Biomarkers and uses thereof for selecting immunotherapy intervention

Abstract

The instant disclosure provides biomarkers and methods for identifying subjects at risk of developing cytokine release syndrome (CRS), neurotoxicity, or both after adoptive immunotherapy to guide preemptive intervention, modified therapy, or the like. For example, adverse event biomarkers may be measured in a subject before pre-conditioning chemotherapy, before immunotherapy (e.g., adoptive immunotherapy infusion comprising a chimeric antigen receptor (CAR)-modified T cell), or shortly after pre-conditioning chemotherapy and/or immunotherapy. Exemplary biomarkers include temperature, cytokine levels and endothelial activation biomarkers, such as angiopoietin-2, von Willebrand factor (vWF), ratio of angiopoietin-2 to angiopoietin-1, and ratio of ADAMTS13 to vWF. Also provided are methods of treating subjects identified as at risk of developing cytokine release syndrome (CRS), neurotoxicity, or both to minimize such potential adverse events.

1 .- 19 . (canceled) 
     
     
         20 . A method for diagnosing or detecting the risk of an adverse event associated with cellular immunotherapy, comprising:
 measuring the level of an adverse event biomarker of endothelial activation in a biological sample from a mammalian subject having a hematologic malignancy within about 12 hours to about 48 hours after cellular immunotherapy, wherein the adverse event biomarker measured comprises the subject's temperature, or comprises the subject's temperature and a cytokine selected from IL-6, CCL2, IFN-γ, IL-10, IL-15, IL-2, or any combination thereof provided that at least IL-6, CCL2 or both cytokine levels are measured; and   (ii) identifying the subject as at risk of developing an adverse event of cytokine release syndrome (CRS), neurotoxicity, or both after cellular immunotherapy when the subject's temperature is at least 38° C. and the level of IL-6 is increased at least 2- to 5-fold and/or the level of CCL2 is increased at least 5- to 20-fold as compared to a normal sample, wherein the at risk subject receives pre-emptive treatment for the adverse event, receives an altered cellular immunotherapy regimen, or both.   
     
     
         21 . (canceled) 
     
     
         22 . The method of  claim 20 , wherein the measuring comprises measuring the level of two, three, four or five adverse event biomarkers. 
     
     
         23 . The method of  claim 20 , wherein the sample is obtained from the subject within 36 hours after cellular immunotherapy. 
     
     
         24 . The method of  claim 20 , wherein the measured adverse event biomarker comprises the subject's temperature of at least 38.5° C. to at least 39° C., or comprises the subject's temperature of at least 38.5° C. to at least 39° C., the level of IL-6 of at least 12 pg/mL to at least 16 pg/mL, and the level of CCL2 of at least 1,300 pg/mL to at least 1,350 pg/mL. 
     
     
         25 . The method of  claim 20 , wherein the method further comprises measuring the level of a biomarker of endothelial activation. 
     
     
         26 . The method of  claim 25 , wherein the biomarker of endothelial activation comprises a component of endothelial Weibel-Palade bodies selected from angiopoietin-2, vWF Ag, IL-8, CCL26, endothelin-1, osteoprotegerin, CD142 tissue factor, P-selectin, P-selectin cofactor CD63/LAMP3, PAI-1, α-fucosyltransferase VI, or any combination thereof. 
     
     
         27 . The method of  claim 25 , wherein the biomarker of endothelial activation measured comprises vWF Ag, angiopoietin-2, angiopoietin-1, VCAM-1, or a combination thereof. 
     
     
         28 . The method of  claim 25 , wherein the method further comprises measuring a co-factor to the biomarker of endothelial activation. 
     
     
         29 . The method of  claim 28 , wherein the method comprises measuring the level of vWF Ag and the co-factor measured comprises ADAMTS13 activity, wherein a ratio of ADAMTS13:vWF Ag that is reduced as compared to a normal sample identifies the subject as at risk of developing an adverse event associated with cellular immunotherapy. 
     
     
         30 . The method of  claim 28 , wherein the method comprises (a) measuring the level of angiopoietin-2 and the co-factor measured comprises measuring angiopoietin-1 level, wherein a ratio of angiopoietin-2:angiopoietin-1 that is increased as compared to a normal sample identifies the subject as at risk of developing an adverse event associated with cellular immunotherapy, and/or (b) measuring the level of VCAM-1 and the co-factor measured comprises measuring angiopoietin-1 level, wherein a ratio of VCAM-1:angiopoietin-1 that is increased as compared to a normal sample identifies the subject as at risk of developing an adverse event associated with cellular immunotherapy. 
     
     
         31 . The method of  claim 20 , wherein the pre-emptive treatment for the adverse event or the altered cellular immunotherapy regimen comprises administering the cellular immunotherapy at a reduced dose, a corticosteroid, an inflammatory cytokine antagonist, an endothelial cell stabilizing agent, or any combination thereof. 
     
     
         32 . The method of  claim 31 , wherein the pre-emptive treatment for the adverse event comprises administering the corticosteroid, the inflammatory cytokine antagonist, or both. 
     
     
         33 .- 40 . (canceled) 
     
     
         41 . The method of  claim 20 , wherein the hematologic malignancy is selected from Hodgkin's lymphoma, non-Hodgkins lymphoma (NHL), primary central nervous system lymphomas, T cell lymphomas, small lymphocytic lymphoma (SLL), B-cell prolymphocytic leukemia, lymphoplasmacytic lymphoma, splenic marginal zone lymphoma, plasma cell myeloma, solitary plasmacytoma of bone, extraosseous plasmacytoma, extra-nodal marginal zone B-cell lymphoma (mucosa-associated lymphoid tissue (MALT) lymphoma), nodal marginal zone B-cell lymphoma, follicular lymphoma, mantle cell lymphoma, diffuse large B-cell lymphoma (DLBCL), mediastinal (thymic) large B-cell lymphoma, intravascular large B-cell lymphoma, primary effusion lymphoma, acute lymphoblastic leukemia (ALL), acute myeloid leukemia (AML), chronic lymphocytic leukemia (CLL), chronic myoblastic leukemia (CML), Hairy cell leukemia (HCL), chronic myelomonocytic leukemia (CMML), juvenile myelomonocytic leukemia (JMML), large granular lymphocytic leukemia (LGL), blastic plasmacytoid dendritic cell neoplasm (BPDCN), Burkitt lymphoma/leukemia, multiple myeloma, Bence-Jones myeloma, non-secretory myeloma, plasmacytoma, amyloidosis, monoclonal gammopathy of unknown significance (MGUS), or Waldenstrom's macroglobulinemia. 
     
     
         42 . The method of  claim 20 , wherein the adverse event is cytokine release syndrome (CRS), neurotoxicity, or both. 
     
     
         43 . A kit for use in diagnosing or detecting the risk of an adverse event associated with cellular immunotherapy in a mammalian subject having a hematologic malignancy, comprising:
 a binding reagent and detectable agent for measuring the level of a plurality of cytokines selected from IL-6, CCL2, IFN-γ, IL-10, IL-15, IL-2, or any combination thereof, provided that reagents for detecting at least IL-6, CCL2 or both are provided;   optionally a device for measuring the subject's temperature;   an optional binding reagent and detectable agent for measuring the level or activity of a biomarker of endothelial activation selected from angiopoietin-2, angiopoietin-1, VCAM-1, vWF Ag, IL-8, CCL26, endothelin-1, osteoprotegerin, CD142 tissue factor, P-selectin, P-selectin cofactor CD63/LAMP3, PAI-1, α-fucosyltransferase VI, ADAMTS13, angiopoietin-1, or any combination thereof, provided that when the binding reagent for angiopoietin-2 or vWF Ag is present, the kit also contains a reagent for detecting activity of ADAMTS13 or detecting angiopoietin-1, respectively; and   optional reagents for performing a binding reaction using the detectable agent,   optional instructions for using the binding reagent and the detectable agent;   wherein the subject is identified as at risk of developing an adverse event associated with cellular immunotherapy when the biomarker of endothelial activation is increased as compared to a normal sample; or   wherein the subject is identified as at risk of developing an adverse event of cytokine release syndrome (CRS), neurotoxicity, or both after cellular immunotherapy when the subject's temperature is at least 38° C. and the level of IL-6 is increased at least 2- to 5-fold and/or the level of CCL2 is increased at least 5- to 20-fold as compared to a normal sample.   
     
     
         44 . The kit of  claim 43 , wherein the binding reagent comprises a nanobody or a binding fragment thereof, an antibody or a binding fragment thereof, or a T cell receptor or a binding fragment thereof. 
     
     
         45 . The kit of  claim 43 , wherein the binding reagent is conjugated to a detectable agent. 
     
     
         46 . The kit of  claim 43 , wherein the detectable agent is detectable by one or more of: a colorimetric assay, fluorescence imaging, an enzymatic assay, spectrophotometry, mass spectroscopy, or radiation imaging. 
     
     
         47 . A method for treating hematologic malignancy in a mammalian subject, the method comprising:
 (a) obtaining a result from the method of  claim 20  to determine the risk of an adverse event associated with cellular immunotherapy in the subject; and   (b) administering to the subject a pre-emptive treatment, an altered cellular immunotherapy regimen, or both to minimize the risk for the potential adverse event.   
     
     
         48 .- 49 . (canceled) 
     
     
         50 . The method of  claim 20 , wherein the therapy comprises chemotherapy, combined chemotherapies, biologic therapy, hormonal therapy, or any combination thereof. 
     
     
         51 . The method of  claim 50 , wherein the biologic therapy comprises an antibody, an scFv, a nanobody, a fusion protein, a tyrosine kinase inhibitor, an immunoreactive T cell, an immunoreactive Natural Killer cell, or any combination thereof. 
     
     
         52 .- 53 . (canceled)