Treating cancer with tumor-infiltrating lymphocytes

Abstract

Provided herein are methods for generating TILs that can be used to treat patients with cancer (eg, pediatric cancer, uveal melanoma, or mesothelioma).

Claims

1. A population of tumor infiltrating lymphocytes (TILs) for use in a method of treating cancer, wherein the cancer is mesothelioma, childhood cancer or uveal melanoma; The population of TILs: a) obtaining a first population of TILs from a tumor excised from said subject by processing a tumor sample obtained from said subject into (i) a plurality of tumor fragments or (ii) a tumor digest. obtained from a process comprising: a first expansion is performed for about 3-11 days to obtain said second population of TILs; A second expansion is performed for about 7-11 days to obtain said third population of TILs.

2. The population of TILs, (a) obtaining a first population of TILs from a tumor excised from a patient or subject by processing a tumor sample obtained from the subject into (i) a plurality of tumor fragments or (ii) a tumor digest; (b) adding the first population of TILs to a closed system; (c) performing a first expansion by culturing the first TIL population in a cell culture medium comprising IL-2 to produce a second TIL population, wherein the first expansion is performed in a closed vessel providing a first gas permeable surface area, and wherein the first expansion is performed for about 3-11 days to obtain the second TIL population, and wherein the transition from step (b) to step (c) occurs without opening the system; (d) performing a second expansion by supplementing the cell culture medium of the second TIL population with additional IL-2, OKT-3, and antigen presenting cells (APCs) to produce a third TIL population, wherein the second expansion is performed for about 7-11 days to obtain the third TIL population, the third TIL population being a therapeutic TIL population, and wherein the second expansion is performed in a closed vessel providing a second gas permeable surface area, and the transition from step (c) to step (d) occurs without opening the system. (e) harvesting the therapeutic TIL population obtained from step (d), wherein the transition from step (d) to step (e) occurs without opening the system; (f) transferring the harvested therapeutic TIL population from step (e) into an infusion bag, wherein the transition from step (e) to (f) occurs without opening the system; (g) cryopreserving the infusion bag containing the harvested therapeutic TIL population from step (f) using a cryopreservation process; (h) administering a therapeutically effective dose of the third population of TILs to the subject from the infusion bag of step (g).

3. A population of tumor infiltrating lymphocytes (TILs) for use in a method of treating cancer in a subject in need thereof, wherein the cancer is mesothelioma, childhood cancer or uveal melanoma; The population of TILs: (a) obtaining and / or receiving a first population of TILs from a surgical resection, needle biopsy, core biopsy, mini-biopsy, or other means for obtaining a sample containing a mixture of tumor cells and TIL cells from said subject; (b) contacting the first population of TILs with a first cell culture medium; (c) performing an initial expansion (or a first expansion by priming) of said first TIL population in said first cell culture medium to obtain a second TIL population, said first cell culture medium comprising IL-2, and optionally said first expansion by priming occurs over a period of 1 to 8 days; (d) performing a rapid expansion of the second TIL population in a second cell culture medium to obtain a third TIL population, wherein the second cell culture medium comprises IL-2, OKT-3 (anti-CD3 antibody), and optionally irradiated allogeneic peripheral blood mononuclear cells (PBMCs), and the rapid expansion is performed for a period of 14 days or less, and optionally the rapid expansion can proceed for 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, 7 days, 8 days, 9 days, or 10 days after initiation of the rapid second expansion to obtain the third TIL population; (e) administering a therapeutically effective portion of the third population of TILs to the subject with cancer.

4. The population of TILs of claim 2 , wherein the second population of TILs in step (c) is at least 50-fold more numerous than the first population of TILs.

5. The mesothelioma is i. from pleural, peritoneal, and pericardial mesothelioma; ii. from epithelioid mesothelioma, sarcomatoid mesothelioma, and biphasic mesothelioma; iii. from epithelioid mesothelioma, sarcomatoid mesothelioma, and biphasic mesothelioma; or iv. from adenomatous mesothelioma, cystic mesothelioma, anaplastic mesothelioma, well-differentiated papillary mesothelioma, and small cell mesothelioma; 5. The population of TILs of any one of claims 1 to 4, selected and optionally wherein the subject has previously undergone treatment for said mesothelioma, and further optionally wherein said previous treatment is surgery, radiation therapy, chemotherapy, immunotherapy, or a combination thereof, and optionally wherein said chemotherapy comprises platinum-based cisplatin, platinum-based carboplatin, or carboplatin, and / or said immunotherapy comprises anti-PD-1 antibody therapy, anti-CTLA-4 antibody therapy, or a combination thereof.

6. 6. The population of TILs of claim 5, wherein the subject is also administered immunotherapy, optionally wherein the immunotherapy is anti-PD-1 antibody therapy, anti-CTLA-4 antibody therapy, or a combination thereof.

7. i. The uveal melanoma is choroidal melanoma, ciliary body melanoma, or iris melanoma; ii. The childhood cancer is neuroblastoma; iii. the childhood cancer is a sarcoma, optionally wherein the sarcoma is osteosarcoma or the sarcoma is a soft tissue sarcoma, further optionally wherein the soft tissue sarcoma is rhabdomyosarcoma, Ewing's sarcoma, or primitive neuroectodermal tumor (PNET); iv. The population of TILs of any one of claims 1-4, wherein the pediatric cancer is a central nervous system (CNS) related cancer, optionally wherein the CNS related cancer is a medulloblastoma, a pineoblastoma, a glioma, or an epithelioma, a glioblastoma.

8. The object is i. is under 2 years of age, between 2 and 12 years of age, or between 12 and 21 years of age; ii. weighs less than 40 kg; or iii. The population of TILs of any one of claims 1 to 7, which is between 8 kg and 40 kg.

9. The childhood cancer is i. neuroblastoma and the subject has previously received dinutuximab treatment; ii. neuroblastoma and the subject has not previously received dinutuximab treatment; iii. Neuroblastoma, and the neuroblastoma is refractory to dinutuximab; iv. neuroblastoma and the subject has previously received vincristine sulfate treatment; v. neuroblastoma and the subject has not previously been treated with vincristine sulfate, and / or vi. neuroblastoma, and said neuroblastoma is refractory to vincristine sulfate treatment; or vii. Ewing's sarcoma and the subject has previously received dactinomycin treatment; viii. Ewing's sarcoma and the subject has not previously received dactinomycin treatment; ix. Ewing's sarcoma, and the Ewing's sarcoma is refractory to dactinomycin; x. rhabdomyosarcoma and the subject has previously received dactinomycin treatment; xi. rhabdomyosarcoma and the subject has not previously received dactinomycin treatment; xii. Rhabdomyosarcoma, and the rhabdomyosarcoma is refractory to dactinomycin; xiii. rhabdomyosarcoma and the subject has previously received vincristine sulfate treatment; xiv. rhabdomyosarcoma and the subject has not previously received vincristine sulfate treatment; and / or xv. The population of TILs of claim 7, which is rhabdomyosarcoma, and said rhabdomyosarcoma is refractory to vincristine sulfate treatment.

10. The first extension comprises: i. takes place over a period of about 11 days; ii. the IL-2 is present in the cell culture medium in the first expansion at an initial concentration of 1000 IU / mL to 6000 IU / mL; iii. in the second expansion step, the IL-2 is present at an initial concentration of 1000 IU / mL to 6000 IU / mL and the OKT-3 antibody is present at an initial concentration of about 30 ng / mL; iv. in the second expansion step, the IL-2 is present at an initial concentration of 1000 IU / mL to 6000 IU / mL and the OKT-3 antibody is present at an initial concentration of about 30 ng / mL; v. the first expansion is performed using a gas permeable container; vi. the second expansion is performed using a gas permeable container; vii. the first cell culture medium further comprises a cytokine selected from the group consisting of IL-4, IL-7, IL-15, IL-21, and combinations thereof; and / or The population of TILs of any one of claims 2, 4-9, wherein the second cell culture medium further comprises a cytokine selected from the group consisting of IL-4, IL-7, IL-15, IL-21, and combinations thereof.

11. i. The initial expansion is performed over a period of about 11 days; ii. the IL-2 is present in the cell culture medium during the initial expansion at an initial concentration of 1000 IU / mL to 6000 IU / mL; iii. in the initial expansion step, the IL-2 is present at an initial concentration of 1000 IU / mL to 6000 IU / mL and the OKT-3 antibody is present at an initial concentration of about 30 ng / mL; iv. the initial expansion is performed using a gas permeable container; v. the rapid expansion is performed using a gas permeable container; vi. the cell culture medium of the first expansion further comprises a cytokine selected from the group consisting of IL-4, IL-7, IL-15, IL-21, and combinations thereof; and / or The population of TILs of any one of claims 3-9, wherein the cell culture medium of the second expansion further comprises a cytokine selected from the group consisting of IL-4, IL-7, IL-15, IL-21, and combinations thereof.

12. and optionally further comprising treating the subject with a non-myeloablative lymphodepletion regimen prior to administering the TILs to the subject. i. the non-myeloablative lymphodepletion regimen comprises cyclophosphamide administered at a dose of 60 mg / m2 / day for 2 days, followed by fludarabine administered at a dose of 25 mg / m2 / day for 5 days; ii. the non-myeloablative lymphodepletion regimen comprises cyclophosphamide at a dose of 60 mg / m2 / day and fludarabine at a dose of 25 mg / m2 / day for 2 days, followed by fludarabine at a dose of 25 mg / m2 / day for 3 days; 12. The population of TILs of any one of claims 1 to 11, optionally wherein the cyclophosphamide is administered with mesna.

13. The method according to claim 1, i. treating said subject with an IL-2 regimen beginning the day after administration of said third population of TILs to said subject; ii. Treating said subject with an IL-2 regimen beginning on the same day as administering said third population of TILs to said subject. and optionally wherein the IL-2 regimen is a high-dose IL-2 regimen comprising 600,000 or 720,000 IU / kg aldesleukin, or a biosimilar or variant thereof, administered as a 15-minute bolus intravenous infusion every 8 hours until tolerated; Optionally, the therapeutically effective population of TILs administered comprises about 2.3×10 10 to about 13.7×10 10 TILs.

14. i. The initial expansion occurs over a period of 21 days or less; ii. the initial expansion occurs over a period of 7 days or less; iii. The population of TILs of any one of claims 3 to 13, wherein said rapid expansion is performed over a period of 7 days or less.

15. i. The first expansion in step (c) and the second expansion in step (d) are each performed separately within a period of 11 days; and / or ii. The population of TILs of any one of claims 2, 4-14, wherein steps (a)-(e) are performed for about 10 days to about 22 days.

16. processing the tumor sample obtained from the subject in step (a) into a tumor digest; i. incubating the tumor sample in an enzyme medium; ii. mechanically disrupting the tumor sample to dissociate the tumor sample; and / or iii. Purifying the dissociated tumor sample using density gradient separation.

3. The population of TILs of claim 1 or 2, optionally comprising the enzyme medium comprising DNase, further optionally comprising 30 units / mL of DNase, and / or the enzyme medium comprising collagenase, further optionally comprising 1.0 mg / mL of collagenase.

17. 17. The population of TILs of any one of claims 1 to 16, wherein the cancer is a cancer with a V600 mutation, optionally wherein the V600 mutation is selected from the group consisting of a V600E mutation, a V600E2 mutation, a V600K mutation, a V600R mutation, a V600M4 mutation, and a V600D mutation, and optionally wherein the method further comprises administering to the subject at least one BRAF and / or MEK inhibitor.

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