CXCR4 inhibitors and / or beta adrenergic receptor inhibitors for mobilization of lymphocytes andimmune cells

Abstract

This invention relates to methods and compositions directed to mobilizing a cell in a subject by blocking CXCR4, a beta-adrenergic receptor, a GPCR, or any combination thereof. In some embodiments, the cell is an immune cell. In some embodiments, the cell is a lymphocyte.

Description

CXCR4 INHIBITORS AND / OR BETA ADRENERGIC RECEPTOR INHIBITORS FOR MOBILIZATION OF LYMPHOCYTES ANDIMMUNE CELLSCROSS-REFERENCE TO RELATED APPLICATION

[0001] This application claims the benefit under 35 U.S C. §119(e) of United States Provisional Application No. 63 / 673,035, filed on July 18, 2024, which is hereby incorporated by reference in its entirety.BACKGROUND OF THE INVENTION

[0002] The invention disclosed herein relates generally to the mobilization of lymphocytes and immune cells.

[0003] CXC Chemokine receptor 4 (CXCR4) belongs to the superfamily of G protein-coupled receptors (GPCR). Binding of the chemokine CXCL12 (also known as SDF-1) to its receptor CXCR4 plays an essential role in homing and retention of hematopoietic stem cells (HSC) in the bone marrow. Blocking the CXCL12 / CXCR4 axis can elicit rapid mobilization of HSC from bone marrow to the peripheral blood (Domingues et al 2017). CXCR4 antagonists like Burixafor (also referred to as GPC-100 or TG-0054), as well as Plerixafor (also referred to as AMD3100 or Mozobil), have been used clinically in combination with granulocyte-colony stimulating factor (G-CSF) for hematopoietic stem cell mobilization and subsequent autologous stem cell transplant in non-Hodgkin’s Lymphoma and multiple myeloma patients.

[0004] CXCR4 is also expressed by immune cells. In our previous preclinical studies in mice, administration of GPC-100 induced rapid mobilization of WBC in peripheral blood within 30 min with a peak mobilization at 2 hours. This effect was further enhanced by propranolol pretreatment for 7 days. Therefore, in the present study, we investigated if propranolol and GPC- 100 combination results in enhanced mobilization of lymphoid immune cell subtypes.SUMMARY

[0005] Disclosed herein, in some embodiments, are methods of mobilizing a lymphocyte or an immune cell in a subject, the method comprising blocking CXCR4 signaling and / or beta- adrenergic receptor signaling in the subject.

[0006] Disclosed herein, in some embodiments, are methods of inducing lymphocyte or immune cell mobilization in a subject, the method comprising blocking CXCR4 signaling and / or beta- adrenergic receptor signaling in the subject.

[0007] Disclosed herein, in some embodiments, are methods of enhancing apheresis by inducing lymphocyte or immune cell mobilization in a subject, the method comprising blocking CXCR4 signaling and / or beta-adrenergic receptor signaling in the subject.

[0008] Disclosed herein, in some embodiments, are methods of enhancing apheresis by mobilizing a lymphocyte or an immune cell in a subject, the method comprising blocking CXCR4 signaling and / or beta-adrenergic receptor signaling in the subject.

[0009] In some embodiments, the blocking beta-adrenergic receptor signaling is performed before the blocking CXCR4 signaling. In some embodiments, the blocking beta-adrenergic receptor signaling continues after the blocking CXCR4 signaling is terminated.

[0010] In some embodiments, the blocking CXCR4 signaling comprises administering a CXCR4 inhibitor to the subject. In some embodiments, the blocking beta-adrenergic receptor signaling comprises administering a beta-adrenergic receptor inhibitor to the subject. In some embodiments, the lymphocyte or immune cell is a lymphocyte. In some embodiments, the blocking CXCR4 signaling comprises administering a CXCR4 inhibitor to the subject and the blocking beta-adrenergic receptor signaling comprises administering a beta-adrenergic receptor inhibitor to the subject. In some embodiments, the lymphocy te or immune cell is a lymphocyte.

[0011] Disclosed herein, in some embodiments, are methods of mobilizing a lymphocyte in a subject, the method comprising administering a beta-adrenergic receptor inhibitor and / or a CXCR4 inhibitor to the subject.

[0012] Disclosed herein, in some embodiments, are methods of inducing lymphocyte mobilization in a subject, the method comprising administering a beta-adrenergic receptor inhibitor and / or a CXCR4 inhibitor to the subject.

[0013] Disclosed herein, in some embodiments, are methods of enhancing apheresis in a subject, the method comprising administering a beta-adrenergic receptor inhibitor and / or a CXCR4 inhibitor to the subject.

[0014] Disclosed herein, in some embodiments, are methods of enhancing apheresis by inducing cell mobilization in a subject, the method comprising administering a beta-adrenergic receptor inhibitor and / or a CXCR4 inhibitor to the subject.

[0015] Disclosed herein, in some embodiments, are methods of enhancing apheresis by mobilizing a cell in a subject, the method comprising administering a beta-adrenergic receptor inhibitor and / or a CXCR4 inhibitor to the subject.

[0016] In some embodiments, the administering the beta-adrenergic receptor inhibitor is performed before the administering the CXCR4 inhibitor. In some embodiments, the administering the beta-adrenergic receptor inhibitor continues after the administering the CXCR4 inhibitor is terminated. In some embodiments, the method further comprises administering G- CSF to the subject. In some embodiments, the administering the beta-adrenergic receptor inhibitor and the CXCR4 inhibitor to the subject is performed in the absence of G-CSF.

[0017] Disclosed herein, in some embodiments, are methods of mobilizing a lymphocyte in a subject, the method comprising administering a CXCR4 inhibitor and G-CSF to the subject, in the absence of a beta-adrenergic receptor inhibitor.

[0018] Disclosed herein, in some embodiments, are methods of inducing lymphocyte mobilization in a subject, the method comprising administering a CXCR4 inhibitor and G-CSF to the subject, in the absence of a beta-adrenergic receptor inhibitor.

[0019] Disclosed herein, in some embodiments, are methods of enhancing apheresis in a subject, the method comprising administering a CXCR4 inhibitor and G-CSF to the subject, in the absence of a beta-adrenergic receptor inhibitor.

[0020] Disclosed herein, in some embodiments, are methods of enhancing apheresis by inducing cell mobilization in a subject, the method comprising administering a CXCR4 inhibitor and G- CSF to the subject, in the absence of a beta-adrenergic receptor inhibitor.

[0021] Disclosed herein, in some embodiments, are methods of enhancing apheresis by mobilizing a cell in a subject, the method comprising administering a CXCR4 inhibitor and G- CSF to the subject, in the absence of a beta-adrenergic receptor inhibitor.

[0022] In some embodiments, the beta-adrenergic receptor inhibitor is an ADRB2 inhibitor. In some embodiments, the beta-adrenergic receptor inhibitor is selected from the group consisting of alprenolol, atenolol, betaxolol, bupranolol, butoxamine, carazolol, carvedilol, CGP 12177, cicloprolol, ICI 118551, ICYP, labetalol, levobetaxolol, levobunolol, LK 204-545, metoprolol, nadolol, NIHP, NIP, propafenone, propranolol, sotalol, SR59230A. and timolol. In some embodiments, the beta-adrenergic receptor inhibitor is selected from the group consisting of propranolol, nadolol, and ICI 118551. In some embodiments, the beta-adrenergic receptor inhibitor is propranolol.

[0023] In some embodiments, the CXCR4 inhibitor is selected from the group consisting of ALX40-4C, AMD070 (AMD11070, X4P-001). AMD3100 (plerixafor), AMD3465. ATI 2341, BKT140 (BL-8040; TF14016; 4F-Benzoyl-TN 14003), CTCE-9908, CX549„D-[Lys3] GHRP-6, FC122, FC131, GMI-1359, GSK812397, GST-NT21MP, isothiourea- la, isothiourea-lt (ITlt), KRH-1636, KRH-3955, LY2510924, MSX-122, N-[llC]Methyl-AMD3465, POL6326, SDF-1 1-9[P2G] dimer, SDF1 P2G. T134, T140, T22, TC 14012, TG-0054 (Burixafor), USL311, viral macrophage inflammatory protein-II (vMIP-II), WZ811, [64Cu]-AMD3100, [64Cu]-AMD3465. [68Ga]pentixafor, [90Y]pentixather, [99mTc]02-AMD3100, [ 177Lu]pentixather, and 508MC1 (Compound 26). In some embodiments, the CXCR4 inhibitor is selected from the group consisting of AD-214, AMD070 (AMD11070, X4P-001), AMD3100 (plerixafor), BKT140 (BL- 8040; TF14016; 4F-Benzoyl-TN 14003). CTCE-9908. LY2510924, LY2624587, T140. TG-0054 (Burixafor), PF-06747143, POL6326, and ulocuplumab (MDX1338 / BMS-936564). In some embodiments, the CXCR4 inhibitor is TG-0054 (burixafor). In some embodiments, the CXCR4 inhibitor is AMD3100 (plerixafor). In some embodiments, the CXCR4 inhibitor is ulocuplumab (MDX1338 / BMS-936564).

[0024] In some embodiments, the administering the CXCR4 inhibitor and beta-adrenergic receptor inhibitor to the subject comprises administering TG-0054 (burixafor) and propranolol. In some embodiments, the administering the CXCR4 inhibitor beta-adrenergic receptor inhibitor to the subject comprises administering AMD3100 (plerixafor) and propranolol. In some embodiments, the administering the CXCR4 inhibitor beta-adrenergic receptor inhibitor to the subject comprises administering ulocuplumab (MDX1338 / BMS-936564) and propranolol.

[0025] In some embodiments, the administering a combination of the CXCR4 inhibitor and the G-CSF induces an enhanced amount of lymphocyte or immune cell mobilization relative to the amount of lymphocyte or immune cell mobilization induced by the CXCR4 inhibitor only. In some embodiments, the administering a combination of the CXCR4 inhibitor and the G-CSF mobilizes a lymphocyte or an immune cell by an amount enhanced relative to the amount of lymphocyte or immune cell mobilization induced by the CXCR4 inhibitor only. In some embodiments, the administering a combination of the CXCR4 inhibitor and the G-CSF induces an enhanced amount of apheresis relative to the amount of apheresis induced by the CXCR4 inhibitor only. In some embodiments, the administering a combination of the CXCR4 inhibitor and the beta-adrenergic receptor inhibitor induces an enhanced amount of lymphocyte or immune cell mobilization relative to the amount of lymphocyte or immune cell mobilization induced bythe CXCR4 inhibitor only. In some embodiments, the administering a combination of the CXCR4 inhibitor and the beta-adrenergic receptor inhibitor mobilizes a lymphocyte or an immune cell by an amount enhanced relative to the amount of lymphocyte or immune cell mobilization induced by the CXCR4 inhibitor only. In some embodiments, the administering a combination of the CXCR4 inhibitor and the beta-adrenergic receptor inhibitor induces an enhanced amount of apheresis relative to the amount of apheresis induced by the CXCR4 inhibitor only. In some embodiments, the administering a combination of the CXCR4 inhibitor, the beta-adrenergic receptor inhibitor, and the G-CSF induces an enhanced amount of lymphocyte or immune cell mobilization relative to the amount of lymphocyte or immune cell mobilization induced by the CXCR4 inhibitor and the beta-adrenergic receptor inhibitor only. In some embodiments, the administering a combination of the CXCR4 inhibitor, the beta-adrenergic receptor inhibitor, and the G-CSF mobilizes a cell by an amount enhanced relative to the amount of cell mobilization induced by the CXCR4 inhibitor and the beta-adrenergic receptor inhibitor only. In some embodiments, the administering a combination of the CXCR4 inhibitor and the beta-adrenergic receptor inhibitor, and the G-CSF induces an enhanced amount of apheresis relative to the amount of apheresis induced by the CXCR4 inhibitor and the beta-adrenergic receptor inhibitor only.

[0026] In some embodiments, the administering a combination of TG-0054 (burixafor) and the G-CSF induces an enhanced amount of lymphocyte or immune cell mobilization relative to the amount of lymphocyte or immune cell mobilization induced by AMD3100 (plerixafor) and the G-CSF. In some embodiments, the administering a combination of the TG-0054 (burixafor) and the G-CSF mobilizes a lymphocyte or an immune cell by an amount enhanced relative to the amount of lymphocyte or immune cell mobilization induced by the AMD3100 (plerixafor) and the G-CSF. In some embodiments, the administering a combination of the TG-0054 (burixafor) and the G-CSF induces an enhanced amount of apheresis relative to the amount of apheresis induced by the AMD3100 (plerixafor) and the G-CSF.

[0027] In some embodiments, an enhanced amount of lymphocyte or immune cell mobilization or apheresis is measured by a method selected from the group consisting of complete blood count (CBC) analysis, flow cytometry, and colony forming unit (CFU) assay. In some embodiments, the enhanced amount of lymphocyte or immune cell mobilization or apheresis is measured by flow cytometry'. In some embodiments, the flow cytometry is performed on (Lin-Scal+c-Kit+)LSK cells and / or CD34+ cells. In some embodiments, the enhanced amount of lymphocyte or immune cell mobilization or apheresis is measured by colony forming unit (CFU) assay.

[0028] In some embodiments, the subject has a CXCR4 protomer in the lymphocyte or immune cell. In some embodiments, the subject has an ADRB2 protomer in the lymphocyte or immune cell. In some embodiments, the subject has a CXCR4 protomer and an ADRB2 protomer in the lymphocyte or immune cell. In some embodiments, the subject has a CXCR4-ADRB2 heteromer in the lymphocyte or immune cell.

[0029] In some embodiments, i) the CXCR4-ADRB2 heteromer has an enhanced amount of downstream calcium mobilization relative to downstream calcium mobilization from a CXCR4 protomer or ADRB2 protomer; and ii) the administered combination of inhibitors suppresses the enhanced downstream calcium mobilization from said CXCR4-ADRB2 heteromer in the lymphocyte.

[0030] In some embodiments, the lymphocyte or immune cell is a lymphocyte. In some embodiments, the lymphocyte is selected from the group consisting of a T cell, B cell, and NK cell. In some embodiments, the lymphocyte is mobilized from bone marrow to peripheral blood. In some embodiments, the lymphocyte is collected for transplantation to a patient having cancer. In some embodiments, the cancer is selected from the group consisting of lymphoma, leukemia, and myeloma. In some embodiments, the cancer is non-Hodgkin lymphoma (NHL), acute myeloid leukemia (AML), acute lymphoblastic leukemia (ALL), or multiple myeloma (MM). In some embodiments, the lymphocyte is mobilized from bone marrow to peripheral blood. In some embodiments, the lymphocyte is mobilized for treatment of a condition selected from the group consisting of neurological disorder, cardiac ischemia, myocardial infarction, diabetes, tissue repair, bone and cartilage disease, autoimmune disease, graft versus host disease, Crohn's disease, multiple sclerosis, systemic lupus erythematosus, and systemic sclerosis. In some embodiments, the lymphocyte is mobilized into blood. In some embodiments, the lymphocyte is mobilized for treatment of a cancer.

[0031] In some embodiments, the cell is an immune cell. In some embodiments, the cell is an immune cell. In some embodiments, the white blood cell is a lymphocyte. In some embodiments, the lymphocyte is selected from the group consisting of a T cell, a B cell, and a natural killer (NK) cell. In some embodiments, the lymphocyte is a T cell. In some embodiments, the lymphocyte is a natural killer (NK) cell. In some embodiments, the white blood cell is a granulocyte. In some embodiments, the granulocyte is selected from the groupconsisting of a neutrophil, an eosinophil, and a basophil. In some embodiments, the granulocyte is a neutrophil. In some embodiments, the white blood cell is a monocyte.

[0032] In some embodiments, the immune cell is mobilized from bone marrow to peripheral blood. In some embodiments, the immune cell is mobilized from lymph node to peripheral blood. In some embodiments, the mobilized immune cell is used for adoptive cell therapy(ACT). In some embodiments, the adoptive cell therapy (ACT) is chimeric antigen receptor (CAR) T cell therapy. In some embodiments, the adoptive cell therapy (ACT) is netural killer(NK) cell therapy. In some embodiments, the adoptive cell therapy (ACT) is engineered T-cell receptor (TCR) therapy. In some embodiments, the adoptive cell therapy (ACT) is tumorinfiltrating lymphocyte (TIL) therapy.

[0033] In some embodiments, the method does not involve the addition of a CXCR2 agonist. In some embodiments, the CXCR2 agonist is MGTA-145.

[0034] In some embodiments, the method is not used to treat a patient having a sickle cell disease.

[0035] Disclosed herein, in some embodiments, are methods of mobilizing a lymphocyte or an immune cell in a subject, the method comprising administering, alone or in combination, a CXCR4 inhibitor and G-CSF to the subject.

[0036] Disclosed herein, in some embodiments, are methods for treatment of cancer comprising administering, alone or in combination, a CXCR4 inhibitor and G-CSF to a subject.

[0037] Disclosed herein, in some embodiments, are methods of enhancing leukapheresis by inducing lymphocyte or immune cell mobilization in a subject, the method comprising administering, alone or in combination, a CXCR4 inhibitor and G-CSF to the subject.

[0038] In some embodiments, the CXCR4 inhibitor and the G-CSF are administered to the subj ect in the absence of a beta-adrenergic receptor inhibitor. In some embodiments, the method comprises administering the G-CSF to the subject before administering the CXCR4 inhibitor to the subject. In some embodiments, the method comprises administering a combination of the CXCR4 inhibitor and G-CSF to the subject, and wherein the administering of the combination of the CXCR4 inhibitor and G-CSF to the subject induces an enhanced amount of lymphocyte mobilization relative to an amount of lymphocyte mobilization induced by administration of G- CSF only to the subject. In some embodiments, the method comprises administering a combination of the CXCR4 inhibitor and G-CSF to the subject, and wherein the administering of the combination of the CXCR4 inhibitor and G-CSF to the subject induces an enhanced amountof lymphocyte mobilization relative to an amount of lymphocyte mobilization induced by administration of the CXCR4 inhibitor only to the subject.

[0039] In some embodiments, the CXCR4 inhibitor is selected from the group consisting of ALX40-4C, AMD070 (AMD11070, X4P-001), AMD3100 (plerixafor), AMD3465, ATI 2341, BKT140 (BL-8040; TF14016; 4F-Benzoyl-TN 14003), CTCE-9908, CX549„D-[Lys3] GHRP-6, FC122. FC131, GMI-1359, GSK812397, GST-NT21MP, isothiourea- la, isothiourea- It (ITlt), KRH-1636, KRH-3955, LY2510924, MSX-122, N-[l lC]Methyl-AMD3465, POL6326. SDF-1 1 -9[P2G] dimer, SDF1 P2G, T134, T140, T22, TC 14012, TG-0054 (Burixafor), USL31 1 , viral macrophage inflammatory protein-II (vMIP-II), WZ811, [64Cu]-AMD3100, [64Cu]-AMD3465, [68Ga]pentixafor, [90Y]pentixather, [99mTc]02-AMD3100, [177Lu]pentixather, and 508MC1 (Compound 26). In some embodiments, the CXCR4 inhibitor is selected from the group consisting of AD-214, AMD070 (AMD11070, X4P-001), AMD3100 (plerixafor), BKT140 (BL- 8040; TF14016; 4F-Benzoyl-TN14003), CTCE-9908, LY2510924, LY2624587, T140, TG-0054 (Burixafor), PF-06747143, POL6326, and ulocuplumab (MDX1338 / BMS-936564). In some embodiments, the CXCR4 inhibitor is TG-0054 (burixafor).

[0040] In some embodiments, the method comprises administering a combination of burixafor and G-CSF to the subject, and wherein the administering of the combination of burixafor and G- CSF to the subject induces an enhanced amount of lymphocyte mobilization relative to an amount of lymphocyte mobilization induced by administration of G-CSF only to the subject. In some embodiments, the method comprises administering a combination of burixafor and G-CSF to the subject, and wherein the administering of the combination of the burixafor and G-CSF to the subject induces an enhanced amount of lymphocyte mobilization relative to an amount of lymphocyte mobilization induced by administration of burixafor only to the subject. In some embodiments, the enhanced amount of lymphocyte mobilization is measured by a method selected from the group consisting of complete blood count (CBC) analysis, flow cytometry, and colony forming unit (CFU) assay. In some embodiments, the enhanced amount of lymphocyte mobilization is measured by CBC analysis.

[0041] In some embodiments, the subject has a CXCR4 protomer in the lymphocy te. In some embodiments, the lymphocyte or immune cell is selected from the group consisting of a T cell, B cell, a natural killer (NK) cell, a neutrophil, eosinophil, and a basophil.

[0042] In some embodiments, the lymphocyte or immune cell is mobilized for treatment of a condition selected from the group consisting of neurological disorder, cardiac ischemia,myocardial infarction, diabetes, tissue repair, bone and cartilage disease, autoimmune disease, graft versus host disease, Crohn's disease, multiple sclerosis, systemic lupus erythematosus, and systemic sclerosis. In some embodiments, the lymphocyte or immune cell is mobilized for treatment of a cancer. In some embodiments, the cancer is non-Hodgkin lymphoma (NHL), acute myeloid leukemia (AML), acute lymphoblastic leukemia (ALL), or multiple myeloma (MM). In some embodiments, the cancer is multiple myeloma (MM). In some embodiments, the lymphocyte or immune cell is mobilized for adoptive cell therapy (ACT). In some embodiments, the adoptive cell therapy (ACT) is chimeric antigen (CAR) T cell therapy. In some embodiments, the cancer is non-Hodgkin lymphoma (NHL), acute myeloid leukemia (AML), acute lymphoblastic leukemia (ALL), or multiple myeloma (MM). In some embodiments, the cancer is multiple myeloma (MM). In some embodiments, the treatment comprises leukapheresis. In some embodiments, the treatment comprises adoptive cell therapy (ACT). In some embodiments, the adoptive cell therapy (ACT) is chimeric antigen (CAR) T cell therapy.

[0043] Disclosed herein, in some embodiments, is a composition or compositions for mobilizing a lymphocyte or an immune cell in a subject comprising, alone or in combination, a CXCR4 inhibitor and G-CSF.

[0044] Disclosed herein, in some embodiments, is a composition or compositions for treatment of cancer comprising, alone or in combination, a CXCR4 inhibitor and G-CSF.

[0045] Disclosed herein, in some embodiments, is a composition or compositions for enhancing leukapheresis by inducing lymphocyte or immune cell mobilization in a subject comprising, alone or in combination, a CXCR4 inhibitor and G-CSF.

[0046] In some embodiments, the composition does not include a beta-adrenergic receptor inhibitor. In some embodiments, the composition further includes a beta-adrenergic receptor inhibitor.

[0047] In some embodiments, the CXCR4 inhibitor is selected from the group consisting of ALX40-4C, AMD070 (AMD11070, X4P-001), AMD3100 (plerixafor), AMD3465, ATI 2341, BKT140 (BL-8040; TF14016; 4F-Benzoyl-TN14003), CTCE-9908, CX549„D-[Lys3] GHRP-6, FC122. FC131, GMI-1359, GSK812397. GST-NT21MP, isothiourea- la, isothiourea- It (ITlt). KRH-1636, KRH-3955, LY2510924, MSX-122, N-[l 1 C] Methyl -AMD3465, POL6326, SDF-1 1-9[P2G] dimer, SDF1 P2G, T134, T140, T22, TC 14012, TG-0054 (Burixafor), USL311, viral macrophage inflammatory protein-II (vMIP-II), WZ811, [64Cu]-AMD3100, [64Cu]-AMD3465,[68Ga]pentixafor, [90Y]pentixather, [99mTc]02-AMD3100, [177Lu]pentixather, and 508MC1 (Compound 26). In some embodiments, the CXCR4 inhibitor is selected from the group consisting of AD-214, AMD070 (AMD11070, X4P-001), AMD3100 (plenxafor), BKT140 (BL- 8040; TF14016; 4F-Benzoyl-TN14003), CTCE-9908, LY2510924, LY2624587, T140, TG-0054 (Burixafor), PF-06747143, POL6326, and ulocuplumab (MDX1338 / BMS-936564). In some embodiments, the CXCR4 inhibitor is TG-0054 (burixafor).

[0048] In some embodiments, the subject has a CXCR4 protomer in the lymphocyte. In some embodiments, the lymphocyte or immune cell is selected from the group consisting of a T cell, a B cell, and a natural killer (NK) cell, a neutrophil, a eosinophil, and a basophil. In some embodiments, the lymphocyte or immune cell is mobilized for treatment of a condition selected from the group consisting of neurological disorder, cardiac ischemia, myocardial infarction, diabetes, tissue repair, bone and cartilage disease, autoimmune disease, graft versus host disease, Crohn's disease, multiple sclerosis, systemic lupus ery thematosus, and systemic sclerosis. In some embodiments, the lymphocyte or immune cell is mobilized for treatment of a cancer. In some embodiments, the cancer is non-Hodgkin lymphoma (NHL), acute myeloid leukemia (AML), acute lymphoblastic leukemia (ALL), or multiple myeloma (MM). In some embodiments, the cancer is multiple myeloma (MM). In some embodiments, the lymphocyte is mobilized for adoptive cell therapy (ACT). In some embodiments, the adoptive cell therapy (ACT) is chimeric antigen (CAR) T cell therapy.

[0049] In some embodiments, the cancer is non-Hodgkin lymphoma (NHL), acute myeloid leukemia (AML), acute lymphoblastic leukemia (ALL), or multiple myeloma (MM). In some embodiments, the cancer is multiple myeloma (MM).

[0050] In some embodiments, the treatment comprises leukapheresis. In some embodiments, the treatment comprises adoptive cell therapy (ACT). In some embodiments, the adoptive cell therapy (ACT) is chimeric and gen (CAR) T cell therapy.

[0051] Disclosed herein, in some embodiments, is a pharmaceutical composition or pharmaceutical compositions for mobilizing a lymphocyte or an immune cell in a subject comprising, alone or in combination, a CXCR4 inhibitor and G-CSF, and a pharmaceutically acceptable excipient.

[0052] Disclosed herein, in some embodiments, is a pharmaceutical composition or pharmaceutical compositions for treatment of cancer comprising, alone or in combination, a CXCR4 inhibitor and G-CSF, and a pharmaceutically acceptable excipient.

[0053] Disclosed herein, in some embodiments, is a pharmaceutical composition or pharmaceutical compositions for enhancing leukapheresis by inducing lymphocyte or immune cell mobilization in a subject comprising, alone or in combination, a CXCR4 inhibitor and G- CSF, and a pharmaceutically acceptable excipient.

[0054] In some embodiments, the pharmaceutical composition does not include a beta- adrenergic receptor inhibitor. In some embodiments, the pharmaceutical composition further includes a beta-adrenergic receptor inhibitor.

[0055] In some embodiments, the CXCR4 inhibitor is selected from the group consisting of ALX40-4C, AMD070 (AMD11070, X4P-001), AMD3100 (plerixafor), AMD3465, ATI 2341, BKT140 (BL-8040; TF14016; 4F-Benzoyl-TN14003), CTCE-9908, CX549„D-[Lys3] GHRP-6, FC122. FC131, GMI-1359, GSK812397, GST-NT21MP, isothiourea- la, isothiourea- It (mt). KRH-1636, KRH-3955, LY2510924, MSX-122, N-[l lC]Methyl-AMD3465, POL6326, SDF-1 1-9[P2G] dimer, SDF1 P2G, T134, T140, T22, TC 14012, TG-0054 (Burixafor), USL311, viral macrophage inflammatory protein-II (vMIP-II), WZ811, [64Cu]-AMD3100, [64Cu]-AMD3465, [68Ga]pentixafor, [90Y]pentixather, [99mTc]02-AMD3100. [177Lu]pentixather, and 508MC1 (Compound 26). In some embodiments, the CXCR4 inhibitor is selected from the group consisting of AD-214, AMD070 (AMD11070, X4P-001), AMD3100 (plerixafor), BKT140 (BL- 8040; TF14016; 4F-Benzoyl-TN14003), CTCE-9908, LY2510924, LY2624587, T140, TG-0054 (Burixafor), PF-06747143, POL6326, and ulocuplumab (MDX1338 / BMS-936564). In some embodiments, the CXCR4 inhibitor is TG-0054 (burixafor).

[0056] In some embodiments, the subject has a CXCR4 protomer in the lymphocyte. In some embodiments, the lymphocyte or immune cell is selected from the group consisting of a T cell, a B cell, and a natural killer (NK) cell, a neutrophil, an eosinophil, and a basophil. In some embodiments, the lymphocyte or immune cell is mobilized for treatment of a condition selected from the group consisting of neurological disorder, cardiac ischemia, myocardial infarction, diabetes, tissue repair, bone and cartilage disease, autoimmune disease, graft versus host disease, Crohn's disease, multiple sclerosis, systemic lupus erythematosus, and systemic sclerosis. In some embodiments, the lymphocyte or immune cell is mobilized for treatment of a cancer. In some embodiments, the cancer is non-Hodgkin lymphoma (NHL), acute myeloid leukemia (AML), acute lymphoblastic leukemia (ALL), or multiple myeloma (MM). In some embodiments, the cancer is multiple myeloma (MM). In some embodiments, the lymphocyte orimmune cell is mobilized for adoptive cell therapy (ACT). In some embodiments, the adoptive cell therapy (ACT) is chimeric antigen (CAR) T cell therapy.

[0057] In some embodiments, the cancer is non-Hodgkin lymphoma (NHL), acute myeloid leukemia (AML), acute lymphoblastic leukemia (ALL), or multiple myeloma (MM). In some embodiments, the cancer is multiple myeloma (MM). In some embodiments, the treatment comprises leukapheresis. In some embodiments, the treatment comprises adoptive cell therapy (ACT). In some embodiments, the adoptive cell therapy (ACT) is chimeric antigen (CAR) T cell therapy.BRIEF DESCRIPTION OF THE DRAWINGS

[0058] FIG. 1 shows that propranolol enhances GPC-100 mobilization of lymphocytes to a level that is higher than the standard of care for mobilization (G-CSF).

[0059] FIGs. 2A-2B show white blood cell (WBC) (FIG. 2A) and lymphocyte (FIG. 2B) mobilization in Balb / c mice treated with vehicle, GPC-100. propranolol, GPC-100+propranolol. AMD3100, or AMD3100+propranolol.

[0060] FIGs. 3A-3H show GPC-100 induced mobilization of immune cells by propranolol treatment compared to AMD3100. The graphs show mobilization of CD45+ cells (FIG. 3A), CD3+ cells (FIG. 3B). CD4+ cells (FIG. 3C), CD8+ cells (FIG. 3D), CDl lb+ cells (FIG. 3E), F4 / 80+ cells (FIG. 3F), CD 19+ cells (FIG. 3G), or NK cells (FIG. 3H).

[0061] FIGs. 4A-4B show the fold change in mobilization of immune cells compared to vehicle controls in mice treated with GPC-100 with or without propranolol (FIG. 4A) and AMD3100 with or without propranolol (FIG. 4B).

[0062] FIGs. 5A-5H show proportions of immune cells in bone marrow from mice treated with GPC-100 with or without introduced mobilization of immune cells by propranolol treatment compared to AMD3100. The graphs show proportions of CD45+ cells (FIG. 5A), CD3+ cells (FIG. 5B), CD4+ cells (FIG. 5C), CD8+ cells (FIG. 5D), CD1 lb+ cells (FIG. 5E), F4 / 80+ cells (FIG. 5F), CD 19+ cells (FIG. 5G), or NK cells (FIG. 5H).

[0063] FIGs. 6A-6C show white blood cell (WBC) (FIG. 6A), lymphocyte (FIG. 6B), and neutrophil (FIG. 6C) mobilization into peripheral blood in mice treated with vehicle, GPC-100, propranolol+GPC-100, or propranolol alone.

[0064] FIGs. 7A-7F show GPC-100 induced mobilization of immune cells by propranolol treatment compared to AMD3100. The graphs show mobilization of CD45+ cells (FIG. 7A).F4 / 80+ macrophages (FIG. 7B), CD 19+ cells (FIG. 7C), CD8+ cells (FIG. 7D), CD4+ cells (FIG. 7E), NKp46+ cells (FIG. 7F).

[0065] FIGs. 8A-8C show GPC100 induced mobilization of WBCs (FIG. 8A), lymphocytes (FIG. 8B), and neutrophils (FIG. 8C) in tumor bearing mice compared to naive mice.

[0066] FIGs. 9A-9H show mobilization of lymphoid cell subsets by GPC-100 in tumor bearing and naive mice. The graphs show mobilization of CD45+ cells (FIG. 9A). CD3+ T-cells (FIG. 9B), NK cells (FIG. 9C), CD19+ B cells (FIG. 9D), CD8+ T-cells (FIG. 9E), CD8+Granzyme+ cells (FIG. 9F), CD4+ T-cells (FIG. 9G), or CD4+foxp3+CD25+ Treg cells (FIG. 9H).

[0067] FIGs. 10A-10F show mobilization of myeloid cell subsets by GPC-100 in tumor bearing and naive mice. The graphs show mobilization of CDl lb+ myeloid cells (FIG. 10A), F4 / 80hi cells (FIG. 10B), CD80+ Ml cells (FIG. IOC). CD 163+ M2 cells (FIG. 10D), Ly-6C+Ly-6G- monocytic myeloid-derived suppressor cells (M-MDSC) (FIG. 10E), or Ly-6C low Ly-6G+ polymorphonuclear myeloid-derived suppressor cells (PMN-MDSC) (FIG. 10F).

[0068] FIGs. 11A-11D show the fold change in GPC-100 induced mobilization of CD3+, CD4+, CD4+FoxP3+, CD8+, Granz+CD8+, PD-1+CD8+, NKp46+, and CD19+ cells (FIG. 11A) and CDl lb+, F4 / 80hi, Ml. M-MDSC cells (FIG. 11B) compared to vehicle controls in naive mice; and GPC-100 induced mobilization of CD3+, CD4+, CD4+FoxP3+, CD8+, Granz+CD8+, PD-1+CD8+, NKp46+, and CD19+ cells (FIG. 11C) and CDl lb+, F4 / 80hi, Ml, M-MDSC cells (FIG. 11D) compared to vehicle controls in tumor bearing mice.

[0069] FIGs. 12A-12E show the changes in the tumor from GPC-100 induced mobilization. The graphs show the density (cells / cm3) of immune cells in tumors from mice treated with vehicle (FIG. 12A) and GPC-100 (FIG. 12B); the density of F4 / 80 cells in tumors from mice treated with vehicle and GPC-100 (FIG. 12C); the density of Ml macrophages in tumors from mice treated with vehicle and GPC-100 (FIG. 12D): and the density of M2 macrophages in tumors from mice treated with vehicle and GPC-100 (FIG. 12E).

[0070] FIGs. 13A-13G show GPC-100 induced mobilization of WBC (FIG. 13A), lymphocytes (FIG. 13B), and neutrophils (FIG. 13C) in lymphoma bearing mice compared to naive mice. FIGs. 13D-13G are pie charts showing the percent of lymphocytes and neutrophils in naive mice treated with phosphate buffered saline (PBS) (FIG. 13D), naive mice treated with GPC-100 (FIG. 13E), tumor bearing mice treated with PBS (FIG. 13F), and tumor bearing mice treated with GPC-100 (FIG. 13G).

[0071] FIGs. 14A-14E show GPC-100 induced mobilization of CD45+ (FIG. 14A), CD3+ (FIG. 14B), F4 / 80+ (FIG. 14C). CD 19+ (FIG. 14D), and NKp46 (FIG. 14E) in naive and lymphoma bearing mice.

[0072] FIGs. 15A-15J show CXCR4 expression on mobilized immune cells by GPC-100. The graphs show the number of CD3+CD4+ cells (FIG. 15A), CD3+ cells (FIG. 15B), CD19+ cells (FIG. 15C), F4 / 80+ cells (FIG. 15D). and NKp46+ cells (FIG. 15E) after treatment with GPC- 100; and the number of CXCR4+CD3+CD4+ cells (FIG. 15F), CXCR4+CD3+ cells (FIG. 15G). CXCR4+CD19+ cells (FIG. 15H), CXCR4+F4 / 80+ cells (FIG. 151), and CXCR4+NKp46+ cells (FIG. 15 J) after treatment with GPC-100.

[0073] FIGs. 16A-16F show WBC (FIG. 16A), lymphocy te (FIG. 16B), and neutrophil (FIG. 16C) mobilization in plasmacytoma tumor models; and WBC (FIG. 16D), lymphocyte (FIG. 16E), and neutrophil (FIG. 16F) mobilization in fibrosarcoma tumor models.

[0074] FIGs. 17A-17M show flow cytometry gating strategies for T cell populations. The flow cytometry' graphs show' ungated cells (FIG. 17A), singlets (FIG. 17B), live cells (FIG. 17C), CD45+ cells (FIG. 17D), NKp46+ cells (FIG. 17E). CD3+CD4+ and CD3+CD8+ cells (FIG. 17F). CD 19+ cells (FIG. 17G). F4 / 80+ cells (FIG. 17H). NKp45+CXCR4+ cells (FIG. 171). CD4+CXCR4+ cells (FIG. 17J), CD19+CXCR4+ cells (FIG. 17K), F4 / 80+CXCR4+ cells (FIG. 17L), and CD8+CXCR4+ cells (FIG. 17M).

[0075] FIGs. 18A-18B show the time course of white blood cell and lymphocyte mobilization. The time post drug graphs show white blood cell mobilization (FIG. 18A) and lymphocyte mobilization (FIG. 18B)

[0076] FIGs. 19A-19B show' the comparison of lymphocyte mobilization in two mouse strains. The lympocyte mobilization is shown for C57 / BL6 (FIG. 19A) and Balc / c (FIG. 19B).

[0077] FIG. 20 shows the mean concentration-time profiles for absolute lymphocyte counts (cells / mm3) after a single IV infusion of TG-0054 (GPC-100) in healthy subjects.

[0078] FIGs. 21A-21D show the break down of clinical data measuring the number of lymphocytes mobilized into the peripheral blood of multiple myeloma patients, where FIG. 21 A shows the absolute (ABS) number of lymphocytes of each patient at screening, pre-burixafor (post-G-CSF), 1 h post-burixafor and 6 h post-burixafor. FIG. 21B shows average lymphocyte count at each time-point tested, FIG. 21C shows fold change in lymphocytes after each treatment, and FIG. 21D shows the cumulative increase in lymphocytes over screening by the G-CSF and burixafor combination.

[0079] FIG. 22 shows the absolute (ABS) neutrophil counts (cells / mm3) after a single IV infusion of varying doses of TG-0054 (GPC-100) in healthy subjects as a function of time.ABBREVIATIONS

[0080] Unless indicated otherwise, the following includes abbreviations for terms disclosed herein: acute myeloid leukemia (AML), Adenosine A3 Receptor (AD0RA3), Adenosine Receptor A2b (ADORA2B), adenovirus high-throughput system (AdHTS), Adenylate Cyclase Activating Polypeptide 1 (Pituitary) Receptor Type I (ADCYAP1R1), Adrenoceptor Alpha 1A (ADRA1A), Adrenoceptor Beta 2 (ADRB2), Apelin Receptor (APLNR), Atypical chemokine receptor 3 (ACKR3). bimolecular fluorescence complementation (BiFC), Bioluminescence Resonance Energy Transfer (BRET), bovine serum albumin (BSA), Calcitonin Receptor (CALCR), Cancer stem cells (CSCs), C-C chemokine receptor type 2 (CCR2), chemerin chemokine-like receptor 1 (CMKLR1), Cholinergic Receptor Muscarinic 1 (CHRM1), chronic lymphocytic leukemia (CLL), chronic myeloid leukemia (CML), chronic obstructive pulmonary disease (COPD), Complement C5a Receptor 1 (C5AR1), C-terminal fragment of Venus (VC), C-X-C Motif Chemokine ligand 12 (CXCL12), CXC receptor 4 (CXCR4), cytotoxic T- lymphocyte-associated antigen 4 (CTLA-4), 5-opioid receptor (OPRD), Endothelin Receptor Type B (EDNRB), enzyme-linked immunosorbent assay (ELISA), formalin-fixed paraffin- embedded (FFPE), fluorescence resonance energy transfer (FRET), G protein-coupled receptor (GPCR), Galanin Receptor 1 (GALR1), glioblastoma multiforme (GBM), Glucagon receptor (GCGR), GPCR heteromer identification technology' (GPCR-HIT), Granulocyte-colony stimulating factor (G-CSF), hematopoietic stem cells (HSCs), hepatocellular carcinoma (HCC). Histamine Receptor Hl (HRH1), human immunodeficiency virus (HIV), International Union of Basic and Clinical Pharmacology Committee on Receptor Nomenclature and Drug Classification (NC-IUPHAR), p,-opioid receptor (MOR), Motilin Receptor (MLNR), Multiple myeloma (MM), multiplicity of infection (MOI). Myelodysplastic Syndromes (MDS). Neurotensin Receptor 1 (NTSR1), non-Hodgkin lymphoma (NHL), non-small-cell lung cancer (NSCLC), N-terminal fragments of Venus (VN), patient derived cell (PDC), Patient-Derived Xenograft (PDX), positron emission tomography (PET), Computed Tomography (CT), programmed cell death ligand 1 (PD- Ll), programmed cell death protein l(PD-l), Prostaglandin E Receptor 2 (PTGER2), Prostaglandin E Receptor 3 (PTGER3), proximity ligation assay (PLA), reverse transcription- quantitative polymerase chain reaction (RT-qPCR), Single-photon emission computedtomography (SPECT), small lymphocytic lymphoma (SLL), small-cell lung cancer (SCLC), Somatostatin Receptor 2 (SSTR2), Stromal cell-derived factor 1 (SDF-1), systemic lupus erythematosus (SLE), Tachykinin Receptor 3 (TACR3), Threshold cycles (Ct), time-resolved FRET (TR-FRET), tumor microenvironment (TME), Vascular endothelial growth factor (VEGF), vascular smooth muscle cells (VSMC), WHIM syndrome (Warts, Hypogammaglobulinemia. Infections, and Myelokathexis), green fluorescence protein (GFP), and yellow fluorescence protein (YFP).DETAILED DESCRIPTION OF THE INVENTION

[0081] Blood cells play a crucial part in maintaining the health and viability of animals, including humans. White blood cells, part of the body’s immune system that help the body fight infection and other diseases, include granulocytes (neutrophils, eosinophils and basophils / mast cells), monocytes / macrophages, as well the lymphocytes (T and B cells) of the immune sy stem. White blood cells are continuously replaced via the hematopoietic system, by the action of colony stimulating factor (CSF) and various cytokines on stem cells and progenitor cells in hematopoietic tissues.

[0082] The chemokine receptor CXCR4 and its natural ligand stromal cell derived factor- 1 (SDF-1) appear to be important in the development and maturation of blood cells (for reviews, see Maekawa, T., et al.. Internal Med. (2000) 39:90-100; Nagasawa, T., et al., Int. J. Hematol. (2000) 72:408-411). This is demonstrated by reports that CXCR4 or SDF-1 knock-out mice exhibit embry onic lethality and hematopoietic defects (Ma, Q., et al., Proc. Natl. Acad. Sci USA (1998) 95:9448-9453; Tachibana, K , et al., Nature (1998) 393:591-594; Zou. Y-R., et al., Nature (1998) 393:595-599).

[0083] SDF-1 is also an important chemoattractant, signaling via the CXCR4 receptor, for several other more committed progenitors and mature blood cells including T-lymphocytes and monocytes (Bleul, C., et al., J. Exp. Med. (1996) 184: 1101-1109), pro-and pre-B lymphocytes (Fedyk, E. R., et al., J. Leukoc. Biol. (1999) 66:667-673; Ma, Q., et al., Immunity (1999) 10:463- 471) and megakaryocytes (Hodohara, K., et al., Blood (2000) 95:769-775; Riviere, C., et al.. Blood (1999) 95: 1511-1523; Majka, M„ et al., Blood (2000) 96:4142-4151; Gear, A., et al., Blood (2001) 97:937-945; Abi-Younes, S„ et al, Circ. Res. (2000) 86: 131-138).

[0084] Thus, it appears that SDF-1 is able to control the positioning and differentiation of cells bearing CXCR4 receptors whether these cells are stem cells (i.e.. cells which are CD34+).progenitor cells (which, being either CD34+ or CD34-, can result in the formation of specified ty pes of colonies in response to particular stimuli), and / or cells that are somewhat more differentiated (i.e., lymphocytes, monocytes, and other immune cells).

[0085] As used herein, an “immune cell” includes all white blood cells, for example, lymphocytes, neutrophils, and monocytes / macrophages. As used herein, a “lymphocyte” includes T cells. B cells, and NK cells. Each immune cell and / or lymphocyte subtype can be identified using cell markers described herein, according to the gating strategy described in FIGs. 17A- 17M, and according to markers and gating strategies known to one of skill in the art.

[0086] The term “CXCR4” as used herein refers to C-X-C Motif Chemokine Receptor 4, also identified by unique database identifiers (IDs) and alternate names as shown in Table 1 (Chatteijee et al., 2014; Debnath et al., 2013; Domanska et al.. 2013; Guo et al.. 2016; Peled et al., 2012; Roccaro et al., 2014; Walenkamp et al., 2017). Table 1 also provides the nomenclature of CXCR4 and GPCRx that form heteromers with CXCR4 and synergistically enhance Ca2+ response upon co-stimulation with both agonists.Table 1*GCID: Genecards identificationHGNC: HUGO Gene Nomenclature Committee

[0087] The terms “GPCRx” as used herein refers to GPCRs that were used in this study to investigate if these GPCRs interact with CXCR4 and show properties distinct from those of individual protomers, including ADCYAP1R1 (ADCYAP Receptor Type I). ADORA2B (Adenosine A2b Receptor), AD0RA3 (Adenosine A3 Receptor), ADRB2 (Adrenoceptor Beta 2), APLNR (Apelin Receptor), C5AR1 (Complement C5a Receptor 1), CALCR (Calcitonin Receptor), CCR5 (Chemokine (C-C Motif) Receptor 5), CHRM1 (Cholinergic Receptor Muscarinic 1), GALR1 (Galanin Receptor 1), EDNRB (Endothelin Receptor Type B). HRH1 (Histamine Receptor Hl), MLNR (Motilin Receptor), NTSR1 (Neurotensin Receptor 1). PTGER2 (Prostaglandin E Receptor 2), PTGER3 (Prostaglandin E Receptor 3), SSTR2 (Somatostatin Receptor 2), and TACR3 (Tachykinin Receptor 3), also identified by unique database identifiers (IDs) and alternate names as shown in Table 1.

[0088] The term "‘inhibitor' as used herein refers to molecule that inhibits or suppresses the enhanced function of a CXCR4, a beta-adrenergic receptor, a GPCR, a heteromer of CXCR4 and a beta-adrenergic receptor, and / or a CXCR4-GPCRx heteromer. Non-limiting examples of theinhibitor of the invention that can be used for mobilization of cells include GPCRx antagonist, GPCRx inverse agonist. GPCRx positive and negative allosteric modulator, CXCR4-GPCRx heteromer-specific antibody or its antigen biding portions including single-domain antibody-like scaffolds, bivalent ligands which have a pharmacophore selective for CXCR4 joined by a spacer arm to a pharmacophore selective for GPCRx, bispecific antibody against CXCR4 and GPCRx, trispecific antibodies such as those targeting CXCR4 and GPCRx and a T-cell antigen (e.g., CD3), radiolabeled CXCR4 ligand linked with GPCRx ligand, and small molecule ligands that inhibit heteromer-selective signaling. Certain examples of inhibitors against GPCRx that form heteromers with CXCR4 and enhance Ca2+ response upon co-stimulation with both agonists are listed in Table 2.

[0089] The term "antagonist" as used herein refers to a type of receptor ligand or drug that blocks or dampens a biological response by binding to and blocking a receptor, also called blockers. Antagonists have affinity but no efficacy for their cognate receptors, and their binding disrupts the interaction and inhibit the function of an agonist or inverse agonist at the cognate receptors. Certain examples of antagonists against GPCRx that form heteromers with CXCR4 and enhance Ca2+ response upon co-stimulation with both agonists are listed in Table 2.Table 2. Examples of inhibitors against CXCR4 and ADRB2

[0090] The term “heteromer” as used herein refers to macromolecular complex composed of at least two GPCR units [protomers] with biochemical properties that are demonstrably different from those of its individual components. Heteromerization can be evaluated by in situ hybridization, immunohistochemistry, RNAseq, Reverse transcription-quantitative PCR (RT- qPCR, realtime PCR). microarray, proximity ligation assay (PLA), time-resolved FRET (TR- FRET), whole-body Single-photon emission computed tomography (SPECT) or Positron Emission Tomography / Computed Tomography (PET / CT).

[0091] The phrase "effective amount" as used herein refers to an amount sufficient to effect beneficial or desired results. An effective amount can be administered in one or more administrations, applications or dosages. Such delivery is dependent on a number of variables including the time period for which the individual dosage unit is to be used, the bioavailability of the agent, the route of administration, etc.

[0092] The phrase “therapeutically effective amount” as used herein refers to the amount of a therapeutic agent (e.g.. an inhibitor, an antagonist, or any other therapeutic agent provided herein) which is sufficient to reduce, ameliorate, and / or prevent the severity and / or duration of a cancer and / or a symptom related thereto. A therapeutically effective amount of a therapeutic agent can be an amount necessary for the reduction, amelioration, or prevention of the advancement or progression of a cancer, reduction, amelioration, or prevention of the recurrence, development or onset of a cancer, and / or to improve or enhance the prophylactic or therapeutic effect of another therapy (e.g., a therapy other than the administration of a inhibitor, an antagonist, or any other therapeutic agent provided herein).

[0093] The phrase “therapeutic agent” refers to any agent that can be used in the treatment, amelioration, prevention, or management of a cancer and / or a symptom related thereto. In certain embodiments, a therapeutic agent refers to an inhibitor of CXCR4-GPCRx heteromer of the invention. A therapeutic agent can be an agent which is well known to be useful for or has been or is currently being used for the treatment, amelioration, prevention, or management of a cancer and / or a symptom related thereto.

[0094] The phrase “intracellular Ca2+ assay,” “calcium mobilization assay,” or their variants as used herein refer to cell-based assay to measure the calcium flux associated with GPCR activation or inhibition. The method utilizes a calcium sensitive fluorescent dye that is taken up into the cytoplasm of most cells. The dye binds the calcium released from intracellular store and itsfluorescence increases. The change in the fluorescence intensity is directly correlated to the amount of intracellular calcium that is released into cytoplasm in response to ligand activation of the receptor of interest.

[0095] The phrase “proximity-based assay” as used herein refers to biophysical and biochemical techniques that are able to monitor proximity and / or binding of two protein molecules in vitro (in cell lysates) and in live cells, including bioluminescence resonance energy transfer (BRET), fluorescence resonance energy transfer (FRET), bimolecular fluorescence complementation (BiFC), Proximity ligation assay (PLA), cysteine crosslinking, and co-immunoprecipitation (Ferre et al., 2009; Gomes et al., 2016).

[0096] Disclosed herein are methods and compositions directed to mobilizing a cell in a subj ect by blocking CXCR4, a beta-adrenergic receptor, a GPCR, or any combination thereof. In some embodiments, the cell is an immune cell. In some embodiments, the cell is a lymphocyte. In some embodiments, the mobilizing a cell in a subject comprises blocking CXCR4. In some embodiments, the mobilizing a cell in a subject comprises blocking a beta-adrenergic receptor. In some embodiments, the mobilizing a cell in a subject comprises blocking a GPCR. In some embodiments, the mobilizing a cell in a subj ect comprises blocking CXCR4 and a beta-adrenergic receptor. In some embodiments, the mobilizing a cell in a subject comprises blocking CXCR4 and a GPCR. In some embodiments, the mobilizing a cell in a subject comprises blocking a CXCR4-GPCR heteromer.

[0097] Disclosed herein are methods of mobilizing a lymphocyte or an immune cell in a subject, the method comprising: blocking CXCR4 signaling and beta-adrenergic receptor signaling in the subject. Also disclosed herein are methods of inducing cell mobilization in a subject, the method comprising: blocking CXCR4 signaling and beta-adrenergic receptor signaling in the subject. In embodiments, the blocking beta-adrenergic receptor signaling is performed before the blocking CXCR4 signaling. In some embodiments, the blocking beta-adrenergic receptor signaling is performed at a first specific time interval before the blocking CXCR4 signaling. In some embodiments, the first specific time interval is between 5 minutes to 10 minutes, 10 minutes to 20 minutes, 20 minutes to 30 minutes, 30 minutes to 40 minutes, 40 minutes to 50 minutes, 50 minutes to 1 hour, 1 hour to 2 hours. 2 hours to 3 hours. 3 hours to 4 hours. 4 hours to 5 hours. 5 hours to 6 hours, 6 hours to 12 hours, 12 hours to 24 hours, 1 day to 2 days, 2 days to 3 days, 3 days to 4 days, 4 days to 5 days, 5 days to 6 days, 6 days to 7 days, 7 days to 8 days, 8 days to 9 days, 9 days to 10 days, 10 days to 11 days, 11 days to 12 days, 12 days to 13 days, 13 days to14 days, or 14 days or more. In embodiments, the blocking beta-adrenergic receptor signaling continues after the blocking CXCR4 signaling is terminated. In some embodiments, the blocking beta-adrenergic receptor signaling continues for a second specific time interval after the blocking CXCR4 signaling is terminated. In some embodiments, the second specific time interval is between 5 minutes to 10 minutes, 10 minutes to 20 minutes, 20 minutes to 30 minutes, 30 minutes to 40 minutes, 40 minutes to 50 minutes, 50 minutes to 1 hour, 1 hour to 2 hours, 2 hours to 3 hours, 3 hours to 4 hours, 4 hours to 5 hours, 5 hours to 6 hours, 6 hours to 12 hours. 12 hours to 24 hours, 1 day to 2 days, 2 days to 3 days, 3 days to 4 days, 4 days to 5 days, 5 days to 6 days, 6 days to 7 days, 7 days to 8 days, 8 days to 9 days, 9 days to 10 days, 10 days to 11 days, 11 days to 12 days, 12 days to 13 days, 13 days to 14 days, or 14 days or more.

[0098] In embodiments, the blocking CXCR4 signaling comprises administering a CXCR4 inhibitor to the subject.

[0099] In embodiments, the blocking beta-adrenergic receptor signaling comprises administering a beta-adrenergic receptor inhibitor to the subject. In embodiments, the blocking CXCR4 signaling comprises administering a CXCR4 inhibitor to the subject and the blocking beta-adrenergic receptor signaling comprises administering a beta-adrenergic receptor inhibitor to the subject. In embodiments, the cell is an immune cell. In some embodiments, the cell is a lymphocyte.

[0100] Disclosed herein are methods of mobilizing a lymphocyte or an immune cell in a subject, the method comprising: administering a beta-adrenergic receptor inhibitor and a CXCR4 inhibitor to the subject. Also disclosed herein are methods of inducing lymphocyte mobilization in a subject, the method comprising: administering a beta-adrenergic receptor inhibitor and a CXCR4 inhibitor to the subject. In some embodiments, the administering the beta-adrenergic receptor inhibitor is performed before the administering the CXCR4 inhibitor. In some embodiments, the administering the beta-adrenergic receptor inhibitor is performed at a first specific time interval before the administering the CXCR4 inhibitor. In some embodiments, the first specific time interval is between 5 minutes to 10 minutes, 10 minutes to 20 minutes, 20 minutes to 30 minutes, 30 minutes to 40 minutes, 40 minutes to 50 minutes, 50 minutes to 1 hour, 1 hour to 2 hours, 2 hours to 3 hours. 3 hours to 4 hours. 4 hours to 5 hours, 5 hours to 6 hours, 6 hours to 12 hours. 12 hours to 24 hours, 1 day to 2 days, 2 days to 3 days, 3 days to 4 days, 4 days to 5 days, 5 days to 6 days, 6 days to 7 days, 7 days to 8 days, 8 days to 9 days, 9 days to 10 days, 10 days to 11 days, 11 days to 12 days, 12 days to 13 days, 13 days to 14 days, or 14 days or more. Inembodiments, the administering the beta-adrenergic receptor inhibitor continues after the administering the CXCR4 inhibitor is terminated. In some embodiments, the administering the beta-adrenergic receptor inhibitor continues for a second specific time interval after the administering the CXCR4 inhibitor is terminated. In some embodiments, the second specific time interval is between 5 minutes to 10 minutes, 10 minutes to 20 minutes, 20 minutes to 30 minutes, 30 minutes to 40 minutes, 40 minutes to 50 minutes, 50 minutes to 1 hour, 1 hour to 2 hours, 2 hours to 3 hours. 3 hours to 4 hours. 4 hours to 5 hours, 5 hours to 6 hours, 6 hours to 12 hours. 12 hours to 24 hours, 1 day to 2 days, 2 days to 3 days, 3 days to 4 days, 4 days to 5 days, 5 days to 6 days, 6 days to 7 days, 7 days to 8 days, 8 days to 9 days, 9 days to 10 days, 10 days to 11 days, 11 days to 12 days, 12 days to 13 days, 13 days to 14 days, or 14 days or more.

[0101] In embodiments, the beta-adrenergic receptor inhibitor is an ADRB2 inhibitor. In embodiments, the beta-adrenergic receptor inhibitor is selected from the group consisting of alprenolol, atenolol, betaxolol, bupranolol, butoxamine, carazolol, carvedilol, CGP 12177, cicloprolol, ICI 118551, ICYP, labetalol, levobetaxolol, levobunolol, LK 204-545, metoprolol, nadolol, NIHP. NIP, propafenone, propranolol, sotalol, SR59230A, and timolol. In embodiments, the beta-adrenergic receptor inhibitor is selected from the group consisting of propranolol, nadolol, and ICI 118551. In embodiments, the beta-adrenergic receptor inhibitor is propranolol.

[0102] In embodiments, the CXCR4 inhibitor is selected from the group consisting of ALX40- 4C. AMD070 (AMD11070, X4P-001), AMD3100 (plerixafor), AMD3465, ATI 2341. BKT140 (BL-8040; TF14016; 4F-Benzoyl-TN14003), CTCE-9908, CX549„D-[Lys3] GHRP-6, FC122. FC131, GMI-1359, GSK812397, GST-NT21MP, isothiourea- la, isothiourea- It (ITlt), KRH- 1636, KRH-3955, LY2510924, MSX-122, N-[l lC]Methyl-AMD3465, POL6326, SDF-1 1- 9[P2G] dimer, SDF1 P2G, T134, T140, T22. TC 14012, TG-0054 (Burixafor), USL311. viral macrophage inflammatory protein-II (vMIP-II), WZ811, [64Cu]-AMD3100, [64Cu]-AMD3465, [68Ga]pentixafor, [90Y]pentixather, [99mTc]02-AMD3100, [177Lu]pentixather, and 508MC1 (Compound 26). Burixafor is also referred to as GPC-100 or TG-0054. Plerixafor is also referred to as AMD3100 or Mozobil. In embodiments, the CXCR4 inhibitor is selected from the group consisting of AD-214, AMD070 (AMD11070, X4P-001), AMD3100 (plerixafor), BKT140 (BL- 8040; TF14016; 4F-Benzoyl-TN14003). CTCE-9908. LY2510924, LY2624587, T140. TG-0054 (Burixafor), PF-06747143, POL6326, and ulocuplumab (MDX1338 / BMS-936564). In embodiments, the CXCR4 inhibitor is TG-0054 (burixafor). In embodiments, the CXCR4inhibitor is AMD3100 (plerixafor). In embodiments, the CXCR4 inhibitor is ulocuplumab (MDX1338 / BMS-936564).

[0103] In embodiments, the administering of the CXCR4 inhibitor and the ADRB2 inhibitor to the subject comprises administering TG-0054 (burixafor) and propranolol. In embodiments, the administering of the CXCR4 inhibitor and the ADRB2 inhibitor to the subject comprises administering AMD3100 (plerixafor) and propranolol. In embodiments, the administering of the CXCR4 inhibitor and the ADRB2 inhibitor to the subject comprises administering ulocuplumab (MDX1338 / BMS-936564) and propranolol.

[0104] In embodiments, the method further comprises administering G-CSF to the subject. In embodiments, the administering the beta-adrenergic receptor inhibitor and the CXCR4 inhibitor to the subject is performed in the absence of G-CSF. Disclosed herein are methods of mobilizing a lymphocyte or an immune cell in a subject, the method comprising: administering a CXCR4 inhibitor and G-CSF to the subject, in the absence of a beta-adrenergic receptor inhibitor. Also disclosed herein are methods of inducing lymphocy te or immune cell mobilization in a subject, the method comprising: administering a CXCR4 inhibitor and G-CSF to the subject, in the absence of a beta-adrenergic receptor inhibitor. In some embodiments, the administering the CXCR4 inhibitor to the subject comprises administering TG-0054 (burixafor). In embodiments, the administering the CXCR4 inhibitor to the subject comprises administering AMD3100 (plerixafor). In embodiments, the administering the CXCR4 inhibitor to the subject comprises administering ulocuplumab (MDX1338 / BMS-936564).

[0105] In embodiments, the administering a combination of the CXCR4 inhibitor and the G- CSF induces an enhanced amount of lymphocyte or immune cell mobilization relative to the amount of lymphocyte or immune cell mobilization induced by the CXCR4 inhibitor only. In embodiments, the administering a combination of the CXCR4 inhibitor and the G-CSF mobilizes a lymphocyte or an immune cell by an amount enhanced relative to the amount of cell mobilization induced by the CXCR4 inhibitor only. In some embodiments, the enhanced amount of lymphocyte or immune cell mobilization relative to the amount of lymphocy te or immune cell mobilization induced by the CXCR4 inhibitor only is between 1.1 -fold to 1.2-fold, 1.2-fold to 1.3-fold, 1.3-fold to 1.4-fold, 1.4-fold to 1.5-fold. 1.5-fold to 1.6-fold, 1.6-fold to 1.7-fold, 1.7- fold to 1.8-fold, 1.8-fold to 1.9-fold, 1.9-fold to 2-fold, 2-fold to 2.5-fold, 2.5-fold to 3-fold, 3- fold to 4-fold, 4-fold to 5-fold, 5-fold to 10-fold, or 10-fold or more. In some embodiments, the enhanced amount of lymphocyte or immune cell mobilization relative to the amount oflymphocyte or immune cell mobilization induced by the CXCR4 inhibitor only is between 5%- 10% more, 10%-20% more, 20%-30% more, 30%-40% more, 40%-50% more, 50%-60% more, 60%-70% more, 70%-80% more, 80%-90% more, 90%-100% more, 100%-120% more, 120%- 140% more, 140%-160% more, 160%-180% more, 180%-200% more, 200%-250% more, 250%- 300% more, 300%-400% more, 400%-500% more, 500%-750% more, 750%-1000% more, or 1000% or more.

[0106] In embodiments, the administering a combination of the CXCR4 inhibitor and the beta- adrenergic receptor inhibitor induces an enhanced amount of lymphocyte or immune cell mobilization relative to the amount of lymphocyte or immune cell mobilization induced by the CXCR4 inhibitor only. In embodiments, the administering a combination of the CXCR4 inhibitor and the beta-adrenergic receptor inhibitor mobilizes a lymphocyte or an immune cell by an amount enhanced relative to the amount of lymphocyte or immune cell mobilization induced by the CXCR4 inhibitor only. In some embodiments, the enhanced amount of lymphocyte or immune cell mobilization relative to the amount of ly mphocyte or immune cell mobilization induced by the CXCR4 inhibitor only is between 1.1-fold to 1.2-fold, 1.2-fold to 1.3-fold, 1.3- fold to 1.4-fold, 1.4-fold to 1.5-fold, 1.5-fold to 1.6-fold. 1.6-fold to 1.7-fold, 1.7-fold to 1.8-fold. 1.8-fold to 1.9-fold, 1.9-fold to 2-fold, 2-fold to 2.5-fold, 2.5-fold to 3-fold, 3-fold to 4-fold, 4- fold to 5-fold, 5-fold to 10-fold, or 10-fold or more. In some embodiments, the enhanced amount of lymphocyte or immune cell mobilization relative to the amount of lymphocyte or immune cell mobilization induced by the CXCR4 inhibitor only is between 5%-10% more. 10%-20% more. 20%-30% more, 30%-40% more, 40%-50% more, 50%-60% more, 60%-70% more, 70%-80% more, 80%-90% more, 90%-100% more, 100%- 120% more, 120%- 140% more, 140%- 160% more, 160%-180% more, 180%-200% more, 200%-250% more, 250%-300% more, 300%-400% more, 400%-500% more, 500%-750% more, 750%-1000% more, or 1000% or more.

[0107] In embodiments, the administering a combination of the CXCR4 inhibitor, the beta- adrenergic receptor inhibitor, and the G-CSF induces an enhanced amount of lymphocyte or immune cell mobilization relative to the amount of lymphocyte or immune cell mobilization induced by the CXCR4 inhibitor and the beta-adrenergic receptor inhibitor only. In embodiments, the administering a combination of the CXCR4 inhibitor, the beta-adrenergic receptor inhibitor, and the G-CSF mobilizes a lymphocyte or an immune cell by an amount enhanced relative to the amount of lymphocyte or immune cell mobilization induced by the CXCR4 inhibitor and the beta-adrenergic receptor inhibitor only. In embodiments, the administering a combination of TG-0054 (burixafor) and the G-CSF induces an enhanced amount of lymphocyte or immune cell mobilization relative to the amount of lymphocyte or immune cell mobilization induced by AMD3100 (plerixafor) and the G-CSF. In embodiments, the administering a combination of the TG-0054 (burixafor) and the G-CSF mobilizes a lymphocyte or an immune cell by an amount enhanced relative to the amount of lymphocyte or immune cell mobilization induced by the AMD3100 (plerixafor) and the G-CSF. In some embodiments, the enhanced amount of lymphocyte or immune cell mobilization relative to the amount of lymphocyte or immune cell mobilization induced by the CXCR4 inhibitor only is between 1.1 -fold to 1.2-fold, 1.2-fold to 1.3-fold, 1.3-fold to 1.4-fold, 1.4-fold to 1.5-fold, 1.5-fold to 1.6-fold, 1.6-fold to 1.7-fold, 1.7- fold to 1.8-fold, 1.8-fold to 1.9-fold, 1.9-fold to 2-fold, 2-fold to 2.5-fold, 2.5-fold to 3-fold, 3- fold to 4-fold, 4-fold to 5-fold, 5-fold to 10-fold, or 10-fold or more. In some embodiments, the enhanced amount of lymphocyte or immune cell mobilization relative to the amount of lymphocyte or immune cell mobilization induced by the CXCR4 inhibitor only is between 5%- 10% more, 10%-20% more, 20%-30% more, 30%-40% more, 40%-50% more, 50%-60% more, 60%-70% more, 70%-80% more, 80%-90% more, 90%-100% more, 100%-120% more. 120%- 140% more, 140%- 160% more, 160%- 180% more, 180%-200% more, 200%-250% more, 250%- 300% more, 300%-400% more, 400%-500% more, 500%-750% more, 750%-1000% more, or 1000% or more. In embodiments, an enhanced amount of lymphocyte or immune cell mobilization or apheresis is measured by a method selected from the group consisting of complete blood count (CBC) analysis, flow cytometry, and colony forming unit (CFU) assay. In embodiments, the enhanced amount of cell mobilization or apheresis is measured by flow cytometry. In embodiments, the flow cytometry' is performed on (Lin-Scal+c-Kit+) LSK cells. In embodiments, the enhanced amount of ly mphocyte or immune cell mobilization or apheresis is measured by colony forming unit (CFU) assay.

[0108] In embodiments, the subject has a CXCR4 protomer in the lymphocyte or immune cell. In embodiments, the subject has an ADRB2 protomer in the lymphocyte or immune cell. In embodiments, the subject has a CXCR4 protomer and an ADRB2 protomer in the lymphocyte or immune cell. In embodiments, the subject has a CXCR4-ADRB2 heteromer in the lymphocyte or immune cell. In embodiments, i) the CXCR4-ADRB2 heteromer has an enhanced amount of downstream calcium mobilization relative to downstream calcium mobilization from a CXCR4 protomer or ADRB2 protomer; and ii) the administered combination of inhibitors suppresses theenhanced downstream calcium mobilization from said CXCR4-ADRB2 heteromer in the lymphocyte.

[0109] In embodiments, the cell is an immune cell. In embodiments, the immune cell is a white blood cell. In embodiments, the white blood cell is a lymphocyte. In embodiments, the lymphocyte is selected from the group consisting of a T cell, a B cell, and a natural killer (NK) cell. In embodiments, the lymphocyte is a T cell. In embodiments, the lymphocyte is a natural killer (NK) cell. In embodiments, the white blood cell is a granulocyte. In embodiments, the granulocyte is selected from the group consisting of a neutrophile, an eosinophile, and a basophile. In embodiments, the granulocyte is a neutrophile. In embodiments, the white blood cell is a monocyte. In embodiments, the immune cell is mobilized from bone marrow to peripheral blood. In embodiments, the immune cell is mobilized from lymph node to peripheral blood. In embodiments, the mobilized immune cell is used for adoptive cell therapy (ACT). In embodiments, the adoptive cell therapy (ACT) is chimeric antigen receptor (CAR) T cell therapy. In embodiments, the adoptive cell therapy (ACT) is netural killer (NK) cell therapy. In embodiments, the adoptive cell therapy (ACT) is engineered T-cell receptor (TCR) therapy. In embodiments, the adoptive cell therapy (ACT) is tumor-infiltrating lymphocyte (TIL) therapy. In embodiments, the adoptive cell therapy (ACT) include cell and gene therapy (CGT). In certain embodiments, the CTG involves the use of vectors, including lentiviral vectors, adeno-associated virus vectors, anellovirus vectors, and the likes, whether engineered or natural. In embodiments, a lentiviral vector is injected into a patient to deliver genetic information into the patient.

[0110] In some embodiments of the present invention, the mobilizing a lymphocyte or an immune cell in a subject comprises blocking CXCR4. Many antiviral agents that inhibit HIV replication via inhibition of CXCR4, the co-receptor required for fusion and entry of T-tropic HIV strains, also inhibit the binding and signaling induced by the natural ligand, the chemokine SDF-1 (also known as CXCL12). While not wishing to be bound by any theory, the agents which inhibit the binding of SDF-1 to CXCR4 can effect an increase in mobilization of lymphocyte or immune cells to the periphery by virtue of such inhibition.[OHl] In some embodiments of the present invention, mobilizing a lymphocyte or an immune cell in a subject comprises blocking a CXCR4-GPCR heteromer. Various CXCR4-GPCR heteromers with distinct physiological and pharmacological properties have been reported, but their roles in lymphocyte or immune cell mobilization or possibilities for developing lymphocyteor immune cell mobilization therapeutics targeting CXCR4-GPCR heteromers have not been clearly understood or appreciated.

[0112] In the art, GPCRs were believed to function as monomers that interact with heterotrimeric G proteins upon ligand binding, and drugs were developed based on monomeric or homomeric GPCRs (Milligan 2008). Recently, this view changed drastically based on discoveries that GPCRs can form heteromers, and that heteromerization is obligatory for some GPCRs. GPCR heteromerization is known to alter GPCR maturation and cell surface delivery, ligand binding affinity, signaling intensity and pathways, as well as receptor desensitization and recycling (Terrillon and Bouvier 2004; Ferre et al., 2010; Rozenfeld and Devi 2010; Gomes et al., 2016; Farran 2017). Different GPCR heteromers display distinct functional and pharmacological properties, and GPCR heteromerization can vary depending on cell types, tissues, and diseases or pathological conditions (Terrillon and Bouvier 2004; Ferre et al., 2010;Rozenfeld and Devi 2010; Gomes et al., 2016; Farran 2017). GPCR heteromerization is currently regarded as a general phenomenon, and deciphering GPCR heteromerization opens new avenues for understanding receptor function, physiology, roles in diseases and pathological conditions. Accordingly, identification of GPCR heteromers and their functional properties offers new opportunity for developing new pharmaceuticals or finding new use of old drugs with fewer side effects, higher efficacy, and increased tissue selectivity (Ferre et al., 2010; Rozenfeld and Devi 2010; Farran 2017).

[0113] Apheresis is a standard practice to obtain a larger number of immune cells as starting material for Adoptive Cell Therapy (ACT), which is a treatment based on transferring cells into a patient (1-3). Apheresis may involve passing the blood of a patient through an apparatus that separates out one particular constituent and returns the remainder to the blood circulation of the patient. Apheresis is thus an extracorporeal therapy. Depending on the substance being removed, different processes are employed in apheresis. If separation by density is required, centrifugation is the most common method. Other methods involve absorption onto beads coated with an absorbent material and filtration. The centrifugation method can be divided into two basic categories: continuous flow centrifugation (CFC) and intermittent flow centrifugation.

[0114] CFC historically required two venipunctures as the “continuous"’ means that the blood was collected, spun, and returned simultaneously. Newer systems can use a single venipuncture. The main advantage of CFC is the low extracorporeal volume (calculated by volume of the apheresis chamber, the donor’s hematocrit, and total blood volume of the donor) used in theprocedure, which may be advantageuous in the elderly and for children. Intermitent flow centrifugation works in cycles, taking blood, spinning / processing the blood, then giving back the unused parts to the donor in a bolus. The main advantage is a single venipuncture site. To stop the blood from coagulating, anticoagulant is automatically mixed with the blood as it is punped from the body into the apheresis machine.

[0115] The various apheresis techniques may be used whenever the removed constituent is causing severe symptoms of disease in a patient. Generally, apheresis has to be performed fairly often and is an invasive procedure. It is therefore generally employed if other means to control a particular disease have failed, or if the symptoms are of such a nature that w aiting for medication to become effective w ould cause suffering or risk of complications. Apheresis techniques include: (1) plasma exchange - removal of the liquid portion of blood to remove harmful substances, where the plasma is replaced with a replacement solution; (2) LDL apheresis - removal of low density lipoprotein in patients with familial hypercholesterolemia; (3) photopheresis - used to treat graft- versus-host disease, cutaneous T-cell lymphoma, and rejection in heart transplantation; (4) immunoadsorbtion with Staphylococcal protein A-agarose column - removal of allo- and autoantibodies (in autoimmune diseases, transplant rejection, hemophilia) by directing plasma through protein A-agarose columns (Protein A is a cell wall component produced by several strains of Staphylococcus aureus which binds to the Fc region of IgG); (5) leukocytapheresis - removal of malignant white blood cells in people with leukemia and very high white blood cell counts causing symptoms; (6) erythrocytapheresis - removal of erythrocytes (red blood cells) in people with iron overload as a result of Hereditary haemochromatosis or transfusional iron overload; (7) thrombocytapheresis - removal of platelets in people with symptoms from extreme elevations in platelet count such as those with essential thrombocythemia or polycythemia vera; and (8) leukapheresis - separates out excess white blood cells of leukemia patients while recycling the remainder of their blood.

[0116] Apheresis is a difficult procedure, inconvenient and expensive. With the rapid growth of ACTs including CAR-T, CAR-NK, Tumor-Infiltrating Lymphocyte (TIL), and engineered T cell receptor (TCR), the need for apheresis technology for the routine production of pure immune cells is increasing (2). The industry that supplies GMP-grade starting materials for ACTs is also growing rapidly (4-5). Thus, immune cell mobilization technologies that can control types of immune cells and improve the yield of apheresis have become important.

[0117] Enhanced lymphocyte or immune cell mobilization methods as disclosed herein, can further augment or facilitate the conventional apheresis procedure. In a specific embodiment, lymphocyte or immune cell mobilization is particularly beneficial for the apheresis technique of leukapheresis. In some embodiments, administering a CXCR4 antagonist to a subject further enhances apheresis by augmenting lymphocyte or immune cell mobilization. In some embodiments, administering a beta-adrenergic receptor antagonist in conjunction with a CXCR4 antagonist to a subject further enhances apheresis by augmenting lymphocyte or immune cell mobilization, and / or replacing the G-CSF component of the treatment regime with anon-selective beta-blocker, such as propranolol. In some embodiments, the augmentation of lymphocyte or immune cell mobilization in turn benefits manufacturing of CAR-T cells for cancer immunotherapy.

[0118] Disclosed herein are methods of enhancing apheresis in a subject, the method comprising: blocking CXCR4 signaling and beta-adrenergic receptor signaling in the subject. Also disclosed herein are methods of enhancing apheresis by inducing lymphocyte or immune cell mobilization in a subject, the method comprising: blocking CXCR4 signaling and beta- adrenergic receptor signaling in the subject. Further disclosed herein are methods of enhancing apheresis by mobilizing a lymphocyte or an immune cell in a subject, the method comprising: blocking CXCR4 signaling and beta-adrenergic receptor signaling in the subject. In embodiments, the blocking beta-adrenergic receptor signaling is performed before the blocking CXCR4 signaling. In some embodiments, the blocking beta-adrenergic receptor signaling is performed at a first specific time interval before the blocking CXCR4 signaling. In some embodiments, the first specific time interval is between 5 minutes to 10 minutes, 10 minutes to 20 minutes, 20 minutes to 30 minutes, 30 minutes to 40 minutes, 40 minutes to 50 minutes, 50 minutes to 1 hour, 1 hour to 2 hours. 2 hours to 3 hours. 3 hours to 4 hours. 4 hours to 5 hours. 5 hours to 6 hours, 6 hours to 12 hours, 12 hours to 24 hours, 1 day to 2 days, 2 days to 3 days, 3 days to 4 days, 4 days to 5 days, 5 days to 6 days, 6 days to 7 days, 7 days to 8 days, 8 days to 9 days, 9 days to 10 days, 10 days to 11 days, 11 days to 12 days, 12 days to 13 days, 13 days to 14 days, or 14 days or more. In embodiments, the blocking beta-adrenergic receptor signaling continues after the blocking CXCR4 signaling is terminated. In some embodiments, the blocking beta-adrenergic receptor signaling continues for a second specific time interval after the blocking CXCR4 signaling is terminated. In some embodiments, the second specific time interval is between 5 minutes to 10 minutes, 10 minutes to 20 minutes, 20 minutes to 30 minutes, 30 minutesto 40 minutes, 40 minutes to 50 minutes, 50 minutes to 1 hour, 1 hour to 2 hours, 2 hours to 3 hours, 3 hours to 4 hours, 4 hours to 5 hours, 5 hours to 6 hours, 6 hours to 12 hours, 12 hours to 24 hours, 1 day to 2 days, 2 days to 3 days, 3 days to 4 days, 4 days to 5 days, 5 days to 6 days, 6 days to 7 days, 7 days to 8 days, 8 days to 9 days, 9 days to 10 days, 10 days to 11 days, 11 days to 12 days, 12 days to 13 days, 13 days to 14 days, or 14 days or more.

[0119] In embodiments, the blocking CXCR4 signaling comprises administering a CXCR4 inhibitor to the subject.

[0120] Disclosed herein are methods of enhancing apheresis in a subject, the method comprising: administering a beta-adrenergic receptor inhibitor and a CXCR4 inhibitor to the subject. Also disclosed herein are methods of enhancing apheresis by inducing lymphocyte or immune cell mobilization in a subject, the method comprising: administering a beta-adrenergic receptor inhibitor and a CXCR4 inhibitor to the subject. Further disclosed herein are methods of enhancing apheresis by mobilizing a lymphocyte or an immune cell in a subject, the method comprising: administering a beta-adrenergic receptor inhibitor and a CXCR4 inhibitor to the subject. In some embodiments, the administering the beta-adrenergic receptor inhibitor is performed at a first specific time interval before the administering the CXCR4 inhibitor. In some embodiments, the first specific time interval is between 5 minutes to 10 minutes, 10 minutes to 20 minutes, 20 minutes to 30 minutes, 30 minutes to 40 minutes, 40 minutes to 50 minutes, 50 minutes to 1 hour, 1 hour to 2 hours. 2 hours to 3 hours, 3 hours to 4 hours, 4 hours to 5 hours, 5 hours to 6 hours. 6 hours to 12 hours, 12 hours to 24 hours, 1 day to 2 days, 2 days to 3 days. 3 days to 4 days, 4 days to 5 days, 5 days to 6 days, 6 days to 7 days, 7 days to 8 days, 8 days to 9 days, 9 days to 10 days, 10 days to 11 days, 11 days to 12 days, 12 days to 13 days, 13 days to 14 days, or 14 days or more. In embodiments, the administering the beta-adrenergic receptor inhibitor continues after the administering the CXCR4 inhibitor is terminated. In some embodiments, the administering the beta-adrenergic receptor inhibitor continues for a second specific time interval after the administering the CXCR4 inhibitor is terminated. In some embodiments, the second specific time interval is between 5 minutes to 10 minutes, 10 minutes to 20 minutes, 20 minutes to 30 minutes, 30 minutes to 40 minutes, 40 minutes to 50 minutes, 50 minutes to 1 hour, 1 hour to 2 hours. 2 hours to 3 hours. 3 hours to 4 hours. 4 hours to 5 hours. 5 hours to 6 hours, 6 hours to 12 hours, 12 hours to 24 hours, 1 day to 2 days, 2 days to 3 days, 3 days to 4 days, 4 days to 5 days, 5 days to 6 days, 6 days to 7 days, 7 days to 8 days, 8 days to 9days, 9 days to 10 days, 10 days to 11 days, 11 days to 12 days, 12 days to 13 days, 13 days to 14 days, or 14 days or more.

[0121] In embodiments, the beta-adrenergic receptor inhibitor is an ADRB2 inhibitor. In embodiments, the beta-adrenergic receptor inhibitor is selected from the group consisting of alprenolol, atenolol, betaxolol, bupranolol, butoxamine, carazolol, carvedilol, CGP 12177, cicloprolol, ICI 118551, ICYP, labetalol, levobetaxolol, levobunolol, LK 204-545, metoprolol, nadolol, NIHP. NIP, propafenone, propranolol, sotalol, SR59230A, and timolol. In embodiments, the beta-adrenergic receptor inhibitor is selected from the group consisting of propranolol, nadolol, and ICI 118551. In embodiments, the beta-adrenergic receptor inhibitor is propranolol.

[0122] In embodiments, the CXCR4 inhibitor is selected from the group consisting of ALX40- 4C. AMD070 (AMD11070, X4P-001), AMD3100 (plerixafor). AMD3465, ATI 2341. BKT140 (BL-8040; TF14016; 4F-Benzoyl-TN 14003), CTCE-9908, CX549„D-[Lys3] GHRP-6, FC122, FC131, GMI-1359, GSK812397, GST-NT21MP, isothiourea- la, isothiourea- It (H t), KRH- 1636, KRH-3955, LY2510924, MSX-122, N-[l lC]Methyl-AMD3465, POL6326, SDF-1 1- 9[P2G] dimer, SDF1 P2G, T134, T140, T22. TC 14012, TG-0054 (Burixafor). USL311, viral macrophage inflammatory protein-II (vMIP-II). WZ811, [64Cu]-AMD3100, [64Cu]-AMD3465. [68Ga]pentixafor, [90Y]pentixather, [99mTc]02-AMD3100, [177Lu]pentixather, and 508MC1 (Compound 26). In embodiments, the CXCR4 inhibitor is selected from the group consisting of AD-214, AMD070 (AMD11070, X4P-001), AMD3100 (plerixafor), BKT140 (BL-8040; TF14016; 4F-Benzoyl-TN14003), CTCE-9908, LY2510924, LY2624587, T140, TG-0054 (Burixafor), PF-06747143, POL6326, and ulocuplumab (MDX1338 / BMS-936564). In embodiments, the CXCR4 inhibitor is TG-0054 (burixafor). In embodiments, the CXCR4 inhibitor is AMD3100 (plerixafor). In embodiments, the CXCR4 inhibitor is ulocuplumab (MDX1338 / BMS-936564).

[0123] In embodiments, the administering the CXCR4 inhibitor to the subject comprises administering TG-0054 (burixafor) and propranolol. In embodiments, the administering the CXCR4 inhibitor to the subject comprises administering AMD3100 (plerixafor) and propranolol. In embodiments, the administering the CXCR4 inhibitor to the subject comprises administering ulocuplumab (MDX1338 / BMS-936564) and propranolol.

[0124] In embodiments, the method further comprises administering G-CSF to the subject. In embodiments, the administering the beta-adrenergic receptor inhibitor and the CXCR4 inhibitor to the subject is performed in the absence of G-CSF. Disclosed herein are methods of enhancingapheresis in a subject, the method comprising: administering a CXCR4 inhibitor and G-CSF to the subject, in the absence of a beta-adrenergic receptor inhibitor. Further disclosed herein are methods of enhancing apheresis by inducing lymphocyte or immune cell mobilization in a subject, the method comprising: administering a CXCR4 inhibitor and G-CSF to the subject, in the absence of a beta-adrenergic receptor inhibitor. Also disclosed herein are methods of enhancing apheresis by mobilizing a lymphocyte or an immune cell in a subject, the method comprising: administering a CXCR4 inhibitor and G-CSF to the subject, in the absence of a beta- adrenergic receptor inhibitor. In embodiments, the administering a combination of the CXCR4 inhibitor and the G-CSF induces an enhanced amount of apheresis relative to the amount of apheresis induced by the CXCR4 inhibitor only. In embodiments, the administering a combination of the CXCR4 inhibitor and the beta-adrenergic receptor inhibitor induces an enhanced amount of apheresis relative to the amount of apheresis induced by the CXCR4 inhibitor only. In embodiments, the administering a combination of the CXCR4 inhibitor and the beta-adrenergic receptor inhibitor, and the G-CSF induces an enhanced amount of apheresis relative to the amount of apheresis induced by the CXCR4 inhibitor and the beta-adrenergic receptor inhibitor only. In embodiments, the administering a combination of the TG-0054 (burixafor) and the G-CSF induces an enhanced amount of apheresis relative to the amount of apheresis induced by the AMD3100 (plerixafor) and the G-CSF. In some embodiments, the enhanced amount of lymphocyte or immune cell mobilization relative to the amount of lymphocyte or immune cell mobilization induced by the CXCR4 inhibitor only is between 1.1- fold to 1.2-fold, 1.2-fold to 1.3-fold, 1.3 -fold to 1.4-fold, 1.4-fold to 1.5-fold, 1.5-fold to 1.6- fold, 1.6-fold to 1.7-fold, 1.7-fold to 1.8-fold, 1.8-fold to 1.9-fold, 1.9-fold to 2-fold, 2-fold to 2.5-fold, 2.5-fold to 3-fold, 3-fold to 4-fold, 4-fold to 5-fold, 5-fold to 10-fold, or 10-fold or more. In some embodiments, the enhanced amount of lymphocyte or immune cell mobilization relative to the amount of lymphocyte or immune cell mobilization induced by the CXCR4 inhibitor only is between 5%-10% more, 10%-20% more, 20%-30% more, 30%-40% more, 40%-50% more, 50%-60% more, 60%-70% more, 70%-80% more, 80%-90% more, 90%-100% more, 100%- 120% more, 120%- 140% more, 140%- 160% more, 160%- 180% more, 180%-200% more, 200%- 250% more, 250%-300% more, 300%-400% more, 400%-500% more, 500%-750% more, 750%- 1 00% more, or 1000% or more. In embodiments, an enhanced amount of lymphocyte or immune cell mobilization or apheresis is measured by a method selected from the group consisting of complete blood count (CBC) analysis, flow cytometry, and colony forming unit (CFU) assay. Inembodiments, the enhanced amount of lymphocyte or immune cell mobilization or apheresis is measured by flow cytometry. In embodiments, the flow cytometry is performed on (Lin-Scal+c- Kit+) LSK cells. In embodiments, the enhanced amount of lymphocyte or immune cell mobilization or apheresis is measured by colony forming unit (CFU) assay.

[0125] Further information regarding the ADRB2, evaluated herein as forming heteromers with CXCR4, are detailed below:

[0126] ADRB2 — The beta-2 adrenergic receptor ([32 adrenoreceptor), also known as ADRB2. is a cell membrane-spanning beta-adrenergic receptor that interacts with epinephrine, a hormone and neurotransmitter (ligand synonym, adrenaline) whose signaling, via a downstream L-type calcium channel interaction, mediates physiologic responses such as smooth muscle relaxation and bronchodilation (Gregorio et al., 2017). ADRB2 functions in muscular system such as smooth muscle relaxation, motor nerve terminals, glycogenolysis and in circulatory system such as heart muscle contraction, cardiac output increase. In the normal eye, beta-2 stimulation by salbutamol increases intraocular pressure via net. In digestive system, the ADRB2 induces glycogenolysis and gluconeogenesis in liver and insulin secretion from pancreas (Fitzpatrick, 2004).

[0127] ADRB2 signaling in the cardiac myocyte is modulated by interactions with CXCR4 (LaRocca et al., 2010). Norepinephrine attenuates CXCR4 expression and the corresponding invasion of MDA-MB-231 breast cancer cells via ADRB2 (Wang et al., 2015a). ADRB2 is expressed in several cancers such as pancreatic, prostate (Braadland et al., 2014; Xu et al., 2017), renal and breast cancer (Choy et al., 2016).

[0128] Alternative methods for detecting heteromer formation include, but are not limited to: immunostaing (Bushlin et al., 2012; Decaillot et al., 2008); immunoelectron microscopy (Fernandez-Duenas et al., 2015); BRET (Pfleger and Eidne, 2006); Time-resolved FRET assays (Fernandez-Duenas et al., 2015); In Situ Hybridization (He et al.. 2011); FRET (Lohse et al., 2012); p-arrestin recruitment assay using GPCR heteromer identification technology (GPCR- HIT, Dimerix Bioscience) (Mustafa and Pfleger, 2011) using BRET, FRET, BiFC, Bimolecular Luminescence Complementation, enzy me fragmentation assay, and Tango Tango GPCR assay system (Thermo Fisher Scientific) (Mustafa, 2010); PRESTO-Tango system (Kroeze et al.. 2015); regulated secretion / aggregation technology (ARIAD Pharmaceuticals) (Hansen et al., 2009); Receptor Selection and Amplification Technology (ACADIA Pharmaceuticals) (Hansen et al., 2009); DimerScreen (Cara Therapeutics) (Mustafa, 2010); Dimer / interacting protein translocation assay (Patobios) (Mustafa, 2010); Co-immunoprecipitation (Abd Alla et al., 2009);GPCR internalization assays using surface enzyme-linked immunosorbent assay (ELISA) (Decaillot et al., 2008) or Flow Cytometry (Law et al., 2005); Whole Cell Phosphorylation Assays (Pfeiffer et al., 2002); and Proximity-ligation assay (PLA) (Fredenck et al., 2015).

[0129] Alternative methods for detecting changes in pharmacological properties, signaling properties, and / or trafficking properties, in cells expressing both CXCR4 and GPCRx include, but are not limited to: Radioligand Binding Assays (Bushlin et al., 2012; Pfeiffer et al., 2002); Cell Surface Biotinylation and Immunoblotting (He et al., 2011); immunostaing (Bushlin et al.. 2012; Decaillot et al., 2008); immunoelectron microscopy (Fernandez-Duenas et al., 2015); [35S]GTP S Binding assays (Bushlin et al., 2012); Calcuim imaging or assays using dyes such as Fura 2-acetomethoxy ester (Molecular Probes), Fluo-4 NW calcium dye (Thermo Fisher Scientific), or FLIPR5 dye (Molecular Devices); cAMP assays using radioimmunoassay kit (Amersham Biosciences); AlphaScreen (PerkinElmer Life Sciences); Parameter Cyclic AMP Assay (R&D Systems); femto cAMP kit (Cisbio); cAMP Direct Immunoassay Kit (Calbiochem) or GloSensor cAMP assay (Promega); GTPase assay (Pello et al., 2008); PKA activation (Stefan et al., 2007); ERK1 / 2 and / or Akt / PKB Phosphorylation Assays (Callen et al, 2012); Src and STAT3 phosphorylation assays (Rios et al. 2006); reporter assays such as cAMP response element (CRE); nuclear factor of activated T-cells response element (NFAT-RE); serum response element (SRE); serum response factor response element (SRF-RE); and NF-KB-response element luciferase reporter assays; Secreted alkaline phosphatase Assay (Decaillot et al, 2011); Measurement of Inositol 1 -Phosphate Production Using TR-FRET or [3H]myo-Inositol (Mustafa et al, 2012); RT-qPCR for measuring downstream target gene expression (Mustafa et al, 2012); and Adenylyl Cyclase Activity' (George et al, 2000); next generation sequencing (NGS); and any other assay that can detect a change in receptor function as a result of receptor heterodimerization.

[0130] The phrase ‘"protein-protein interaction inhibitor,” “PPI inhibitor.” or their variants as used herein refer to any molecules that can interfere with protein-protein interactions. Proteinprotein interaction, unlike enzyme-substrate interaction involving well-defined binding pockets, is a transient interaction or association between proteins over relatively large areas and is often driven by electrostatic interactions, hydrophobic interactions, hydrogen bonds, and / or Van der Waals forces. PPI inhibitors may include, but not limited to, membrane-permeable peptides or lipid fused to a peptide sequence that disrupts the GPCR heteromeric interface, for example, transmembrane helix, intracellular loop, or C-terminal tail of GPCRx. The PPI inhibitor of the CXCR4-GPCRx heteromer. for example, may be a membrane-permeable peptide or cell-penetrating peptide (CPP) conjugated with peptide that targets the CXCR4-GPCRx heteromeric interface(s), or may be a cell-penetrating lipidated peptide targeting the CXCR4-GPCRx heteromeric interface(s).

[0131] For example, the membrane-permeable peptide or cell-penetrating peptide includes: HIV-1 TAT peptides, such as TAT48-60 and TAT49-57; Penetratins, such as pAntp(43-58); Poly arginines (Rn such as R5 to R12); Diatos peptide vector 1047 (DPV1047, Vectocell®); MPG (HIV gp41 fused to the nuclear localization signal (NLS) of the SV40 large T antigen); Pep-1 (tryptophan-rich cluster fused to the NLS of SV40 large T antigen); pVEC peptide (vascular endothelial cadherin); p!4 alternative reading frame (ARF) protein-based ARF(l-22); N- terminus of the unprocessed bovine prion protein BPrPr(l-28); Model amphipathic peptide (MAP); Transportans; Azurin-derived p28 peptide; amphipathic p-sheet peptides, such as VT5; proline-rich CPPs, such as Bac 7 (Bacl-24); hydrophobic CPPs, such as C105Y derived from al -Antitrypsin; PFVYLI derived from synthetic C105Y; Pep-7 peptide (CHL8 peptide phage clone); and modified hydrophobic CPPs, such as stapled peptides and prenylated peptides (Guidotti et al., 2017; Kristensen et al., 2016). The membrane-permeable peptide or cellpenetrating peptide can further include, for example, TAT-derived cell-penetrating peptides, signal sequence-based (e.g., NLS) cell-penetrating peptides, hydrophobic membrane translocating sequence (MTS) peptides, and arginine-rich molecular transporters. The cellpenetrating lapidated peptide includes, for example, pepducins, such as ICL 1 / 2 / 3, C-tail- short palmitoylated peptides (Covic et al., 2002; O'Callaghan et al., 2012).

[0132] The peptide(s) that target the CXCR4-GPCRx heteromeric interface may be, for example, a transmembrane domain of CXCR4, transmembrane domain of GPCRx, intracellular loop of CXCR4, intracellular loop of GPCRx, C-terminal domain of CXCR4, or C-terminal domain of GPCRx., extracellular loop of CXCR4, extracellular loop of GPCRx, N-terminal region of CXCR4, or N-terminal region of GPCRx.CXCR4 Inhibitor and G-CSF

[0133] Disclosed herein, in some embodiments, are methods of mobilizing a lymphocyte or an immune cell in a subject, the method comprising administering, alone or in combination, a CXCR4 inhibitor and G-CSF to the subject.

[0134] Disclosed herein, in some embodiments, are methods for treatment of cancer comprising administering, alone or in combination, a CXCR4 inhibitor and G-CSF to a subject.

[0135] Disclosed herein, in some embodiments, are methods of enhancing leukapheresis byinducing lymphocyte or immune cell mobilization in a subject, the method comprising administering, alone or in combination, a CXCR4 inhibitor and G-CSF to the subject.

[0136] In some embodiments, the CXCR4 inhibitor and the G-CSF are administered to the subj ect in the absence of a beta-adrenergic receptor inhibitor. In some embodiments, the method comprises administering the G-CSF to the subject before administering the CXCR4 inhibitor to the subject. In some embodiments, the method comprises administering a combination of the CXCR4 inhibitor and G-CSF to the subject, and wherein the administering of the combination of the CXCR4 inhibitor and G-CSF to the subject induces an enhanced amount of lymphocyte mobilization relative to an amount of lymphocyte mobilization induced by administration of G- CSF only to the subject. In some embodiments, the method comprises administering a combination of the CXCR4 inhibitor and G-CSF to the subject, and wherein the administering of the combination of the CXCR4 inhibitor and G-CSF to the subject induces an enhanced amount of lymphocyte mobilization relative to an amount of lymphocy te mobilization induced by administration of the CXCR4 inhibitor only to the subject.

[0137] In some embodiments, the CXCR4 inhibitor is selected from the group consisting of ALX40-4C, AMD070 (AMD11070, X4P-001), AMD3100 (plerixafor), AMD3465, ATI 2341, BKT140 (BL-8040; TF14016; 4F-Benzoyl-TN 14003), CTCE-9908, CX549„D-[Lys3] GHRP-6, FC122. FC131, GMI-1359, GSK812397, GST-NT21MP, isothiourea- la, isothiourea- It (ITlt), KRH-1636, KRH-3955, LY2510924, MSX-122, N-[HC]Methyl-AMD3465, POL6326. SDF-1 1-9[P2G] dimer, SDF1 P2G, T134, T140, T22, TC 14012, TG-0054 (Burixafor), USL311, viral macrophage inflammatory protein-II (vMIP-II), WZ811, [64Cu]-AMD3100, [64Cu]-AMD3465, [68Ga]pentixafor, [90Y]pentixather, [99mTc]02-AMD3100, [177Lu]pentixather, and 508MC1 (Compound 26). In some embodiments, the CXCR4 inhibitor is selected from the group consisting of AD-214, AMD070 (AMD11070, X4P-001), AMD3100 (plerixafor), BKT140 (BL- 8040; TF14016; 4F-Benzoyl-TN14003), CTCE-9908, LY2510924, LY2624587, T140, TG-0054 (Burixafor), PF-06747143, POL6326, and ulocuplumab (MDX1338 / BMS-936564). In some embodiments, the CXCR4 inhibitor is TG-0054 (burixafor).

[0138] In some embodiments, the method comprises administering a combination of burixafor and G-CSF to the subject, and wherein the administering of the combination of burixafor and G- CSF to the subject induces an enhanced amount of lymphocyte mobilization relative to an amount of lymphocyte mobilization induced by administration of G-CSF only to the subject. In someembodiments, the method comprises administering a combination of burixafor and G-CSF to the subject, and wherein the administering of the combination of the burixafor and G-CSF to the subject induces an enhanced amount of lymphocyte mobilization relative to an amount of lymphocyte mobilization induced by administration of burixafor only to the subject. In some embodiments, the enhanced amount of lymphocyte mobilization is measured by a method selected from the group consisting of complete blood count (CBC) analysis, flow cytometry, and colony forming unit (CFU) assay. In some embodiments, the enhanced amount of lymphocyte mobilization is measured by CBC analysis.

[0139] In some embodiments, the subject has a CXCR4 protomer in the lymphocyte. In some embodiments, the lymphocyte or immune cell is selected from the group consisting of a T cell, B cell, a natural killer (NK) cell, a neutrophil, eosinophil, and a basophil.

[0140] In some embodiments, the lymphocyte or immune cell is mobilized for treatment of a condition selected from the group consisting of neurological disorder, cardiac ischemia, myocardial infarction, diabetes, tissue repair, bone and cartilage disease, autoimmune disease, graft versus host disease, Crohn's disease, multiple sclerosis, systemic lupus erythematosus, and systemic sclerosis. In some embodiments, the lymphocyte or immune cell is mobilized for treatment of a cancer. In some embodiments, the cancer is non-Hodgkin lymphoma (NHL), acute myeloid leukemia (AML), acute lymphoblastic leukemia (ALL), or multiple myeloma (MM). In some embodiments, the cancer is multiple myeloma (MM). In some embodiments, the lymphocyte or immune cell is mobilized for adoptive cell therapy (ACT). In some embodiments, the adoptive cell therapy (ACT) is chimeric antigen (CAR) T cell therapy. In some embodiments, the cancer is non-Hodgkin lymphoma (NHL), acute myeloid leukemia (AML), acute lymphoblastic leukemia (ALL), or multiple myeloma (MM). In some embodiments, the cancer is multiple myeloma (MM). In some embodiments, the treatment comprises leukapheresis. In some embodiments, the treatment comprises adoptive cell therapy (ACT). In some embodiments, the adoptive cell therapy (ACT) is chimeric antigen (CAR) T cell therapy.

[0141] Disclosed herein, in some embodiments, is a composition or compositions for mobilizing a lymphocyte or an immune cell in a subject comprising, alone or in combination, a CXCR4 inhibitor and G-CSF.

[0142] Disclosed herein, in some embodiments, is a composition or compositions for treatment of cancer comprising, alone or in combination, a CXCR4 inhibitor and G-CSF.

[0143] Disclosed herein, in some embodiments, is a composition or compositions for enhancing leukapheresis by inducing lymphocyte or immune cell mobilization in a subject comprising, alone or in combination, a CXCR4 inhibitor and G-CSF.

[0144] In some embodiments, the composition does not include a beta-adrenergic receptor inhibitor. In some embodiments, the composition further includes a beta-adrenergic receptor inhibitor.

[0145] In some embodiments, the CXCR4 inhibitor is selected from the group consisting of ALX40-4C, AMD070 (AMD11070, X4P-001), AMD3100 (plerixafor), AMD3465, ATI 2341, BKT140 (BL-8040; TF14016; 4F-Benzoyl-TN 14003), CTCE-9908, CX549„D-[Lys3] GHRP-6, FC122, FC131, GMI-1359, GSK812397, GST-NT21MP, isothiourea- la, isothiourea- It (ITlt), KRH-1636, KRH-3955, LY2510924, MSX-122, N-[HC]Methyl-AMD3465, POL6326. SDF-1 1-9[P2G] dimer, SDF1 P2G, T134, T140, T22, TC 14012, TG-0054 (Burixafor), USL311, viral macrophage inflammatory protein-II (vMIP-II), WZ811, [64Cu]-AMD3100, [64Cu]-AMD3465, [68Ga]pentixafor, [90Y]pentixather, [99mTc]02-AMD3100, [177Lu]pentixather, and 508MC1 (Compound 26). In some embodiments, the CXCR4 inhibitor is selected from the group consisting of AD-214, AMD070 (AMD ! 1070, X4P-001). AMD3100 (plenxafor), BKT140 (BL- 8040; TF14016; 4F-Benzoyl-TN14003), CTCE-9908, LY2510924, LY2624587, T140, TG-0054 (Burixafor), PF-06747143, POL6326, and ulocuplumab (MDX1338 / BMS-936564). In some embodiments, the CXCR4 inhibitor is TG-0054 (burixafor).

[0146] In some embodiments, the subject has a CXCR4 protomer in the lymphocyte. In some embodiments, the lymphocyte or immune cell is selected from the group consisting of a T cell, a B cell, and a natural killer (NK) cell, a neutrophil, a eosinophil, and a basophil. In some embodiments, the lymphocyte or immune cell is mobilized for treatment of a condition selected from the group consisting of neurological disorder, cardiac ischemia, myocardial infarction, diabetes, tissue repair, bone and cartilage disease, autoimmune disease, graft versus host disease, Crohn's disease, multiple sclerosis, systemic lupus erythematosus, and systemic sclerosis. In some embodiments, the lymphocyte or immune cell is mobilized for treatment of a cancer. In some embodiments, the cancer is non-Hodgkin lymphoma (NHL), acute myeloid leukemia (AML), acute lymphoblastic leukemia (ALL), or multiple myeloma (MM). In some embodiments, the cancer is multiple myeloma (MM). In some embodiments, the lymphocyte is mobilized for adoptive cell therapy (ACT). In some embodiments, the adoptive cell therapy (ACT) is chimeric antigen (CAR) T cell therapy.

[0147] In some embodiments, the cancer is non-Hodgkin lymphoma (NHL), acute myeloid leukemia (AML), acute lymphoblastic leukemia (ALL), or multiple myeloma (MM). In some embodiments, the cancer is multiple myeloma (MM).

[0148] In some embodiments, the treatment comprises leukapheresis. In some embodiments, the treatment comprises adoptive cell therapy (ACT). In some embodiments, the adoptive cell therapy (ACT) is chimeric antigen (CAR) T cell therapy.

[0149] Disclosed herein, in some embodiments, is a pharmaceutical composition or pharmaceutical compositions for mobilizing a lymphocyte or an immune cell in a subject comprising, alone or in combination, a CXCR4 inhibitor and G-CSF, and a pharmaceutically acceptable excipient.

[0150] Disclosed herein, in some embodiments, is a pharmaceutical composition or pharmaceutical compositions for treatment of cancer comprising, alone or in combination, a CXCR4 inhibitor and G-CSF, and a pharmaceutically acceptable excipient.

[0151] Disclosed herein, in some embodiments, is a pharmaceutical composition or pharmaceutical compositions for enhancing leukapheresis by inducing lymphocyte or immune cell mobilization in a subject comprising, alone or in combination, a CXCR4 inhibitor and G- CSF, and a pharmaceutically acceptable excipient.

[0152] In some embodiments, the pharmaceutical composition does not include a beta- adrenergic receptor inhibitor. In some embodiments, the pharmaceutical composition further includes a beta-adrenergic receptor inhibitor.

[0153] In some embodiments, the CXCR4 inhibitor is selected from the group consisting of ALX40-4C, AMD070 (AMD11070, X4P-001), AMD3100 (plenxafor), AMD3465, ATI 2341, BKT140 (BL-8040; TF14016; 4F-Benzoyl-TN14003), CTCE-9908, CX549„D-[Lys3] GHRP-6, FC122. FC131, GMI-1359, GSK812397, GST-NT21MP, isothiourea- la, isothiourea- It (ITlt), KRH-1636, KRH-3955, LY2510924, MSX-122, N-[HC]Methyl-AMD3465, POL6326, SDF-1 1-9[P2G] dimer, SDF1 P2G, T134, T140, T22, TC 14012, TG-0054 (Burixafor), USL311, viral macrophage inflammatory7protein-II (vMIP-II), WZ811, [64Cu]-AMD3100, [64Cu]-AMD3465, [68Ga]pentixafor, [90Y]pentixather, [99mTc]02-AMD3100. [177Lu]pentixather, and 508MC1 (Compound 26). In some embodiments, the CXCR4 inhibitor is selected from the group consisting of AD-214, AMD070 (AMD1 1070, X4P-001 ), AMD3100 (plerixafor), BKT140 (BL- 8040; TF14016; 4F-Benzoyl-TN14003), CTCE-9908, LY2510924, LY2624587, T140, TG-0054(Burixafor), PF-06747143, POL6326, and ulocuplumab (MDX1338 / BMS-936564). In some embodiments, the CXCR4 inhibitor is TG-0054 (burixafor).

[0154] In some embodiments, the subject has a CXCR4 protomer in the lymphocyte. In some embodiments, the lymphocyte or immune cell is selected from the group consisting of a T cell, a B cell, and a natural killer (NK) cell, a neutrophil, an eosinophil, and a basophil. In some embodiments, the lymphocyte or immune cell is mobilized for treatment of a condition selected from the group consisting of neurological disorder, cardiac ischemia, myocardial infarction, diabetes, tissue repair, bone and cartilage disease, autoimmune disease, graft versus host disease, Crohn's disease, multiple sclerosis, systemic lupus erythematosus, and systemic sclerosis. In some embodiments, the lymphocyte or immune cell is mobilized for treatment of a cancer. In some embodiments, the cancer is non-Hodgkin lymphoma (NHL), acute myeloid leukemia (AML), acute lymphoblastic leukemia (ALL), or multiple myeloma (MM). In some embodiments, the cancer is multiple myeloma (MM). In some embodiments, the lymphocyte or immune cell is mobilized for adoptive cell therapy (ACT). In some embodiments, the adoptive cell therapy (ACT) is chimeric antigen (CAR) T cell therapy.

[0155] In some embodiments, the cancer is non-Hodgkin lymphoma (NHL), acute myeloid leukemia (AML), acute lymphoblastic leukemia (ALL), or multiple myeloma (MM). In some embodiments, the cancer is multiple myeloma (MM). In some embodiments, the treatment comprises leukapheresis. In some embodiments, the treatment comprises adoptive cell therapy (ACT). In some embodiments, the adoptive cell therapy (ACT) is chimeric antigen (CAR) T cell therapy.Applications

[0156] In some embodiments of the present disclosure, the lymphocyte or immune cell is mobilized for use in adoptive cell therapy (ACT). The adoptive cell therapy may be allogeneic (off-the-shelf) or autologous adoptive cell therapy. In some embodiments, the adoptive cell therapy may be CAR T cell therapy, CAR-NK cell therapy, tumor-infiltrating lymphocyte (TIL) therapy, yo T cell (CAR- y5 T cell) therapy, macrophage (CAR-M) therapy, or dendritic cell therapy.

[0157] In some embodiments of the present disclosure, the immune cell is mobilized for use in stem cell therapy, such as hematopoietic stem cell therapy. The stem cell therapy may be used for applications such as, but not limited to, cancer treatment, regenerative medicine, treatment of blood disorders, treatment of immunodeficiencies, or treatment of infectious diseases.

[0158] In some embodiments of the present disclosure, the lymphocyte or immune cell is mobilized for use in immunotherapy. The immunotherapy may be used for treatment of various disorders such as, but not limited to, immunodeficiencies, autoimmune diseases, and solid and blood cancers. Types of solid and blood cancers may include, but are not limited to, lung cancer, melanoma, breast cancer, bladder cancer, kidney cancer, colorectal cancer, head and neck cancer, cervical cancer, esophageal cancer, liver cancer, stomach cancer, brain cancer, leukemia, lymphoma, multiple myeloma, pancreatic cancer, prostate cancer, sarcoma, and skin cancer.

[0159] In some embodiments of the present disclosure, the immune cell is mobilized for treatment of an autoimmune disease. Autoimmune diseases may include, but are not limited to, Crohn’s disease, ulcerative colitis, multiple sclerosis, Guillain-Barre syndrome, lupus, rheumatoid arthritis, type 1 diabetes, psoriasis, celiac disease. Graves’ disease, inflammatory bowel disease (IBD), chronic inflammatory demyelinating polyneuropathy (CIPD), vitiligo, scleroderma, Hashimoto’s thyroiditis, Addison’s disease, alopecia areata, pernicious anemia, systemic lupus erythematosus, Warts, Hypogammaglobulinemia, Immunodeficiency, and Myelokathexis (WHIM) syndrome, and Goodpasture syndrome.

[0160] In some embodiments of the present disclosure, the immune cell is mobilized for the treatment of neurological disorders. The neurological disorders may include, but are not limited to, Alzheimer’s disease, Parkinson’s disease, Huntington’s disease, and multiple sclerosis. In some embodiments, the immune cell is mobilized for treatment of amyloidosis, including those treatments involving immunotherapy, adoptive cell therapy (ACT), or stem cell transplant such as autologous stem cell transplantation (ASCT).

[0161] In some embodiments of the present disclosure, the immune cell is mobilized for the treatment of a blood disorder. Blood disorders may include, but are not limited to, sickle cell disease, anemia, hemophila, von Willebrand disease, clotting disorders, and blood cancers.

[0162] In some embodiments of the present disclosure, the immune cell is mobilized for treatment of neutropenia or conditions in which neutropenia is problematic. Such conditions may include, without limitation, aplastic anemia, myelodysplastic syndrome, and autoimmune diseases. Autoimmune diseases may include, but are not limited to, Crohn’s disease, ulcerative colitis, multiple sclerosis, Guillain-Barre syndrome, lupus, rheumatoid arthritis, type 1 diabetes, psoriasis, celiac disease, Graves’ disease, inflammatory bowel disease (IBD), chronic inflammatory demyelinating polyneuropathy (CIPD), vitiligo, scleroderma, Hashimoto’s thyroiditis, Addison's disease, alopecia areata, pernicious anemia, systemic lupuserythematosus, Warts, Hypogammaglobulinemia, Immunodeficiency, and Myelokathexis (WHIM) syndrome, and Goodpasture syndrome. In some embodiments of the present disclosure, the immune cell is mobilized for replenishing immune cells in patients undergoing chemotherapy or radiation.

[0163] In some embodiments, the immune cell is mobilized from healthy volunteers. For example, the immune cell may be mobilized from a healthy volunteer in a blood donation process to provide a “leukopak” which is a sterile pack of apheresis product enriched with leukocytes for various applications including research, clinical applications, immunotherapy, drug development, and cell therapy with the benefit of mobilization without the side effects (e.g., bone pain) that may result with G-CSF use.

[0164] In some embodiments of the present disclosure, the immune cell is mobilized for use in apheresis. The apheresis may include, for example, plasmapheresis, plateletpheresis, erythrocytapheresis, or leukapheresis. In some embodiments of the present disclosure, the immune cell is mobilized for tissue repair and regeneration, treatment of bone and cartilage diseases, or treatment of cardiac disorders such as. but not limited to, cardiac repair, heart failure, cardiac fibrosis, or cardiac ischemia. In some embodiments of the present disclosure, the immune cell is mobilized for treatment of an inflammatory disease such as, but not limited to, encephalitis, neuritis, or meningitis. In some embodiments of the present disclosure, the immune cell is mobilized for treatment of an infectious disease. Examples of infectious diseases include, but are not limited to, HIV, tuberculosis, malaria, ziki vims, or COVID-19. In some embodiments of the present disclosure, the immune cell is mobilized for use in organ transplantation, bone marrow transplantation, or blood transfusion. In some embodiments of the present disclosure, the immune cell is mobilized for use in immunomodulation.

[0165] In some embodiments of the present disclosue. the immune cell is mobilized for use in scientific research and development. In some embodiments of the present disclosure, the immune cell is mobilized for use in diagnostics or diagnostic imaging. Diagnostic imaging may involve, for example, monitoring immune cell trafficking or signaling.

[0166] It is understood that modifications which do not substantially affect the activity of the various embodiments of this invention are also provided within the definition of the invention provided herein. Accordingly, the following examples are intended to illustrate but not limit the invention disclosed herein.EXAMPLESExample 1. In vivo pharamacology studies evaluating combination of GPC-100 and propranolol for lymphocyte and immune cell mobilization.

[0167] The chemokine receptor 4 (CXCR4) signaling is involved in mobilization and retention of hematopoietic stem cells (HSC) in the bone marrow. GPC-100, a potent CXCR4 antagonist, also known as TG-0054 or Burixafor, was shown to be a safe stem cell mobilizer in healthy human volunteers (clinical trial: NCT00822341). When combined with G-CSF in multiple myeloma and non-Hodgkin’s lymphoma patients (clinical trial: NCT01458288), GPC-100 more quickly mobilized HSC than the standard of care treatment with G-CSF and Plerixafor. In our previous preclinical studies in mice, administration of GPC-100 induced rapid mobilization of WBC in peripheral blood within 30 min with a peak mobilization at 2 hours. This effect was further enhanced by propranolol pretreatment for 7 days. CXCR4 is also expressed by immune cells. Therefore, in the present study, whether propranolol and GPC-100 combination results in enhanced mobilization of lymphoid immune cell subtypes was investigated. Lymphoid cell mobilizationwas further compared with that of myeloid cells. Combination treatment of propranolol and GPC-100 with and without G-CSF will be administered clinically for stem cell mobilization in aphase II clinical trial (NCT05561751). The purpose of this study is to understand the immune cell subtypes mobilized by the combination of GPC-100 and propranolol to elucidate their potential role in autologous and allogenic transplants as well as adoptive cell therapies.Materials and Methods

[0168] Animals: C57BL / 6 or BALB / c mice (female, 6-9 weeks old) were purchased from Jackson Laboratory and maintained on a 12-h light / dark cycle with free access to food and water. Mice were housed 4 per cage at the laboratory animal facility' (Explora Biolabs, San Carlos), which is accredited by AAALAC (Association for Assessment and Accreditation of Laboratory Animal Care International) and the IACUC (Institutional Animal Care and Use Committee). During the mobilization study, mice were weighed each day and prior to the treatment, randomized for equal weight distribution.

[0169] Mouse Tumor models: Effect of GPC-100 on mobilization of WBC or immune cells was further investigated in syngeneic mouse tumor models with intact immune system along with naive mice. After the habituation period of 1 week, mice were shaved in the flank area and cancer cells were injected subcutaneously under the flank skin in the volume of 100 uL. Tumor volume was measured every 3-4 days until 1000 mm3 with electronic calipers and data was captured inthe cloud-based data management system called Benchling Studies (Overwatch Research). After 1000 mm3, tumor volume was measured every 2 days. The maximum tumor volume was allowed to be 2000 mm3 as per the animal research guidelines. Following mouse tumor models were used: C57 / BL6 T-cell lymphoma model EL4, Balb / c colon adenocarcinoma model CT26, Balb / c plasmacytoma model J558 and C57 / BL6 fibrosarcoma model MCA205. Following cell number was injected for each model. CT26: 1x106 cells, EL4: 2 x 104 cells. MCA205: 8 x 105 cells. J558: 1x106 cells.

[0170] Compounds and treatment: Propranolol (HY-B0573, Lot# 26496, MedChem Express, Princeton, NJ) was intraperitoneally (IP) administered at 20 mg / kg for seven days once daily. GPC-100 was administered intravenously (IV) once on day 7 at 30 mg / kg following propranolol. Vehicle controls received PBS IV or IP. The mice were divided into the following groups (1) vehicle control (2) GPC-100 (3) Propranolol alone (4) GPC-100 plus propranolol and treated as given in Table 3. To compare mobilization with standards of care, G-CSF (Peprotech, NJ) was administered for 5 days twice daily at 0.1 mg / kg / dose. Some mice also received AMD3100 (5 mg / kg SC) (MedChemExpress, NJ) alone or 12 hours after G-CSF (G-CSF plus AMD3100 combination treatment) or co-administered with propranolol (20 mg / kg IP in balb / c mice) on day 7 (propranolol plus AMD3100). All compounds were reconstituted in PBS. G-CSF was reconstituted once and stored at 4C for up to one week. Other compounds w ere reconstituted fresh daily.Table 3: Dosing regimen

[0171] Sample collection: Peripheral blood was collected via cardiac puncture on day 7 or on the day of single treatment in K3EDTA tubes 2 hours after GPC-100 or PBS administration, 12 hours after G-CSF administration or 1 hour after AMD3100 administration. Approximately 70 uL of blood was used for complete blood count (CBC) analysis. The remaining blood was used for flow cytometry analysis on the same day. In tumor bearing mice, bone marrow and tumor were also collected in addition to the blood. The tumor volumes at the time of sample collections were ~ 1200 mm3 EL4, 150 mm3 CT26, MCA205: 360mm3, J558:500 mm3.

[0172] CBC analysis: Blood samples were processed for CBC analysis using the Abaxis VetScan HM5 hematology analyzer, which reports 18-parameters including WBC, lymphocytes, monocytes and neutrophils.

[0173] Evaluation of immune cells in samples using flow cytometry: Blood was processed with RBC lysis buffer (14 parts ammonium chloride solution, eBioscience to 1 part blood) followed by pre-staining with a Fc-Block and Live / Dead cell stain. This mixture consists of l,000ng / mL Mouse TruStain FcX blocker (BioLegend) and a fixable viability dye eFluor780 (eBioscience) at 10.000X dilution. The cells were then stained with a cocktail of antibodies mentioned in Tables 4 and 5, finally fixed with 2% paraformaldehyde. Samples were acquired with a Cytek Aurora spectral flow cytometer and data analysis was performed with CellEngine cloud-based analysissoftware platform (https: / / cellengine.com). Gating was determined using FMO controls. Cells were gated to remove doublets and non-viable cells. The percentage of CD3, NKp46. F4 / 80, CD l ib and CXCR4 were analyzed as a subset of parent CD45+ cells, whereas the percent of CD4+ and CD8+ were analyzed as a subset of the CD3+ parent population. The total number of mobilized cells was calculated based on the white blood count (WBC) data through a complete blood profile by Abaxis VetScan HM5 Hematology Analyzer. A similar process was followed also for bone marrow and tumor samples. Cells from these samples were extracted and cell clumps were removed using cell strainers (70pm, VWR). Cell suspensions were collected and processed using the same procedure listed above on the same day. PD-1 indicating T cell exhaustion and granzyme B indicating T cell cytotoxicity were analyzed as a subset of CD8 parent population. Foxp3 was analyzed as a subset of CD4 indicating regulatory T cells. Other antibodies used in this study, as well as the gating strategy is elaborated in Table 6 and FIG. 17A- 17M. The total number of mobilized cells was calculated based on the absolute WBC count for that sample. For example, CD45 count / mL = WBC count x % CD45 and so on.Table 4: Lymphoid PanelTable 5: Myeloid PanelTable 6: Gating strategy

[0174] Statistical analyses: Data analyses were performed using Prism (GraphPad) and all data are presented as mean values (mean ± SEM). Comparison across multiple groups was made using ordinary one-way ANOVA followed by Turkey's multiple comparison test. Comparisons of data across two dosing conditions were made using the Mann-Whitney test. P < 0.05 was considered statistically significant for all tests. * P < 0.5, ** P =< 0.01, *** P =< 0.001, **** P =< 0.0001.Results

[0175] Propranolol enhances GPC-100 induced mobilization of lymphocytes that is significantly greater than the standards of care for mobilization.

[0176] All treatment mobilized significantly greater lymphocytes compared to the vehicle (F=24.06, PO.OOOl) (FIG. 1). Mice that received propranolol pretreatment for 7 days and coadministration of GPC- 100 on the 7th day showed lymphocyte mobilization that was significantly higher than the standards of care G-CSF alone or G-CSF plus AMD3100 as well as GPC-100 and AMD3100 alone (all p values < 0.0001). Propranolol induced a 53% increase in GPC-100 dependent lymphocyte mobilization. This warranted further investigation of the subset of lymphocytes mobilized by propranolol and GPC-100 combination by flow cytometric analysis.

[0177] Propranolol significantly improved mobilization of all immune cell subtypes by GPC-100 in BALB / c mice with preference for mobilization of CD8+ T cells.

[0178] The total number of mobilized CD45+ cells was determined by normalization based on the total WBC count (FIG. 2A-2B). Propranolol and GPC-100 combination mobilized significantly greater WBCs (FIG. 2A) (F=44.64 P<0.0001) and lymphocytes (FIG. 2B) (F=31.44 P<0.0001) compared to GPC-100 alone as well as other treatment groups. Propranolol did not improve AMD3100 induced mobilization significantly. Additionally, propranolol alone did not mobilize either WBC or lymphocytes.

[0179] GPC-100 by itself significantly mobilized leukocytes as assessed by CD45 expression and the CD3 T cells, CD4+T cells, CD8+T cells as well as CDl lb+ myeloid cells and F4 / 80 macrophages (FIG. 3A-3F). Pretreatment with propranolol resulted in a further and moresignificant increase in all these populations while also significantly increasing the CD 19+B cell and NK cell mobilization (FIG. 3G-3H). GPC-100 alone mobilized all cell populations at least by 2-fold when compared to the vehicle (FIG. 4A). The increase in CD8+ T cells with GPC-100 was 3-fold whereas that in CD1 lb+ myeloid cells and F4 / 80+ macrophages was almost 4-fold. Propranolol pretreatment increased GPC-100 induced mobilization of all populations with almost 7-fold increase in CD8+ T cells, CD1 lb+ myeloid cells and F4 / 80+ macrophages (FIG. 4A). In contrast, AMD3100 only produced a 2-fold increase in CD8+T cell mobilization with a slightly more impact of propranolol on mobilization of NK cells (FIG. 4B).

[0180] Bone marrow analyses showed no change in proportions of of immune cells except CDl lb+ population slightly decreasing and CD19+ B cell proportions slightly increasing with single GPC-100 dosing (FIG. 5A-5H). Further analyses will benefit from normalizing the bone marrow total cells, which will reflect the true change in the bone marrow cells.

[0181] GPC-100 and propranolol combination treatment mobilized CD8+ T cells, B cells, and macrophages to peripheral blood in C57 / BL6 mice.

[0182] The total number of mobilized CD45+ cells was determined by normalization based on the total WBC count (FIG. 6A-6C). GPC-100 alone and with propranolol mobilized significantly greater number of CD45+ cells (F=18.2, PO.OOOl) compared to PBS treated mice. This increase was also reflected in the mobilization of F4 / 80+ macrophages (F=85.3, P<0.0001) as well as CD19+ B cells (F=9.5, P=0.0004). GPC-100 mobilized significantly greater CD8+ T cells when combined with propranolol pretreatment (F=5.4, p=0.007) (FIG. 7A-7F). Mann-Whitney T-test. but not AN OVA, indicated that GPC-100 and propranolol mobilized significantly more NK cells (p=0.04). Propranolol without GPC-100 did not have any effect on the immune cell mobilization.

[0183] Propranolol and GPC-100 combination mobilized WBC and immune cells more effectively in tumor bearing mice.

[0184] To determine GPC-100 induced mobilization of immune cells in tumor-bearing mice, CT26 colon cancer model (solid tumor) was utilized. At the time of sample collection, the average tumor volume was 150 mm3. In addition to the cell types described above, this study further analyzed sub-populations of CD8+ T cells such as those expressing the exhaustion marker PD-1 and cytotoxicity marker granzyme B. Mobilization of regulatory T cells was also determined as CD4+T cells expressing Foxp3 and CD25. To determine comparison of lymphoid cell mobilization with myeloid cell mobilization, granulocytic and monocytic myeloid-derived suppressor cells (PMN-MDSC and M-MDSC, respectively) were also analyzed. MDSCs weredefined as CDl lb+ F4 / 80- populations with M-MDSCs further defined as Ly6C+Ly6G- cells and PMN-MDSCs as Ly6ClowLy6G+ cells. Macrophages were characterized as CD1 lb+F4 / 80hi with Ml macrophages expressing CD80 and CD86 whereas M2 macrophages expressing CD 163 and CD206.

[0185] Parallel comparison with naive mice revealed that GPC-100 mobilized WBC, lymphocytes, and neutrophils in both naive and tumor bearing mice, although the baseline circulating cells in the vehicle treated tumor bearing mice appeared to be slightly lower (FIG. 8A-8C). All T cell subtypes; B cells and NK cells were mobilized by GPC-100 in both tumor bearing and naive mice (FIG. 9A-9H). In tumor mice, CD8+T cells were preferentially mobilized over CD4+T cells (although the absolute number of CD4 was higher). Moreover, GPC-100 significantly mobilized CDl lb+ myeloid cells, F4 / 80+ macrophages and M-MDSCs in both naive and tumor mice and Ml macrophages in tumor mice (FIG. 10A-10F). In both naive and tumor-bearing mice, there were more Ml than M2 macrophages and more M-MDSC than PMN- MDSC. This ratio was maintained after GPC-100 treatment.

[0186] Overall, GPC-100 induced mobilization of both myeloid and lymphoid cells was greater in tumor-bearing mice than naive mice (FIG. 11A-11D). While there was a 2-fold change in regulatory T cells, cytotoxic T cells and exhausted T cells in naive mice compared to the vehicle treated naive mice, this was increased to almost 3 -fold change from tumor vehicle controls in mobilization of CD8+T cells (both PD1+ and granzyme B+) and NK cells (FIG. 11A). There was also a 4-fold increase in myeloid cell mobilization (FIG. 11B). Additionally, changes in the tumor from GPC-100 were also analyzed (FIG. 12A-12E). However, whether CXCR4 signaling inhibition by GPC-100 directs immune cells towards the tumor or mobilizes them out of the tumor needs further investigation in a larger study. Effect of propranolol on proliferation, function, differentiation, and mobilization of immune cells will next be determined in this model.

[0187] GPC-100 induced mobilization of immune cells in a mouse lymphoma model (hematological malignancy).

[0188] GPC-100 (30 mg / kg, IV) or PBS was administered to C57 / BL6 mice carry ing a T-cell lymphoma tumor. At the time of sample collection, the average tumor volume was 1300 mm3. GPC-100 induced mobilization of WBC, neutrophils, and lymphocytes in both naive and tumor bearing mice. Lymphoma bearing mice had higher WBC count than naive mice (FIG. 13A). There was an increase in the baseline neutrophil count in peripheral blood of lymphoma mice whereas lymphocytes dominated in the naive mice (FIG. 13B).

[0189] GPC-100 mobilized CD45+ leukocytes and the subsets in both lymphoma-bearing and naive mice. There were more macrophages present in the peripheral blood of lymphoma bearing mice. GPC-100 increased number of circulating macrophages more significantly in lymphoma bearing mice compared to the naive mice. Some tumor-bearing mice had higher CD3+T and CD19+ B cells (FIG. 14A-14E). Furthermore, mobilized immune cells (FIG. 15A-15E) revealed CXCR4 expression (FIG. 15F-15J). WBC, differential, or immune cell counts in peripheral blood were independent of the tumor volume.

[0190] Overall, tumor bearing mice showed greater increase in macrophage and NK mobilization from GPC-100 treatment compared to non-tumor mice (Table 7). For example, GPC-100 induced a 2.7-fold increase in F4 / 80 mobilization in naive mice, but 4.2-fold increase in tumor mice. Tumor mice had higher number of circulating CD3, CD8, CD 19 cells and significantly more macrophages (even with no treatment). For example, Tumor bearing mice had 7 times more F4 / 80 in their PB and more than double of CD3+CD8+ T cells. Therefore, it can be concluded that baseline levels of immune cells may depend on the type and volume of a tumor and GPC-100 mobilization may vary among the models.Table 7: Fold-change comparing lymphoma bearing and naive mice

[0191] Lymphocyte mobilization in mouse models of plasmacytoma and fibrosarcoma.

[0192] A preliminary’ study was performed to evaluate GPC-100 induced lymphocyte mobilization in non-epithelial tumors, a plasmacytoma model J558 and fibrosarcoma model MCA205. At the time of treatment, average plasmacytoma tumor volume was 500 mm3 and average fibrosarcoma tumor volume was 400 mm3. In the plasmacytoma model (FIG. 16A-16C), single dose of GPC-100 resulted in a 3-fold increase in total circulating WBCs and over 3-fold increase in lymphocytes. In a fibrosarcoma model (FIG. 16D-16F), GPC-100 induced a 2-fold increase in peripheral blood WBCs and about 2.5-fold increase lymphocytes. This indicates ability’ of GPC-100 to mobilization cells in several tumor models, mouse strains and tumor volumes. Further studies will determine if the subsets of immune cells mobilized by GPC-100 and GPC-100 with propranolol differ among tumor types.Example 2. Time course of white blood cell and lymphocyte mobilizationMaterials and Methods

[0193] C57 / B6 mice were pre-bled by submental bleeding for baseline WBC counts and randomized into treatment groups based on weights and blood cell counts. Recovery’ was allowed for one week. On the day of the experiment. AMD3100, GPC-100 or BL8040 were administered at doses and routes mentioned above. Each mouse was bled twice or for two time points. For the first time point, blood was collected by non-terminal submental method. For the second time point, terminal cardiac punctures were performed. Blood was then processed for hematology analysis for complete blood counts.Results

[0194] Time dependent increase in WBC counts (FIG. ISA) and lymphocyte counts was observed post treatment (FIG. 18B). Peak mobilization for AMD3100, BL-8040 and GPC-100 occurred within one to two hours. The percent increase in WBC counts and lymphocyte counts was greatest following GPC-100 administration as compared to BL-8040 or AMD3100. Elevation of lymphocyte counts continued for the entire 6-hour duration that the study was conducted and was significantly elevated compared to baseline.Example 3. Lymphocyte mobilization in two mouse strains.Materials and Methods

[0195] C57 / BL6 or Balb / c female mice (6-9 weeks old) were used for the study. BL8040 or AMD3100 were administered once at 5 mg / kg subcutaneously. GPC-100 was administered once at 30 mg / kg intravenously. Blood was collected by cardiac puncture and processed on ahematology analyzer for complete blood count at 2 hours-post drug administration. Total white blood cell counts and differential was obtained.Results

[0196] A comparison of mobilization induced by GPC-100, BL-8040 and Plerixafor were conducted in two strains of mice, C57 / BL6 and Balb / c. Following a two-hour injection, lymphocyte counts were elevated by all three CXCR4 mobilizers. The efficacy was highest with GPC-100, when compared to BL-8040 or AMD3100. This pattern was observed with both mouse strains (FIGs. 19A-19B).Example 4. Evaluating the Safety, Tolerability, Pharmacokinetics and Efficacy of TG-0054 With Single IV Doses Escalation in Healthy Volunteers.

[0197] Evaluating the Safety, Tolerability, Pharmacokinetics and Efficacy of TG-0054 With Single IV Doses Escalation in Healthy Volunteers (ClinicalTrials.gov ID NCT00822341) is a Phase I randomized, double-blind, placebo-controlled, sequential, ascending single IV dose study of the first in human (FIH) administration of TG-0054 (GPC-100) in healthy male and female subjects.Materials and Methods

[0198] Eight groups were studied, each including 8 subjects per group (6:2 ratio to receive TG- 0054 / placebo), for a total of 64 subjects. Eligible subjects were randomized within each dose group and sequentially received a single dose of 0.10, 0.14. 0.28, 0.56, 1.12, 2.24, 3.14, and 4.4 mg / kg of TG-0054 / placebo via a 15-min IV infusion. All doses were infused within 15 minutes.

[0199] The total volume of blood drawn from each subject was approximately 100 mL. Blood samples for pharmacodynamic assessments of various cell types, including lymphocytes, were obtained at pre-dose, 1 hr, 2 hr, 4 hr, 6 hr, 9 hr, 12 hr, 24 hr. and 36 hr after infusion.

[0200] Based on the cell number vs. time data of individual subjects, using the actual sampling time, the pharmacodynamic parameters for lymphocyte counts after TG-0054 administration were determined. Changes from pre-dose were analyzed descriptively for each time point after study drug administration. A one-way analysis of variance (ANOVA) was used to examine the peak differences from baseline observed for each cell type at each TG-0054 dose level examined. All calculations were performed using SAS® software Version 9.1.Results:

[0201] In healthy subjects, statistically significant increases in peak baseline corrected lymphocyte count were generally observed at each dose level of TG-0054 (GPC-100) (FIG. 20).Peak increase of lymphocyte counts (>3-fold change from baseline) was observed at the 1.12 mg / kg TG-0054 (GPC-100) dose.Example 5. GPC-100 Significantly Enhances G-CSF-Induced Mobilization of Lymphocytes in Multiple Myeloma Patients (Clinical Data)

[0202] In recent years, B cell maturation antigen-targeted CAR-T cell therapy has emerged as a promising treatment in multiple myeloma, particularly for the relapsed / refractory disease. The first step in CAR-T manufacturing is collection of lymphocytes, specifically T cells, from patient’s peripheral blood through leukapheresis. Successful leukapheresis and high lymphocyte counts are linked to better CAR T-cell expansion and potentially improved treatment outcomes (Qayed, M. et al. Leukapheresis guidance and best practices for optimal chimeric antigen receptor T-cell manufacturing. Cytotherapy. 2022 Sep; 24(9):869-878. doi:10. 1016 / j.jcyt.2022.05.003. Epub 2022 Jun 17. PMID: 35718701). However, success is limited due to challenges arising from prior treatments negatively impacting blood cell counts as well as scalability and in vivo persistence (Zhang, X. et al. CAR-T cell therapy in multiple myeloma: Current limitations and potential strategies. Front Immunol. 2023 Feb 20; 14:1101495. doi: 10.3389 / fimmu.2023. 1 101495. PMID: 36891310; PMCID: PMC9986336). GPC-100, also known as burixafor, was show n to mobilize lymphocytes in a phase 1 study in healthy volunteers, at single doses from 0.10 to 3.14 mg / kg with peak levels at approximately 4 hours after administration. In multiple myeloma patients eligible for autologous hematopoietic stem cell (HSC)-transplant, GPC-100 was administered in combination with G-CSF (NCT01458288). Although the primary objective of the study was to determine the total number of HSCs collected within the first 4 leukapheresis sessions, impact of burixafor infusion post-G-CSF on lymphocyte mobilization was also evaluated at several timepoints.Methods

[0203] The clinical study was conducted according to the ethical principles of “good clinical practice” and the Declaration of Helsinki after obtaining a written informed consent from each patient. The protocol and its amendment, written study subject information, informed consents, and other appropriate study-related material ere reviewed and approved by Western Institutional Review Board and Chesapeake IRB. Key eligibility criteria included patients aged 18 to 75 years with a confirmed diagnosis of multiple myeloma (MM), having an Eastern Cooperative Oncology Group performance status of 0-1, and being eligible for hematopoietic stem cell transplantation (HSCT). From day 1, patients received subcutaneous (SC) injectionsof 10 pg / kg / day G-CSF daily at 4:00 PM (±1 hour) as needed to reach the target HSC collection goal. Patients were allowed to receive G-CSF for a maximum of 8 consecutive days. Beginning at 8:30 AM (±1 hour) on day 5 patients received intravenous burixafor infusion (3. 14 mg / kg) over 1 minutes. Leukapheresis was performed generally 2 ±1 hours after burixafor administration each day as needed to reach the target HSC collection goal. Peripheral blood samples for circulating lymphocytes were collected prior to each dose of burixafor (labeled pre- burixafor, collected within 30 minutes before the start of administration), approximately 1-hour post-burixafor (after completion of the burixafor infusion and before the start of leukapheresis) and 6 hours post-burixafor.Results:

[0204] The number of lymphocytes mobilized into the peripheral blood with the study drugs were measured. All 9 multiple myeloma patients received G-CSF for at least 4 days. Four of the five patients continued to receive G-CSF on the fifth day. In these 4 patients, burixafor was administered on days 5 and 6, whereas in the remaining 5 patients, burixafor was only administered on day 5. FIG. 21 shows lymphocyte counts and fold change over screening values at several time points. The pre-burixafor time-point shows lymphocyte levels after 4- days of G-CSF treatment, but prior to the burixafor infusion on day 5. FIG. 21A shows the absolute (ABS) number of lymphocytes per patient at screening, pre-burixafor, 1 h post- burixafor and 6 h post-burixafor. FIG. 21B shows average lymphocyte count at each time-point tested, which indicates an average increase at 1 h post-burixafor. FIG. 21C shows fold change in lymphocytes after each treatment. G-CSF induced a mild increase in circulating lymphocytes in 6 of the 9 patients with an average increase of 2.5-fold. Lymphocyte counts were unavailable in 2 patients at 1 h post-burixafor, but the remaining 7 patients showed a median increase of 4.4-fold. In most patients, lymphocyte count decreased from 1 h levels at 6 h post-burixafor. On average, at 6 hours post burixafor, there is still a 2.6-fold higher lymphocyte count than at screening. In 2 of the 3 patients that did not mobilize lymphocytes from G-CSF treatment, burixafor induced an 8-fold and 4-fold increase over screening, respectively. In the third patient, there was a small 1.5 -fold and 1.9-fold increase by burixafor alone at 1 h and 6 h. In combination, the total yield of lymphocytes at pre-, 1 h and 6 h post-burixafor showed 3 to 11- fold increase, which was mainly driven by burixafor as shown in FIG. 21D.Discussion:

[0205] The data showed that combining burixafor with G-CSF can significantly enhance circulating lymphocyte count. As seen in 3 patients that did not mobilize lymphocytes in response to G-CSF, burixafor alone was sufficient. The primary endpoint of this study was to collect >5.0 x 106CD34+ cells / kg, which was met in 8 of the 9 patients (Setia, G. et al., A Phase II, Open-Label Pilot Study to Evaluate the Hematopoietic Stem Cell Mobilization of TG- 0054 Combined with G-CSF in 12 Patients with Multiple Myeloma, Non-Hodgkin Lymphoma or Hodgkin Lymphoma - an Interim Analysis. Blood. Volume 126, Issue 23, 2015. Page 515, ISSN 0006-4971, https: / / doi.org / 10.1182 / blood.V126.23.515.515). The one patient (patient 7) who did not meet the primary endpoint had been recently treated with lenalidomide, which is known to reduce the HSC yield (He X et al.. Predictive factors for peripheral blood stem cell mobilization in multiple myeloma in the era of novel therapies: A single-center experience. Cancer Med. Jun 2024:13(1 l):e7356. doi: 10. I002 / cam4.7356; Pozotrigo M. et al., Factors impacting stem cell mobilization failure rate and efficiency in multiple myeloma in the era of novel therapies: experience at Memorial Sloan Kettering Cancer Center. Bone Marrow Transplant . Aug 2013:48(8): 1033-9). This patient did not mobilize lymphocytes from G-CSF treatment but showed a cumulative increase of 2.9-fold at I h and 6 h post-burixafor infusion. In other patients, HSC mobilization did not correlate directly with circulating lymphocyte counts. Overall, the data indicates that burixafor makes a critical contribution in mobilization of lymphocytes, which is important for several reasons in the context of multiple myeloma. Some studies have reported association between higher lymphocyte counts in the graft and better immune recover}', as well as improved progression-free and overall survival in MM (Turunen A. et al., Autograft cellular composition and outcome in myeloma patients: Results of the prospective multicenter GOA study. Transfusion. 2021 Jun;61(6): 1830-1844. doi: 10.1111 / trf. 16424. Epub 2021 May 6. PMID: 33955591; Jantunen E. et al.. Mobilization Strategies in Myeloma Patients Intended for Autologous Hematopoietic Cell Transplantation. Transfus Med Hemother. 2023 Aug 24;50(5):438-447. doi: 10.1159 / 000531940. PMID: 37899993; PMCID: PMC10603622). Specific functionality7of graft-derived T cells, however, remains undetermined. As described previously, prior therapies can also negatively impact blood cell counts. For instance, melphalan, a conditioning treatment given prior to HSC transplant, can lead to T-cell depletion or increase T-cell exhaustion diminishing their antitumor effects (Mika T. et al., Altered T-Lymphocyte Biology7Following High-Dose Melphalan and Autologous Stem Cell Transplantation With Implications for Adoptive T-Cell Therapy.Front Oncol. 2020 Dec ll;10:568056. doi: 10.3389 / fonc.2020.568056. PMID: 33363008;PMCID: PMC7759611). Low counts of absolute lymphocytes in these treated patients can lead to insufficient T-cell yield and increased cost or delayed CAR-T manufacturing (Jo T. et al.. Risk factors for CAR-T cell manufacturing failure among DLBCL patients: A nationwide survey in Japan. Br J Haematol. 2023 Jul;202(2):256-266. doi: 10. 1111 / bjh. 18831. Epub 2023 Apr 25. PMID: 37096915). Lasty, CAR-T cells may not show persistent activity when injected into heavily pretreated patients (Wittibschlager V. et al., CAR T-Cell Persistence Correlates with Improved Outcome in Patients with B-Cell Lymphoma. Int J Mol Sci. 2023 Mar 16;24(6):5688. doi: 10.3390 / ijms24065688. PMID: 36982764; PMCID: PMC10056741). Furthermore, circulating lymphocytes are smaller and denser compared to HSPCs and make the isolation from red blood cells more challenging (Fesnak A. et al., CAR-T Cell Therapies From the Transfusion Medicine Perspective. Transfius Med Rev. 2016 Jul;30(3): 139-45. doi: 10. 1016 / j.tmrv.2016.03.001. Epub 2016 Mar 28. PMID: 27067907; PMCID: PMC4914456). Higher composition of naive or memory T-cells compared to exhausted T-cells were shown to be beneficial for success of CAR-T (Tao Z. et al. Impact of T cell characteristics on CAR-T cell therapy in hematological malignancies. Blood Cancer J. 2024 Dec 3; 14(1 ):213. doi: 10.1038 / s41408-024-01193-6. PMID: 39627220; PMCID: PMC11615218). Therefore, boosting peripheral lymphocyte count prior to apheresis could be a critical step toward increasing T-cell fitness and potentially leading to a better clinical outcome. A phase 2 clinical trial (NCT05561751) is currently underway where exhaustive or activated markers on mobilized T lymphocytes will be evaluated. Understanding the lymphocyte subsets mobilized by burixfaor will provide further evidence of its potential utility in CAR-T therapies.Example 6: Evaluating the Safety, Tolerability, Pharmacokinetics and Efficacy of TG- 0054 (Burixafor) with Single IV Dose Escalation in Healthy Volunteers

[0206] Evaluating the Safety, Tolerability, Pharmacokinetics and Efficacy of TG-0054 With Single IV Doses Escalation in Healthy Volunteers (ClinicalTrials.gov ID NCT00822341) is a Phase I randomized, double-blind, placebo-controlled, sequential, ascending single IV dose study of the first in human (FIH) administration of TG-0054 (GPC-100) in healthy male and female subjects.Materials and Methods

[0207] Eight groups were studied, each including 8 subjects per group (6:2 ratio to receive TG- 0054 / placebo), for a total of 64 subjects. Eligible subjects were randomized within each dosegroup and sequentially received a single dose of 0.10, 0.14, 0.28, 0.56, 1.12, 2.24, 3.14, and 4.4 mg / kg of TG-0054 / placebo via a 15-min IV infusion. All doses were infused within 15 minutes.

[0208] The total volume of blood drawn from each subject was approximately 100 mL. Blood samples for pharmacodynamic assessments of various cell types, including lymphocytes and neutrophils, were obtained at pre-dose, 1 hr, 2 hr, 4 hr, 6 hr, 9 hr, 12 hr, 24 hr, and 36 hr after infusion.

[0209] Based on the cell number vs. time data of individual subjects, using the actual sampling time, the pharmacodynamic parameters for lymphocyte and neutrophil counts after TG-0054 administration were determined. Changes from pre-dose were analyzed descriptively for each time point after study drug administration. A one-way analysis of variance (ANOVA) was used to examine the peak differences from baseline observed for each cell type at each TG-0054 dose level examined. All calculations were performed using SAS® software Version 9.1.Results:

[0210] Statistically significant differences in peak baseline corrected neutrophil count were generally observed at the 1.12 mg / kg TG-0054 (GPC-100) dose level (FIG. 22). Compared to baseline, neutrophil counts were elevated within 2 hours post-burixafor and maximum mobilization was observed at 4-6 h post-burixafor infusion. Peak increase of neutrophil counts was observed at the 2.24 mg / kg TG-0054 (GPC-100).Conclusion, application, and future direction

[0211] Studies presented in this report analyzed mobilization of several lymphoid and myeloid cell populations. Single intravenous injection of GPC-100 was shown to mobilize immune cell subsets with greater mobilization in tumor bearing mice compared to the naive mice, and in Balb / c mice compared to C57 / BL6 mice. In general, GPC-100 more preferentially mobilized CD8+T cells and myeloid cells in both mouse strains. Propranolol pretreatment significantly improved GPC-100 induced mobilization. Therefore, it may be helpful to understand the potential clinical applications of GPC-100 plus propranolol induced mobilization of cells.

[0212] There are research efforts to develop an effective adoptive T-cell transfer product with an early memory phenoty pe that is absent of exhaustion markers. For CAR-T cell therapies, large number of CAR-T cells can be generated ex vivo from patient blood. However, efficacy depends on ability to localize to sites, persist and exert function (often in an immunosuppressive microenvironment). Therefore, phenotype of these cells determines efficacy of this therapy. So far, GPC-100 and propranolol mobilizes cytotoxic CD8+ T cells which have preserved theirimmune function. Some of the mobilized CD8+T cells also expressed the exhaustion marker PD-1. Therefore, additional analysis of mobilized cells expressing exhaustion markers such as TIM-3 and LAG3, or stem T cell markers SCA-1, CCR7, TCF-1 will be conducted.

[0213] The bispecific T cell engager antibodies are dependent on endogenous T cells and directing them to killing tumor cells. Their short half-life needs continuous administration, which can lead to T cell exhaustion. Therefore, it will be important to determine if enhanced mobilization of T cells into peripheral blood by GPC-100 and propranolol will make them more accessible for the T cell engagers and improve their efficacy.

[0214] In general, in vivo persistence of T cells leads to a better anti-tumor response. T cells with stem cell memory (TSCM) phenotype are known to have best in vivo persistence and ability to differentiate. For example, in B cell lymphoma patients treated with CAR.CD19 T cells, in vivo expansion and response correlated with TSCM like phenotype. Therefore, further research is ongoing exploring the type of T cells mobilized by GPC-100 alone and with propranolol, role of propranolol in T-cell differentiation and function, and ability to demonstrate this in vivo.

[0215] All publications and patent applications mentioned in this specification are herein incorporated by reference to the same extent as if each individual publication or patent application was specifically and individually indicated to be incorporated by reference.

[0216] While preferred embodiments have been shown and described herein, those skilled in the art that such embodiments would recognize that the embodiments are provided by way of example only. It is intended that the following claims define the scope of the invention and that methods and structures within the scope of these claims and their equivalents be covered thereby.Exemplary Embodiments

[0217] In an embodiment, disclosed herein is a method of mobilizing a cell in a subject, the method comprising: blocking CXCR4 signaling and beta-adrenergic receptor signaling in the subject.

[0218] In an embodiment, disclosed herein is a method of inducing cell mobilization in a subject, the method comprising: blocking CXCR4 signaling and beta-adrenergic receptor signaling in the subject.

[0219] In an embodiment, disclosed herein is a method of enhancing apheresis in a subject, the method comprising: blocking CXCR4 signaling and beta-adrenergic receptor signaling in the subject.

[0220] In an embodiment, disclosed herein is a method of enhancing apheresis by inducing cell mobilization in a subject, the method comprising: blocking CXCR4 signaling and beta- adrenergic receptor signaling in the subject.

[0221] In an embodiment, disclosed herein is a method of enhancing apheresis by mobilizing a cell in a subject, the method comprising: blocking CXCR4 signaling and beta-adrenergic receptor signaling in the subject.

[0222] In an embodiment, the blocking beta-adrenergic receptor signaling is performed before the blocking CXCR4 signaling.

[0223] In an embodiment, the blocking beta-adrenergic receptor signaling continues after the blocking CXCR4 signaling is terminated.

[0224] In an embodiment, the blocking CXCR4 signaling comprises administering a CXCR4 inhibitor to the subject.

[0225] In an embodiment, the blocking beta-adrenergic receptor signaling comprises administering a beta-adrenergic receptor inhibitor to the subject.

[0226] In an embodiment, the cell is a lymphocyte.

[0227] In an embodiment, the blocking CXCR4 signaling comprises administering a CXCR4 inhibitor to the subject and the blocking beta-adrenergic receptor signaling comprises administering a beta-adrenergic receptor inhibitor to the subject.

[0228] In an embodiment, the cell is a lymphocyte.

[0229] In an embodiment, disclosed herein is a method of mobilizing a lymphocyte in a subject, the method comprising: administering a beta-adrenergic receptor inhibitor and a CXCR4 inhibitor to the subject.

[0230] In an embodiment, disclosed herein is a method of inducing lymphocyte mobilization in a subject, the method comprising: administering a beta-adrenergic receptor inhibitor and a CXCR4 inhibitor to the subject.

[0231] In an embodiment, disclosed herein is a method of enhancing apheresis in a subject, the method comprising: administering a beta-adrenergic receptor inhibitor and a CXCR4 inhibitor to the subject.

[0232] In an embodiment, disclosed herein is a method of enhancing apheresis by inducing cell mobilization in a subject, the method comprising: administering a beta-adrenergic receptor inhibitor and a CXCR4 inhibitor to the subject.

[0233] In an embodiment, disclosed herein is a method of enhancing apheresis by mobilizing a cell in a subject, the method comprising: administering a beta-adrenergic receptor inhibitor and a CXCR4 inhibitor to the subject.

[0234] In an embodiment, the administering the beta-adrenergic receptor inhibitor is performed before the administering the CXCR4 inhibitor.

[0235] In an embodiment, the administering the beta-adrenergic receptor inhibitor continues after the administering the CXCR4 inhibitor is terminated.

[0236] In an embodiment, the method further comprises administering G-CSF to the subject.

[0237] In an embodiment, the administering the beta-adrenergic receptor inhibitor and the CXCR4 inhibitor to the subject is performed in the absence of G-CSF.

[0238] In an embodiment, disclosed herein is a method of mobilizing a lymphocyte in a subject, the method comprising: administering a CXCR4 inhibitor and G-CSF to the subject, in the absence of a beta-adrenergic receptor inhibitor.

[0239] In an embodiment, disclosed herein is a method of inducing lymphocyte mobilization in a subject, the method comprising: administering a CXCR4 inhibitor and G-CSF to the subject, in the absence of a beta-adrenergic receptor inhibitor.

[0240] In an embodiment, disclosed herein is a method of enhancing apheresis in a subject, the method comprising: administering a CXCR4 inhibitor and G-CSF to the subject, in the absence of a beta-adrenergic receptor inhibitor.

[0241] In an embodiment, disclosed herein is a method of enhancing apheresis by inducing cell mobilization in a subject, the method comprising: administering a CXCR4 inhibitor and G-CSF to the subject, in the absence of a beta-adrenergic receptor inhibitor.

[0242] In an embodiment, disclosed herein is a method of enhancing apheresis by mobilizing a cell in a subject, the method comprising: administering a CXCR4 inhibitor and G-CSF to the subject, in the absence of a beta-adrenergic receptor inhibitor.

[0243] In an embodiment, the beta-adrenergic receptor inhibitor is an ADRB2 inhibitor.

[0244] In an embodiment, the beta-adrenergic receptor inhibitor is selected from the group consisting of alprenolol, atenolol, betaxolol, bupranolol, butoxamine, carazolol, carvedilol, CGP 12177, cicloprolol, ICI 118551, ICYP, labetalol, levobetaxolol,levobunolol, LK 204-545, metoprolol, nadolol. NIHP, NIP, propafenone, propranolol, sotalol, SR59230A, and timolol.

[0245] In an embodiment, the beta-adrenergic receptor inhibitor is selected from the group consisting of propranolol, nadolol, and ICI 118551.

[0246] In an embodiment, the beta-adrenergic receptor inhibitor is propranolol.

[0247] In an embodiment, the CXCR4 inhibitor is selected from the group consisting of ALX40- 4C. AMD070 (AMD11070, X4P-001), AMD3100 (plerixafor), AMD3465. ATI 2341. BKT140 (BL-8040; TF14016; 4F-Benzoyl-TN14003), CTCE-9908, CX549„D-[Lys3] GHRP-6, FC122, FC131, GMI-1359, GSK812397, GST-NT21MP, isothiourea- la, isothiourea- It (ITlt). KRH-1636, KRH-3955. LY2510924. MSX-122, N-[llC]Methyl- AMD3465. POL6326, SDF-1 1-9[P2G] dimer, SDF1 P2G, T134. T140, T22. TC 14012. TG-0054 (Burixafor), USL311, viral macrophage inflammatory protein-II (vMIP-II), WZ81I, [64Cu]-AMD3100, [64Cu]-AMD3465, [68Ga]pentixafor, [90Y]pentixather, [99mTc]02-AMD3100, [177Lu]pentixather, and 508MC1 (Compound 26).

[0248] In an embodiment, the CXCR4 inhibitor is selected from the group consisting of AD-214, AMD070 (AMD! 1070. X4P-001), AMD3100 (plenxafor), BKT140 (BL-8040; TF14016; 4F-Benzoyl-TN14003), CTCE-9908, LY2510924, LY2624587, T140, TG- 0054 (Burixafor), PF-06747143, POL6326, and ulocuplumab (MDX1338 / BMS-936564).

[0249] In an embodiment, the CXCR4 inhibitor is TG-0054 (burixafor).

[0250] In an embodiment, the CXCR4 inhibitor is AMD3100 (plerixafor).

[0251] In an embodiment, the CXCR4 inhibitor is ulocuplumab (MDX1338 / BMS-936564).

[0252] In an embodiment, the administering the CXCR4 inhibitor to the subject comprises administering TG-0054 (burixafor) and propranolol.

[0253] In an embodiment, the administering the CXCR4 inhibitor to the subject comprises administering AMD3100 (plerixafor) and propranolol.

[0254] In an embodiment, the administering the CXCR4 inhibitor to the subject comprises administering ulocuplumab (MDX1338 / BMS-936564) and propranolol.

[0255] In an embodiment, the administering a combination of the CXCR4 inhibitor and the G- CSF induces an enhanced amount of cell mobilization relative to the amount of cell mobilization induced by the CXCR4 inhibitor only.

[0256] In an embodiment, the administering a combination of the CXCR4 inhibitor and the G- CSF mobilizes a cell by an amount enhanced relative to the amount of cell mobilization induced by the CXCR4 inhibitor only.

[0257] In an embodiment, the administering a combination of the CXCR4 inhibitor and the G- CSF induces an enhanced amount of apheresis relative to the amount of apheresis induced by the CXCR4 inhibitor only.

[0258] In an embodiment, the administering a combination of the CXCR4 inhibitor and the beta- adrenergic receptor inhibitor induces an enhanced amount of cell mobilization relative to the amount of cell mobilization induced by the CXCR4 inhibitor only.

[0259] In an embodiment, the administering a combination of the CXCR4 inhibitor and the beta- adrenergic receptor inhibitor mobilizes a cell by an amount enhanced relative to the amount of cell mobilization induced by the CXCR4 inhibitor only.

[0260] In an embodiment, the administering a combination of the CXCR4 inhibitor and the beta- adrenergic receptor inhibitor induces an enhanced amount of apheresis relative to the amount of apheresis induced by the CXCR4 inhibitor only.

[0261] In an embodiment, the administering a combination of the CXCR4 inhibitor, the beta- adrenergic receptor inhibitor, and the G-CSF induces an enhanced amount of cell mobilization relative to the amount of cell mobilization induced by the CXCR4 inhibitor and the beta-adrenergic receptor inhibitor only.

[0262] In an embodiment, the administering a combination of the CXCR4 inhibitor, the beta- adrenergic receptor inhibitor, and the G-CSF mobilizes a cell by an amount enhanced relative to the amount of cell mobilization induced by the CXCR4 inhibitor and the beta- adrenergic receptor inhibitor only.

[0263] In an embodiment, the administering a combination of the CXCR4 inhibitor and the beta- adrenergic receptor inhibitor, and the G-CSF induces an enhanced amount of apheresis relative to the amount of apheresis induced by the CXCR4 inhibitor and the beta- adrenergic receptor inhibitor only.

[0264] In an embodiment, the administering a combination of TG-0054 (burixafor) and the G- CSF induces an enhanced amount of cell mobilization relative to the amount of cell mobilization induced by AMD3100 (plerixafor) and the G-CSF.61

[0265] In an embodiment, the administering a combination of the TG-0054 (burixafor) and the G-CSF mobilizes a cell by an amount enhanced relative to the amount of cell mobilization induced by the AMD3100 (plenxafor) and the G-CSF.

[0266] In an embodiment, the administering a combination of the TG-0054 (burixafor) and the G-CSF induces an enhanced amount of apheresis relative to the amount of apheresis induced by the AMD3100 (plerixafor) and the G-CSF.

[0267] In an embodiment, an enhanced amount of cell mobilization or apheresis is measured by a method selected from the group consisting of complete blood count (CBC) analysis, flow cytometry, and colony forming unit (CFU) assay.

[0268] In an embodiment, the enhanced amount of cell mobilization or apheresis is measured by flow cytometry.

[0269] In an embodiment, the flow cytometry is performed on (Lin-Scal+c-Kit+) LSK cells.

[0270] In an embodiment, the enhanced amount of cell mobilization or apheresis is measured by colony forming unit (CFU) assay.

[0271] In an embodiment, the subject has a CXCR4 protomer in the cell.

[0272] In an embodiment, the subject has an ADRB2 protomer in the cell.

[0273] In an embodiment, the subject has a CXCR4 protomer and an ADRB2 protomer in the cell.

[0274] In an embodiment, the subject has a CXCR4-ADRB2 heteromer in the cell.

[0275] In an embodiment, i) the CXCR4-ADRB2 heteromer has an enhanced amount of downstream calcium mobilization relative to downstream calcium mobilization from a CXCR4 protomer or ADRB2 protomer; and ii) the administered combination of inhibitors suppresses the enhanced downstream calcium mobilization from said CXCR4-ADRB2 heteromer in the lymphocyte.

[0276] In an embodiment, the cell is a lymphocyte.

[0277] In an embodiment, the lymphocyte is selected from the group consisting of a T cell, B cell, and NK cell.

[0278] In an embodiment, the lymphocyte is mobilized from bone marrow to peripheral blood.

[0279] In an embodiment, the mobilized lymphocyte is collected for transplantation to a patient having cancer.

[0280] In an embodiment, the cancer is selected from the group consisting of lymphoma, leukemia, and myeloma.

[0281] In an embodiment, the cancer is non-Hodgkin lymphoma (NHL), acute myeloid leukemia (AML), acute lymphoblastic leukemia (ALL), or multiple myeloma (MM).

[0282] In an embodiment, the lymphocyte is mobilized for treatment of a condition selected from the group consisting of neurological disorder, cardiac ischemia, myocardial infarction, diabetes, tissue repair, bone and cartilage disease, autoimmune disease, graft versus host disease, Crohn's disease, multiple sclerosis, systemic lupus erythematosus, and systemic sclerosis.

[0283] In an embodiment, the lymphocyte is mobilized into blood.

[0284] In an embodiment, the lymphocyte is mobilized for treatment of a cancer.

[0285] In an embodiment, the cell is an immune cell.

[0286] In an embodiment, the immune cell is a white blood cell.

[0287] In an embodiment, the white blood cell is a lymphocyte.

[0288] In an embodiment, the lymphocyte is selected from the group consisting of a T cell, a B cell, and a natural killer (NK) cell.

[0289] In an embodiment, the lymphocyte is a T cell.

[0290] In an embodiment, the lymphocyte is a natural killer (NK) cell.

[0291] In an embodiment, the white blood cell is a granulocyte.

[0292] In an embodiment, the granulocyte is selected from the group consisting of a neutrophil, an eosinophil, and a basophil.

[0293] In an embodiment, the granulocyte is a neutrophil.

[0294] In an embodiment, the white blood cell is a monocyte.

[0295] In an embodiment, the immune cell is mobilized from bone marrow to peripheral blood.

[0296] In an embodiment, the immune cell is mobilized from lymph node to peripheral blood.

[0297] In an embodiment, the mobilized immune cell is used for adoptive cell therapy (ACT).

[0298] In an embodiment, the adoptive cell therapy (ACT) is chimeric antigen receptor (CAR) T cell therapy.

[0299] In an embodiment, the adoptive cell therapy (ACT) is netural killer (NK) cell therapy.

[0300] In an embodiment, the adoptive cell therapy (ACT) is engineered T-cell receptor (TCR) therapy.

[0301] In an embodiment, the adoptive cell therapy (ACT) is tumor-infiltrating lymphocyte (TIL) therapy.

[0302] In an embodiment, disclosed herein is a method of mobilizing a lymphocyte or an immune cell in a subject, the method comprising administering, alone or in combination, a CXCR4 inhibitor and G-CSF to the subject.

[0303] In an embodiment, disclosed herein is a method for treatment of cancer comprising administering, alone or in combination, a CXCR4 inhibitor and G-CSF to a subject.

[0304] In an embodiment, disclosed herein is a method of enhancing leukapheresis by inducing lymphocyte or immune cell mobilization in a subject, the method comprising administering, alone or in combination, a CXCR4 inhibitor and G-CSF to the subject.

[0305] In an embodiment, the CXCR4 inhibitor and the G-CSF are administered to the subject in the absence of a beta-adrenergic receptor inhibitor.

[0306] In an embodiment, the method comprises administering the G-CSF to the subject before administering the CXCR4 inhibitor to the subject.

[0307] In an embodiment, the method comprises administering a combination of the CXCR4 inhibitor and G-CSF to the subject, and wherein the administering of the combination of the CXCR4 inhibitor and G-CSF to the subject induces an enhanced amount of lymphocyte mobilization relative to an amount of lymphocyte mobilization induced by administration of G-CSF only to the subject.

[0308] In an embodiment, the method comprises administering a combination of the CXCR4 inhibitor and G-CSF to the subject, and wherein the administering of the combination of the CXCR4 inhibitor and G-CSF to the subject induces an enhanced amount of lymphocyte mobilization relative to an amount of lymphocyte mobilization induced by administration of the CXCR4 inhibitor only to the subject.

[0309] In an embodiment, the CXCR4 inhibitor is selected from the group consisting of ALX40-4C, AMD070 (AMD11070, X4P-001), AMD3100 (plerixafor), AMD3465, ATI 2341, BKT140 (BL-8040; TF14016; 4F-Benzoyl-TNI4003), CTCE-9908, CX549„D- [Lys3] GHRP-6, FC122, FC131, GMI-1359, GSK812397, GST-NT21MP, isothiourea- la, isothiourea- It (ITlt), KRH-1636, KRH-3955, LY2510924, MSX-122, N- [HC]Methyl-AMD3465, POL6326, SDF-1 1-9[P2G] dimer, SDF1 P2G, T134, T140, T22, TC 14012, TG-0054 (Burixafor). USL311. viral macrophage inflammatory protein-II (vMIP-II), WZ811, [64Cu]-AMD3100, [64Cu]-AMD3465, [68Ga]pentixafor, [90Y]pentixather, [99mTc]02-AMD3100, [177Lu]pentixather, and 508MC1 (Compound 26).

[0310] In an embodiment, the CXCR4 inhibitor is selected from the group consisting of AD- 214, AMD070 (AMD11070, X4P-001), AMD3100 (plerixafor), BKT140 (BL-8040; TF14016; 4F-Benzoyl-TN 14003), CTCE-9908, LY2510924, LY2624587, T140, TG- 0054 (Burixafor), PF-06747143, POL6326, and ulocuplumab (MDX1338 / BMS- 936564).

[0311] In an embodiment, the CXCR4 inhibitor is TG-0054 (burixafor).

[0312] In an embodiment, the method comprises administering a combination of burixafor and G-CSF to the subject, and wherein the administering of the combination of burixafor and G-CSF to the subject induces an enhanced amount of lymphocyte mobilization relative to an amount of lymphocyte mobilization induced by administration of G-CSF only to the subject.

[0313] In an embodiment, the method comprises administering a combination of burixafor and G-CSF to the subject, and wherein the administering of the combination of the burixafor and G-CSF to the subject induces an enhanced amount of ly mphocyte mobilization relative to an amount of lymphocyte mobilization induced by administration of burixafor only to the subject.

[0314] In an embodiment, the enhanced amount of lymphocyte mobilization is measured by a method selected from the group consisting of complete blood count (CBC) analysis, flow cytometry, and colony forming unit (CFU) assay.

[0315] In an embodiment, the enhanced amount of lymphocyte mobilization is measured by CBC analysis.

[0316] In an embodiment, the subject has a CXCR4 protomer in the lymphocyte.

[0317] In an embodiment, the ly mphocyte or immune cell is selected from the group consisting of a T cell, B cell, a natural killer (NK) cell, a neutrophil, eosinophil, and a basophil.

[0318] In an embodiment, the lymphocyte or immune cell is mobilized for treatment of a condition selected from the group consisting of neurological disorder, cardiac ischemia, myocardial infarction, diabetes, tissue repair, bone and cartilage disease, autoimmune disease, graft versus host disease, Crohn's disease, multiple sclerosis, systemic lupus erythematosus, and systemic sclerosis.

[0319] In an embodiment, the lymphocyte or immune cell is mobilized for treatment of a cancer.

[0320] In an embodiment, the cancer is non-Hodgkin lymphoma (NHL), acute my eloid leukemia (AML), acute lymphoblastic leukemia (ALL), or multiple myeloma (MM).

[0321] In an embodiment, the cancer is multiple myeloma (MM).

[0322] In an embodiment, the lymphocyte or immune cell is mobilized for adoptive cell therapy (ACT).

[0323] In an embodiment, the treatment comprises leukapheresis.

[0324] In an embodiment, the treatment comprises adoptive cell therapy (ACT).

[0325] In an embodiment, the adoptive cell therapy (ACT) is chimeric antigen (CAR) T cell therapy.

[0326] In an embodiment, disclosed herein is a composition or compositions for mobilizing a lymphocyte or an immune cell in a subject comprising, alone or in combination, a CXCR4 inhibitor and G-CSF.

[0327] In an embodiment, disclosed herein is a composition or compositions for treatment of cancer comprising, alone or in combination, a CXCR4 inhibitor and G-CSF.

[0328] In an embodiment, disclosed herein is a composition or compositions for enhancing leukapheresis by inducing lymphocyte or immune cell mobilization in a subject comprising, alone or in combination, a CXCR4 inhibitor and G-CSF.

[0329] In an embodiment, the composition does not include a beta-adrenergic receptor inhibitor.

[0330] In an embodiment, the composition further includes a beta-adrenergic receptor inhibitor.

[0331] In an embodiment, the CXCR4 inhibitor is selected from the group consisting ofALX40-4C, AMD070 (AMD11070, X4P-001), AMD3100 (plerixafor), AMD3465, ATI 2341, BKT140 (BL-8040; TF14016; 4F-Benzoyl-TN14003), CTCE-9908, CX549„D- [Lys3] GHRP-6, FC122, FC13L GMI-1359, GSK812397, GST-NT21MP, isothiourea- la, isothiourea- It (ITlt), KRH-1636, KRH-3955, LY2510924, MSX-122, N- [HC]Methyl-AMD3465, POL6326, SDF-1 1-9[P2G] dimer, SDF1 P2G, T134, T140, T22, TC 14012, TG-0054 (Burixafor), USL311, viral macrophage inflammatory protein-II (vMIP-II), WZ811, [64Cu]-AMD3100, [64Cu]-AMD3465. [68Ga]pentixafor, [90Y]pentixather, [99mTc|02-AMD3100. [177Lu]pentixather, and 508MC1 (Compound 26).

[0332] In an embodiment, the CXCR4 inhibitor is selected from the group consisting of AD- 214, AMD070 (AMD11070, X4P-001), AMD3100 (plerixafor), BKT140 (BL-8040;TF14016; 4F-Benzoyl-TN 14003), CTCE-9908, LY2510924, LY2624587, T I40. TG- 0054 (Burixafor), PF-06747143, POL6326, and ulocuplumab (MDX1338 / BMS- 936564).

[0333] In an embodiment, the CXCR4 inhibitor is TG-0054 (burixafor).

[0334] In an embodiment, the subject has a CXCR4 protomer in the lymphocyte.

[0335] In an embodiment, the lymphocyte or immune cell is selected from the group consisting of a T cell, a B cell, and a natural killer (NK) cell, a neutrophil, a eosinophil, and a basophil.

[0336] In an embodiment, the lymphocyte or immune cell is mobilized for treatment of a condition selected from the group consisting of neurological disorder, cardiac ischemia, myocardial infarction, diabetes, tissue repair, bone and cartilage disease, autoimmune disease, graft versus host disease, Crohn's disease, multiple sclerosis, systemic lupus erythematosus, and systemic sclerosis.

[0337] In an embodiment, the lymphocyte or immune cell is mobilized for treatment of a cancer.

[0338] In an embodiment, the cancer is non-Hodgkin lymphoma (NHL), acute myeloid leukemia (AML), acute lymphoblastic leukemia (ALL), or multiple myeloma (MM).

[0339] In an embodiment, the cancer is multiple myeloma (MM).

[0340] In an embodiment, the lymphocyte or immune cell is mobilized for adoptive cell therapy (ACT).

[0341] In an embodiment, the adoptive cell therapy (ACT) is chimeric antigen (CAR) T cell therapy.

[0342] In an embodiment, the cancer is non-Hodgkin lymphoma (NHL), acute myeloid leukemia (AML), acute lymphoblastic leukemia (ALL), or multiple myeloma (MM).

[0343] In an embodiment, the cancer is multiple myeloma (MM).

[0344] In an embodiment, the treatment comprises leukapheresis.

[0345] In an embodiment, the treatment comprises adoptive cell therapy (ACT).

[0346] In an embodiment, the adoptive cell therapy (ACT) is chimeric antigen (CAR) T cell therapy.

[0347] In an embodiment, disclosed herein is a pharmaceutical composition or pharmaceutical compositions for mobilizing a lymphocyte or an immune cell in a subject comprising,alone or in combination, a CXCR4 inhibitor and G-CSF, and a pharmaceutically acceptable excipient.

[0348] In an embodiment, disclosed herein is a pharmaceutical composition or pharmaceutical compositions for treatment of cancer comprising, alone or in combination, a CXCR4 inhibitor and G-CSF, and a pharmaceutically acceptable excipient.

[0349] In an embodiment, disclosed herein is a pharmaceutical composition or pharmaceutical compositions for enhancing leukapheresis by inducing lymphocyte or immune cell mobilization in a subject comprising, alone or in combination, a CXCR4 inhibitor and G-CSF, and a pharmaceutically acceptable excipient.

[0350] In an embodiment, the pharmaceutical composition does not include a beta-adrenergic receptor inhibitor.

[0351] In an embodiment, the pharmaceutical composition further includes a beta-adrenergic receptor inhibitor.

[0352] In an embodiment, the CXCR4 inhibitor is selected from the group consisting of ALX40-4C, AMD070 (AMD11070, X4P-001), AMD3100 (plerixafor), AMD3465, ATI 2341. BK.T140 (BL-8040; TF14016; 4F-Benzoyl-TN 14003), CTCE-9908, CX549.,D- [Lys3] GHRP-6, FC122, FC131, GMI-1359, GSK812397, GST-NT21MP, isothiourea- la, isothiourea- It (ITlt), KRH-1636, KRH-3955, LY2510924, MSX-122, N- [HC]Methyl-AMD3465, POL6326, SDF-1 1-9[P2G] dimer, SDF1 P2G, T134, 140, T22, TC 14012, TG-0054 (Burixafor). USL311. viral macrophage inflammatory protein-II (vMIP-II), WZ811, [64Cu]-AMD3100, [64Cu]-AMD3465, [68Ga]pentixafor, [90Y]pentixather, [99mTc]02-AMD3100, [177Lu]pentixather, and 508MC1 (Compound 26).

[0353] In an embodiment, the CXCR4 inhibitor is selected from the group consisting of AD- 214, AMD070 (AMD11070, X4P-001), AMD3100 (plenxafor), BKT140 (BL-8040; TF14016; 4F-Benzoyl-TN 14003), CTCE-9908, LY2510924, LY2624587, T140, TG- 0054 (Burixafor), PF-06747143, POL6326, and ulocuplumab (MDX1338 / BMS- 936564).

[0354] In an embodiment, the CXCR4 inhibitor is TG-0054 (burixafor).

[0355] In an embodiment, the subject has a CXCR4 protomer in the lymphocyte.

[0356] In an embodiment, wherein the lymphocyte or immune cell is selected from the group consisting of a T cell, a B cell, and a natural killer (NK) cell, a neutrophil, an eosinophil, and a basophil.

[0357] In an embodiment, the lymphocyte or immune cell is mobilized for treatment of a condition selected from the group consisting of neurological disorder, cardiac ischemia, myocardial infarction, diabetes, tissue repair, bone and cartilage disease, autoimmune disease, graft versus host disease, Crohn's disease, multiple sclerosis, systemic lupus erythematosus, and systemic sclerosis.

[0358] In an embodiment, the lymphocyte or immune cell is mobilized for treatment of a cancer.

[0359] In an embodiment, the cancer is non-Hodgkin lymphoma (NHL), acute myeloid leukemia (AML), acute lymphoblastic leukemia (ALL), or multiple myeloma (MM).

[0360] In an embodiment, the cancer is multiple myeloma (MM).

[0361] In an embodiment, the lymphocyte or immune cell is mobilized for adoptive cell therapy (ACT).

[0362] In an embodiment, the adoptive cell therapy (ACT) is chimeric antigen (CAR) T cell therapy.

[0363] In an embodiment, the cancer is non-Hodgkin lymphoma (NHL), acute myeloid leukemia (AML), acute lymphoblastic leukemia (ALL), or multiple myeloma (MM).

[0364] In an embodiment, the cancer is multiple myeloma (MM).

[0365] In an embodiment, the treatment comprises leukapheresis.

[0366] In an embodiment, the treatment comprises adoptive cell therapy (ACT).

[0367] In an embodiment, the adoptive cell therapy (ACT) is chimeric antigen (CAR) T cell therapy.

Claims

WHAT IS CLAIMED IS:

1. A method of mobilizing a lymphocyte or an immune cell in a subject, the method comprising: blocking CXCR4 signaling and / or beta-adrenergic receptor signaling in the subject.

2. A method of inducing lymphocyte or immune cell mobilization in a subject, the method comprising: blocking CXCR4 signaling and / or beta-adrenergic receptor signaling in the subject.

3. A method of enhancing apheresis in a subject, the method comprising: blocking CXCR4 signaling and / or beta-adrenergic receptor signaling in the subject.

4. A method of enhancing apheresis by inducing lymphocyte or immune cell mobilization in a subject, the method comprising: blocking CXCR4 signaling and / or beta-adrenergic receptor signaling in the subject.

5. A method of enhancing apheresis by mobilizing a lymphocyte or an immune cell in a subject, the method comprising: blocking CXCR4 signaling and / or beta-adrenergic receptor signaling in the subject.

6. The method of any one of claims 1-5, wherein the blocking beta-adrenergic receptor signaling is performed before the blocking CXCR4 signaling.

7. The method of any one of claims 1-5, wherein the blocking beta-adrenergic receptor signaling continues after the blocking CXCR4 signaling is terminated.

8. The method of any one of claims 1-7, wherein the blocking CXCR4 signaling comprises administering a CXCR4 inhibitor to the subject.

9. The method of any one of claims 1-7, wherein the blocking beta-adrenergic receptor signaling comprises administering a beta-adrenergic receptor inhibitor to the subject.

10. The method of any one of claims 1-2 and 4-9, wherein the lymphocyte or immune cell is a lymphocyte.

11. The method of any one of claims 1-7, wherein the blocking CXCR4 signaling comprises administering a CXCR4 inhibitor to the subject and the blocking beta-adrenergic receptor signaling comprises administering a beta-adrenergic receptor inhibitor to the subject.

12. The method of claim 11, wherein the lymphocyte or immune cell is a lymphocyte.

13. A method of mobilizing a lymphocyte in a subject, the method comprising: administering a beta-adrenergic receptor inhibitor and / or a CXCR4 inhibitor to the subject.

14. A method of inducing lymphocyte mobilization in a subject, the method comprising: administering a beta-adrenergic receptor inhibitor and / or a CXCR4 inhibitor to the subject.

15. A method of enhancing apheresis in a subject, the method comprising: administering a beta-adrenergic receptor inhibitor and / or a CXCR4 inhibitor to the subject.

16. A method of enhancing apheresis by inducing cell mobilization in a subject, the method comprising: administering a beta-adrenergic receptor inhibitor and / or a CXCR4 inhibitor to the subject.

17. A method of enhancing apheresis by mobilizing a cell in a subject, the method comprising: administering a beta-adrenergic receptor inhibitor and / or a CXCR4 inhibitor to the subject.

18. The method of any one of claims 13-17, wherein the administering the beta-adrenergic receptor inhibitor is performed before the administering the CXCR4 inhibitor.

19. The method of any one of claims 13-17, wherein the administering the beta-adrenergic receptor inhibitor continues after the administering the CXCR4 inhibitor is terminated.

20. The method of any one of claims 13-19, the method further comprising: administering G-CSF to the subject.

21. The method of any one of claims 13-19, wherein the administering the beta-adrenergic receptor inhibitor and the CXCR4 inhibitor to the subject is performed in the absence of G- CSF.

22. A method of mobilizing a ly mphocyte in a subject, the method comprising: administering a CXCR4 inhibitor and G-CSF to the subject, in the absence of a beta- adrenergic receptor inhibitor.

23. A method of inducing lymphocyte mobilization in a subject, the method comprising: administering a CXCR4 inhibitor and G-CSF to the subject, in the absence of a beta- adrenergic receptor inhibitor.

24. A method of enhancing apheresis in a subject, the method comprising: administering a CXCR4 inhibitor and G-CSF to the subject, in the absence of a beta- adrenergic receptor inhibitor.

25. A method of enhancing apheresis by inducing cell mobilization in a subject, the method comprising: administering a CXCR4 inhibitor and G-CSF to the subject, in the absence of a beta- adrenergic receptor inhibitor.

26. A method of enhancing apheresis by mobilizing a cell in a subject, the method comprising:administering a CXCR4 inhibitor and G-CSF to the subject, in the absence of a beta- adrenergic receptor inhibitor.

27. The method of any one of claims 9 and 11-21, wherein the beta-adrenergic receptor inhibitor is an ADRB2 inhibitor.

28. The method of any one of claims 9 and 11-21, wherein the beta-adrenergic receptor inhibitor is selected from the group consisting of alprenolol, atenolol, betaxolol, bupranolol, butoxamine, carazolol, carvedilol, CGP 12177, cicloprolol, ICI 118551, ICYP, labetalol, levobetaxolol, levobunolol, LK 204-545, metoprolol, nadolol, NIHP, NIP, propafenone, propranolol, sotalol, SR59230A. and timolol.

29. The method of claim 28, wherein the beta-adrenergic receptor inhibitor is selected from the group consisting of propranolol, nadolol, and ICI 118551.

30. The method of claim 29, wherein the beta-adrenergic receptor inhibitor is propranolol.

31. The method of any one of claims 8, 11-30, wherein the CXCR4 inhibitor is selected from the group consisting of ALX40-4C, AMD070 (AMD11070, X4P-001), AMD3100 (plerixafor), AMD3465, ATI 2341, BKT140 (BL-8040; TF14016; 4F-Benzoyl-TN14003). CTCE-9908, CX549„D-[Lys3] GHRP-6, FC122, FC131, GMI-1359, GSK812397, GST- NT21MP, isothiourea- la, isothiourea-lt (ITlt), KRH-1636, KRH-3955, LY2510924, MSX- 122, N-[l lC]Methyl-AMD3465, POL6326, SDF-1 1-9[P2G] dimer, SDF1 P2G, T134, T140, T22, TC 14012, TG-0054 (Burixafor). USL311. viral macrophage inflammatory protein-II (vMIP-II), WZ811, [64Cu]-AMD3100, [64Cu]-AMD3465, [68Ga]pentixafor, [90Y]pentixather, [99mTc]02-AMD3100, [177Lu]pentixather, and 508MC1 (Compound 26).

32. The method of claim 31, wherein the CXCR4 inhibitor is selected from the group consisting of AD-214, AMD070 (AMD11070, X4P-001). AMD3100 (plerixafor), BKT140 (BL-8040; TF14016; 4F-Benzoyl-TN14003), CTCE-9908, LY2510924, LY2624587, T140, TG-0054 (Burixafor), PF-06747143, POL6326, and ulocuplumab (MDX1338 / BMS-936564).

33. The method of claim 32, wherein the CXCR4 inhibitor is TG-0054 (burixafor).

34. The method of claim 32, wherein the CXCR4 inhibitor is AMD3100 (plerixafor).

35. The method of claim 32, wherein the CXCR4 inhibitor is ulocuplumab (MDX1338 / BMS-936564).

36. The method of any one of claims 8, 1 1 -21 , and 27-30, wherein the administering the CXCR4 inhibitor and beta-adrenergic receptor inhibitor to the subject comprises administering TG-0054 (burixafor) and propranolol.

37. The method of any one of claims 8, 11-21, and 27-30, wherein the administering the CXCR4 inhibitor beta-adrenergic receptor inhibitor to the subject comprises administering AMD3100 (plerixafor) and propranolol.

38. The method of any one of claims 8, 11-21, and 27-30, wherein the administering the CXCR4 inhibitor beta-adrenergic receptor inhibitor to the subject comprises administering ulocuplumab (MDX1338 / BMS-936564) and propranolol.

39. The method of any one of claims 20 and 22-26, wherein the administering a combination of the CXCR4 inhibitor and the G-CSF induces an enhanced amount of lymphocyte or immune cell mobilization relative to the amount of lymphocyte or immune cell mobilization induced by the CXCR4 inhibitor only.

40. The method of any one of claims 20 and 22-26. wherein the administering a combination of the CXCR4 inhibitor and the G-CSF mobilizes a lymphocyte or an immune cell by an amount enhanced relative to the amount of lymphocyte or immune cell mobilization induced by the CXCR4 inhibitor only.

41. The method of any one of claims 20 and 22-26. wherein the administering a combination of the CXCR4 inhibitor and the G-CSF induces an enhanced amount of apheresis relative to the amount of apheresis induced by the CXCR4 inhibitor only.

42. The method of any one of claims 11-21 and 27-38, wherein the administering a combination of the CXCR4 inhibitor and the beta-adrenergic receptor inhibitor induces an enhanced amount of lymphocyte or immune cell mobilization relative to the amount of lymphocyte or immune cell mobilization induced by the CXCR4 inhibitor only.

43. The method of any one of claims 11-21 and 27-38, wherein the administering a combination of the CXCR4 inhibitor and the beta-adrenergic receptor inhibitor mobilizes a lymphocyte or an immune cell by an amount enhanced relative to the amount of lymphocyte or immune cell mobilization induced by the CXCR4 inhibitor only.

44. The method of any one of claims 11-21 and 27-38, wherein the administering a combination of the CXCR4 inhibitor and the beta-adrenergic receptor inhibitor induces an enhanced amount of apheresis relative to the amount of apheresis induced by the CXCR4 inhibitor only.

45. The method of claim 20, wherein the administering a combination of the CXCR4 inhibitor, the beta-adrenergic receptor inhibitor, and the G-CSF induces an enhanced amount of lymphocyte or immune cell mobilization relative to the amount of lymphocyte or immune cell mobilization induced by the CXCR4 inhibitor and the beta-adrenergic receptor inhibitor only.

46. The method of claim 20, wherein the administering a combination of the CXCR4 inhibitor, the beta-adrenergic receptor inhibitor, and the G-CSF mobilizes a cell by an amount enhanced relative to the amount of cell mobilization induced by the CXCR4 inhibitor and the beta-adrenergic receptor inhibitor only.

47. The method of claim 20, wherein the administering a combination of the CXCR4 inhibitor and the beta-adrenergic receptor inhibitor, and the G-CSF induces an enhanced amount of apheresis relative to the amount of apheresis induced by the CXCR4 inhibitor and the beta-adrenergic receptor inhibitor only.

48. The method of claim 33, wherein the administering a combination of TG-0054 (burixafor) and the G-CSF induces an enhanced amount of lymphocyte or immune cell mobilization relative to the amount of lymphocyte or immune cell mobilization induced by AMD3100 (plerixafor) and the G-CSF.

49. The method of claim 33, wherein the administering a combination of the TG-0054 (burixafor) and the G-CSF mobilizes a lymphocyte or an immune cell by an amount enhanced relative to the amount of lymphocyte or immune cell mobilization induced by the AMD31 0 (plerixafor) and the G-CSF.

50. The method of claim 33, wherein the administering a combination of the TG-0054 (burixafor) and the G-CSF induces an enhanced amount of apheresis relative to the amount of apheresis induced by the AMD3100 (plerixafor) and the G-CSF.

51. The method of any one of claims 1-50, wherein an enhanced amount of lymphocyte or immune cell mobilization or apheresis is measured by a method selected from the group consisting of complete blood count (CBC) analysis, flow cytometry, and colony forming unit (CFU) assay.

52. The method of claim 51, wherein the enhanced amount of lymphocyte or immune cell mobilization or apheresis is measured by flow cytometry.

53. The method of claim 51, wherein the flow cytometry is performed on (Lin-Scal+c- Kit+) LSK cells and / or CD34+ cells.

54. The method of claim 51, wherein the enhanced amount of lymphocyte or immune cell mobilization or apheresis is measured by colony forming unit (CFU) assay.

55. The method of any one of claims 1-54, wherein the subject has a CXCR4 protomer in the lymphocyte or immune cell.

56. The method of any one of claims 1-54, wherein the subject has an ADRB2 protomer in the lymphocyte or immune cell.

57. The method of any one of claims 1-54, wherein the subject has a CXCR4 protomer and an ADRB2 protomer in the lymphocyte or immune cell.

58. The method of claim 57, wherein the subject has a CXCR4-ADRB2 heteromer in the lymphocyte or immune cell.

59. The method of claim 58, wherein: i) the CXCR4-ADRB2 heteromer has an enhanced amount of downstream calcium mobilization relative to downstream calcium mobilization from a CXCR4 protomer or ADRB2 protomer; and ii) the administered combination of inhibitors suppresses the enhanced downstream calcium mobilization from said CXCR4-ADRB2 heteromer in the lymphocyte.

60. The method of any one of claims 1, 2, and 4-11, wherein the lymphocyte or immune cell is a lymphocyte.

61. The method of claim 60, wherein the lymphocyte is selected from the group consisting of a T cell, B cell, and NK cell.

62. The method of claim 61, wherein the lymphocyte is mobilized from bone marrow to peripheral blood.

63. The method of claim 62, wherein the lymphocyte is collected for transplantation to a patient having cancer.

64. The method of claim 63, wherein the cancer is selected from the group consisting of lymphoma, leukemia, and myeloma.

65. The method of claim 64, wherein the cancer is non-Hodgkin lymphoma (NHL), acute myeloid leukemia (AML), acute lymphoblastic leukemia (ALL), or multiple myeloma (MM).

66. The method of claim 65, wherein the lymphocyte is mobilized from bone marrow to peripheral blood.

67. The method of claim 66, wherein the lymphocyte is mobilized for treatment of a condition selected from the group consisting of neurological disorder, cardiac ischemia, myocardial infarction, diabetes, tissue repair, bone and cartilage disease, autoimmune disease, graft versus host disease, Crohn's disease, multiple sclerosis, systemic lupus ery thematosus, and systemic sclerosis.

68. The method of claim 67, wherein the lymphocyte is mobilized into blood.

69. The method of claim 68, wherein the lymphocyte is mobilized for treatment of a cancer.

70. The method of any one of claims 1, 2, 4-11, 16, 17, 25, and 26, wherein the cell is an immune cell.

71. The method of claim 70, wherein the immune cell is a white blood cell.

72. The method of claim 71, wherein the white blood cell is a lymphocyte.

73. The method of claim 72, wherein the lymphocyte is selected from the group consisting of a T cell, a B cell, and a natural killer (NK) cell.

74. The method of claim 73, wherein the lymphocyte is a T cell.

75. The method of claim 73, wherein the lymphocyte is a natural killer (NK) cell.

76. The method of claim 71, wherein the white blood cell is a granulocyte.

77. The method of claim 76, wherein the granulocyte is selected from the group consisting of a neutrophil, an eosinophil, and a basophil.

78. The method of claim 77, wherein the granulocyte is a neutrophil.

79. The method of claim 71, wherein the white blood cell is a monocyte.

80. The method of any one of claims 70-79, wherein the immune cell is mobilized from bone marrow to peripheral blood.

81. The method of any one of claims 70-79, wherein the immune cell is mobilized from lymph node to peripheral blood.

82. The method of any one of claims 70-79, wherein the mobilized immune cell is used for adoptive cell therapy (ACT).

83. The method of claim 82, wherein the adoptive cell therapy (ACT) is chimeric antigen receptor (CAR) T cell therapy.

84. The method of claim 82, wherein the adoptive cell therapy (ACT) is netural killer (NK) cell therapy.

85. The method of claim 82, wherein the adoptive cell therapy (ACT) is engineered T-cell receptor (TCR) therapy.

86. The method of claim 82, wherein the adoptive cell therapy (ACT) is tumor-infiltrating lymphocyte (TIL) therapy.

87. The method of any one of claims 1-82, wherein the method does not involve the addition of a CXCR2 agonist.

88. The method of claim 87, wherein the CXCR2 agonist is MGTA-145.

89. The method of any one of claims 1-88, wherein the method is not used to treat a patient having a sickle cell disease.

90. A method of mobilizing a lymphocyte or an immune cell in a subject, the method comprising administering, alone or in combination, a CXCR4 inhibitor and G-CSF to the subject.

91. A method for treatment of cancer comprising administering, alone or in combination, a CXCR4 inhibitor and G-CSF to a subject.

92. A method of enhancing leukapheresis by inducing lymphocyte or immune cell mobilization in a subject, the method comprising administering, alone or in combination, a CXCR4 inhibitor and G-CSF to the subject.

93. The method of any one of claims 90 to 92, wherein the CXCR4 inhibitor and the G- CSF are administered to the subject in the absence of a beta-adrenergic receptor inhibitor.

94. The method of any one of claims 90 to 93, wherein the method comprises administering the G-CSF to the subject before administering the CXCR4 inhibitor to the subject.

95. The method of any one of claims 90 to 94, wherein the method comprises administering a combination of the CXCR4 inhibitor and G-CSF to the subject, and wherein the administering of the combination of the CXCR4 inhibitor and G-CSF to the subject induces an enhanced amount of lymphocyte mobilization relative to an amount of lymphocyte mobilization induced by administration of G-CSF only to the subject.

96. The method of any one of claims 90 to 94, wherein the method comprises administering a combination of the CXCR4 inhibitor and G-CSF to the subject, and wherein the administering of the combination of the CXCR4 inhibitor and G-CSF to the subject induces anenhanced amount of lymphocyte mobilization relative to an amount of ly mphocy te mobilization induced by administration of the CXCR4 inhibitor only to the subject.

97. The method of any one of claims 90 to 96, wherein the CXCR4 inhibitor is selected from the group consisting of ALX40-4C, AMD070 (AMD11070, X4P-001), AMD3100 (plerixafor), AMD3465, ATI 2341, BKT140 (BL-8040; TF14016; 4F-Benzoyl-TN14003). CTCE-9908. CX549„D-[Lys3] GHRP-6, FC122, FC131. GMI-1359. GSK812397. GST- NT21 MP, isothiourea- la, isothiourea- It (ITlt), KRH-1636, KRH-3955, LY2510924, MSX- 122, N-[l lC]Methyl-AMD3465, POL6326, SDF-1 1-9[P2G] dimer, SDF1 P2G, T134, T140, T22, TC 14012, TG-0054 (Burixafor), USL311, viral macrophage inflammatory protein-II (vMIP-II), WZ811, [64Cu]-AMD3100, [64Cu]-AMD3465, [68Ga]pentixafor, [90Y]pentixather, [99mTc]02-AMD3100, [177Lu]pentixather, and 508MC1 (Compound 26).

98. The method of any one of claims 90 to 96, wherein the CXCR4 inhibitor is selected from the group consisting of AD-214, AMD070 (AMD11070, X4P-001). AMD3100 (plerixafor), BKT140 (BL-8040; TF14016; 4F-Benzoyl -TN 14003). CTCE-9908. LY2510924. LY2624587, T140, TG-0054 (Burixafor), PF-06747143, POL6326, and ulocuplumab (MDX1338 / BMS-936564).

99. The method of any one of claims 90 to 96, wherein the CXCR4 inhibitor is TG-0054 (burixafor).

100. The method of any one of claims 90 to 99, wherein the method comprises administering a combination of burixafor and G-CSF to the subject, and wherein the administering of the combination of burixafor and G-CSF to the subject induces an enhanced amount of lymphocyte mobilization relative to an amount of lymphocyte mobilization induced by administration of G-CSF only to the subject.

101. The method of any one of claims 90 to 99, wherein the method comprises administering a combination of burixafor and G-CSF to the subject, and wherein the administering of the combination of the burixafor and G-CSF to the subject induces anenhanced amount of lymphocyte mobilization relative to an amount of ly mphocy te mobilization induced by administration of burixafor only to the subject.

102. The method of claims 100 or 101, wherein the enhanced amount of lymphocyte mobilization is measured by a method selected from the group consisting of complete blood count (CBC) analysis, flow cytometry, and colony forming unit (CFU) assay.

103. The method of claims 100 or 1 1 , wherein the enhanced amount of lymphocyte mobilization is measured by CBC analysis.

104. The method of any one of claim 90 and 92 to 103, wherein the subject has a CXCR4 protomer in the lymphocyte.

105. The method of any one of claims 90 and 92 to 104, wherein the lymphocyte or immune cell is selected from the group consisting of a T cell, B cell, a natural killer (NK) cell, a neutrophil, eosinophil, and a basophil.

106. The method of any one of claims 90 and 92 to 105, wherein the lymphocyte or immune cell is mobilized for treatment of a condition selected from the group consisting of neurological disorder, cardiac ischemia, myocardial infarction, diabetes, tissue repair, bone and cartilage disease, autoimmune disease, graft versus host disease, Crohn's disease, multiple sclerosis, systemic lupus erythematosus, and systemic sclerosis.

107. The method of any one of claims 90 and 92 to 105, wherein the lymphocyte or immune cell is mobilized for treatment of a cancer.

108. The method of claim 107, wherein the cancer is non-Hodgkin lymphoma (NHL), acute myeloid leukemia (AML), acute lymphoblastic leukemia (ALL), or multiple myeloma (MM).

109. The method of claim 107, wherein the cancer is multiple myeloma (MM).

110. The method of any one of claims 90 and 92 to 109, wherein the lymphocyte or immune cell is mobilized for adoptive cell therapy (ACT).1 11. The method of claim 110, wherein the adoptive cell therapy (ACT) is chimeric antigen (CAR) T cell therapy.

112. The method of any one of claims 91 and 93 to 105. wherein the cancer is non-Hodgkin lymphoma (NHL), acute myeloid leukemia (AML), acute lymphoblastic leukemia (ALL), or multiple myeloma (MM).

113. The method of any one of claims 91 and 93 to 105. wherein the cancer is multiple myeloma (MM).

114. The method of any one of claims 91, 93 to 105, and 112 to 113, wherein the treatment comprises leukapheresis.1 15. The method of any one of claims 91 and 93 to 105, and 112 to 113, wherein the treatment comprises adoptive cell therapy (ACT).

116. The method of claim 115. wherein the adoptive cell therapy (ACT) is chimeric antigen (CAR) T cell therapy.

117. A composition or compositions for mobilizing a lymphocyte or an immune cell in a subject comprising, alone or in combination, a CXCR4 inhibitor and G-CSF.1 18. A composition or compositions for treatment of cancer comprising, alone or in combination, a CXCR4 inhibitor and G-CSF.

119. A composition or compositions for enhancing leukapheresis by inducing lymphocyte or immune cell mobilization in a subject comprising, alone or in combination, a CXCR4 inhibitor and G-CSF.

120. The composition or compositions of any one of claims 117 to 119, wherein the composition does not include a beta-adrenergic receptor inhibitor.

121. The composition or compositions of any one of claims 117 to 119, wherein the composition further includes a beta-adrenergic receptor inhibitor.

122. The composition or compositions of any one of claims 117 to 121. wherein the CXCR4 inhibitor is selected from the group consisting of ALX40-4C, AMD070 (AMD 11070, X4P- 001), AMD3100 (plerixafor), AMD3465, ATI 2341, BKT140 (BL-8040; TF14016; 4F- Benzoyl-TN14003), CTCE-9908, CX549„D-[Lys3] GHRP-6, FC122, FC131, GMI-1359, GSK812397. GST-NT21MP, isothiourea- la, isothiourea- It (ITlt), KRH-1636. KRH-3955, LY2510924, MSX-122, N-[l lC]Methyl-AMD3465, POL6326, SDF-1 1-9[P2G] dimer, SDF1 P2G, T134, T140, T22, TC 14012, TG-0054 (Burixafor), USL311, viral macrophage inflammatory protein-II (vMIP-II), WZ811, [64Cu]-AMD3100, [64Cu]-AMD3465, [68Ga]pentixafor, [90Y]pentixather. [99mTc]02-AMD3100, [177Lu]pentixather, and 508MC1 (Compound 26).

123. The composition or compositions of any one of claims 117 to 121, wherein the CXCR4 inhibitor is selected from the group consisting of AD-214, AMD070 (AMD11070, X4P-001), AMD3100 (plerixafor), BKT140 (BL-8040; TF14016; 4F-Benzoyl-TN14003). CTCE-9908. LY2510924, LY2624587, T140, TG-0054 (Burixafor), PF-06747143, POL6326, and ulocuplumab (MDX1338 / BMS-936564).

124. The composition or compositions of any one of claims 117 to 121, wherein the CXCR4 inhibitor is TG-0054 (burixafor).

125. The composition or compositions of any one of claims 117 and 119 to 124, wherein the subject has a CXCR4 protomer in the lymphocyte.

126. The composition or compositions of any one of claims 117 and 119 to 125, wherein the lymphocyte or immune cell is selected from the group consisting of a T cell, a B cell, and a natural killer (NK) cell, a neutrophil, a eosinophil, and a basophil.

127. The composition or compositions of any one of claims 117 and 119 to 126, wherein the lymphocyte or immune cell is mobilized for treatment of a condition selected from the group consisting of neurological disorder, cardiac ischemia, myocardial infarction, diabetes, tissue repair, bone and cartilage disease, autoimmune disease, graft versus host disease, Crohn's disease, multiple sclerosis, systemic lupus erythematosus, and systemic sclerosis.

128. The composition or compositions of any one of claims 1 17 and 119 to 126, wherein the lymphocyte or immune cell is mobilized for treatment of a cancer.

129. The composition or compositions of claim 128. the cancer is non-Hodgkin lymphoma (NHL), acute myeloid leukemia (AML), acute lymphoblastic leukemia (ALL), or multiple myeloma (MM).

130. The composition or compositions of claim 128. wherein the cancer is multiple myeloma (MM).

131. The composition or compositions of any one of claims 117 and 119 to 130, wherein the lymphocyte is mobilized for adoptive cell therapy (ACT).

132. The composition or compositions of claim 131, wherein the adoptive cell therapy (ACT) is chimeric antigen (CAR) T cell therapy.

133. The composition or compositions of any one of claims 118 and 120 to 124, wherein the cancer is non-Hodgkin lymphoma (NHL), acute myeloid leukemia (AML), acute lymphoblastic leukemia (ALL), or multiple myeloma (MM).

134. The composition or compositions of any one of claims 118 and 120 to 124, wherein the cancer is multiple myeloma (MM).

135. The composition or compositions of any one of claims 118, 120 to 124, and 133 to 134, wherein the treatment comprises leukapheresis.

136. The composition or compositions of any one of claims 118, 120 to 124, and 133 to 135, wherein the treatment comprises adoptive cell therapy (ACT).

137. The composition or compositions of claim 136, wherein the adoptive cell therapy (ACT) is chimeric antigen (CAR) T cell therapy.

138. A pharmaceutical composition or pharmaceutical compositions for mobilizing a lymphocyte or an immune cell in a subject comprising, alone or in combination, a CXCR4 inhibitor and G-CSF, and a pharmaceutically acceptable excipient.

139. A pharmaceutical composition or pharmaceutical compositions for treatment of cancer comprising, alone or in combination, a CXCR4 inhibitor and G-CSF, and a pharmaceutically acceptable excipient.

140. A pharmaceutical composition or pharmaceutical compositions for enhancing leukapheresis by inducing lymphocyte or immune cell mobilization in a subject comprising, alone or in combination, a CXCR4 inhibitor and G-CSF, and a pharmaceutically acceptable excipient.

141. The pharmaceutical composition or pharmaceutical compositions of any one of claims 138 to 140, wherein the pharmaceutical composition does not include a beta-adrenergic receptor inhibitor.

142. The pharmaceutical composition or pharmaceutical compositions of any one of claims 138 to 140, wherein the pharmaceutical composition further includes a beta-adrenergic receptor inhibitor.

143. The pharmaceutical composition or pharmaceutical compositions of any one of claims 138 to 142, wherein the CXCR4 inhibitor is selected from the group consisting of ALX40-4C, AMD070 (AMD11070, X4P-001), AMD3100 (plerixafor), AMD3465, ATI 2341, BKT140 (BL-8040; TF14016: 4F-Benzoyl-TN14003), CTCE-9908, CX549„D-[Lys3] GHRP-6, FC122,FC131, GMI-1359, GSK812397, GST-NT21MP, isothiourea- la, isothiourea-lt (ITlt), KRH- 1636, KRH-3955, LY2510924, MSX-122, N-[l lC]Methyl-AMD3465. POL6326, SDF-1 1- 9[P2G] dimer, SDF1 P2G, T134, T140, T22, TC 14012, TG-0054 (Bunxafor), USL31 1, viral macrophage inflammatory protein-II (vMIP-II), WZ811, [64Cu]-AMD3100, [64Cu]- AMD3465, [68Ga]pentixafor, [90Y]pentixather, [99mTc]02-AMD3100, [177Lu]pentixather, and 508MC1 (Compound 26).

144. The pharmaceutical composition or pharmaceutical compositions of any one of claims 138 to 142, wherein the CXCR4 inhibitor is selected from the group consisting of AD-214, AMD070 (AMD 11070, X4P-001), AMD3100 (plerixafor), BKT140 (BL-8040; TF14016; 4F- Benzoyl-TN 14003), CTCE-9908, LY2510924, LY2624587, T140, TG-0054 (Burixafor), PF- 06747143, POL6326, and ulocuplumab (MDX1338 / BMS-936564).

145. The pharmaceutical composition or pharmaceutical compositions of any one of claims 138 to 142. wherein the CXCR4 inhibitor is TG-0054 (burixafor).

146. The pharmaceutical composition or pharmaceutical compositions of any one of claims 138 and 140 to 145, wherein the subject has a CXCR4 protomer in the lymphocyte.

147. The pharmaceutical composition or pharmaceutical compositions of any one of claims 138 and 140 to 146, wherein the lymphocyte or immune cell is selected from the group consisting of a T cell, a B cell, and a natural killer (NK) cell, a neutrophil, an eosinophil, and a basophil.

148. The pharmaceutical composition or pharmaceutical compositions of any one of claims 138 and 140 to 147, wherein the lymphocyte or immune cell is mobilized for treatment of a condition selected from the group consisting of neurological disorder, cardiac ischemia, myocardial infarction, diabetes, tissue repair, bone and cartilage disease, autoimmune disease, graft versus host disease, Crohn's disease, multiple sclerosis, systemic lupus erythematosus, and systemic sclerosis.

149. The pharmaceutical composition or pharmaceutical compositions of any one of claims 138 and 140 to 147, wherein the lymphocyte or immune cell is mobilized for treatment of a cancer.

150. The pharmaceutical composition or pharmaceutical compositions of claim 149, wherein the cancer is non-Hodgkin lymphoma (NHL), acute myeloid leukemia (AML), acute lymphoblastic leukemia (ALL), or multiple myeloma (MM).

151. The pharmaceutical composition or pharmaceutical compositions of claim 149, wherein the cancer is multiple myeloma (MM).

152. The pharmaceutical composition or pharmaceutical compositions of any one of claims138 and 140 to 151, wherein the lymphocyte or immune cell is mobilized for adoptive cell therapy (ACT).

153. The pharmaceutical composition or pharmaceutical compositions of claim 152, wherein the adoptive cell therapy (ACT) is chimeric antigen (CAR) T cell therapy.

154. The pharmaceutical composition or pharmaceutical compositions of any one of claims139 and 141 to 145, wherein the cancer is non-Hodgkin lymphoma (NHL), acute myeloid leukemia (AML), acute lymphoblastic leukemia (ALL), or multiple myeloma (MM).

155. The pharmaceutical composition or pharmaceutical compositions of any one of claims 139 and 141 to 145, wherein the cancer is multiple myeloma (MM).

156. The pharmaceutical composition or pharmaceutical compositions of any one of claims 139, 141 to 145, and 154 to 155, wherein the treatment comprises leukapheresis.

157. The pharmaceutical composition or pharmaceutical compositions of any one of claims 139, 141 to 145, and 154 to 156, wherein the treatment comprises adoptive cell therapy (ACT).

158. The pharmaceutical composition or pharmaceutical compositions of claim 157, wherein the adoptive cell therapy (ACT) is chimeric antigen (CAR) T cell therapy.

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