Method of improving car-t cell therapies

Abstract

The instant disclosure relates to methods of treating cancer, including methods of improving CAR-T cell therapies.

Description

METHOD OF IMPROVING CAR-T CELL THERAPIESCROSS-REFERENCE TO RELATED APPLICATIONS

[0001] The present application claims the benefit of U.S. Provisional Patent Applications No. 63 / 548,685, filed February 1, 2024, and No. 63 / 684,158, filed August 16, 2024, each of which is incorporated herein by reference in its entirety.GOVERNMENT LICENSE RIGHTS

[0002] This invention was made with government support under Award # 5R01CA252940- 04 awarded by the National Institutes of Health. The government has certain rights in the invention.FIELD

[0003] The instant disclosure relates to methods of treating cancer, including methods of improving CAR-T cell therapies.BACKGROUND

[0004] CAR-T cells are frequently used in the treatment of hematological malignancies such as non-Hodgkin lymphoma.

[0005] Non-Hodgkin lymphoma is a heterogeneous group of cancers originating in B lymphocytes, T lymphocytes or natural killer cells. In the United States, B cell lymphomas represent 80-85% of cases reported. Non-Hodgkin lymphoma is the most prevalent hematological malignancy and is the seventh leading site of new cancers among men and women and account for 4% of all new cancer cases and 3% of deaths related to cancer. Large B-cell lymphomas represent the most common sub-group of NHL. Diffuse large B-cell lymphoma (DLBCL) is the most common subtype of large B-cell lymphoma, accounting for approximately 30% of NHL cases. Patients with chemotherapy-refractory DLBCL following treatment under the current standards of care still have a particularly dire prognosis, with no curative treatment options. These consistently discouraging results demonstrate that new treatment options are urgently needed for patients whose tumors have demonstrated a lack of response to chemotherapy.

[0006] Primary mediastinal B-cell lymphoma (PMBCL) is thought to arise from thymic (medullary) B cells and represents approximately 3% of patients diagnosed with large B-cell lymphoma. Follicular lymphoma (FL), a B cell lymphoma, is the most common indolent (slow-growing) form of NHL, accounting for approximately 20% to 30% of all NHLs. Somepatients with FL will transform (TFL) histologically to DLBCL which is more aggressive and associated with a poor outcome. Treatment options for relap sed / refractory PMBCL and TFL are similar to those in DLBCL. Given the low prevalence of these diseases, no large prospective randomized studies in these patient populations have been conducted. Patients with chemotherapy refractory disease have a similar or worse prognosis to those with refractory DLBCL.

[0007] Despite being dispensable for direct cytotoxicity of CAR-T cells, IFNy plays two major roles in this context: (i) IFNy stimulates T-cell activity proliferation, especially in CAR-T cells containing a CD28 costimulatory domain, and (ii) IFNy drives macrophage activation and subsequent production of proinflammatory cytokines / chemokines. See, e.g., Bailey et. al., B Blood Cancer Discov. 2022 Mar 1; 3(2): 136-153.

[0008] The primary acute toxicities observed to date with CAR-T cells have been CRS and ICANS. CRS is defined as a constellation of symptoms which may include (but are not limited to) fever, chills, hypotension, hypoxia, and when extreme, macrophage activation syndrome. Manifestations of ICANS vary and include confusion, obtundation, seizures, hallucinations, aphasia, ataxia, and more rarely, profound cerebral edema. The future success and application of commercial product to a broader population of patients is limited by the development of these toxicities. Therefore, there remains an unmet need for methods of improving existing CAR-T cell therapies to decrease toxicities.SUMMARY

[0009] In one aspect, provided herein is a method of improving a CAR-T cell therapy by decreasing a CAR-T cell associated toxicity in a subject receiving CAR-T cell therapy, comprising administering to the subject an effective dose of emapalumab. In some embodiments, the toxicity is cytokine-release syndrome. In some embodiments, the toxicity is immune effector cell-associated neurotoxicity syndrome (ICANS). In some embodiments, the toxicity is prolonged cytopenia.

[0010] In some embodiments, the emapalumab is administered at a dose of 0.1 to 10 mg / kg intravenously. In some embodiments, the emapalumab is administered at a dose of 1 to 10 mg / kg intravenously. In some embodiments, the emapalumab is administered at a dose of 3 mg / kg intravenously. In some embodiments, the dose is a one-time dose.

[0011] In some embodiments, the CAR-T cell therapy has IFNy mediated toxicities. In some embodiments, the CAR-T cell therapy is axicabtagene ciloleucel. In some embodiments, the axicabtagene ciloleucel is administered at a target dose of 2 x 106anti-CD19 CAR-T cells / kg, administered intravenously. In some embodiments, the axicabtagene ciloleucel is administered as a one-time dose. In some embodiments, the axicabtagene ciloleucel is administered concurrently, before or after emapalumab administration. In some embodiments, the axicabtagene ciloleucel is administered one day before emapalumab administration. In some embodiments, the axicabtagene ciloleucel is administered one to five days before emapalumab administration. In some embodiments, the axicabtagene ciloleucel is administered about 24 hours after emapalumab administration.

[0012] In some embodiments, the emapalumab is administered about 1-5 days after the lymphodepleting regimen. In some embodiments, the CAR-T cell therapy is administered concurrently with or about one to five days after the emapalumab.

[0013] In some embodiments, the subject is further administered at least one additional therapy. In some embodiments, the at least one additional therapy is a lymphodepleting regimen. In some embodiments, the lymphodepleting regimen is administered for 1, 2, 3, 4, 6, 7, or more days. In some embodiments, the lymphodepleting regimen comprises fludarabine and cyclophosphamide or bendamustine. In some embodiments, the lymphodepleting regimen comprises radiation In some embodiments, the lymphodepleting chemotherapy regimen is administered prior to the administration of the CAR-T cell therapy.

[0014] In some embodiments, the method results in a prolongation of the duration of response to the CAR-T cell therapy of at least 10% as compared to the duration of response to the CAR-T cell therapy in the absence of emapalumab. In some embodiments, the method results in an increase in the overall response rate to the CAR-T cell therapy of at least 10% as compared to the overall response rate to the CAR-T cell therapy in the absence of emapalumab. In some embodiments, the method results in an increase in progression-free survival after administration of the CAR-T cell therapy of at least 10% as compared to the progression-free survival after administration of the CAR-T cell therapy in the absence of emapalumab. In some embodiments, the method results in an increase in overall survival after administration of the CAR-T cell therapy of at least 10% as compared to the overall survival after administration of the CAR-T cell therapy in the absence of emapalumab. In some embodiments, the method results in at least 500 CAR-T cells per microliter blood of the subject.

[0015] In some embodiments, the patient has a large B-cell lymphoma that is refractory to first-line chemoimmunotherapy. In some embodiments, the patient has a large B-cell lymphoma that relapses within 12 months of first-line chemoimmunotherapy. In some embodiments, the patient has a relapsed or refractory large B-cell lymphoma after two ormore lines of systemic therapy. In some embodiments, the patient has non-Hodgkin lymphoma. In some embodiments, the patient has diffuse large B-cell lymphoma. In some embodiments, the patient has primary mediastinal B-cell lymphoma. In some embodiments, the patient has diffuse large B-cell lymphoma arising from follicular lymphoma. In some embodiments, the patient has high grade B-cell lymphoma.

[0016] In another aspect, provided herein is a method for treating cancer in a subject in need thereof, the method comprising: (a) administering to the subject a lymphodepleting regimen; (b) administering to the subject emapalumab; and (c) administering to the subject a CAR-T cell therapy.

[0017] In some embodiments, the emapalumab is administered at a dose of 0.1 to 10 mg / kg intravenously. In some embodiments, the emapalumab is administered at a dose of 1 to 10 mg / kg intravenously. In some embodiments, the emapalumab is administered at a dose of 3 mg / kg intravenously. In some embodiments, the dose is a one-time dose.

[0018] In some embodiments, the CAR-T cell therapy has IFNy mediated toxicities.

[0019] In some embodiments, the CAR-T cell therapy is axicabtagene ciloleucel. In some embodiments, the axicabtagene ciloleucel is administered at a target dose of 2 x 106anti-CD19 CAR-T cells / kg, administered intravenously. In some embodiments, the axicabtagene ciloleucel is administered as a one-time dose. In some embodiments, the axicabtagene ciloleucel is administered concurrently, before or after emapalumab administration. In some embodiments, the axicabtagene ciloleucel is administered one day before emapalumab administration. In some embodiments, the axicabtagene ciloleucel is administered one to five days before emapalumab administration. In some embodiments, the emapalumab is administered about 1-5 days after the lymphodepleting regimen. In some embodiments, the CAR-T cell therapy is administered concurrently with or about one to five days after the emapalumab.

[0020] In some embodiments, the subject is further administered at least one additional therapy. In some embodiments, the at least one additional therapy is a lymphodepleting regimen. In some embodiments, the lymphodepleting regimen is administered for 1, 2, 3, 4, 6, 7, or more days. In some embodiments, the lymphodepleting regimen comprises fludarabine and cyclophosphamide or bendamustine. In some embodiments, the lymphodepleting regimen comprises radiation In some embodiments, the lymphodepleting chemotherapy regimen is administered prior to the administration of the CAR-T cell therapy.

[0021] In some embodiments, the method results in a prolongation of the duration of response to the CAR-T cell therapy of at least 10% as compared to the duration of response to the CAR-T cell therapy in the absence of emapalumab. In some embodiments, the method results in an increase in the overall response rate to the CAR-T cell therapy of at least 10% as compared to the overall response rate to the CAR-T cell therapy in the absence of emapalumab. In some embodiments, the method results in an increase in progression-free survival after administration of the CAR-T cell therapy of at least 10% as compared to the progression-free survival after administration of the CAR-T cell therapy in the absence of emapalumab. In some embodiments, the method results in an increase in overall survival after administration of the CAR-T cell therapy of at least 10% as compared to the overall survival after administration of the CAR-T cell therapy in the absence of emapalumab.

[0022] In some embodiments, the patient has a large B-cell lymphoma that is refractory to first-line chemoimmunotherapy. In some embodiments, the patient has a large B-cell lymphoma that relapses within 12 months of first-line chemoimmunotherapy. In some embodiments, the patient has a relapsed or refractory large B-cell lymphoma after two or more lines of systemic therapy. In some embodiments, the patient has non-Hodgkin lymphoma. In some embodiments, the patient has diffuse large B-cell lymphoma. In some embodiments, the patient has primary mediastinal B-cell lymphoma. In some embodiments, the patient has diffuse large B-cell lymphoma arising from follicular lymphoma. In some embodiments, the patient has high grade B-cell lymphoma.

[0023] In some embodiments, the method further comprises monitoring the levels of CAR-T cells in the blood of the subject, such as by in vitro measuring the level of CAR-T cells in a blood sample obtained from the subject. In some embodiments, the levels of CAR-T cells in the blood of the subject, such as in a blood sample obtained from the subject, is measured every other day. In some embodiments, the levels of CAR-T cells in the blood of the subject, such as in a blood sample obtained from the subject, is measured once every 5 days. In some embodiments, the levels of CAR-T cells in the blood of the subject, such as in a blood sample obtained from the subject, is measured once a week.BRIEF DESCRIPTION OF DRAWINGS

[0024] FIG. 1 shows a schematic illustrating the design of a clinical trial to assess the impact of emapalumab on CAR-T related cytokine release syndrome in patients with non-Hodgkin’s lymphoma (NHL).DETAILED DESCRIPTION

[0025] Provided herein are methods of improving CAR-T cell therapies by administering emapalumab, as well as methods of treating cancer comprising administering a CAR-T cell therapy and emapalumab.

[0026] In one aspect, provided herein is a method of decreasing a CAR-T cell associated toxicity in a subject receiving CAR-T cell therapy, comprising administering to the subject an effective amount of emapalumab. In another aspect, provided herein is a method for treating cancer in a subject in need thereof, the method comprising: (a) administering to the subject a lymphodepleting chemotherapy regimen; (b) administering to the subject emapalumab; and (c) administering to the subject a CAR-T cell therapy.

[0027] In some embodiments, a method of improving a CAR-T cell therapy further comprises a step of monitoring the levels of CAR-T cells in the blood of the subject. The level of CAR-T cells is typically measured in a blood sample obtained from the subject. Methods for measuring the levels of CAR-T cells in the blood, such as in a blood sample, are well known in the art an include, for example, flow cytometry. The levels of CAR-T cells may be monitored at any suitable interval. In some embodiments, the levels of CAR-T cells are measured daily. In some embodiments, the levels of CAR-T cells are measured every other day. In some embodiments, the levels of CAR-T cells are measured every 3 days. In some embodiments, the levels of CAR-T cells are measured every five days. In some embodiments, the levels of CAR-T cells are measured once a week. In some embodiments, the levels of CAR-T cells are measured every other week. In some embodiments, the levels of CAR-T cells are measured once a month. In some embodiments, the levels of CAR-T cells are measured on day 3, 7, 14, 21, and 28 after administration of the CAR-T cells. In some embodiments, the levels of CAR-T cells are measured on day 3 and 7 after administration of the CAR-T cells and then once a week.

[0028] A person of skill in the art will appreciate that monitoring the levels of CAR-T cells in the blood of a patient may be useful to determine whether further doses of CAR-T cell therapy should be administered.Improvement of Toxicities

[0029] Without wishing to be bound by theory, it is believed that at least part of the toxicities associated with CAR-T cell therapies are mediated by interferon gamma (IFNy). Therefore, it was hypothesized that inhibiting IFNY signaling could improve the toxicities associated with CAR-T cell therapies.Cytokine Release Syndrome

[0030] In some embodiments, the toxicity being improved by the methods described herein is cytokine-release syndrome (CRS). CRS occurs when large numbers of immune cells (e.g., B cells, T cells, natural killer cells, macrophages, dendritic cells, and monocytes) become activated and release inflammatory cytokines. Such inflammatory cytokines can include Tumor necrosis factor (TNF), IFNy, IL-ip, IL-2, IL-6, IL-8, and IL-10. Symptoms of CRS are essentially those of an exaggerated inflammatory response and include fever, chills, skin rashes, myalgia, nausea, vomiting, diarrhea, hypoxia, tachycardia, hypotension, confusion, seizures, as well as kidney and liver function abnormalities. The clinical manifestation can range from mild to life-threatening (Grade 1-4) and ultimately death (Grade 5). See, e.g., Shimabukuro-Vornhage et al., J Immunother Cancer. 2018; 6: 56. Grading of CRS is also described further in the examples, e.g., in Table 2.

[0031] Low grade CRS is usually treated symptomatically, using, for example, antihistamines, antipyretics and fluids. High grade CRS caused by agents that are administered repeatedly may require dose adjustments or treatment interruptions. However, this is not feasible for CAR-T cell therapies which are generally administered once and only reach their full dose after expansion in the blood. High grade CRS after the administration of CAR-T cells has limited treatment options, which include IL-6 blockade, TNF inhibition, and steroids.

[0032] In some embodiments, a method of improving a CAR-T cell therapy or a method of treatment described herein results in a reduction in CRS of 1, 2, 3 or 4 grades compared to the CRS induced by administration of the CAR-T cell therapy in the absence of emapalumab. In some embodiments, a method of improving a CAR-T cell therapy or a method of treatment described herein results in a reduction in CRS of at least 1 grade compared to the CRS induced by administration of the CAR-T cell therapy in the absence of emapalumab. In some embodiments, a method of improving a CAR-T cell therapy or a method of treatment described herein results in a reduction in CRS of at least 2 grades compared to the CRS induced by administration of the CAR-T cell therapy in the absence of emapalumab. In some embodiments, a method of improving a CAR-T cell therapy or a method of treatment described herein results in a reduction in CRS of at least 3 grades compared to the CRS induced by administration of the CAR-T cell therapy in the absence of emapalumab.

[0033] In some embodiments, a method of improving a CAR-T cell therapy or a method of treatment described herein results in a reduction in CRS of at least 10% compared to the CRS induced by administration of the CAR-T cell therapy in the absence of emapalumab. In someembodiments, a method of improving a CAR-T cell therapy or a method of treatment described herein results in a reduction in CRS of at least 20% compared to the CRS induced by administration of the CAR-T cell therapy in the absence of emapalumab. In some embodiments, a method of improving a CAR-T cell therapy or a method of treatment described herein results in a reduction in CRS of at least 30% compared to the CRS induced by administration of the CAR-T cell therapy in the absence of emapalumab. In some embodiments, a method of improving a CAR-T cell therapy or a method of treatment described herein results in a reduction in CRS of at least 40% compared to the CRS induced by administration of the CAR-T cell therapy in the absence of emapalumab. In some embodiments, a method of improving a CAR-T cell therapy or a method of treatment described herein results in a reduction in CRS of at least 50% compared to the CRS induced by administration of the CAR-T cell therapy in the absence of emapalumab.Immune Effector Cell Associated Neurotoxicity Syndrome

[0034] In some embodiments, the toxicity being improved by the methods described herein is Immune Effector Cell Associated Neurotoxicity Syndrome (ICANS). ICANS is associated with disruption of the blood-brain barrier and increased cytokine levels in the cerebrospinal fluid. For example, white blood cell counts, proteins, IFNy, IL-6, IL- 10, and granzyme B may be increased in the CSF and / or IFNy, IL-10, granzyme B, granulocyte-macrophage colonystimulating factor (GM-CSF), MIP-la, IL-2, ferritin and TNF may be increased in the serum. See, e.g., Sterner and Sterner, Fron. Immunol. 2022; 13: 879608.

[0035] Symptoms of ICANS include confusion, headache, attention deficits, word finding difficulties, focal neurological deficits, or encephalopathy but may be as severe as cerebral edema, transient coma, or seizures. ICANS is mostly treated symptomatically, using mainly steroids. Similar to CRS, ICANS is classified by grade (Grade 1 being the mildest and Grade 4 being the most severe, and Grade 5 being death). ICANS grading is also described further in the examples, e.g., in Table 3 and Table 4.

[0036] In some embodiments, a method of improving a CAR-T cell therapy or a method of treatment described herein results in a reduction in ICANS of 1, 2, or 3 grades compared to the CRS induced by administration of the CAR-T cell therapy in the absence of emapalumab. In some embodiments, a method of improving a CAR-T cell therapy or a method of treatment described herein results in a reduction in ICANS of at least 1 grade compared to the ICANS induced by administration of the CAR-T cell therapy in the absence of emapalumab. In some embodiments, a method of improving a CAR-T cell therapy or a method of treatmentdescribed herein results in a reduction in ICANS of at least 2 grades compared to the ICANS induced by administration of the CAR-T cell therapy in the absence of emapalumab. In some embodiments, a method of improving a CAR-T cell therapy or a method of treatment described herein results in a reduction in ICANS of at least 3 grades compared to the ICANS induced by administration of the CAR-T cell therapy in the absence of emapalumab.

[0037] In some embodiments, a method of improving a CAR-T cell therapy or a method of treatment described herein results in a reduction in ICANS of at least 10% compared to the ICANS induced by administration of the CAR-T cell therapy in the absence of emapalumab. In some embodiments, a method of improving a CAR-T cell therapy or a method of treatment described herein results in a reduction in ICANS of at least 20% compared to the ICANS induced by administration of the CAR-T cell therapy in the absence of emapalumab. In some embodiments, a method of improving a CAR-T cell therapy or a method of treatment described herein results in a reduction in ICANS of at least 30% compared to the ICANS induced by administration of the CAR-T cell therapy in the absence of emapalumab. In some embodiments, a method of improving a CAR-T cell therapy or a method of treatment described herein results in a reduction in ICANS of at least 40% compared to the ICANS induced by administration of the CAR-T cell therapy in the absence of emapalumab. In some embodiments, a method of improving a CAR-T cell therapy or a method of treatment described herein results in a reduction in ICANS of at least 50% compared to the ICANS induced by administration of the CAR-T cell in the absence of emapalumab.Reduction in Inflammatory Cytokines

[0038] In some embodiments, a method of improving a CAR-T cell therapy or a method of treatment described herein results in a reduction in the release of at least one inflammatory cytokine compared to the levels of the cytokine after administration of the CAR-T cell therapy in the absence of emapalumab.

[0039] In some embodiments, a method of improving a CAR-T cell therapy or a method of treatment described herein results in a reduction in the release of at least one inflammatory cytokine of at least 10% compared to the levels of the cytokine after administration of the CAR-T cell therapy in the absence of emapalumab. In some embodiments, a method of improving a CAR-T cell therapy or a method of treatment described herein results in a reduction in the release of at least one inflammatory cytokine of at least 20% compared to the levels of the cytokine after administration of the CAR-T cell therapy in the absence of emapalumab. In some embodiments, a method of improving a CAR-T cell therapy or amethod of treatment described herein results in a reduction in the release of at least one inflammatory cytokine of at least 30% compared to the levels of the cytokine after administration of the CAR-T cell therapy in the absence of emapalumab. In some embodiments, a method of improving a CAR-T cell therapy or a method of treatment described herein results in a reduction in the release of at least one inflammatory cytokine of at least 40% compared to the levels of the cytokine after administration of the CAR-T cell therapy in the absence of emapalumab. In some embodiments, a method of improving a CAR-T cell therapy or a method of treatment described herein results in a reduction in the release of at least one inflammatory cytokine of at least 50% compared to the levels of the cytokine after administration of the CAR-T cell therapy in the absence of emapalumab.

[0040] The inflammatory cytokine may be INFy, IL-ip, IL-2, IL-5, IL-6, IL-8, IL-10, IL-13, GM-CSF, granzyme B, MIP-la, ferritin or TNF.

[0041] The levels of the inflammatory cytokine may be determined using any suitable method known in the art or described herein (e.g., ELISA or RNA sequencing). The levels of the inflammatory cytokine may be determined in any suitable sample from the patient, such as blood, serum, or CSF.Cytopenia

[0042] In some embodiments, the toxicity being improved by the methods described herein is prolonged cytopenia. Cytopenias are a group of conditions characterized by lower than normal blood cell counts. Examples of cytopenias include anemia (low red blood cell counts), leukopenia (low white blood cell counts), neutropenia (low neutrophil counts) and thrombocytopenia (low platelet counts).

[0043] Patients receiving CAR-T cell therapies are generally lymphodepleted prior to administration of the CAR-T cells. This lymphodepletion can cause cytopenia, however, prolonged or recurrent cytopenia may also occur after CAR-T cell therapy, sometimes more than 30 days after. The cause of these prolonged cytopenias is not fully understood, however, is has been suggested that cytokines play a role. See Sharma et al., Cancers (Basel). 2022 Mar; 14(6): 1501.

[0044] Cytopenia increases the risk of infection and bleeding in a patient. Treatment of prolonged cytopenias is largely symptomatic with use of transfusion products and supportive care with colony stimulating factor. Similar to CRS and ICANS, cytopenias are classified by grade (Grade 1 being the mildest and Grade 4 being the most severe).

[0045] In some embodiments, a method of improving a CAR-T cell therapy or a method of treatment described herein results in a reduction in at least one cytopenia of 1, 2, or 3 grades compared to the cytopenia induced by administration of the CAR-T cell in the absence of emapalumab. In some embodiments, a method of improving a CAR-T cell therapy or a method of treatment described herein results in a reduction in at least one cytopenia of at least 1 grade compared to the cytopenia induced by administration of the CAR-T cell therapy in the absence of emapalumab. In some embodiments, a method of improving a CAR-T cell therapy or a method of treatment described herein results in a reduction in at least one cytopenia of at least 2 grades compared to the cytopenia induced by administration of the CAR-T cell therapy in the absence of emapalumab. In some embodiments, a method of improving a CAR-T cell therapy or a method of treatment described herein results in a reduction in at least one cytopenia of at least 3 grades compared to the cytopenia induced by administration of the CAR-T cell therapy in the absence of emapalumab. In some embodiments, a method of improving a CAR-T cell therapy or a method of treatment described herein results in a reduction in at least one cytopenia of at least 4 grades compared to the cytopenia induced by administration of the CAR-T cell therapy in the absence of emapalumab. The at least one cytopenia may be anemia, leukopenia, neutropenia, or thrombocytopenia.

[0046] In some embodiments, a method of improving a CAR-T cell therapy or a method of treatment described herein results in a reduction in at least cytopenia of at least 10% compared to the cytopenia induced by administration of the CAR-T cell therapy in the absence of emapalumab. In some embodiments, a method of improving a CAR-T cell therapy or a method of treatment described herein results in a reduction in at least cytopenia of at least 20% compared to the cytopenia induced by administration of the CAR-T cell therapy in the absence of emapalumab. In some embodiments, a method of improving a CAR-T cell therapy or a method of treatment described herein results in a reduction in at least cytopenia of at least 30% compared to the cytopenia induced by administration of the CAR-T cell therapy in the absence of emapalumab. In some embodiments, a method of improving a CAR-T cell therapy or a method of treatment described herein results in a reduction in at least cytopenia of at least 40% compared to the cytopenia induced by administration of the CAR-T cell therapy in the absence of emapalumab. In some embodiments, a method of improving a CAR-T cell therapy or a method of treatment described herein results in a reduction in at least cytopenia of at least 50% compared to the cytopenia induced by administration of the CAR-Tcell therapy in the absence of emapalumab. The at least one cytopenia may be anemia, leukopenia, neutropenia, or thrombocytopenia.Improvement of Efficacy

[0047] Without wishing to be bound by theory, it is hypothesized that the methods of improving a CAR-T cell therapy and the methods of treatment described herein result in decreased toxicities than conventional methods of CAR-T cell therapy (i.e., CAR-T cell therapies administered in the absence of emapalumab). A person of skill in the art will appreciate that decreases in toxicity may result in better efficacy of a therapy, for example, improvements in response rates and survival. This is because reduction of toxicities may allow higher doses of CAR-T cells to be administered, thereby potentially improving efficacy.Duration of Response

[0048] In some embodiments, a method of improving a CAR-T cell therapy or a method of treatment described herein results in a prolongation of the duration of response to the CAR-T cell therapy compared to the duration of response to the CAR-T cell therapy in the absence of emapalumab. The duration of response is generally defined as the interval from response initiation (when the treated subject first shows a complete or partial response to therapy) to the earlier of disease progression or death. Disease relapse after CAR-T cell therapy in B-cell malignancies is generally measured as the recurrence of tumor cells after the subject has achieved a complete response. Generally, the relapse can be categorized as the bone marrow relapse, the extramedullary relapse (e.g., in central nervous system), or a combined relapse according to anatomical. Relapse may further be categorized as antigen-positive or antigennegative based on the expression profile of initially targeted antigens of tumor cells.

[0049] In some embodiments, a method of improving a CAR-T cell therapy or a method of treatment described herein results in a prolongation of the duration of response to the CAR-T cell therapy of at least 10% as compared to the duration of response to the CAR-T cell therapy in the absence of emapalumab. In some embodiments, a method of improving a CAR-T cell therapy or a method of treatment described herein results in a prolongation of the duration of response to the CAR-T cell therapy of at least 20% as compared to the duration of response to the CAR-T cell therapy in the absence of emapalumab. In some embodiments, a method of improving a CAR-T cell therapy or a method of treatment described herein results in a prolongation of the duration of response to the CAR-T cell therapy of at least 30% ascompared to the duration of response to the CAR-T cell therapy in the absence of emapalumab. In some embodiments, a method of improving a CAR-T cell therapy or a method of treatment described herein results in a prolongation of the duration of response to the CAR-T cell therapy of at least 40% as compared to the duration of response to the CAR-T cell therapy in the absence of emapalumab. In some embodiments, a method of improving a CAR-T cell therapy or a method of treatment described herein results in a prolongation of the duration of response to the CAR-T cell therapy of at least 50% as compared to the duration of response to the CAR-T cell therapy in the absence of emapalumab.

[0050] In some embodiments, a method of improving a CAR-T cell therapy or a method of treatment described herein results in a prolongation of the duration of response to the CAR-T cell therapy of at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 weeks as compared to the duration of response to the CAR-T cell therapy in the absence of emapalumab. In some embodiments, a method of improving a CAR-T cell therapy or a method of treatment described herein results in a prolongation of the duration of response to the CAR-T cell therapy of at least 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 months as compared to the duration of response to the CAR-T cell therapy in the absence of emapalumab. In some embodiments, a method of improving a CAR-T cell therapy or a method of treatment described herein results in a prolongation of the duration of response to the CAR-T cell therapy of at least 1, 2, 3, 4, or 5 years as compared to the duration of response to the CAR-T cell therapy in the absence of emapalumab.Number of CAR-T Cells

[0051] In some embodiments, a method of improving a CAR-T cell therapy or a method of treatment described herein results in an increase of the number CAR-T cells in the blood of a subject compared to the number of CAR-T cells in the absence of emapalumab. The number of CAR-T cells in the blood can be determined using any suitable method known in the art or described herein, including for example flow cytometry.

[0052] In some embodiments, a method of improving a CAR-T cell therapy or a method of treatment described herein results in at least 400 CAR-T cells per microliter in the blood of the subject. In some embodiments, a method of improving a CAR-T cell therapy or a method of treatment described herein results in at least 450 CAR-T cells per microliter in the blood of the subject. In some embodiments, a method of improving a CAR-T cell therapy or a method of treatment described herein results in at least 500 CAR-T cells per microliter in the blood of the subject. In some embodiments, a method of improving a CAR-T cell therapy or a method of treatment described herein results in at least 550 CAR-T cells per microliter in the blood ofthe subject. In some embodiments, a method of improving a CAR-T cell therapy or a method of treatment described herein results in at least 600 CAR-T cells per microliter in the blood of the subject.Overall Response Rate

[0053] In some embodiments, a method of improving a CAR-T cell therapy or a method of treatment described herein results in an increase in overall response rate after administration of the CAR-T cell therapy compared to the overall response rate after administration of the CAR-T cell therapy in the absence of emapalumab. Overall response rate is generally defined as the proportion of patients in a treatment group who have a partial response (a reduction in tumor burden) or complete response (complete clearance of the tumor) to a therapy (not including stable disease). Overall response rate is a measure of tumoricidal activity.

[0054] In some embodiments, a method of improving a CAR-T cell therapy or a method of treatment described herein results in an increase in overall response rate after administration of the CAR-T cell therapy of at least 10% as compared to the overall response rate after administration of the CAR-T cell therapy in the absence of emapalumab. In some embodiments, a method of improving a CAR-T cell therapy or a method of treatment described herein results in an increase in overall response rate after administration of the CAR-T cell therapy of at least 20% as compared to the overall response rate after administration of the CAR-T cell therapy in the absence of emapalumab. In some embodiments, a method of improving a CAR-T cell therapy or a method of treatment described herein results in an increase in overall response rate after administration of the CAR-T cell therapy of at least 30% as compared to the overall response rate after administration of the CAR-T cell therapy in the absence of emapalumab. In some embodiments, a method of improving a CAR-T cell therapy or a method of treatment described herein results in an increase in overall response rate after administration of the CAR-T cell therapy of at least 40% as compared to the overall response rate after administration of the CAR-T cell therapy in the absence of emapalumab. In some embodiments, a method of improving a CAR-T cell therapy or a method of treatment described herein results in an increase in overall response rate after administration of the CAR-T cell therapy of at least 50% as compared to the overall response rate after administration of the CAR-T cell therapy in the absence of emapalumab.Progression-Free Survival

[0055] In some embodiments, a method of improving a CAR-T cell therapy or a method of treatment described herein results in an increase in progression-free survival after administration of the CAR-T cell therapy compared to the progression-free survival after administration of the CAR-T cell therapy in the absence of emapalumab. Progression-free survival is generally defined as the time between randomization (or treatment) in a clinical trial to the earlier of disease progression or death.

[0056] In some embodiments, a method of improving a CAR-T cell therapy or a method of treatment described herein results in an increase in progression-free survival after administration of the CAR-T cell therapy of at least 10% as compared to the progression-free survival after administration of the CAR-T cell therapy in the absence of emapalumab. In some embodiments, a method of improving a CAR-T cell therapy or a method of treatment described herein results in an increase in progression-free survival after administration of the CAR-T cell therapy of at least 20% as compared to the progression-free survival after administration of the CAR-T cell therapy in the absence of emapalumab. In some embodiments, a method of improving a CAR-T cell therapy or a method of treatment described herein results in an increase in progression-free survival after administration of the CAR-T cell therapy of at least 30% as compared to the progression-free survival after administration of the CAR-T cell therapy in the absence of emapalumab. In some embodiments, a method of improving a CAR-T cell therapy or a method of treatment described herein results in an increase in progression-free survival after administration of the CAR-T cell therapy of at least 40% as compared to the progression-free survival after administration of the CAR-T cell therapy in the absence of emapalumab. In some embodiments, a method of improving a CAR-T cell therapy or a method of treatment described herein results in an increase in progression-free survival after administration of the CAR-T cell therapy of at least 50% as compared to the progression-free survival after administration of the CAR-T cell therapy in the absence of emapalumab.

[0057] In some embodiments, a method of improving a CAR-T cell therapy or a method of treatment described herein results in an increase in progression-free survival after administration of the CAR-T cell therapy of at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 weeks as compared to the progression-free survival after administration of the CAR-T cell therapy in the absence of emapalumab. In some embodiments, a method of improving a CAR-T cell therapy or a method of treatment described herein results in an increase in progression-free survival after administration of the CAR-T cell therapy of at least 4, 5, 6, 7,8, 9, 10, 11, or 12 months as compared to the progression-free survival after administration of the CAR-T cell therapy in the absence of emapalumab. In some embodiments, a method of improving a CAR-T cell therapy or a method of treatment described herein results in an increase in progression-free survival after administration of the CAR-T cell therapy of at least 1, 2, 3, 4 or 5 year as compared to the progression-free survival after administration of the CAR-T cell therapy in the absence of emapalumab.Overall Survival

[0058] In some embodiments, a method of improving a CAR-T cell therapy or a method of treatment described herein results in an increase in overall survival after administration of the CAR-T cell therapy compared to the overall survival after administration of the CAR-T cell therapy in the absence of emapalumab. Overall survival is generally defined as the time between treatment and death irrespective of disease progression.

[0059] In some embodiments, a method of improving a CAR-T cell therapy or a method of treatment described herein results in an increase in overall survival after administration of the CAR-T cell therapy of at least 10% as compared to the overall survival after administration of the CAR-T cell therapy in the absence of emapalumab. In some embodiments, a method of improving a CAR-T cell therapy or a method of treatment described herein results in an increase in overall survival after administration of the CAR-T cell therapy of at least 20% as compared to the overall survival after administration of the CAR-T cell therapy in the absence of emapalumab. In some embodiments, a method of improving a CAR-T cell therapy or a method of treatment described herein results in an increase in overall survival after administration of the CAR-T cell therapy of at least 30% as compared to the overall survival after administration of the CAR-T cell therapy in the absence of emapalumab. In some embodiments, a method of improving a CAR-T cell therapy or a method of treatment described herein results in an increase in overall survival after administration of the CAR-T cell therapy of at least 40% as compared to the overall survival after administration of the CAR-T cell therapy in the absence of emapalumab. In some embodiments, a method of improving a CAR-T cell therapy or a method of treatment described herein results in an increase in overall survival after administration of the CAR-T cell therapy of at least 50% as compared to the overall survival after administration of the CAR-T cell therapy in the absence of emapalumab.

[0060] In some embodiments, a method of improving a CAR-T cell therapy or a method of treatment described herein results in an increase in overall survival after administration of theCAR-T cell therapy of at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 weeks as compared to the overall survival after administration of the CAR-T cell therapy in the absence of emapalumab.

[0061] In some embodiments, a method of improving a CAR-T cell therapy or a method of treatment described herein results in an increase in overall survival after administration of the CAR-T cell therapy of at least 4, 5, 6, 7, 8, 9, 10, 11, or 12 months as compared to the overall survival after administration of the CAR-T cell therapy in the absence of emapalumab. In some embodiments, a method of improving a CAR-T cell therapy or a method of treatment described herein results in an increase in overall survival after administration of the CAR-T cell therapy of at least 1, 2, 3, 4 or 5 years as compared to the overall survival after administration of the CAR-T cell therapy in the absence of emapalumab.Indications and Patients

[0062] The CAR-T cells being improved using the methods described herein or being used in the methods of treatment described herein may be used for any approved indication (see, e.g., Table 1). Generally, CAR-T cell therapies are being used for cancers, mainly hematological malignancies such as leukemias and lymphomas. In some embodiments, the cancer is a relapsed or refractory cancer.

[0063] In some embodiments, a subject being treated in accordance with a method described herein has received prior treatment for their cancer. In some embodiments, the subject has a cancer that has relapsed after prior therapy. In some embodiments, the subject has a cancer that is refractory to first-line chemoimmunotherapy. In some embodiments, the subject relapses within 12 months of first-line chemoimmunotherapy. In some embodiments, the subject has relapsed or refractory disease after two or more lines of systemic therapy. In some embodiments, the subject is not eligible for hematopoietic stem cell transplantation.

[0064] In some embodiments, a subject being treated in accordance with a method described herein has a B-cell leukemia or lymphoma.

[0065] In some embodiments, the subject has a B-cell precursor acute lymphoblastic leukemia (ALL). In some embodiments, the subject has a B-cell precursor acute lymphoblastic leukemia (ALL) that is refractory or in second or later relapse.

[0066] In some embodiments, the subject has diffuse large B-cell lymphoma (DLBCL). In some embodiments, the subject has a high grade B-cell lymphoma. In some embodiments, the subject has DLBCL arising from follicular lymphoma. In some embodiments, the subject has primary mediastinal large B-cell lymphoma.

[0067] In some embodiments, a subject being treated in accordance with a method described herein has a large B-cell lymphoma. In some embodiments, a subject being treated in accordance with a method described herein has a relapsed or refractory large B-cell lymphoma after two or more lines of systemic therapy. In some embodiments, a subject being treated in accordance with a method described herein has a large B-cell lymphoma that is refractory to first-line chemoimmunotherapy or that relapses within 12 months of first-line chemoimmunotherapy .

[0068] In some embodiments, a subject being treated in accordance with a method described herein has follicular lymphoma. In some embodiments, the subject has relapsed or refractory follicular lymphoma after two or more lines of systemic therapy.

[0069] In some embodiments, a subject being treated in accordance with a method described herein has mantle cell lymphoma. In some embodiments, the subject has mantle cell lymphoma.

[0070] In some embodiments, a subject being treated in accordance with a method described herein has multiple myeloma. In some embodiments, the subject has relapsed or refractory multiple myeloma. In some embodiments, the subject has relapsed or refractory multiple myeloma after four or more prior lines of therapy (e.g., therapy including an immunomodulatory agent, a proteasome inhibitor, and / or an anti-CD38 monoclonal antibody).

[0071] In some embodiments, the subject has non-Hodgkin lymphoma.

[0072] The terms “patient” and “subject” are used interchangeably herein. In some embodiments, the patient is a human adult. In some embodiments, the patient is a human child.Emapalumab

[0073] The methods described herein comprise the administration of emapalumab. Emapalumab (GAMIFANT®) is a fully human anti-IFNy monoclonal antibody that was FDA approved in 2018 for the treatment of adult and pediatric primary hemophagocytic lymphohistiocytosis (HLH).

[0074] In some embodiments of the methods of improving a CAR-T cell therapy or the methods of treatment described herein, emapalumab is administered at a dose of about 1 to about 10 mg / kg. In some embodiments of the methods of improving a CAR-T cell therapy or the methods of treatment described herein, emapalumab is administered at a dose of about 1 mg / kg. In some embodiments of the methods of improving a CAR-T cell therapy or themethods of treatment described herein, emapalumab is administered at a dose of about 2 mg / kg. In some embodiments of the methods of improving a CAR-T cell therapy or the methods of treatment described herein, emapalumab is administered at a dose of about 3 mg / kg. In some embodiments of the methods of improving a CAR-T cell therapy or the methods of treatment described herein, emapalumab is administered at a dose of about 3 mg / kg. In some embodiments of the methods of improving a CAR-T cell therapy or the methods of treatment described herein, emapalumab is administered at a dose of about 5 mg / kg. In some embodiments of the methods of improving a CAR-T cell therapy or the methods of treatment described herein, emapalumab is administered at a dose of about 6 mg / kg. In some embodiments, emapalumab is administered as a one-time dose of 1 mg / kg intravenously. In some embodiments, emapalumab is administered as a one-time dose of 2 mg / kg intravenously. In some embodiments, emapalumab is administered as a one-time dose of 3 mg / kg intravenously. In some embodiments, emapalumab is administered as a one-time dose of 4 mg / kg intravenously. In some embodiments, emapalumab is administered as a onetime dose of 5 mg / kg intravenously. In some embodiments, emapalumab is administered as a one-time dose of 6 mg / kg intravenously.

[0075] The emapalumab may be administered at any suitable time. For example, the emapalumab is administered concurrently (i.e. same day) with the administration of the CAR- T cell therapy, after the administration of the CAR-T therapy or before the administration CAR-T therapy.

[0076] The emapalumab may be administered at any suitable time prior to the administration of the CAR-T cell therapy. In some embodiments, emapalumab is administered 1-5 days before the CAR-T cell administration. In some embodiments, emapalumab is administered 1- 3 days before the CAR-T cell administration. In some embodiments, emapalumab is administered 1 or 2 days before the CAR-T cell administration. In some embodiments, emapalumab is administered the day before the CAR-T cell administration.

[0077] The emapalumab may be administered at any suitable time after to the administration of the CAR-T cell therapy. In some embodiments, emapalumab is administered 1-5 days after the CAR-T cell administration. In some embodiments, emapalumab is administered 1-3 days after the CAR-T cell administration. In some embodiments, emapalumab is administered 1 or 2 days after the CAR-T cell administration. In some embodiments, emapalumab is administered the day after the CAR-T cell administration.

[0078] In some embodiments, a one-time dose of 1 mg / kg emapalumab is administered intravenously the day before the CAR-T cell therapy. In some embodiments, a one-time doseof 2 mg / kg emapalumab is administered intravenously the day before the CAR-T cell therapy. In some embodiments, a one-time dose of 3 mg / kg emapalumab is administered intravenously the day before the CAR-T cell therapy. In some embodiments, a one-time dose of 4 mg / kg emapalumab is administered intravenously the day before the CAR-T cell therapy. In some embodiments, a one-time dose of 5 mg / kg emapalumab is administered intravenously the day before the CAR-T cell therapy. In some embodiments, a one-time dose of 6 mg / kg emapalumab is administered intravenously the day before the CAR-T cell therapy.

[0079] In some embodiments, the methods described herein further comprise the administration of a lymphodepleting chemotherapy regimen. In such embodiments, the emapalumab may be administered at any suitable time between the administration of the lymphodepleting chemotherapy regimen and the administration of the CAR-T cell therapy. In some embodiments, the emapalumab is administered 1-3 days after the lymphodepleting chemotherapy regimen. In some embodiments, the emapalumab is administered about 2 days after the lymphodepleting chemotherapy regimen.

[0080] In some embodiments, the emapalumab is administered on day 1, with the CAR-T cell therapy being administered on day 0.CAR-T cell Therapies

[0081] The methods described herein may be used to improve existing CAR-T cell therapies and the methods of treatment described herein comprise the administration of CAR-T cell therapies.

[0082] CAR-T cells are autologous cells modified to express a chimeric antigen receptor (CAR). Generally, a CAR comprises an antigen recognition domain (often an scFv), a hinge domain (often from CD8, CD28 or IgG), a transmembrane domain (often from CD4, CD8 or CD28), and a signaling domain (often CD3Q. Second and third generation CARs further comprise one and two co-stimulatory domains (e.g., CD28, 4- IBB, 0X40, CD27 or ICOS), respectively. Upon binding to the antigen on the tumor cell, the CAR-T cell becomes activated attacks the tumor cell.

[0083] To date, six CAR-T cell therapies are approved by the FDA, in each case for the treatment of blood cancer. Each of these six therapies targets either CD 19 or BCMA. See Table 1 and Chen etal., Cancers (Basel). 2023 Feb; 15(3): 663. for more details.Table 1: FDA-Approved CAR-T Cell Therapies

[0084] Any approved CAR-T cell therapy may be improved by or used in a method described herein.

[0085] In some embodiments, the CAR-T cell therapy is a CD- 19 targeting CAR-T cell therapy. In some embodiments, the CAR-T cell therapy is a CD-20 targeting CAR-T cell therapy. In some embodiments, the CAR-T cell therapy is a CD-22 targeting CAR-T cell therapy. In some embodiments, the CAR-T cell therapy is a BCMA targeting CAR-T cell therapy.

[0086] In some embodiments, the CAR-T cell therapy is axicabtagene ciloleucel. In some embodiments, the CAR-T cell therapy is tisagenlecleucel. In some embodiments, the CAR-T cell therapy is brexucabtagene autoleucal. In some embodiments, the CAR-T cell therapy is lisocabtagene maraleucel. In some embodiments, the CAR-T cell therapy is idecabtagene vicleucel. In some embodiments, the CAR-T cell therapy is ciltacabtagene autoleucel.

[0087] In some embodiments, the CAR-T cell therapy being improved by or used in the methods described herein may be administered at the dosage indicated on the drug label.

[0088] In some embodiments, the CAR-T cell therapy is axicabtagene ciloleucel administered at a dose of 2 * 106CAR-positive viable T cells per kg body weight with a maximum of 2 * 108CAR-positive viable T cells per kg body weight (e.g., for the treatment of relapsed or refractory large B-cell lymphoma or relapsed or refractory follicular lymphoma).

[0089] In some embodiments, the CAR-T cell therapy is tisagenlecleucel administered at a dose of 0.2 to 5.0 x 106CAR-positive viable T cells per kg body weight intravenously (e.g., for the treatment of Pediatric and Young Adult B-cell ALL in patients of 50 kg or less). In some embodiments, the CAR-T cell therapy is tisagenlecleucel administered at a dose of 0.1 to 2.5 x 108total CAR-positive viable T cells (non-weight based) intravenously for patients above 50 kg (e.g., for the treatment of B-cell ALL in patients of over 50 kg). In some embodiments, the CAR-T cell therapy is tisagenlecleucel administered at a dose of 0.6 to 6.0 x 108CAR-positive viable T cells intravenously (e.g., for the treatment of Adult Relapsed or Refractory Diffuse Large B-cell Lymphoma and Follicular Lymphoma).

[0090] In some embodiments, the CAR-T cell therapy is brexucabtagene autoleucal administered at a dose of 2 * 106CAR-positive viable T cells per kg body weight, with a maximum of 2 * 108CAR-positive viable T cells (e.g., for the treatment of mantle cell lymphoma). In some embodiments, the CAR-T cell therapy is brexucabtagene autoleucal administered at a dose of 1 x 106CAR-positive viable T cells per kg body weight, with a maximum of 1 x 108CAR-positive viable T cells (e.g., for the treatment of ALL).

[0091] In some embodiments, the CAR-T cell therapy is lisocabtagene maraleucel administered at a dose of 90 to 110 x io6CAR-positive viable T cells (e.g., for the treatment of LBCL after one line of therapy). In some embodiments, the CAR-T cell therapy is lisocabtagene maraleucel administered at a dose of 50 to 110 x io6CAR-positive viable T cells (e.g., for the treatment of LBCL after two lines of treatment).

[0092] In some embodiments, the CAR-T cell therapy is idecabtagene vicleucel 300 to 460 x 106CAR-positive T cells (e.g., for the treatment of relapsed or refractory multiple myeloma).

[0093] In some embodiments, the CAR-T cell therapy is ciltacabtagene autoleucel administered at a dose of 0.5-1.0x l06CAR-positive viable T cells per kg of body weight, with a maximum dose of 1 x 108 CAR-positive viable T cells per single-dose infusion (e.g., for the treatment of relapsed or refractory multiple myeloma).

[0094] In some embodiments, the CAR-T cell therapy being improved by or used in the methods described herein may be administered at a higher dosage indicated on the drug label.Examples of the dosages indicated on drug labels of FDA-approved CAR-T cell therapies are:• Axicabtagene ciloleucel: 2 * 106CAR-positive viable T cells per kg body weight, with a maximum of 2 * 108 CAR-positive viable T cells.• Tisagenlecleucel: o For Pediatric and Young Adult B-cell ALL (up to 25 years of age):• For patients 50 kg or less, 0.2 to 5.0 x 106CAR-positive viable T cells per kg body weight intravenously.• For patients above 50 kg, 0.1 to 2.5 x 108total CAR-positive viable T cells (non-weight based) intravenously. o For Adult Relapsed or Refractory Diffuse Large B-cell Lymphoma and Follicular Lymphoma: 0.6 to 6.0 x 108CAR-positive viable T cells intravenously.• Brexucabtagene autoleucal: o MCL: 2 * 106CAR-positive viable T cells per kg body weight, with a maximum of 2 * 108CAR-positive viable T cells. o ALL: 1 * 106CAR-positive viable T cells per kg body weight, with a maximum of 1x108CAR-positive viable T cells.• Lisocabtagene maraleucel: o For LBCL after one line of therapy: 90 to 110 x io6CAR-positive viable T cells. o For LBCL after two or more lines of therapy: 50 to 110 x 106CAR-positive viable T cells.• Idecabtagene vicleucel: 300 to 460 x io6CAR-positive T cells.• Ciltacabtagene autoleucel: 0.5-1 ,0x 106CAR-positive viable T cells per kg of body weight, with a maximum dose of 1 x 108CAR-positive viable T cells per single-dose infusion.

[0095] In some embodiments, the dose of the CAR-T cell therapy being improved by or used in the methods described herein (e.g., axicabtagene ciloleucel, tisagenlecleucel, brexucabtagene autoleucal, lisocabtagene maraleucel, idecabtagene vicleucel or ciltacabtagene autoleucel) is administered at a dose that is up to 25% higher than the dosage indicated on the drug label. In some embodiments, the dose of the CAR-T cell therapy being improved by or used in the methods described herein (e.g., axicabtagene ciloleucel,tisagenlecleucel, brexucabtagene autoleucal, lisocabtagene maraleucel, idecabtagene vicleucel or ciltacabtagene autoleucel) is administered at a dose that is 25%-50% higher than the dosage indicated on the drug label. In some embodiments, the dose of the CAR-T cell therapy being improved by or used in the methods described herein (e.g., axicabtagene ciloleucel, tisagenlecleucel, brexucabtagene autoleucal, lisocabtagene maraleucel, idecabtagene vicleucel or ciltacabtagene autoleucel) is administered at a dose that is 50%- 75%% higher than the dosage indicated on the drug label. In some embodiments, the dose of the CAR-T cell therapy being improved by or used in the methods described herein (e.g., axicabtagene ciloleucel, tisagenlecleucel, brexucabtagene autoleucal, lisocabtagene maraleucel, idecabtagene vicleucel or ciltacabtagene autoleucel) is administered at a dose that is 75%-100% higher than the dosage indicated on the drug label.

[0096] In some embodiments, the dose of the CAR-T cell therapy being improved by or used in the methods described herein (e.g., axicabtagene ciloleucel, tisagenlecleucel, brexucabtagene autoleucal, lisocabtagene maraleucel, idecabtagene vicleucel or ciltacabtagene autoleucel) is administered at a dose that about 1-1.5 times the dosage indicated on the drug label. In some embodiments, the dose of the CAR-T cell therapy being improved by or used in the methods described herein (e.g., axicabtagene ciloleucel, tisagenlecleucel, brexucabtagene autoleucal, lisocabtagene maraleucel, idecabtagene vicleucel or ciltacabtagene autoleucel) is administered at a dose that about 1.5-2 times the dosage indicated on the drug label. In some embodiments, the dose of the CAR-T cell therapy being improved by or used in the methods described herein (e.g., axicabtagene ciloleucel, tisagenlecleucel, brexucabtagene autoleucal, lisocabtagene maraleucel, idecabtagene vicleucel or ciltacabtagene autoleucel) is administered at a dose that about 2-2.5 times the dosage indicated on the drug label. In some embodiments, the dose of the CAR-T cell therapy being improved by or used in the methods described herein (e.g., axicabtagene ciloleucel, tisagenlecleucel, brexucabtagene autoleucal, lisocabtagene maraleucel, idecabtagene vicleucel or ciltacabtagene autoleucel) is administered at a dose that about 2.5-5 times the dosage indicated on the drug label.

[0097] In some embodiments, the dose of the CAR-T cell therapy being improved by or used in the methods described herein (e.g., axicabtagene ciloleucel, tisagenlecleucel, brexucabtagene autoleucal, lisocabtagene maraleucel, idecabtagene vicleucel or ciltacabtagene autoleucel) is administered at a dose that is about 50% higher than the dosage indicated on the drug label. In some embodiments, the dose of the CAR-T cell therapy being improved by or used in the methods described herein (e.g., axicabtagene ciloleucel,tisagenlecleucel, brexucabtagene autoleucal, lisocabtagene maraleucel, idecabtagene vicleucel or ciltacabtagene autoleucel) is administered at a dose that is about 100% higher than the dosage indicated on the drug label.

[0098] In some embodiments, the dose of the CAR-T cell therapy being improved by or used in the methods described herein (e.g., axicabtagene ciloleucel, tisagenlecleucel, brexucabtagene autoleucal, lisocabtagene maraleucel, idecabtagene vicleucel or ciltacabtagene autoleucel) is administered at a dose that is about twice the dosage indicated on the drug label. In some embodiments, the dose of the CAR-T cell therapy being improved by or used in the methods described herein (e.g., axicabtagene ciloleucel, tisagenlecleucel, brexucabtagene autoleucal, lisocabtagene maraleucel, idecabtagene vicleucel or ciltacabtagene autoleucel) is administered at a dose that is about three times the dosage indicated on the drug label.Additional Therapies

[0099] The methods of improving a CAR-T cell therapy and the methods of treatment described herein may further comprise the administration of one or more additional therapies. These may be, for example, therapies that are usually administered prior to or after CAR-T cell therapy.

[0100] For example, in one aspect, the subject may undergo a lymphodepleting chemotherapy regimen prior to administration of the CAR-T cell therapy. Such chemotherapy regimens are usually administered to patients prior to adoptive T cell therapies (such as CAR- T cell therapies) to improve CAR-T cell expansion and persistence. See, e.g., Owen et al., C Cancer Immunology, Immunotherapy (2023) 72:805-814. Most commonly, lymphodepleting chemotherapy regimens comprise fludarabine and cyclophosphamide, but a person of skill in the art will appreciate that any suitable lymphodepleting regimen may be used.Thus, in some embodiments, a method of improving a CAR-T cell therapy or a method of treatment described herein further comprises administering to the subject a lymphodepleting chemotherapy regimen. In some embodiments, the lymphodepleting chemotherapy regimen comprises fludarabine (e.g., at a dose of about 30 mg / m2 / day) and / or cyclophosphamide (e.g. at a dose of about 500 mg / m2 / day). In some embodiments, the lymphodepleting chemotherapy is administered for three days.

[0101] The lymphodepleting chemotherapy may be administered at any suitable time prior to the administration of the emapalumab and / or the CAR-T cell therapy. In some embodiments, the lymphodepleting chemotherapy is administered 3-5 days before the CAR-T cell therapy(e.g., from day -5 to day -3, with the CAR-T cell therapy being administered on day 0). In some embodiments, the lymphodepleting chemotherapy regimen is administered about 1-3 days before the emapalumab. In some embodiments, the administration of the lymphodepleting chemotherapy regimen ends about 2 days before the emapalumab.

[0102] In another aspect, the subject is administered a bridging therapy. A bridging therapy is administered to stabilize or debulk disease between the leukapheresis (to obtain blood cells for generating CAR-T cells) and CAR-T cell administration. Bridging therapy usually comprises one or more chemotherapy agent, steroids, or a combination thereof. Examples of agents that may be used for bridging therapy include rituximab, gemcitabine, etoposide, steroids, cisplatin, cytarabine, ibrutinib, lenalidomide, oxaliplatin, brentuximab vedotin, acalabrutinib, dexamethasone, cyclophosphamide, daratumumab, carfilzomib, bortezomib, pomalidomide and any combination thereof. See, e.g., Bhaskar et al., E EJHaem. 2022 Jan; 3 (Suppl 1): 39-45.EXAMPLESExample 1: Phase 2 multi-site, open-label study evaluating the safety and efficacy of emapalumab in subjects with received 2L or 3L axicabtagene ciloleucel for large B cell lymphoma.

[0103] This example describes a study to assess the impact of emapalumab as preventative management of CAR-T related cytokine release syndrome in patients with non-Hodgkin’s lymphoma (NHL), as measured by the incidence and maximal grade, following axicabtagene ciloleucel in subjects receiving 2L and 3L axicabtagene ciloleucel.Study Design

[0104] This is a Phase 2 multi-site, open-label study evaluating the safety and efficacy of emapalumab in subjects with received 2L or 3L axicabtagene ciloleucel for large B cell lymphoma.

[0105] 28 subjects with aggressive refractory NHL, including DLBCL, PMBCL, high grade lymphoma or TFL will be enrolled to evaluate the safety of emapalumab in combination with axicabtagene ciloleucel. If the initial regimen is determined to be safe, the study may be expanded beyond the initial 28 patient cohort or other subgroups may be added to better define the safety and efficacy profile. The first 3 patients will be staggered by 28 days to assess for initial safety.

[0106] Each subject will proceed through the following study periods as shown in FIG. 1.:• Screening• Enrollment / Leukapheresis period• Bridging therapy (if applicable)• Lymphodepleting chemotherapy period• Investigational Product (IP) treatment period• Post treatment assessment period• Long term follow-up periodObjectives

[0107] The primary objective of this study is to assess the impact of emapalumab as preventative management of CAR-T related cytokine release syndrome in patients with nonHodgkin’ s lymphoma (NHL).

[0108] Secondary objectives include:• To assess the impact of emapalumab as preventative management of CAR-T related ICANS.• To evaluate the anti -turn or efficacy of axicabtagene ciloleucel in combination with preventative emapalumab in terms of DOR, ORR, PFS, and OS.

[0109] Exploratory objectives include:• To assess the function, expansion and phenotype of axicabtagene ciloleucel in patients receiving preventative emapalumab.• To assess the cytokine profiles in peripheral blood in patients receiving emapalumab in combination with axicabtagene ciloleucel.• To assess the incidence of other CAR-T associated toxi cities such as prolonged cytopenias.• To evaluate emapalumab drug concentrations during the course of the study.• To evaluate the PD profile of emapalumab during the course of the study.• To explore the impact of the CAR-T therapy on biomarkers of emapalumab (e.g., CXCL9, IFNy, sIL-2Ra and ferritin).Inclusion Criteria:

[0110] Inclusion criteria for the study include:Adult patients with large B-cell lymphoma that is refractory to first-line chemoimmunotherapy or that relapses within 12 months of first-line chemoimmunotherapy .ORAdult patients with relapsed or refractory large B-cell lymphoma after two or more lines of systemic therapy, including diffuse large B-cell lymphoma (DLBCL) not otherwise specified, primary mediastinal large B-cell lymphoma, high grade B-cell lymphoma, and DLBCL arising from follicular lymphoma. At least 1 measurable lesion per Lugano at time of screening.1At least 2 weeks or 5 half-lives, whichever is shorter, must have elapsed since any prior systemic therapy at the time the subject is planned for leukapheresis, except for systemic inhibitory / stimulatory immune checkpoint therapy however steroids only require a 7-day washout. At least 3 half-lives must have elapsed from any prior systemic inhibitory / stimulatory immune checkpoint molecule therapy at the time the subject is planned for leukapheresis (e.g. ipilimumab, nivolumab, pembrolizumab, atezolizumab, 0X40 agonists, 4- IBB agonists, etc.). Age 18 or older. Eastern Cooperative Oncology Group (ECOG) performance status of 0 to 2. Adequate renal, hepatic, pulmonary and cardiac function defined as: ANC >1000 / pL Platelet count >50, OOO / pL Absolute lymphocyte count > 100 / pL o Creatinine clearance (as estimated by Cockcroft Gault or CKD-EPI) > 30 mL / min o Serum ALT / AST <2.5 per institutional ULN o Total bilirubin <1.5 mg / dl, except in subjects with Gilbert’s syndrome o Cardiac ejection fraction > 40%, no clinically significant pericardial effusion, and no clinically significant ECG findings o Baseline oxygen saturation >92% on room air Females of childbearing potential must have a negative serum or urine pregnancy test (females who have undergone surgical sterilization or who have been postmenopausal for at least 2 years are not considered to be of childbearing potential).11. Ability to understand and the willingness to sign a written informed consent document.Exclusion Criteria[OHl] Exclusion criteria for the study include:1. History of malignancy other than nonmelanoma skin cancer or carcinoma in situ (e.g. cervix, bladder, breast) or follicular lymphoma unless disease free for at least 3 years.2. History of Richter’s transformation of CLL.3. Autologous stem cell transplant within 6 weeks of planned axicabtagene ciloleucel infusion.4. History of allogeneic stem cell transplantation.5. Presence of uncontrolled fungal, bacterial, viral, or other infection at time of screening.6. Known history of acute or chronic active hepatitis B or C infection. Subjects with history of hepatitis infection must have cleared their infection as determined by standard serological and genetic testing per current Infectious Diseases Society of America (IDSA) guidelines. o Patients should also be negative for latent Tb, CMV (NAT), EBV (NAT) and adenovirus (NAT) by PCR testing.7. No clinical evidence of active CNS disease, regardless of prior CNS history.8. History or presence of CNS disorder such as seizure disorder, cerebrovascular ischemia / hemorrhage within 6 months of enrollment.9. History of myocardial infarction, cardiac angioplasty or stenting, unstable angina, or other clinically significant cardiac disease within 12 months of enrollment.10. History of symptomatic pulmonary embolism within 3 months of enrollment; ongoing anticoagulation is allowed if beyond 3 months.11. Any medical condition likely to interfere with assessment of safety or efficacy of study treatment.12. History of allergic reaction or severe immediate hypersensitivity reaction to any of the agents used in this study or compounds of similar chemical or biologic composition.13. Females who are pregnant or breastfeeding or female or male participants who are not willing to practice birth control from the time of consent through 6 months after the completion of axicabtagene ciloleucel.14. In the investigator’s judgment, the subject is unlikely to complete all protocol- required study visits or procedures, including follow-up visits, or comply with the study requirements for participation.15. History of autoimmune disease requiring ongoing systemic immunosuppression. Steroids are allowed up to 5mg predinosine-equivalent for adrenal insufficiency.16. Participants who are receiving any other investigational agents for this condition.Treatment RegimenChemotherapy General Instructions

[0112] Subjects will receive a non-myeloablative lymphodepleting regimen consisting of cyclophosphamide and fludarabine in order to induce lymphocyte depletion and create an optimal environment for expansion of axicabtagene ciloleucel in vivo. Subjects will initiate lymphodepleting chemotherapy with cyclophosphamide and fludarabine beginning on Day -5 through Day -3. The 3-day lymphodepleting chemotherapy regimen may be administered in an outpatient setting.

[0113] As per the FDA approved label for axicabtagene ciloleucel, the lymphodepleting chemotherapy dose in will be cyclophosphamide (500 mg / m2) and fludarabine (30 mg / m2) both given for 3 concurrent days with the target dose of 2 x 106anti-CD19 CAR-T cells / kg. Cyclophosphamide (500 mg / m2) and fludarabine (30 mg / m2) both given for 3 concurrent days has been studied and tolerated in subjects with B cell malignancies37.

[0114] Subjects will receive the 3-day lymphodepleting chemotherapy regimen followed by 2-day rest period (48 hrs) as per institutional guidelines, recommendations are as follows:• IV hydration with IL of 0.9% NaCl (or isotonic [crystalloid] fluid) given prior to cyclophosphamide on the day of infusion followed by:• Cyclophosphamide 500 mg / m2IV over 60 minutes on Day -5, Day -4, and Day -3 followed by:• Fludarabine 30 mg / m2IV over 30 minutes on Day -5, Day -4, and Day -3 followed by:• An additional IL of 0.9% NaCl (or isotonic [crystalloid] fluid) at the completion of the fludarabine infusion.• Add mesna (sodium 2-mercaptoethanesulfonate) per institutional guidelines.

[0115] Subjects with creatinine clearance 50 to 60 mL / min should have a 20% dose reduction of each daily fludarabine dose. Subjects with creatinine clearance of 30 to 49 mL / min should have a 40% dose reduction of each daily fludarabine dose.Axicabtagene Ciloleucel General instructions

[0116] All subjects will receive axicabtagene ciloleucel infusion at a healthcare facility, followed by daily monitoring at a healthcare facility for at least 7 days to monitor for signs and symptoms of CRS and neurologic toxicities.

[0117] Patient may receive their treatment as an outpatient assuming daily follow-up for the first 14 days following axicabtagene ciloleucel infusion. Alternatively, subjects may be hospitalized to receive their axicabtagene ciloleucel infusion and be observed for CRS and neurologic toxicities in the hospital setting, if deemed appropriate by the investigator.

[0118] If subjects are hospitalized, subjects should not be discharged from the hospital until all axicabtagene ciloleucel-related non-hematological toxicities resolve to < Grade 1 or return to baseline. Subjects may be discharged with non-critical and clinically stable or improving toxicities (e.g., renal insufficiency) even if > Grade 1, if deemed appropriate by the investigator. Subjects should remain in a hospital for ongoing axicabtagene ciloleucel-related fever, hypotension, hypoxia, or ongoing neurologic toxicities > Grade 1, or if deemed necessary by the investigator.

[0119] Axicabtagene ciloleucel will be administered at a target dose of 2 x 106anti-CD19 CAR-T cells / kg. Subjects who receive doses between 1-2 x 106anti-CD19 CAR-T cells / kg will be included in the mITT analysis set. For subjects weighing greater than 100 kg, a maximum flat dose of 2 x 108anti-CD19 CAR-T cells will be administered.Emapalumab General Instructions:

[0120] All subjects will receive emapalumab IV injections at a healthcare facility. Patients will receive their first dose on day -1 prior to CAR-T cell infusion on day 0.

[0121] Emapalumab will be dosed at 3 mg / kg as an intravenous infusion on day -1 of cell infusion with cell infusion being on day 0. Emapalumab is an intravenous infusion that should be given over 1 hour in the inpatient setting.

[0122] If it is deemed in the patient’s best interest for safety, additional doses may be given after discussion with the site specific PI. It is acceptable to give tocilizumab, steroids or other 3rdline agents for the management of CRS and ICANS in combination with emapalumab perinstitutional guidelines. Patients should not be treated with prophylactic agents for CRS / ICANS or for grade 1 CRS or ICANS.General Concomitant Medication and Supportive Care Guidelines

[0123] During the course of the study, investigators may prescribe any concomitant medications or treatment deemed necessary to provide adequate supportive care except those medications listed below. This includes following institutional guidelines for standard management of CRS and ICANS, including use of tocilizumab and / or high dose steroids as per appendix C & D. Prophylactic steroids, anakinra or tocilizumab should not be used while on study. Patients should not receive anti-cytokine or steroid intervention for grade 1 CRS and / or ICANS. Patients who are prematurely treated prior to grade 2 will be censored.Excluded Medications

[0124] Corticosteroid therapy at a pharmacologic dose (> 5 mg / day of prednisone or equivalent doses of other corticosteroids) and other immunosuppressive drugs must be avoided for 7 days prior to leukapheresis, and 5 days prior to axicabtagene ciloleucel administration. The package insert for Emapalumab administration advises to give dexamethasone prior to administration, but no steroids will be administered prior to Emapalumab administration during this trial. Steroid prophylaxis will be re-instituted per emapalumab product label in the event of any single grade 3 anaphylaxis or if greater than 1 / 3 of the first 6 patients experience any other G2 infusion related reaction.

[0125] Corticosteroids and other immunosuppressive drugs should also be avoided for 3 months after axicabtagene ciloleucel administration, unless used to manage axicabtagene ciloleucel related toxicities. Other medications that might interfere with the evaluation of the axicabtagene ciloleucel in combination with the investigational product, such as non-steroidal anti-inflammatory agents should also be avoided for the same time period unless medically necessary.

[0126] Treatment for lymphoma such as chemotherapy, immunotherapy, targeted agents, radiation, and high dose corticosteroid, other than defined / allowed in this protocol, and other investigational agents are prohibited, except as needed for treatment of disease progression after the axicabtagene ciloleucel infusion.

[0127] Prophylactic steroids, anakinra or tocilizumab should not be used while on study. Patients should not receive anti-cytokine or steroid intervention for grade 1 CRS and / orICANS. Patients who are prematurely treated prior to grade 2 will be replaced on study to accrue the analysis cohort of 28 patients.

[0128] Long acting growth factors should not be given within two weeks of axicabtagene infusion or within 3 days following infusion. G-CSF may be given after day +5 if CRS has resolved with down trending inflammatory markers. TPO mimetics are allowed following day +30 of infusion. No GM-CSF is allowed to be given on study unless approved by sponsor.Criteria for Taking a Participant off Protocol Therapy

[0129] Reasons for removal from protocol required investigational products or procedures include any of the following:• Unacceptable Adverse Event• Consent withdrawal• Participant becomes pregnant• Product not available• Lost to Follow-up• Death• Decision by sponsor• General or specific changes in the participant's condition that render the participant unacceptable for further treatment in the judgment of the treating investigator

[0130] Participants will be removed from the protocol therapy when any of these criteria apply.Duration of Follow Up

[0131] During the post-treatment follow-up period, treatment will be followed daily for a minimum of 7 days up to 13 days, weekly to month 1 then monthly for Months 2 and 3.

[0132] All enrolled subjects will be followed in the long-term follow-up period for survival and disease status, if applicable. Subjects will begin the long-term follow-up period after they have completed the Month 3 visit of the post treatment assessment period (whether they have responded to treatment or went straight to the Month 3 visit due to disease progression) as follows:Every 3 months (± 2 weeks) through Month 6Every 6 months (± 1 month) through Month 24Expected Toxicities

[0133] To date, the following important risks have been identified with axicabtagene ciloleucel: CRS, ICANS, infections, and cytopenias.

[0134] Emapalumab has been associated with an increased incidence of infections. All patients should be screened for TB by either a purified protein derivative (PPD) test result or IFNy release assay as part of their infectious disease screening. Prophylaxis for TB should be administered to patients who are at risk for TB or known to have a positive purified protein derivative (PPD) test result or positive IFNy release assay. During emapalumab treatment (first 30 days after CAR-T), patients should be monitored for TB, adenovirus, Epstein-Barr virus (EBV), and cytomegalovirus (CMV) every 2 weeks and as clinically indicated. Patients should be administered prophylaxis for herpes zoster, Pneumocystis jirovecii, and fungal infections per institutional standards, recommendations include:• HSV prophylaxis (acyclovir, famvir, etc) for 12 months following CAR-T.• PCP (Bactrim, mepron, pentamidine IV, etc) for 6 months following infusions.• Fungal prophylaxis (fluconazole, posaconazole, isaconazole, etc.) for patients with neutropenia >10 days and patients requiring a steroid dose greater than lOmg dexamethasone, or equivalent.

[0135] Infusion-related reactions, including drug eruption, pyrexia, rash, erythema, and hyperhidrosis, were reported with emapalumab treatment in 27% of patients. In one-third of these patients, the infusion-related reaction occurred during the first infusion. In the pivotal trial, the most commonly reported adverse reactions (>10%) for emapalumab included infection (56%), hypertension (41%), infusion-related reactions (27%), pyrexia (24%), hypokalemia (15%), constipation (15%), rash (12%), abdominal pain (12%), CMV-infection (12%), diarrhea (12%), lymphocytosis (12%), cough (12%), irritability (12%), tachycardia (12%), and tachypnea (12%).Adverse Events

[0136] Safety data will be reviewed and discussed with an external Data and Safety Monitoring Board (DSMB) per the study-specific DSMB charter. An adverse event is defined as any untoward medical occurrence in a clinical trial subject. The event does not necessarily have a relationship with study treatment.

[0137] The definition of adverse events includes worsening of a pre-existing medical condition. Worsening indicates that the pre-existing medical condition has increased in severity, frequency, and / or duration or has an association with a worse outcome. A pre-existing condition that has not worsened during the study or involves an intervention such as elective cosmetic surgery or a medical procedure while on study, is not considered an adverse event.

[0138] Interventions for pretreatment conditions (such as elective cosmetic surgery) or medical procedures that were planned before study participation are not considered adverse events. Hospitalization for study treatment infusions or precautionary measures per institutional policy are not considered adverse events.

[0139] The term “disease progression” as assessed by measurement of malignant lesions on radiographs or other methods should not be reported as adverse events. Death due to disease progression in the absence of signs and symptoms should be reported as the primary tumor type (e.g., B-Cell Lymphoma).

[0140] For situations when an adverse event or serious adverse event is due to the disease under investigation report the signs and symptoms. Worsening of signs and symptoms of the malignancy under study should also be reported as adverse events in the appropriate section of the CRF.Serious Adverse Event Definition

[0141] An SAE is any AE occurring at any dose and regardless of causality that:• Results in death.• Is immediately life-threatening-; i.e. the subject was at immediate risk of death at the time of the event. It does not include an adverse event that, had it occurred in a more severe form, might have caused death.• Requires in-patient hospitalization or prolongation of existing in-patient hospitalization. Hospitalization admissions occurring during the study period that are for procedures planned prior to study entry do not meet these criteria, unless there is a complication resulting from procedure that prolongs hospitalization.• Results in persistent or significant disability / incapacity. Disability is defined as a substantial disruption of a subject’s ability to conduct normal life functions.• Is a congenital anomaly / birth defect.• Is an important medical event. An important medical event is an event that may not result in death, be life-threatening, or require hospitalization, but may be considered serious when, based upon appropriate medical judgment, it may jeopardize the subject and may require medical or surgical intervention to prevent 1 of the outcomes listed above.

[0142] The investigator is responsible for reporting adverse events directly related to study related procedures following study consent. Following initiation of emapalumab and cellular therapy infusion, the investigator is responsible for ensuring that all adverse events observed by the investigator or reported by the subject that occur starting at infusion of emapalumab through 1 month after treatment with emapalumab, are monitored and reported regardless of disease progression. After 30 days, only targeted adverse events (listed below) attributable to emapalumab will be monitored and reported until 24 months post-study product infusion, disease progression, or initiation of new subsequent anticancer therapy, whichever comes first. All ongoing adverse events at 30 days will be monitored until resolution, return to baseline, or patient discontinuation of study participation due to completion of study procedures, patient withdrawal of consent, or patient removal from trial per investigator assessment.Targeted Adverse EventsCytopenias deemed related to LDC, cellular therapy infusion, or study product- New malignancies- New clinically significant incidence of hematologic, autoimmune, or rheumatologic disorderAny > Grade 3 incidence of Infection, Hemophagocytic Lymphohistiocytosis (HLH), Macrophage Activation Syndrome (MAS), or Tumor Lysis Syndrome (TLS)- New bacterial, viral or fungal infections Any SAE attributable to emapalumab

[0143] Following completion of study procedures at Month 24 or disease progression, whichever comes first, patients will be transitioned to the CIBMTR long term follow-up registry for AE reporting as is standard with commercial axicabtagene ciloleucel.

[0144] For subjects who sign consent, but do not receive axicabtagene ciloleucel and / or the investigational product, adverse event reporting ends 30 days after the last study-specific procedure (e.g., leukapheresis, lymphodepleting chemotherapy).Description of Study PeriodsScreening

[0145] The screening period begins on the date the subject signs the IRB / IEC approved ICF and continues through confirmation of enrollment. Informed consent must be obtained before completion of any non-standard of care study specific procedures. Procedures that arepart of standard of care are not considered study specific procedures and may be performed prior to obtaining consent and used to confirm eligibility. Confirmation of this data must occur within the time allowance as outlined below and in the SOA.

[0146] After written informed consent has been obtained, subjects will be screened to confirm study eligibility and participation. Only subjects who meet the eligibility criteria listed in Section 3 and who commence leukapheresis will be enrolled in the study. If at any time prior to enrollment the subject fails to meet the eligibility criteria, the subject should be designated as a screen failure on the subject screening log with the reasons for failing screening. Information regarding demographics, reason for screening failure, failed eligibility criteria, and any SAEs.

[0147] The following assessments / procedures are to be completed during the screening period at the time points outlined in the SOA:• Medical history and disease assessment• Physical examination including height and weight• Vital signs, including blood pressure, heart rate, oxygen saturation, and temperature• ECOG performance status• ECG• ECHO / MUGA for LVEF and pericardial effusion assessment o An ECHO / MUGA performed following the subj ects last chemotherapy treatment and within the institutions commercial SOP for commercial CAR-T screening is allowed.• Imaging Studies o Baseline PET of the neck, chest, abdomen and pelvis.■ PET performed following the subjects last line of therapy and prior to signing the consent may be used for confirmation of eligibility.■ If PET is performed > 28 days prior to the initiation of lymphodepleting chemotherapy or if subject receives any anti-cancer therapy between screening and lymphodepleting chemotherapy, the scans must be repeated to establish a new baseline. PET CT should be performed as close to enrollment as possible. If a PET is not accessible due to insurance reasons, CT imaging is allowable after discussion with the study PI.• Labs o Chemistry panel. o CBC with differential. o P-HCG pregnancy test (serum or urine) on all women of child-bearing potential. o Infectious disease testing for HBV (HBsAg, HBcAb, HBV NAT), HCV (IgG, HCVNAT), HIV screening, adenovirus (PCR), CMV (PCR and IgG), EBV (PCR and IgG) and Tb screening for latent Tb.• Serious Adverse Event reporting (refer to Section 7 for safety reporting guidelines)• Concomitant medications documentation and previous cancer treatment history• Once eligibility confirmed, collection of archived or fresh tumor sampleRescreening

[0148] Subjects who are unable to complete or meet the eligibility criteria during the 28-day screening period will be permitted to rescreen. Subjects will retain the same subject identification number assigned at the original screening. If rescreening occurs within 28 days of the signing of the original informed consent, only the procedure(s) / assessment(s) that did not originally meet the eligibility criteria needs to be repeated; all other initial screening procedures / assessments do not need to be repeated. If rescreening occurs, or leukapheresis is delayed, more than 28 days from the signing of the original informed consent, subjects must be reconsented and repeat all screening procedures / assessments.Enrollment / Leukapheresis

[0149] Before leukapheresis commences, the following criteria must be met. If criteria are not met, leukapheresis must be delayed until the event resolves.• No evidence or suspicion of an infection.• Corticosteroid therapy at a pharmacologic dose (>5 mg / day of prednisone or equivalent doses of other corticosteroids) and other immunosuppressive drugs must be avoided for 7 days prior to leukapheresis.

[0150] Once a subject has completed leukapheresis and meets trial eligibility, the subject will be considered enrolled into the study.

[0151] The following procedures / requirements will occur within 7 days of leukapheresis collection day and as outlined in the SOA:• Labs (to be drawn prior to leukapheresis, on the day of or day before leukapheresis) o Chemistry panel (including CMP, LDH, Uric Acid) o CBC with differential o C-reactive protein (CRP) and ferritin o Lymphocyte subsets o IgG levels o Translational lab collectionBridging Therapy

[0152] If prescribed, bridging therapy must be administered after enrollment and completed prior to initiating lymphodepleting chemotherapy per the specifications outlined in Section 5.1.1 for bridging therapy.Lymphodepleting Chemotherapy Period

[0153] If PET will be older than 28 days at the initiation of lymphodepleting chemotherapy or if the subject receives any anti-cancer therapy with therapeutic intent (e.g., radiation, supraphysiologic doses of steroids, chemotherapy) between the last PET and initiation of lymphodepleting chemotherapy, the PET must be repeated to establish a new baseline.

[0154] Before lymphodepleting chemotherapy commences, the following criteria must be met. If these criteria are not met, then lymphodepleting chemotherapy must be delayed until these events resolve: No evidence or suspicion of uncontrolled infection.

[0155] In addition, if any of the following are known to occur, a delay in lymphodepleting chemotherapy may be required: Temperature is > 38.0°C within 48 hours prior to lymphodepleting chemotherapy or during lymphodepletion. Unexplained fever requires panculture, respiratory viral panel, chest computed tomography (CT), and any additional symptom-directed workup to rule out occult infection. Primary PI or sponsor is required to approve initiation of lymphodepletion if this occurs.

[0156] The following procedures will be completed on day -5 at the time points outlined in the SOA:• Vital signs, including blood pressure, heart rate, oxygen saturation, and temperature• Labs (to be drawn prior to chemotherapy) o Chemistry Panel (including CMP, LDH, Uric Acid) o CBC with differential o C-reactive protein (CRP) and ferritino Lymphocyte subsets o Translational lab collection (VCN, flow cytometry, cytokines)• Fludarabine and cyclophosphamide administration• Adverse / Serious Adverse Event reporting• Concomitant medications documentationTreatment Period

[0157] Corticosteroid therapy at a pharmacologic dose (> 5 mg / day of prednisone or equivalent doses of other corticosteroids) and other immunosuppressive drugs should be avoided unless defined in the protocol.

[0158] Patients will be treated with a single dose of emapalumab on day -1.

[0159] Before axicabtagene ciloleucel infusion commences, the following criteria must be met: No evidence or suspicion of uncontrolled infection. Should an event not meet these criteria immediately prior to receiving axicabtagene ciloleucel, the axicabtagene ciloleucel infusion must be delayed until the event resolves. If the axicabtagene ciloleucel infusion is delayed > 2 weeks, lymphodepleting chemotherapy must be repeated. If CAR-T treatment is delayed more than 7 days, a new emapalumab dose will be given the day before CAR-T treatment.

[0160] All subjects will receive emapalumab and axicabtagene ciloleucel infusion at a healthcare facility followed by daily monitoring at a healthcare facility for at least 7 days to monitor for signs and symptoms of CRS and neurologic toxi cities. Alternatively, subjects may be hospitalized to receive their emapalumab and axicabtagene ciloleucel infusion and be observed for CRS and neurologic toxicities in the hospital setting.

[0161] If subjects are hospitalized, subjects should not be discharged from the hospital until all axicabtagene ciloleucel related non-hematological toxicities return to < Grade 1 or return to baseline. Subjects may be discharged with non-critical and clinically stable or improving toxicities (e.g., renal insufficiency) even if > Grade 1, if deemed appropriate by the investigator. Subjects should remain in a hospital for ongoing axicabtagene ciloleucel-related fever, hypotension, hypoxia, or ongoing central neurologic toxicities > Grade 1, or if deemed necessary by the investigator.

[0162] Subjects should be instructed to remain within proximity of the clinical study site for at least 4 weeks following axicabtagene ciloleucel infusion. During this period, the following procedures will be completed at the time points outlined in the SOA:• Neurological assessment for subjects to include ICANS (ICE scores)• Vital signs including blood pressure, heart rate, oxygen saturation, and temperature, daily at a health care facility for at least 7 days• Labs (before axicabtagene ciloleucel infusion, as described in the SOA) o Chemistry Panel o CBC with differential o PT / INR, PTT, Fibrinogen o C-reactive protein (CRP) and ferritin o IgG level o Lymphocyte subsets o Translational lab collection (VCN, flow cytometry, cytokines)• Infusion of axicabtagene ciloleucel• Administration of weight based emapalumab• Adverse / Serious Adverse Even reporting• Concomitant medications documentation

[0163] Monitoring of CRP, ferritin, and LDH (only if LDH is elevated at baseline) levels may assist with the diagnosis and define the clinical course in regard to CRS / ICANS. It is, therefore, mandatory that CRP, ferritin, and LDH (if elevated at baseline) be monitored daily starting at Day 0 and for at least 7 days at a healthcare facility. In addition, lactate should be monitored as clinically indicated.Post-treatment Assessment Period

[0164] After completing axicabtagene ciloleucel infusion and completing the minimum 7-day observation period, all subjects will be followed in the post-treatment assessment period. Counting from Day 0 (axicabtagene ciloleucel infusion), subjects will return to the clinic at the following intervals.• Week 2 (± 2 days)• Week 3 (± 2 days)• Week 4 (± 3 days)• Month 2 (± 1 week)• Month 3 (± 1 week)

[0165] Subject will allow key sponsor contacts to continue to access medical records so that information related to subjects health condition and initial treatment response may be obtained. The following procedures will be completed for subjects as outlined in the SO A:• Neurological assessment for subjects to include ICANS consensus criteria (ICE scoring)• PET for disease assessment: If the PET is not approved by the patients insurer CT will be allowed• Physical exam• Vital signs, including blood pressure, heart rate, oxygen saturation, and temperature• Labs o Chemistry Panel o CBC with differential o PT / INR, PTT, fibrinogen o CRP / Ferritin o Lymphocyte subsets o IgG Levels o Translational lab collection (VCN, flow cytometry, cytokines)• Discontinue levetiracetam as clinically indicated after 4 weeks• Adverse / Serious Adverse Event reporting (refer to Section 7 for safety reporting guidelines)• Concomitant medications documentation

[0166] If a subject is re-admitted to the hospital with any axicabtagene ciloleucel or emapalumab related adverse event(s), the following labs will be collected on the day of hospital re-admission and then weekly through and including on the day of discharge: Translational lab collection.

[0167] At any time during the post treatment assessment period, if a subject progresses the subject will proceed directly to the Month 3 visit and be followed for survival, subsequent therapy and disease outcomes in the long term follow-up period.Long-term Follow-up Period

[0168] All enrolled subjects will be followed in the long-term follow-up period for survival and disease status, if applicable. Subjects will begin the long-term follow-up period afterthey have completed the Month 3 visit of the post treatment assessment period (whether they have responded to treatment or went straight to the Month 3 visit due to disease progression).• Every 3 months (± 2 weeks) through Month 6• Every 6 months (± 1 month) through Month 24

[0169] The following procedures will be completed for subjects who are enrolled and receive axicabtagene ciloleucel at the time points outlined in the SOA:• Physical exam• PET / Disease assessment through 24 months or until disease progression, whichever occurs first. If subject’s disease has not progressed by Month 24, disease assessments will continue to be performed per institutional standard of care• Survival status• Labs o CBC with differential o Lymphocyte subsets o IgG Levels o Translational lab collection• Subsequent therapy for the treatment of NHL• Targeted adverse / serious adverse event reporting o Including neurological, hematological, infections, autoimmune disorders, and secondary malignancies• Targeted concomitant medication documentation (for 24 months or until disease progression, whichever occurs first) o Including gammaglobulins, immunosuppressive drugs, anti-infectives, and vaccinations

[0170] If a subject progresses in the LTFU phase, the subject will be transitioned to the CIBMTR long term follow-up registry and be taken off of the active protocol.

[0171] The following procedures / assessments will be completed for subjects who are enrolled, but do not receive axicabtagene ciloleucel or emapalumab, at the time points outlined in the SOA:• Subsequent therapy for the treatment of NHL• Survival statusDisease assessment per standard of care• Adverse / Serious Adverse Event reporting and concomitant medication documentation until 30 days after last procedure (e.g., leukapheresis, lymphodepleting chemotherapy)

[0172] Should the subject fail to return to the clinic for a scheduled protocol specific visit, sites will need to make 2 attempts by a combination of telephone and mail to contact the subject. Sites must document both attempts to contact the subject. If a subject does not respond within 1 month after the second contact the subject will be considered lost to followup and no additional contact will be required.

[0173] All subjects will be consented to the CIBMTR immune effector cell therapy registry at the same time as consent for this protocol and data will be reported on standard schedules immediately following infusion as required by FDA. Following Month 24 patients, protocol long-term follow-up will continue through the CIBMTR immune effector cell therapy registry as part of the FDA mandated 15-year follow-up period. Patients may be transitioned to the CIBMTR long term follow-up before Month 24 at time of disease progression, initiation of anti-cancer therapy, withdrawal of consent or at the investigators discretion.Measurement Of Effected & ICANS Assessment

[0174] CRS & ICANS will be assessed by ASTCT grading standards, including ICE scores, which can be found in Table 2-Table 4.Table 2: CRS Grading System - ASTCT CRS Consensus GradingCPAP: Continuous positive airway pressure; BiPAP: Bilevel positive airway pressure f Fever is defined as temperature >38°C not attributable to any other cause. In patients who have CRS then receive antipyretics or anti-cytokine therapy such as tocilizumab or steroids, fever is no longer required to grade subsequent CRS severity. In this case, CRS grading is driven by hypotension and / or hypoxia.{ CRS grade is determined by the more severe event: hypotension or hypoxia not attributable to any other cause. For example, a patient with temperature of 39.5°C, hypotension requiring one vasopressor and hypoxia requiring low- flow nasal cannula is classified as having Grade 3 CRS. Organ toxi cities associated with CRS may be graded according to CTCAE v5.0 but they do not influence CRS grading.ALow-flow nasal cannula is defined as oxygen delivered at < 6 liters / minute. Low flow also includes blow-by oxygen delivery, sometimes used in pediatrics. High-flow nasal cannula is defined as oxygen delivered at > 6 liters / minute.Table 3: ICANS GradingTable 4: ICE ScoreScoring: 10, no impairment;7-9, grade 1 ICANS;3-6, grade 2 ICANS;0-2, grade 3 ICANS;0 due to patient unarousable and unable to perform ICE assessment, grade 4 ICANS.Disease Response Assessment

[0175] Subjects will be evaluated for disease response by the site investigator at times indicated in the SOA. Disease assessments will be evaluated per Lugano criteria. Flow cytometric, molecular or cytogenetic studies will not be used to determine response.

[0176] Baseline PET-CT scans of the neck, chest, abdomen and pelvis, along with the appropriate imaging of all other sites of disease are required. Subjects will undergo additional PET or CT tumor assessments after their axicabtagene ciloleucel infusion. While PET is preferred, CT assessment may be used in place of PET in the event of failure to obtain insurance coverage. Month 1, 6 and 12 should be assessed via PET, CT will be allowed if denied by patients insurance. The first of these post-treatment PET tumor assessments will occur 4 weeks after infusion; subsequent assessments will occur at regular intervals throughout the post-treatment and long-term follow-up portions of the study, as highlighted in the SOA.

[0177] After axicabtagene ciloleucel and emapalumab administration, disease assessments will be used to determine the time when progressive disease occurs. Subjects with symptoms suggestive of disease progression should be evaluated for progression at the time symptoms occur even if it is off schedule as per the SOA.

[0178] For subjects who discontinue the study due to an assessment of progressive disease which was not subsequently confirmed by a central radiology reviewer, any additional imaging data, subsequent to the image in question will be submitted to the central reviewer to confirm disease response.Study Endpoints

[0179] Primary Endpoint: Incidence of grade 2+ CRS per ASTCT.

[0180] Secondary Endpoints:• Rate and severity of ICANS as per ASTCT criteria within the first 30 days of infusion.• Objective response rate (ORR): ORR is defined as the incidence of either a complete response or a partial response by Lugano. All subjects that do not meet the criteria for an objective response by the analysis data cutoff date will be considered nonresponders.

[0181] Exploratory Endpoints:• Duration of response (DOR): Among subjects who experience an objective response, DOR is defined as the date of their first objective response to disease progression per Lugano or death regardless of cause. Subjects not meeting the criteria for progression or death by the analysis data cutoff date will be censored at their last evaluable disease assessment date and their response will be noted as ongoing.• Progression-free Survival (PFS): PFS is defined as the time from the axicabtagene ciloleucel infusion date to the date of disease progression per Lugano or death from any cause. Subjects not meeting the criteria for progression by the analysis data cutoff date will be censored at their last evaluable disease assessment date.• Overall survival (OS): OS is defined as the time from axicabtagene ciloleucel infusion to the date of death. Subjects who have not died by the analysis data cutoff date will be censored at their last contact date.• Levels of anti-CD19 CAR-T cells in blood.• Levels of cytokines in serum and CSF.• Investigation of potential biomarker development based on assessment of blood cells, tumor cells and the proposed actions of the investigational product.• Other genetic factors (including DNA / RNA) that are associated with efficacy and / or toxicity.• Serum concentrations of emapalumab (PK).• Levels of circulating free IFN-y at predose, and total IFN-y (free IFN-y + bound to emapalumab) after initiation of the study drug.• Levels of the main IFN-y-induced chemokine CXCL9.• Levels of sIL2Ra.

[0182] Graphical data exploration might be conducted on an exploratory basis between the different biomarkers, laboratory markers and emapalumab concentrations. Further evaluations, such a development of population PK and / or PD analyses might be considered.

Claims

CLAIMSWhat is claimed is:

1. A method of improving a CAR-T cell therapy by decreasing a CAR-T cell associated toxicity in a subject receiving CAR-T cell therapy, comprising administering to the subject an effective dose of emapalumab.

2. A method for treating cancer in a subject in need thereof, the method comprising:(a) administering to the subject a lymphodepleting regimen;(b) administering to the subject emapalumab; and(c) administering to the subject a CAR-T cell therapy.

3. The method of claim 1, wherein the toxicity is cytokine-release syndrome.

4. The method of claim 1, wherein the toxicity is immune effector cell-associated neurotoxicity syndrome (ICANS).

5. The method of claim 1, wherein the toxicity is prolonged cytopenia.

6. The method of any one of claims 1-5, wherein the emapalumab is administered at a dose of 0.1 to 10 mg / kg intravenously.

7. The method of any one of claims 1-5, wherein the emapalumab is administered at a dose of 1 to 10 mg / kg intravenously.

8. The method of any one of claims 1-5, wherein the emapalumab is administered at a dose of 3 mg / kg intravenously.

9. The method of any one of claims 1-8, wherein the dose is a one-time dose.

10. The method of any one of claims 1-9, wherein the CART-T cell therapy has IFNy mediated toxicities.

11. The method of any one of claims 1-10, wherein the CAR-T cell therapy is axicabtagene ciloleucel.

12. The method of claim 11, wherein the axicabtagene ciloleucel is administered at a target dose of 2 x 106anti-CD19 CAR-T cells / kg, administered intravenously.

13. The method of any one of claims 11 or 12 wherein the axicabtagene ciloleucel is administered as a one-time dose.

14. The method of any one of claims 11-13, wherein the axicabtagene ciloleucel is administered concurrently, before or after emapalumab administration.

15. The method claim 14 wherein the axicabtagene ciloleucel is administered one day before emapalumab administration.

16. The method claim 14 wherein the axicabtagene ciloleucel is administered one to five days before emapalumab administration.

17. The method claim 14 wherein the axicabtagene ciloleucel is administered about 24 hours after emapalumab administration.

18. The method of any one of claims 1, 3-14, wherein the subject is further administered at least one additional therapy.

19. The method of claim 15, wherein the at least one additional therapy is a lymphodepleting regimen.

20. The method of claims 2 and 15, wherein the lymphodepleting regimen is administered for 1, 2, 3, 4, 6, 7, or more days.

21. The method of claims 2, or 19, wherein the lymphodepleting regimen comprises fludarabine and cyclophosphamide or bendamustine.

22. The method of claims 2 or 19, wherein the lymphodepleting regimen comprises radiation.

23. The method of claims 2 or 19, wherein the lymphodepleting chemotherapy regimen is administered prior to the administration of the CART-T cell therapy.

24. The method of any one of claims 1-23, wherein the method results in a prolongation of the duration of response to the CAR-T cell therapy of at least 10% as compared to the duration of response to the CAR-T cell therapy in the absence of emapalumab.

25. The method of any one of claims 1-23, wherein the method results in an increase in the overall response rate to the CAR-T cell therapy of at least 10% as compared to the overall response rate to the CAR-T cell therapy in the absence of emapalumab.

26. The method of any one of claims 1-23, wherein the method results in an increase in progression-free survival after administration of the CAR-T cell therapy of at least 10% as compared to the progression-free survival after administration of the CAR-T cell therapy in the absence of emapalumab.

27. The method of any one of claims 1-23, wherein the method results in an increase in overall survival after administration of the CAR-T cell therapy of at least 10% as compared to the overall survival after administration of the CAR-T cell therapy in the absence of emapalumab.

28. The method of any one of claims 1-23, wherein the method results in at least 500 CAR-T cells per microliter blood of the subject.

29. The method of any one of claims 1-23, wherein the patient has a large B-cell lymphoma that is refractory to first-line chemoimmunotherapy.

30. The method of any one of claims 1-23, wherein the patient has a large B-cell lymphoma that relapses within 12 months of first-line chemoimmunotherapy.

31. The method of any one of claims 1-23, wherein the patient has a relapsed or refractory large B-cell lymphoma after two or more lines of systemic therapy.

32. The method of any one of claims 1-23, wherein the patient has non-Hodgkin lymphoma.

33. The method of any one of claims 1-23, wherein the patient has diffuse large B-cell lymphoma.

34. The method of any one of claims 1-23, wherein the patient has primary mediastinal B- cell lymphoma.

35. The method of any one of claims 1-23, wherein the patient has diffuse large B-cell lymphoma arising from follicular lymphoma.

36. The method of any one of claims 1-23, wherein the patient has high grade B-cell lymphoma.

37. The method of claim 1, wherein the emapalumab is administered about 1-5 days after the lymphodepleting regimen.

38. The method of claim 1, wherein the CAR-T cell therapy is administered concurrently with or about one to five days after the emapalumab.

39. The method of claim 2, wherein the emapalumab is administered about 1-5 days after the lymphodepleting regimen.

40. The method of claim 2, wherein the CAR-T cell therapy is administered concurrently with or about one to five days after the emapalumab.

41. The method of any one of claims 1-40, wherein the method further comprises monitoring the levels of CAR-T cells in the blood of the subject.

42. The method of claim 41, wherein the levels of CAR-T cells in the blood of the subject is measured every other day.

43. The method of claim 41, wherein the levels of CAR-T cells in the blood of the subject is measured once every 5 days.

44. The method of claim 41, wherein the levels of CAR-T cells in the blood of the subject is measured once a week.

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