platinum -based chemotherapy. Moreover, targeted therapies such as larotrectinib and entrectinib for NTRK fusion-positive solid tumors, dabrafenib and trametinib for solid tumors with BRAFV600E mutations, and selpercatinib for RET fusion-positive solid tumors have received FDA approval (Wang, Yang et al. 2024).

[0616] Each of the above forms and stages of pancreatic cancer constitutes an embodiment of each and any aspect of the invention, including any embodiment thereof. Also, each of the above methods for treating pancreatic cancer may be applied to the subject in accordance with the present invention as defined in the aspects of the invention, including any embodiment thereof. Finally, each of the above methods for diagnosing pancreatic cancer may be applied to the subject in accordance with the present invention as defined in the aspects of the invention, including any embodiment thereof.

[0617] In an embodiment of each and any aspect of the invention, including any embodiment thereof, an interferon is administered to the subject.

[0618] Interferons (IFNs) comprise an extensive family of cytokines that not only possess antiviral activities, but also play a fundamental role in innate and adaptive immune regulation. There are three major types of IFNs, type I, II and III. Type I IFNs, especially IFNa and IFNp, can be produced by most cell types in the body in response to a wide range of stimuli and, in simplified terms, act as the initiators of many components of immune activation. In addition, they can kill tumor cells directly, sensitize cancer cells to chemotherapy and inhibit angiogenesis. Type II interferon, namely IFNy, produced by cytotoxic T cells and T helper cells, plays an essential role their function. Type III interferons are less well studied but play a specific role in antiviral defense (Yusuf, Allie et al. 2023).An important function of all interferons is the upregulation of major histocompatibility complex (MHC) molecules MHC I and MHC II, and an increase of immunoproteasome activity. All interferons significantly enhance the presentation of MHC I dependent antigens, and IFNy also significantly stimulates the MHC Il-dependent presentation of antigens. Higher MHC I expression increases presentation of viral and abnormal peptides from cancer cells to cytotoxic T cells, while the immunoproteasome processes these peptides for loading onto the MHC I molecule, thereby increasing the recognition and killing of infected or malignant cells. Higher MHC II expression increases presentation of these peptides to helper T cells; these cells release cytokines (such as more interferons and interleukins, among others) that signal to and co-ordinate the activity of other immune cells (Ikeda, Old et al. 2002, Dunn, Bruce et al. 2005, Borden, Sen et al. 2007).

[0619] Type I interferons can induce an anti-bacterial state in infected cells, controlling the spread of infectious and inflammatory agents, particularly viral pathogens (Ivashkiv and Donlin 2014, Duncan, Randall et al. 2021, Pasrija and Naime 2021). In addition, they regulate the innate immune response by facilitating antigen presentation and natural killer cell function (Meyer 2009), while mediating inflammatory pathways and cytokines (Lopez de Padilla and Niewold 2016, Mayer-Barber and Yan 2017, Gallucci, Meka et al. 2021). Finally, type I interferons can trigger the adaptive immune system, prompting high-affinity antigen-immune cell responses and the development of immune memory (McNab, Mayer-Barber et al. 2015, Schreiber 2020).

[0620] IFNy, the only type II interferon, can enhance expression of surface major histocompatibility complex (MHC) molecules and increase the processing and presentation of tumor-specific antigens (Shankaran, Ikeda et al. 2001), thus facilitating T-cell recognition and cytotoxicity (Gao, Shi et al. 2016, Ayers, Lunceford et al. 2017, Manguso, Pope et al. 2017, Patel, Sanjana et al. 2017). However, even though IFNy is one of the best-studied cytokines in animal models and has undergone extensive clinical testing, little is known regarding the impact of IFNy on the immune cells of the human tumor microenvironment (TME). Most of the clinical studies focus on the efficacy of IFNy, although one study shows increased MHC expression on tumor cells in fine needle aspirates from melanoma patients treated with IFNy (Propper, Chao et al. 2003).

[0621] Clearly, the functions of type I and II IFNs suggest that they would be beneficial if harnessed as anticancer agents and, indeed, the interaction between IFNs, tumor cells and immune cells has been extensively studied as components of cancer immunotherapies.Preclinical and clinical research have confirmed the essential role of interferons for effective host immunological responses to malignant cells. Type I interferons (IFNa and IFNP) directly regulate transcription of >100 downstream genes, which results in a myriad of direct (on cancer cells) and indirect (through immune effector cells and vasculature) effects on the tumor (Borden 2019). New insights into endogenous and exogenous activation of type I interferons in the tumor and its microenvironment have given impetus to drug discovery and patient evaluation of interferon-directed strategies.

[0622] Interferons are useful for treatment of few cancer types for their ability to stimulate the immune system, directly inhibit tumor growth, and improve the effectiveness of other treatments. However, their use is typically limited to certain types of cancer, and the therapy is often associated with side effects that need to be managed carefully.

[0623] Two IFNa forms have been approved for treatment of tumors, INTRON A (Interferon alfa-2b (FDA 2021)) and ROFERON-A (interferon alfa-2b).

[0624] IFNa recombinant (INTRON A) is indicated for treatment of few cancer forms,

[0625] • Hairy cell leukemia at 2 million IU / m2administered intramuscularly or subcutaneously 3 times a week for up to 6 months;

[0626] • Malignant melanoma adjuvant to surgical treatment at 20 million IU / m2 as an intravenous infusion, on 5 consecutive days per week, for 4 weeks during induction and at 10 million IU / m2as subcutaneous injection three times per week for 48 weeks for maintenance;

[0627] • Clinically aggressive follicular non-Hodgkin’s lymphoma at 5 million IU subcutaneously three times per week for up to 18 months in conjunction with anthracycline-containing chemotherapy regimen and following completion of the chemotherapy regimen;

[0628] • AIDS-related Kaposi's sarcoma at 30 million IU / m2 / dose administered subcutaneously or intramuscularly three times a week until disease progression or maximal response has been achieved after 16 weeks of treatment.

[0629] In addition, INTRON A is approved for the following non-cancer indications

[0630] • Condylomata acuminata involving external surfaces of the genital and perianal areas at 1 million IU per lesion in a maximum of 5 lesions in a single course. The lesions are directlyinjected three times weekly on alternate days for 3 weeks. An additional course may be administered at 12 to 16 weeks;

[0631] • Chronic hepatitis C at 3 million IU three times a week (TIW) administered subcutaneously or intramuscularly; in patients tolerating therapy with normalization of ALT at 16 weeks of treatment, the therapy should be extended to 18 to 24 months at 3 million IU three times per week to improve the sustained response rate;

[0632] • Chronic hepatitis B at 30 to 35 million IU per week, administered subcutaneously or intramuscularly, either as 5 million IU daily or as 10 million IU three times a week for 16 weeks.

[0633] ROFERON-A (interferon alfa-2b) was another approved IFNa preparation but is no longer marketed. It was approved for the treatment of chronic myelogenous leukemia (CME), renal cell carcinoma (RCC), melanoma and hepatitis C.

[0634] PEGylated IFNa (PEGASYS (FDA 2023)) is FDA-approved for as antiviral agent, but not for cancer indications. It is indicated for treatment of (i) chronic hepatitis C (CHC) in patients > 5 years of age, and (ii) of adult patients with chronic hepatitis B infection. Adult patients receive a subcutaneous dose of 180 pg per week. Other approved PEGylated IFNa preparations are PEGINTRON and SYLATRON.

[0635] The amino acid sequence of INFa 2b (INTRON A, ROFERON-A, KEGG DRUG: Interieron alfa-2b, accessed Mat ) is:

[0636] CDLPQTHSLG SRRTLMLLAQ MRRISLFSCL KDRHDFGFPQ EEFGNQFQKA ETIPVLHEMI QQIFNLFSTK DSSAAWDETL LDKFYTELYQ QLNDLEACVI QGVGVTETPL MKEDSILAVR KYFQRITLYL KEKKYSPCAW EVVRAEIMRS FSLSTNLQES LRSKE (SEQ ID NO: 242)

[0637] The IFNa family has 13 subtypes and 23 different variants. The individual proteins have molecular masses between 19-26 kDa and consist of proteins with lengths of 156-166 and 172 amino acids. All IFNa subtypes possess a common conserved sequence region between amino acid positions 115-151 while the amino-terminal ends are variable. Many IFNa subtypes differ in their sequences at only one or two positions. Naturally occurring variants also include proteins truncated by 10 amino acids at the carboxy-terminal end.The aa sequence of IFNa 2a (PEGASYS, KE(.>.

[0638]

[0639] alfa-2a, accessed Mar 19, 2025) is:

[0640] CDLPQTHSLG SRRTLMLLAQ MRKISLFSCL KDRHDFGFPQ EEFGNQFQKA ETIPVLHEMI QQIFNLFSTK DSSAAWDETL LDKFYTELYQ QLNDLEACVI QGVGVTETPL MKEDSILAVR KYFQRITLYL KEKKYSPCAW EVVRAEIMRS FSLSTNLQES LRSKE (SEQ ID NO: 243)

[0641] No form of IFNP has been approved for treatment of cancer. EXTAVIA is a branded version of interferon beta- lb, a first-line disease-modifying therapy that has been a standard-of-care for multiple sclerosis for many years. EXTAVIA is indicated for the treatment of MS patients with relapsing forms of the disease and for newly diagnosed patients.

[0642] The amino acid sequence of interferon beta- IB (EXTAVIA, http:

[0643]

[0644] tg.jp / entry / D00746 accessed Mai ) is:

[0645] SYNLLGFLQR SSNFQSQKLL WQLNGRLEYC LKDRMNFDIP EEIKQLQQFQ KEDAALTIYE MLQNIFAIFR QDSSSTGWNE TIVENLLANV YHQINHLKTV LEEKLEKEDF TRGKLMSSLH LKRYYGRILH YLKAKEYSHC AWTIVRVEIL RNFYFINRLT GYLRN (SEQ ID NO: 244)

[0646] No form of IFNy has been approved for cancer, but ACTIMMUNE (interferon gamma- IB) is an FDA approved cytokine therapy used as a standard of care treatment for osteopetrosis and chronic granulomatous disease where it is administered subcutaneously three times per week at 50 pg / m2(patients with body surface area > 0.5 m2) or 1.5 pg / kg (patients with body surface area < 0.5 m2). From the 1990s, IFNy was evaluated as a single agent cancer immunotherapy in several prospective randomized trials (Windbichler, Hausmaninger et al. 2000, Giannopoulos, Constantinides et al. 2003, Alberts, Marth et al. 2008). These trials found no benefit in cancer and thus IFNy was largely written off as an ineffective treatment.

[0647] The amino acid sequence of interferon gamma- IB (ACTIMMUNE, https: > > > / ■ ■i.jp / entry / D00747 accessed Mai ■

[0648]

[0649] ) is:

[0650] MQDPYVKEAE NLKKYFNAGH SDVADNGTLF LGILKNWKEE SDRKIMQSQI VSFYFKLFKN FKDDQSIQKS VETIKEDMNV KFFNSNKKKR DDFEKLTNYS VTDLNVQRKA IHELIQVMAE LSPAAKTGKR KRSQMLFRGR (SEQ ID NO: 245)Each of the above interferons constitutes an embodiment of each and any aspect of the invention, including each and any embodiment thereof.

[0651] In an embodiment of each and any aspect, including any embodiment thereof, a CXCL12 antagonist is an antagonist targeting the CXCL12 / CXCR4 / CXCR7 axis and more specifically CXCL12, CXCR4 and / or CXCR7. In an embodiment of each and any one of the aspects of the present invention, including any embodiment thereof, the CXCL12 / CXCR4 / CXCR7 axis refers to the system of interaction of CXCL12 with its receptor CXCL4 and with its receptor CXCL12. It will be appreciated by a person skilled in the art that interfering with this system of interaction is possible by targeting CXCL12 or any one of its receptors, i.e. CXCR4 and CXCR7.

[0652] In a more preferred embodiment, the targeting is such that a targeting agent is therapeutically active by targeting CXCL12, CXCR4 and / or CXCR7, preferably is targeting CXCL12, CXCR4 or CXCR7. In a further preferred embodiment, targeting means direct interaction, preferably physical interaction, between the targeting agent and CXCL12, CXCR4 and / or CXCR7. Due to such targeting a physiological and / or biochemical effect arising from the interaction between CXCL12 and CXCR4 and / or between CXCL12 and CXCR7 is affected and results in a therapeutic effect of the targeting agent. Preferably, the targeting agent is also referred to as CXCL12 antagonist herein. It will therefore be appreciated by a person skilled in the art that a CXCL12 antagonist used in accordance with the present invention may make use of different modes of action. Such modes of action include that the CXCL12 antagonist inhibits signaling of CXCL12 via one or both of CXCL12 receptor C-X-C chemokine receptor type 4 (CXCR4) and CXCL12 receptor C-X-C chemokine receptor type 7 (CXCR7).

[0653] It will be appreciated by a person skilled in the art that such inhibition of the signaling of CXCL12 via one or both of CXCL12 receptor CXCR4 and CXCR7 can be achieved by (a) providing a molecule binding to CXCL12 so as to inhibit the binding of CXCL12 to CXCR4 and / or CXCR7, providing a molecule binding to CXCR4 so as to inhibit the binding of CXCL12 to CXCR4, and (c) providing a molecule binding to CXCR7 so as to inhibit the binding of CXCL12 to CXCR7. Such molecule which is binding to CXCL12 is preferably one selected from the group comprising an aptamer, preferably a CXCL12 binding aptamer, a spiegelmer, preferably a CXCL12-binding spiegelmer”, an antibody, preferably a CXCL12 binding antibody, a protein binding to CXCL12, a CXCL12 binding anticalin, a CXCL12 binding peptide, a fusion protein comprising a CXCL12 binding moiety and a CXCL12 binding small molecule.Such molecule which is binding to CXCR4 to inhibit the signaling of CXCL12 is preferably one selected from the group comprising an aptamer, preferably a CXCR4 binding aptamer, a spiegelmer, preferably a CXCR4-binding Spiegelmer, an antibody, preferably a CXCR4 binding antibody, a protein binding to CXCR4, a CXCR4 binding anticalin, a CXCR4 binding peptide, a fusion protein comprising a CXCR4 binding moiety and a CXCR4 binding small molecule.

[0654] Such molecule which is binding to CXCR7 is preferably one selected from the group comprising an aptamer, preferably a CXCR7 binding aptamer, a spiegelmer, preferably a CXCR7 -binding spiegelmer, an antibody, preferably a CXCR7 binding antibody, a protein binding to CXCR4, a CXCR7 binding anticalin, a CXCR7 binding peptide, a fusion protein comprising a CXCR7 binding moiety and a CXCR7 binding small molecule.

[0655] It will also be appreciated by a person skilled in the art that that such inhibition of the signaling of CXCL12 via one or both of CXCL12 receptor CXCR4 and CXCR7 can be achieved by a compound binding CXCR4 and / or CXCR7, whereby such binding of the compound interferes with signaling, preferably with the signal transducing characteristics of CXCR 4 and / or CXCR7 rather than interfering with the binding of CXCL12 to CXCR4 and / or CXCR7.

[0656] A molecule which is binding to CXCR4 and because of its binding to CXCR4 is interfering with the signal transduction characteristics of CXCR4, preferably signal transduction triggered by the binding of CXCL12 to CXCR4, is preferably one selected from the group comprising an aptamer, preferably a CXCR4 binding aptamer, a spiegelmer, preferably a CXCR4 binding spiegelmer, a protein binding to CXCR4, an antibody or antigen-binding moiety of the antibody, preferably a CXCR4 binding antibody or a CXCR4 binding fragment of the CXCR4 binding antibody, an anticalin, preferably a CXCR4 binding anticalin, a CXCR4 binding peptide, a fusion protein comprising a CXCR4 binding moiety and a CXCR4 binding small molecule.

[0657] The generation and identification, respectively, of such aptamer, spiegelmer, antibody, binding protein, anticalin, peptide, fusion protein and small molecule is known to a person skilled in the art and also disclosed herein.

[0658] In a preferred embodiment, CXCL12 is human CXCL12, CXCR4 is human CXCR4, and CXCR7 is human CXCR7.

[0659] In connection with administering to the subject suffering from the tumor a CXCL12 antagonist, it is to be acknowledged that animal studies showed that radiation-induced devascularization is efficientlyrestored by vasculogenesis after recruitment of bone marrow-derived cells which leads to rapid recurrence of the tumor (Kioi, Vogel et al. 2010). It is assumed that CXCL12 plays an important role in neovascularization by recruiting endothelial and other bone marrow-derived pro-angiogenic cells through a CXCR4- and CXCR7 -dependent mechanism. Notably, irradiation further increases CXCL12 expression (Kioi, Vogel et al. 2010, Kozin, Kamoun et al. 2010, Liu, Alomran et al. 2014).

[0660] In an embodiment of each and any aspect of the present invention, including any embodiment thereof, the CXCL12 antagonist inhibits signaling of CXCL12 via one or both of CXCL12 receptor C-X-C chemokine receptor type 4 (CXCR4) and CXCL12 receptor C-X-C chemokine receptor type 7 (CXCR7). In a preferred embodiment thereof, the CXCL12 antagonist inhibits the binding of CXCL12 to CXCL12 receptor C-X-C chemokine receptor type 4 (CXCR4) and / or to CXCL12 receptor C-X-C chemokine receptor type 7 (CXCR7). The interaction between CXCL12 and its receptor CXCR4 is also referred to as CXCL12 - CXCR4 axis. Such CXCL12 - CXCR4 axis may be targeted by a CXCL12 antagonist. Similarly, interaction between CXCL12 and its receptor CXCR7 is also referred to as CXCL12 - CXCR7 axis. Such CXCL12 - CXCR7 axis may be targeted by a CXCL12 antagonist. In an even more preferred embodiment, such binding of CXCL12 to said receptors is effected by the CXCL12 antagonist binding to CXCL12.

[0661] CXCL12 antagonists binding to CXCL12 include, among others, a nucleic acid molecule binding to CXCL12. Such CXCL12 binding nucleic acid molecule can be a CXCL12 binding aptamer or a CXCL12 binding spiegelmer.

[0662] Aptamers are nucleic acid molecules made of D-nucleotides as building blocks which specifically bind to a target molecule through a mechanism different from Watson Crick base pairing. Spiegelmers are nucleic acid molecules made of L-nucleotides as building blocks which specifically bind to a target molecule through a mechanism different from Watson Crick base pairing. Both aptamers and spiegelmers can be made of ribonucleotides, of deoxyribonucleotides are a combination of both ribonucleotides and deoxyribonucleotides. Aptamers and spiegelmers as such are known to a person skilled in the art and are, among others, described in ‘The Aptamer Handbook’ (eds. Klussmann, 2006) and spiegelmers comprising both ribonucleotides and deoxyribonucleotides are disclosed in international patent application WO 2012 / 095303.

[0663] In an embodiment of each and any aspect of the present invention, including any embodiment thereof, the CXCL12 antagonist is an L-nucleic acid molecule binding to CXCL12, wherein the L-nucleic acid molecule is preferably selected from the group comprising an CXCL12 binding nucleic acid moleculeof type B, an CXCL12 binding nucleic acid molecule of type C, an CXCL12 binding nucleic acid molecule of type A and an CXCL12 binding nucleic acid molecule of type D as defined in more detail herein.

[0664] The CXCL12 binding L-nucleic acid molecules can be characterized in terms of stretches of nucleotides which are also referred to herein as Boxes (see, Example 1).

[0665] The different types of CXCL12 binding nucleic acid molecules comprise three different stretches of nucleotides: the first terminal stretch of nucleotides, the central stretch of nucleotides and second terminal stretch of nucleotides. In general, CXCL12 binding nucleic acid molecules of the present invention comprise at their 5 ’-end and the 3 ’-end the terminal stretches of nucleotides: the first terminal stretch of nucleotides and the second terminal stretch of nucleotides (also referred to as 5’-terminal stretch of nucleotides and 3’-terminal stretch of nucleotides). The first terminal stretch of nucleotides and the second terminal stretch of nucleotides can, in principle due to their base complementarity, hybridize to each other, whereby upon hybridization a double-stranded structure is formed. However, such hybridization is not necessarily realized in the molecule under physiological and / or non-physiological conditions. The three stretches of nucleotides of CXCL12 binding nucleic acid molecules - the first terminal stretch of nucleotides, the central stretch of nucleotides and second terminal stretch of nucleotides - are arranged to each other in 5’ -> 3 ’-direction: the first terminal stretch of nucleotides - the central stretch of nucleotides - the second terminal stretch of nucleotides. However, alternatively, the second terminal stretch of nucleotides, the central stretch of nucleotides and the terminal first stretch of nucleotides are arranged to each other in 5’ -> 3 ’-direction.

[0666] The differences in the sequences of the defined boxes or stretches between the different CXCL12 binding nucleic acid molecules influence the binding affinity to CXCL12. Based on binding analysis of the different CXCL12 binding nucleic acid molecules used in accordance with the present invention the central stretch and the nucleotides forming the same are individually and more preferably in their entirety essential for binding to human CXCL12.

[0667] The terms ‘stretch’ and ‘stretch of nucleotide’ are used herein in a synonymous manner if not indicated to the contrary.

[0668] In an embodiment of each and any aspect of the present invention, including any embodiment thereof, the nucleic acid used in accordance the present invention is a single nucleic acid molecule. In a further embodiment, the single nucleic acid molecule is present as a multitude of the single nucleic acid molecule or as a multitude of the single nucleic acid molecule species.It will be acknowledged by the ones skilled in the art that the nucleic acid molecule used in accordance with the present invention as subject to each and any of aspect, including any embodiment thereof, preferably consists of nucleotides which are covalently linked to each other, preferably through phosphodiester links or linkages.

[0669] It is within the present invention as subject to each and any of aspect, including any embodiment thereof, that the nucleic acids comprise two or more stretches or part(s) thereof can, in principle, hybridise with each other. Upon such hybridisation a double-stranded structure is formed. It will be acknowledged by the ones skilled in the art that such hybridisation may or may not occur, particularly under in vitro and / or in vivo conditions. Also, in case of such hybridisation, it is not necessarily the case that the hybridisation occurs over the entire length of the two stretches where, at least based on the rules for base pairing, such hybridisation and thus formation of a double-stranded structure may, in principle, occur. As preferably used herein, a double-stranded structure is a part of a nucleic acid molecule or a structure formed by two or more separate strands or two spatially separated stretches of a single strand of a nucleic acid molecule, whereby at least one, preferably two or more base pairs exist which are base pairing preferably in accordance with the Watson-Crick base pairing rules. It will also be acknowledged by the one skilled in the art that other base pairing such as Hoogsten base pairing may exist in or form such double-stranded structure. It is also to be acknowledged that the feature that two stretches hybridize preferably indicates that such hybridization is assumed to happen due to base complementarity of the two stretches.

[0670] In a preferred embodiment of each and any aspect of the invention, including any embodiment thereof, the term arrangement as used herein, means the order or sequence of structural or functional features or elements described herein in connection with the nucleic acids disclosed herein.

[0671] It will be acknowledged by the person skilled in the art that the nucleic acids used in accordance with the present invention as subject to each and any of aspect, including any embodiment thereof, are capable of binding to CXCL12. Without wishing to be bound by any theory, the CXCL12 binding results from a combination of three-dimensional structural traits or elements of the nucleic acid molecule which are caused by orientation and folding patterns of the primary sequence of nucleotides forming such traits or elements, whereby preferably such traits or elements are the first terminal stretch of nucleotides, the central stretch of nucleotides and the second terminal stretch of nucleotides of CXCL12 binding nucleic acid molecules. It is evident that the individual trait or element may be formed by various different individual sequences the degree of variation of which may vary depending on the three-dimensional structure such element or trait has to form. The overall binding characteristicof the claimed nucleic acid results from the interplay of the various elements and traits, respectively, which ultimately results in the interaction of the claimed nucleic acid with its target, i. e. CXCL12. Again without being wished to be bound by any theory, the central stretch of nucleotides that is characteristic for CXCL12 binding nucleic acids seems to be important for mediating the binding of the claimed nucleic acid molecules with CXCL12. Accordingly, the nucleic acids according to the present invention are suitable for the interaction with CXCL12. Also, it will be acknowledged by the person skilled in the art that the nucleic acids according used in accordance with the present invention are antagonists to CXCL12. Because of this, the nucleic acids according to the present invention are suitable for the treatment and prevention of brain cancer in accordance with the method of the present invention.

[0672] The nucleic acid molecules used in accordance with the present invention as subject to each and any of aspect, including any embodiment thereof, shall also comprise nucleic acids which are essentially homologous to the particular sequences disclosed herein. The term substantially homologous shall be understood such as the homology is at least 75%, preferably 85%, more preferably 90% and most preferably more that 95 %, 96 %, 97 %, 98 % or 99%.

[0673] The actual percentage of homologous nucleotides present in the nucleic acid used in accordance with the present invention as subject to each and any of aspect, including any embodiment thereof, will depend on the total number of nucleotides present in the nucleic acid. The percent modification can be based upon the total number of nucleotides present in the nucleic acid.

[0674] The homology between two nucleic acid molecules can be determined as known to the person skilled in the art. More specifically, a sequence comparison algorithm may be used for calculating the percent sequence homology for the test sequence(s) relative to the reference sequence, based on the designated program parameters. The test sequence is preferably the sequence or nucleic acid molecule which is said to be homologous or to be tested whether it is homologous, and if so, to what extent, to a different nucleic acid molecule, whereby such different nucleic acid molecule is also referred to as the reference sequence. In an embodiment, the reference sequence is a nucleic acid molecule as described herein, preferably a nucleic acid molecule having a sequence according to any one of SEQ ID NO: 5 to SEQ ID NO: 238, SEQ ID NO: 258 and SEQ ID NO: 259, more preferably a nucleic acid molecule having a sequence according to any one of SEQ ID NO: 67, SEQ ID NO: 132, SEQ ID NO: 102, SEQ ID NO: 110, SEQ ID NO: 111, SEQ ID NO: 116, SEQ ID NO: 117, SEQ ID NO: 134, SEQ ID NO: 135, SEQ ID NO: 31, SEQ ID NO: 133, SEQ ID NO: 142, SEQ ID NO: 143, and SEQ ID NO: 144. Optimal alignment of sequences for comparison can be conducted, e.g., by the local homology algorithm ofSmith & Waterman (Smith & Waterman, 1981) by the homology alignment algorithm of Needleman & Wunsch (Needleman & Wunsch, 1970) by the search for similarity method of Pearson & Lipman (Pearson & Lipman, 1988), by computerized implementations of these algorithms (GAP, BESTFIT, FASTA, and TFASTA in the Wisconsin Genetics Software Package, Genetics Computer Group, 575 Science Dr., Madison, Wis.), or by visual inspection.

[0675] One example of an algorithm that is suitable for determining percent sequence identity is the algorithm used in the basic local alignment search tool (hereinafter " BLAST "), see, e.g. Altschul et al (Altschul, Gish et al. 1990, Altschul, Madden et al. 1997). Software for performing BLAST analyses is publicly available through the National Center for Biotechnology Information (hereinafter " NCBI"). The default parameters used in determining sequence identity using the software available from NCBI, e.g., BLASTN (for nucleotide sequences) and BLASTP (for amino acid sequences) are described in McGinnis et al (McGinnis and Madden 2004).

[0676] The nucleic acid molecules used in accordance with the present invention as subject to each and any of aspect, including any embodiment thereof, shall also comprise nucleic acid molecules which have a certain degree of identity relative to the nucleic acid molecules disclosed herein and defined by their nucleotide sequence. More preferably, the instant invention also comprises those nucleic acid molecules which have an identity of at least 75%, preferably 85%, more preferably 90% and most preferably more than 95 %, 96 %, 97 %, 98 % or 99% relative to the nucleic acid molecules disclosed herein and defined by their nucleotide sequence or a part thereof.

[0677] The terms nucleic acid(s) and nucleic acid molecule(s) are used interchangeably herein. Also, the nucleic acid molecules used in accordance with the present invention shall also comprise those nucleic acid molecules comprising the nucleic acids sequences disclosed herein or part thereof, such as, e.g., a metabolite or derivative of the nucleic acid molecules used in accordance with the present invention, preferably to the extent that the nucleic acid molecules or said parts are involved in the or capable of binding to CXCL12. Such a nucleic acid molecule may be derived from the ones disclosed herein, e.g., by truncation. Truncation may be related to either or both of the ends of the nucleic acid molecules as disclosed herein. Also, truncation may be related to the inner sequence of nucleotides, i.e. it may be related to the nucleotide(s) between the 5’ and the 3’ terminal nucleotide, respectively. Moreover, truncation shall comprise the deletion of as little as a single nucleotide from the sequence of the nucleic acids disclosed herein. Truncation may also be related to more than one stretch of the nucleic acid molecules used in accordance with the present invention, whereby the stretch can be as little as one nucleotide long. The binding of a nucleic acid molecules used in accordance with thepresent invention can be determined by the ones skilled in the art using routine experiments or by using or adopting a method as described herein, preferably as described herein in the example part.

[0678] In addition, it is within that present invention as subject to each and any of aspect, including any embodiment thereof, that one or several parts of the nucleic acid molecule used in accordance with the present invention are present as D-nucleic acids or at least one or several parts of the nucleic acid molecules are L-nucleic acids. The term “part” of the nucleic acid molecules shall mean as little as one nucleotide. Such nucleic acid molecules are generally referred to herein as D- and L-nucleic acids, respectively. Therefore, in a particularly preferred embodiment, the nucleic acid molecules used in accordance with the present invention consist of L-nucleotides and comprise at least one D-nucleotide. Such D-nucleotide is preferably attached to a part different from the stretches defining the nucleic acid molecules used in accordance with the present invention, preferably those parts thereof, where an interaction with other parts of the nucleic acid molecule is involved. Preferably, such D-nucleotide is attached at a terminus of any of the stretches and of any nucleic acid molecule used in accordance with the present invention, respectively. In a further preferred embodiment, such D-nucleotides may act as a spacer or a linker, preferably attaching modifications such as PEG and HES to the nucleic acids according to the present invention.

[0679] It is also within the present invention as subject to each and any of aspect, including any embodiment thereof, that the nucleic acid molecules used in accordance with the present invention as subject to each and any of aspect, including any embodiment thereof, are part of a longer nucleic acid molecule, whereby this longer nucleic acid molecule comprises several parts, whereby at least one such part is a nucleic acid molecule, or a part thereof, used in accordance with the present invention. The other part(s) of these longer nucleic acid molecules can be either one or several D-nucleic acid(s) or L-nucleic acid(s). Any combination may be used in connection with the present invention. These other part(s) of the longer nucleic acid molecule can exhibit a function which is different from binding, preferably from binding to CXCL12. One possible function is to allow interaction with other molecules, whereby such other molecules preferably are different from CXCL12, such as, e.g., for immobilization, cross-linking, detection or amplification. In a further embodiment of the present invention the nucleic acid molecules used in accordance with the invention comprise, as individual or combined moieties, several of the nucleic acid molecules used in accordance with the present invention. Such nucleic acid molecule comprising several of the nucleic acid molecules used in accordance with the present invention as subject to each and any of aspect, including any embodiment thereof, is also encompassed by the term longer nucleic acid.L-nucleic acid molecules as used herein are nucleic acid consisting of consisting of L-nucleotides, preferably consisting completely of L-nucleotides.

[0680] D-nucleic acid molecules as used herein are nucleic acid molecules consisting of D-nucleotides, preferably consisting completely of D-nucleotides.

[0681] The terms nucleic acid and nucleic acid molecule are used herein in an interchangeable manner if not explicitly indicated to the contrary.

[0682] Also, if not indicated to the contrary, any nucleotide sequence is set forth herein in 5’ -> 3’ direction.

[0683] As preferably used herein, any position of a nucleotide is determined or referred to relative to the 5 ’ end of a sequence, a stretch or a substretch. Accordingly, a second nucleotide is the second nucleotide counted from the 5’ end of the sequence, stretch and substretch, respectively. Also, in accordance therewith, a penultimate nucleotide is the second nucleotide counted from the 3’ end of a sequence, stretch and substretch, respectively.

[0684] It is also within the present invention as subject to each and any of aspect, including any embodiment thereof, that the nucleic acid molecules used in accordance with the present invention as subject to each and any of aspect, including any embodiment thereof, regardless whether they are present as D-nucleic acids, L-nucleic acids or D, L-nucleic acids or whether they are DNA or RNA, may be present as single stranded or double stranded nucleic acids. Typically, the nucleic acid molecules used in accordance with the present invention are single stranded nucleic acids which exhibit defined secondary structures due to the primary sequence and may thus also form tertiary structures. The nucleic acid molecules used in accordance with the present invention, however, may also be double stranded in the meaning that two strands which are complementary or partially complementary to each other, are hybridised to each other or at least may be hybridised to each other in light of their base complementarity.

[0685] The nucleic acid molecules used in accordance with the present invention as subject to each and any of aspect, including any embodiment thereof, may be modified. Such modifications may be related to a single nucleotide of the nucleic acid and are well known in the art. Examples for such modification are described by, among others, Venkatesan et al. (Venkatesan, Kim et al. 2003) and Kusser (Kusser 2000). Such modification can be a H atom, an F atom or O-CH3group or NH2-group at the 2’ position of the individual nucleotide of which the nucleic acid consists. Also, the nucleic acid molecule used inaccordance with the present invention can comprises at least one LNA nucleotide. In an embodiment the nucleic acid according to the present invention consists of LNA nucleotides.

[0686] In an embodiment of each and any aspect of the invention, including any embodiment thereof, the nucleic acid molecules used in accordance with the present invention as subject to each and any of aspect, including any embodiment thereof, may be a multipartite nucleic acid molecule. A multipartite nucleic acid molecule as used herein, is a nucleic acid molecule which consists of at least two separate nucleic acid strands. These at least two nucleic acid strands form a functional unit whereby the functional unit is a ligand to a target molecule. The at least two nucleic acid strands may be derived from any of the nucleic acid molecules used in accordance with the present invention by either cleaving the nucleic acid molecule to generate two strands or by synthesising one nucleic acid corresponding to a first part of the overall nucleic acid molecule and another nucleic acid molecule corresponding to the second part of the overall nucleic acid molecule. It is to be acknowledged that both the cleavage and the synthesis may be applied to generate a multipartite nucleic acid molecule where there are more than two strands as exemplified above. In other words, the at least two separate nucleic acid strands are typically different from two strands being complementary and hybridising to each other although a certain extent of complementarity between said at least two separate nucleic acid strands may exist and whereby such complementarity may result in the hybridisation of said separate strands.

[0687] Finally, it is also within the present invention as subject to each and any of aspect, including any embodiment thereof, that a fully closed, i.e. circular structure for the nucleic acid molecules used in accordance with the present invention is realized, i.e. that the nucleic acid molecules are closed in an embodiment, preferably through a covalent linkage, whereby more preferably such covalent linkage is made between the 5’ end and the 3’ end of the nucleic acid sequences as disclosed herein or any derivative thereof.

[0688] An option for determining the binding constants of a nucleic acid molecule used in accordance with the present invention is the use of the methods as described in Examples 5 and 6 which confirms the above finding that the nucleic acid molecules used in accordance with the present invention according to the present invention exhibit a favourable KD value range. An appropriate measure in order to express the intensity of the binding between the individual nucleic acid molecule and the target which is in the present case CXCL12, is the so-called KD value which as such as well the method for its determination are known to the one skilled in the art.Preferably, the KD value shown by the nucleic acid molecules used in accordance with the present invention as subject to each and any of aspect, including any embodiment thereof, is below 1 pM. A KD value of about 1 pM is said to be characteristic for a non-specific binding of a nucleic acid to a target. As will be acknowledged by the ones skilled in the art, the KD value of a group of compounds such as the nucleic acid molecules used in accordance with th...

Claims

1. Claims1. A CXCL12 antagonist for use in a method for treating a tumor in a subject, wherein the method comprises administering to the subject the CXCL12 antagonist and an interferon, wherein the tumor is an immunologically cold tumor.

2. An interferon for use in a method for treating a tumor in a subject, wherein the method comprises administering to the subject the interferon and a CXCL12 antagonist, wherein the tumor is an immunologically cold tumor.

3. A CXCL12 antagonist for use in a method for converting an immunologically cold tumor in a subject into an immunologically hot tumor in the subject, wherein the method comprises administering to the subject the CXCL12 antagonist and an interferon, wherein upon administration of the CXCL12 antagonist and the interferon the immunologically cold tumor is or will be converted into an immunologically hot tumor.

4. An interferon for use in a method for converting an immunologically cold tumor in a subject into an immunologically hot tumor in the subject, wherein the method comprises administering to the subject the interferon and a CXCL12 antagonist, wherein upon administration of the interferon and the CXCL12 antagonist the immunologically cold tumor is or will be converted into an immunologically hot tumor.

5. The CXCL12 antagonist for use of any one of claims 1 and 3 and the interferon for use of any one of claims 2 and 4, wherein the immunologically cold tumor is refractory to an immunotherapy.

6. A CXCL12 antagonist for use in a method for treating a tumor in a subject, wherein the method comprises administering to the subject the CXCL12 antagonist and an interferon, wherein the tumor is refractory to immunotherapy.

7. An interferon for use in a method for treating a tumor in a subject, wherein the method comprises administering to the subject the interferon and a CXCL12 antagonist, wherein the tumor is refractory to immunotherapy.

8. A CXCL12 antagonist for use in a method for converting an immunotherapy refractory tumor in a subject into an immunotherapy non-refractory tumor in the subject, wherein the method comprises administering to the subject the CXCL12 antagonist and an interferon, wherein upon administration of the CXCL12 antagonist and the interferon the immunotherapy refractory tumor is or will be converted into an immunotherapy nonrefractory tumor.

9. An interferon for use in a method for converting an immunotherapy refractory tumor in a subject into an immunotherapy non-refractory tumor in the subject, wherein the method comprises administering to the subject the interferon and a CXCL12 antagonist, wherein upon administration of the interferon and the CXCL12 antagonist the immunotherapy refractory tumor is or will be converted into an immunotherapy non-refractory tumor.

10. The CXCL12 antagonist for use of any one of claims 5, 6, and 8 and the interferon for use of any one of claims 5, 7 and 9, wherein the tumor refractory to an immunotherapy is an immunologically cold tumor.

11. The CXCL12 antagonist for use of any one of claims 5, 6, 8 and 10 and the interferon for use of any one of claims 5, 7, 9 and 10, wherein the immunotherapy is selected from the group consisting of checkpoint inhibition, adoptive cell therapy and tumor vaccines.

12. The CXCL12 antagonist for use of any one of claims 1, 3, 5, 6, 8, 10 and 11 and the interferon for use of any one of claims 2, 4, 5, 7, 9, 10 and 11, wherein the cold tumorand the tumor refractory to an immunotherapy is characterized by an immunosuppressive tumor microenvironment.

13. The CXCL12 antagonist for use of any one of claims 1, 3, 5, 6, 8, 10, 11 and 12 and the interferon for use of any one of claims 2, 4, 5, 7, 9, 10, 11 and 12, wherein the cold tumor and the tumor refractory to immunotherapy have at least one characteristic selected from the group comprising reduced or lacking expression of PD-L1, reduced number of tumor-infiltrating lymphocytes, genomic stability and reduced or lacking preexisting antitumor immune response.

14. The CXCL12 antagonist for use of claim 8 and the interferon for use of claim 9, wherein the tumor non-refractory to an immunotherapy is a tumor which is sensitive to an immunotherapy, preferably the immunotherapy is selected from the group consisting of checkpoint inhibition, adoptive cell therapy and tumor vaccines.

15. The CXCL12 antagonist for use of any one of claims 8 and 14 and the interferon for use of any one of claims 9 and 14, wherein the tumor non-refractory to an immunotherapy is a tumor which is a hot tumor, preferably the tumor non-refractory to an immunotherapy and the immunologically hot tumor are characterized by a nonimmunosuppressive tumor microenvironment and / or one characteristic selected from the group comprising expression of PD-L1, non-reduced number of tumor-infiltrating lymphocytes, genomic instability and non-reduced or preexisting antitumor immune response.

16. The CXCL12 antagonist for use of any one of claims 1, 3, 5, 6, 8 and 10 to 15 and 12 and the interferon for use of any one of claims 2, 4, 5, 7, 9 and 10 to 15, wherein the tumor is a solid tumor, preferably, the tumor is selected from the group comprising pancreatic cancer, triple-negative breast cancer, prostate cancer, ovarian cancer, soft tissue sarcomas and osteosarcoma, more preferably the tumor is selected from the group consisting of synovial sarcoma, pancreatic cancer and osteosarcoma.

17. The CXCL12 antagonist for use of any one of claims 1, 3, 5, 6, 8 and 10 to 16 and the interferon for use of any one of claims 2, 4, 5, 7, 9 and 10 to 16, wherein the method comprises administering to the subject an adoptive cell therapy, preferably the adoptive cell therapy comprises administering a CAR T cell and / or a TCR-T cell to the subject.

18. The CXCL12 antagonist for use of any one of claims 1, 3, 5, 6, 8 and 10 to 17 and the interferon for use of any one of claims 2, 4, 5, 7, 9 and 10 to 17, wherein the CXCL12 antagonist and the interferon are administered concomitantly to the subject or the CXCL12 antagonist is administered to the subject after the administration of the interferon to the subject.

19. The CXCL12 antagonist for use of any one of claims 1, 3, 5, 6, 8 and 10 to 18 and the interferon for use of any one of claims 2, 4, 5, 7, 9 and 10 to 18, wherein the interferon is a type I interferon, preferably the interferon is interferon alpha, more preferably the interferon is interferon alpha comprising an amino acid sequence of SEQ ID NO: 242 or comprising an amino acid sequence of SEQ ID NO: 243.

20. The CXCL12 antagonist for use of any one of claims 1, 3, 5, 6, 8 and 10 to 18 and the interferon for use of any one of claims 2, 4, 5, 7, 9 and 10 to 18, wherein the interferon is a type II interferon, preferably the interferon is interferon gamma, more preferably the interferon is interferon gamma comprising an amino acid sequence of SEQ ID NO: 245.

21. The CXCL12 antagonist for use of any one of claims 1, 3, 5, 6, 8 and 10 to 20 and the interferon for use of any one of claims 2, 4, 5, 7, 9 and 10 to 20, wherein the CXCL12 antagonist is a C-X-C motif chemokine 12 (CXCL12) antagonist, wherein the CXCL12 antagonist inhibits the binding of CXCL12 to CXCL12 receptor C-X-C chemokine receptor type 4 (CXCR4) and / or to CXCL12 receptor C-X-C chemokine receptor type 7 (CXCR7), preferably the CXCL12 antagonist is binding to CXCL12.

22. The CXCL12 antagonist for use of any one of claims 1, 3, 5, 6, 8 and 10 to 21 and the interferon for use of any one of claims 2, 4, 5, 7, 9 and 10 to 21, wherein the CXCL12 antagonist is a nucleic acid molecule binding to CXCL12, wherein the nucleic acid molecule binding to CXCL12 comprises a central stretch of nucleotides, whereby the central stretch of nucleotides comprises the following nucleotide sequence:5’ GUGUGAUCUAGAUGUADWGGCUGWUCCUAGUYAGG 3’ (SEQ ID NO: 57).

23. The CXCL12 antagonist for use of any one of claims 1, 3, 5, 6, 8 and 10 to 22, preferably of claim 22 and the interferon for use of claim 2, 4, 5, 7, 9 and 10 to 22, preferably of 22, wherein the CXCL12 antagonist is a nucleic acid molecule binding to CXCL12, wherein the nucleic acid molecule binding to CXCL12 comprises a nucleotide sequence according to any one of SEQ ID NO: 67 to SEQ ID NO: 72, SEQ ID NO: 132, SEQ ID NO: 46 to SEQ ID NO: 56 and SEQ ID NO: 61 to SEQ ID NO: 65, or a nucleotide sequence at least 80 % identical thereto,preferably a nucleotide sequence according to any one of SEQ ID NO: 67, SEQ ID NO: 132, SEQ ID NO: 46, SEQ ID NO: 47, SEQ ID NO: 48 and SEQ ID NO: 61, or a nucleotide sequence at least 80 % identical thereto,more preferably a nucleotide sequence according to any one of SEQ ID NO: 67 and SEQ ID NO: 132, or a nucleotide sequence at least 80 % identical thereto.

24. The CXCL12 antagonist for use of any one of claims 1, 3, 5, 6, 8 and 10 to 23, preferably of claim 23 and the interferon for use of any one of 2, 4, 5, 7, 9 and 10 to 23, preferably of claim 23, wherein the nucleic acid molecule binding to CXCL12 is an L-ribonucleic acid molecule comprising a nucleotide sequence ofGCGUGGUGUGAUCUAGAUGUAUUGGCUGAUCCUAGUCAGGUACGC (SEQ ID NO: 67), preferably the L-ribonucleic acid molecule comprises a 40 kDa polyethylene glycolmoiety, wherein the 40 kDa polyethylene glycol moiety is attached to the 5’end of the nucleotide sequence, preferably the CXCL12 antagonist is NOX-A12.

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