Methods for the detection and treatment of hematologic malignancy associated hyperinflammation

Abstract

Methods of diagnosing and treating hematologic malignancy-associated hyperinflammation in individuals having a hematologic malignancy are disclosed. The method includes determining an optimized Hemophagocytic lymphohistiocytosis (HLH) inflammatory (OHI) index status of the individual, and based on the index status, administering to the individual an effective amount of an anti-inflammatory or checkpoint blockade therapy. The OHI index status is based on measured levels of soluble CD25 (sCD25) and ferritin in a biological sample of the individual.

Description

METHODS FOR THE DETECTION AND TREATMENT OF HEMATOLOGIC MALIGNANCY ASSOCIATED HYPERINFLAMMATIONCROSS-REFERENCE TO RELATED APPLICATIONS

[0001] The present patent application is an International application that claims priority to and the benefit of U.S. Provisional Application No. 63 / 607,338 filed December 7, 2023, the entire content of which is incorporated herein by reference for all purposes.STATEMENT REGARDING FEDERALLY-SPONSORED RESEARCH

[0002] This invention was made with government support under CA256390 awarded by the National Institutes of Health. The government has certain rights in the invention.BACKGROUND

[0003] Hemophagocytic lymphohistiocytosis (HLH) is a life-threatening hyperinfl ammatory condition characterized by an exaggerated and dysregulated immune response. Traditionally, HLH is clinically defined by diagnostic criteria for infants with familial HLH (FHL) - which is typically attributed to genetic mutations affecting the perforin pathway of lymphocyte cytotoxic function. In adults, HLH is mostly observed in the context of hematologic malignancies (HM- HLH), and many of the conventional diagnostic criteria may reflect the underlying malignancy rather than an abnormal inflammatory state. This leads to under- or mis-diagnosis of HLH in individuals with hematologic malignancies (HMs).

[0004] HM-HLH occurs most commonly in individuals with newly diagnosed HMs (most commonly lymphomas) who do not have genetic lesions associated with FHL. Although HM- HLH has been recognized, its pathophysiology is still not understood. This knowledge gap has likely contributed to poor patient outcomes - a survival rate as low as 10-20% - and has left this field without a rational-based therapeutic approach to improve patient outcomes. Therefore, there is a need to improve the identification and treatment of patients with HM-HLH.BRIEF SUMMARY

[0005] According to aspects of this disclosure, methods of diagnosing and treating hematologic malignancy-associated hyperinflammation in an individual having a hematologic malignancy include determining an optimized Hemophagocytic lymphohistiocytosis (HLH) inflammatory (OHI) index status of the individual, and based on the index status, administering to the individual an effective amount of an anti-inflammatory or checkpoint blockade therapy. The OHI index status is based on measured levels of soluble CD25 (sCD25) and ferritin in a biological sample of the individual.

[0006] In some aspects, methods of diagnosing and treating lymphoma-associated hyperinflammation in an individual having a lymphoma include determining the OHI index status of the individual, and based on the OHI index status of the individual, administering to the individual an effective amount of anti-inflammatory or checkpoint blockade therapy.

[0007] In some aspects, methods of identifying an individual with a hematologic malignancy that is likely to respond to or benefit from treatment with a checkpoint blockade therapy includes determining the OHI index status of the individual, and based on the OHI index status of the individual, identifying the individual as likely to benefit from a checkpoint blockade therapy.

[0008] In some aspects, methods of identifying an individual with a hematologic malignancy that is likely to respond to or benefit from treatment with an anti-inflammatory therapy includes determining the OHI index status of the individual, and based on the OHI index status of the individual, identifying the individual as likely to benefit from an anti-inflammatory therapy.

[0009] In some aspects, methods for identifying an individual having a hematologic malignancy that is likely to respond to or benefit from a treatment in a clinical trial includes determining the OHI index status of the individual, and based on the OHI index status of the individual, identifying the individual as a candidate for the clinical trial.BRIEF DESCRIPTION OF THE DRAWINGS

[0010] This application file contains at least one drawing executed in color. Copies of this patent or patent application publication with color drawing(s) will be provided by the Office upon request and payment of the necessary fee.

[0011] Those of skill in the art will understand that the drawings, described below, are for illustrative purposes only. The drawings are not intended to limit the scope of the present teachings in any way.

[0012] FIG. 1 A is a distribution plot of the ages of the patients in the study of the Experimental Example.

[0013] FIG. IB depicts side-by-side graphs of the survival rates of FHL patients and OHI+ patients in the study of the Experimental Example, showing that the survival rates of both pairwise comparisons between groups are statistically significant using the chi-square test, p=0.04.

[0014] FIG. 1C depicts side-by-side pie charts of types of malignancies of the OHI- (left) and OHI+ (right) patients assayed in the study of the Experimental Example.

[0015] FIG. ID depicts germline mutations in the FHL patients of the study of the Experimental Example. Out of the MUNC13+ group, 2 were homozygous, and 1 was heterozygous. The PRF+ patients were homozygous. None of the OHI+ patients were identified as having germline mutations.

[0016] FIG. IE depicts side-by-side distribution plots of ferritin (left) and sCD25 (right) values closest to the time of sample draw from the patients in the study of the Experimental Example. The statistical difference between quantitative variables was calculated using the Mann-Whitney U test, p<0.0001.

[0017] FIG. 2A is a graph of the unsupervised primary component analysis, showing the OHI+ OHI+ adult patients of the Experimental Example clustered with FHL infants, and showing healthy controls clustered with OHI- patients.

[0018] FIG. 2B is a heatmap of the patient samples of the Experimental Example sorted by hierarchical clustering (q<0.05 comparing OHI+ vs. OHI-, 1456 proteins).

[0019] FIG. 2C depicts side-by-side Hallmark gene set enrichment analyses of FHL vs. pediatric controls (left) and OHI+ vs. OHI- (right). The pathways shown are significantly enriched (q<0.25, p<0.05).

[0020] FIG. 3A is a Venn diagram depicting the under-expressed proteins (P-adjusted<0.05) of OHI+ vs. OHI- as compared to FHL vs. control.

[0021] FIG. 3B is a Venn diagram depicting the over-expressed proteins (P-adjusted<0.05) of OHI+ vs. OHI- as compared to FHL vs. control.

[0022] FIG. 3C is a scatter plot of the significantly differential intersecting overexpressed proteins (P-adjusted<0.01) comparing OHI+ vs. OHI- (x-axis) and FHL vs. control (y-axis).

[0023] FIG. 3D is a scatter plot of the significantly Hallmark enriched pathways (q<0.25) within the intersecting proteins comparing OHI+ vs. OHI- (x-axis) and FHL vs. control (y-axis).

[0024] FIG. 4A is a volcano plot of significantly expressed proteins in the T cell activation pathway of OHI+ patients in the Experimental Example.

[0025] FIG. 4B is a volcano plot of significantly expressed proteins in the IFN-y response pathway of OHI+ patients in the Experimental Example.

[0026] FIG. 4C is a volcano plot of significantly expressed proteins in the IL6 JAK STAT3 signaling pathway of OHI+ patients in the Experimental Example.

[0027] FIG. 4D is a volcano plot of significantly expressed proteins in the complement activation pathway of OHI+ patients in the Experimental Example.

[0028] FIG. 5A is a graph of the Hallmark enriched pathways in OHI+ vs. OHI- patients identified by GSEA.

[0029] FIG. 5B is a graph of the Reactome enriched pathways in OHI+ vs. OHI- patients identified by GSEA.

[0030] FIG. 5C is a graph of the Gene Ontogeny (GO) enriched pathways in OHI+ vs. OHI- patients identified by GSEA.

[0031] FIG. 5D depicts side-by-side heat maps depicting the Hallmark process analysis of selected classifiers for identifying IL6 JAK STAT3 activation, the IFN-y response, and complement activation.

[0032] FIG. 5E depicts side-by-side heat maps depicting the Reactome process analysis of selected classifiers for identifying IFN-y signaling and PD1 signaling.

[0033] FIG. 5F depicts side-by-side heat maps depicting the GO process analysis of selected classifiers for identifying T cell mediated immunity and leukotriene activation.

[0034] FIG. 6A is a graph of GSEA analysis of immune evasion related proteins in OHI+ vs. OHI- (Normalized Enrichment Score (NES) = 1.72 and q=0.008).

[0035] FIG. 6B is a waterfall plot of each individual patient’s enrichment of immune evasion.

[0036] FIG. 6C depicts side-by-side scatter plots of checkpoint ligands and receptors in OHI+ patients (red) and OHI- patients (blue) in DLBCL patients.

[0037] FIG. 6D depicts side-by-side scatter plots of checkpoint ligands and receptors in OHI+ patients (red) and OHI- patients (blue) in CLL patients.

[0038] FIG. 7A depicts side-by-side ELISA results for sCD8, 4-1BB, Granzyme B (GZMB), and Granzyme A (GZMA) in the T cell cytotoxicity and T cell receptor pathway.

[0039] FIG. 7B depicts side-by-side ELISA results for CXCL9, GBP2, CXCL10, and IL18BP in the IFN-y response and signaling pathway.

[0040] FIG. 7C depicts side-by-side ELISA results for sCD14 and HO-1 in the IL6 JAK STAT3 signaling pathway.

[0041] FIG. 7D depicts side-by-side ELISA results for MIF and sVSIG4 in the macrophage activation pathway.

[0042] FIG. 7E depicts the ELISA results for iC3b in the complement regulation activation and cytotoxicity activation pathway.

[0043] FIG. 7F depicts side-by-side ELISA results for the checkpoint proteins, sPDl, TIM3, sPDLl, GAL9, and HLA-G.DETAILED DESCRIPTIONDEFINITIONS

[0044] Unless otherwise noted, terms are to be understood according to conventional usage by those of ordinary skill in the relevant art. In case of conflict, the present document, including definitions, will control. Exemplary methods and materials are described below, although methods and materials similar or equivalent to those described herein may be used in practice or testing of the present disclosure. All publications, patent applications, patents and other references mentioned herein are incorporated by reference in their entirety. The materials, methods, and examples disclosed herein are illustrative only and not intended to be limiting. The methods may comprise, consist of, or consist essentially of the elements of the compositions and / or methods as described herein, as well as any additional or optional element described herein or otherwise useful in the identification and / or treatment of patients having HM-HLH.

[0045] As used herein and in the appended claims, the singular forms “a,” “and,” and “the” include plural referents unless the context clearly dictates otherwise. Thus, for example, reference to “a method” includes a plurality of such methods and reference to “a dose” includes reference to one or more doses and equivalents thereof known to those skilled in the art, and so forth.

[0046] The term “about” or “approximately” means within an acceptable error range for the particular value as determined by one of ordinary skill in the art, which will depend in part on how the value is measured or determined, e.g., the limitations of the measurement system. For example, “about” may mean within 1 or more than 1 standard deviation, per the practice in the art. Alternatively, “about” may mean a range of up to 20%, or up to 10%, or up to 5%, or up to 1% of a given value. Alternatively, particularly with respect to biological systems or processes, the term may mean within an order of magnitude, preferably within 5-fold, and more preferably within 2- fold, of a value. Where particular values are described in the application and claims, unlessotherwise stated the term “about” meaning within an acceptable error range for the particular value should be assumed.

[0047] As used herein, the term “effective amount” means the amount of one or more active components that is sufficient to show a desired effect. This includes both therapeutic and prophylactic effects. When applied to an individual active ingredient, administered alone, the term refers to that ingredient alone. When applied to a combination, the term refers to combined amounts of the active ingredients that result in the therapeutic effect, whether administered in combination, serially or simultaneously.

[0048] The terms “individual,” “host,” “subject,” and “patient” are used interchangeably to refer to an animal that is the object of treatment, observation and / or experiment. Generally, the term refers to a human patient, but the methods and compositions may be equally applicable to non-human subjects such as other mammals. In some aspects, the terms refer to humans. In further aspects, the terms may refer to adults or children, depending on the context.HEMATOLOGIC MALIGNANCY RELATED HYPERINFLAMMATION

[0049] HLH is increasingly being recognized in patients with hematologic malignancies (HM- HLH). While HLH is observed in both adults (HM-HLH) and in children (FLH), and while the mechanism in children is understood to be genetic mutations in lymphocyte cytotoxic pathways, the underlying mechanism of HM-HLH in adults is unclear. However, both FHL and HM-HLH are characterized by fever, splenomegaly, blood cytopenia, abnormal coagulation tests, and heightened markers of inflammation. Despite this, and despite that HM-HLH has been recognized for some time, neither its pathophysiology nor its similarity to FHL is understood - nor whether HM-HLH patients would benefit from immunosuppressive therapies.

[0050] In a previous work, an “Optimized HLH Inflammatory (OHI) index” was introduced. This OHI index enhances the identification of hyperinflammation and improves prognostication in patients with HMs. Zoref-Lorenz et al., “An improved index for diagnosis and mortality prediction in malignancy-associated hemophagocytic lymphohistiocytosis,” Blood, 139(7), pgs. 1098-1110 (2022) (doi: 10.1182 / blod.2021012764), the entire content of which, including the Supplemental Materials, is incorporated herein by reference and attached hereto as Appendix A.The OHT index combines measurements of soluble CD25 (sCD25) and ferritin levels to more accurately diagnose adult patients having HMs with HLH. Indeed, the OHI index proved to be a more effective diagnostic tool than the conventional FHL-based criteria for identifying HM patients having HLH, and for predicting mortality of such patients. Id.

[0051] Syndromic HLH developing in the context of malignancy may be caused by: A) the malignancy directly inducing diagnostic features of HLH; B) abnormal persistent activation of T cells; or C) as yet undefined forms of dysregulated inflammation. In A, the malignancy acts as a mimic of FHL, with little pathophysiologic overlap or rationale for therapeutic overlap. In B, persistent T cell activation , perhaps in response to malignancy, is similar to FHL and may suggest use of targeted therapies developed for FHL (such as, for example, emapalumab) and the potential to therapeutically exploit anti-cancer immunity (if toxicity can be managed). In C, dysregulated inflammation, typically in response to concurrent infection and potentially related to immune compromise, drives HLH features in ways that are not yet understood, and its translational implications are unclear. In this case, inflammation may be associated with direct mononuclear phagocyte activation (MnPs), within the tumor microenvironment.

[0052] Based on these clinical similarities, according to aspects of the present disclosure, the OHI index may be used to identify HM-HLH patients having immune pathophysiological mechanisms similar to those observed in FHL patients.

[0053] The OHI index characterizes patients as either OHI+ or OHI-, based on the measured levels of sCD25 and ferritin. The OHI index status of a patient provides increased accuracy in the prediction or detection of patients having high risk of mortality, OHI+ patients having high risk of mortality. As discussed further herein, the OHI index is based on the measurement of sCD25 and ferritin. OHI+ patients are defined as those having an sCD25 level greater than a predetermined threshold sCD25 level and a ferritin level greater than a predefined threshold ferritin level. Patients with either a sCD25 or ferritin level lower than the predefined threshold levels are identified as OHL. In some embodiments, for example, OHI+ patients are those having an sCD25 level greater than 3900 U / mL, and a ferritin level greater than 1000 ng / mL, and OHI- patients are those having either an sCD25 level lower than 3900 U / mL or a ferritin level lower than 1000 ng / mL.

[0054] The analysis detailed in the EXPERIMENTAL section of this disclosure confirms that patients identified as OHI+ have immune pathophysiological mechanisms similar to those observed in FHL patients. For example, as discussed further herein, the hyperinflammatory response in OHI+ patients is characterized by T cell activation, macrophage activation, interferon (IFN)-y-mediated response, IL6 JAK STAT 3 signaling, and IL2 STAT 5 signaling. OHI+ patients are also characterized by macrophage, complement and leukotriene activation. As such, according to aspects of the present disclosure, OHI+ patients can benefit from the administration of antiinflammatory therapies targeting these pathways.

[0055] OHI+ patients may also be characterized by immune evasion with elevated and correlated checkpoint ligand-receptor pairs, for example, correlated increases in PD-1-PD-L1, LGALS9-TIM3, and HLA-G-LILRB2. As such, according to aspects of the present disclosure, OHI+ patients can benefit from the administration of checkpoint blockade therapy, for example checkpoint blockade therapies targeting these proteins.

[0056] Drawing from this, according to aspects of the present disclosure, methods of diagnosing and treating hematologic malignancy related (or associated) hyperinflammation in an individual having a hematologic malignancy include determining the OHI index status of the individual, and based on the OHI index status, administering an effective amount of an antiinflammatory or checkpoint blockade therapy. In some aspects, the anti-inflammatory or checkpoint blockade therapy is delivered when the individual is determined to have OHI+ index status.

[0057] In aspects, OHI+ status is defined as an individual having a measured sCD25 level greater than a predefined threshold sCD25 level and a ferritin level greater than a predefined threshold ferritin level. In aspects, the predefined threshold sCD25 level is 3900U / mL. In aspects, the predefined threshold ferritin level is 1000 ng / mL. For example, in some aspects, the individual may be determined to have OHI+ status when the measured sCD25 level is greater than 3900 U / mL, and the ferritin level is greater than a threshold ferritin level. In some aspects, the individual may be determined to have OHI+ status when the measured ferritin level is greater than 1000 ng / mL, and the sCD25 level is greater than a threshold sCD25 level. In some aspects, for example,the individual may be determined to have OHI+ status when the measured sCD25 level is greater than 3900 U / mL and the ferritin level is greater than 1000 ng / mL.

[0058] For purposes of determining status, those individuals that are not OHI+ are determined to be OHI-. For example, OHI- status is defined as an individual having a measured sCD25 level lower than the predefined threshold sCD25 level and / or a ferritin level lower than the predefined threshold ferritin level. In some aspects, the predefined threshold sCD25 level is 3900U / mL. In some aspects, the predefined threshold ferritin level is 1000 ng / mL. For example, in some aspects, the individual may be determined to have OHI- status when the measured sCD25 level is lower than 3900 U / mL, or when the ferritin level is lower than 1000 ng / mL.

[0059] Accordingly, in aspects, determining the OHI index status of the individual includes measuring an amount of sCD25 and an amount of ferritin in serum from a biological sample, and comparing the measured amounts of sCD25 and ferritin to predefined (or predetermined) threshold levels. The OHI index status is then determined from the comparison, where, as discussed herein, OHI+ status is determined when both the measured sCD25 and ferritin levels are above the predefined threshold levels, and OHI- status is determined when one or both of the measured sCD25 and ferritin levels are below the predefined threshold levels. Methods of measuring sCD25 and ferritin in serum are known to those skilled in the art, and any suitable method of such measurement may be used.

[0060] The hematologic malignancy underlying the hyperinflammation that is treated by the methods according to aspects of this disclosure may be any hematologic malignancy, or blood cancer. For example, in some aspects, the hematologic malignancy may be a leukemia, a lymphoma, or a myeloma. In aspects, the hematologic malignancy may be a lymphoma. For example, in embodiments in which the hematologic malignancy is a lymphoma, the method may be for diagnosing and treating a lymphoma related (or associated) hyperinflammation in an individual having lymphoma, and may include determining the OHI index status of the individual, and based on the OHI index status, administering an effective amount of an anti-inflammatory or checkpoint blockade therapy.

[0061] Anti-inflammatory therapy may be any therapy or therapeutic effective to treat or reduce inflammation. In some aspects, the anti-inflammatory therapy may be a therapeutic thattargets the IFN-y-mediated response pathway, the IL6 JAK STAT 3 signaling pathway, or the IL2 STAT 5 signaling pathway. In aspects, the anti-inflammatory therapy is selected from anti-IFN-y agents; JAK inhibitors; complement inhibitors; leukotriene inhibitors; anti-IL-6 agents and antibodies; anti-TNF (anti-tumor necrosis factor) agents and antibodies; anti-MIF (antimacrophage migration inhibitory factor) agents and antibodies; and combinations thereof. In aspects, the anti-inflammatory therapy is selected from anti-IFN-y agents; JAK inhibitors; and combinations thereof.

[0062] Some nonlimiting examples of suitable therapeutics that target the IL6 JAK STAT3 pathway include JAK inhibitors and anti-IL-6 agents. Some non-limiting examples of suitable anti-IL-6 agents and antibodies include tocilizumab, sarilumab, siltuximab, sirukumab, olokizumab, clazakizumab, vobarilizumba, ziltivekimab, etc.. Some non-limiting examples of suitable JAK inhibitors include barcitinib, tofacitinib, ruxolitinib, upadacitinib, fedratinib, abrocitinib, ritlecitinib, momelotinib, pacritinib, etc.

[0063] Some nonlimiting examples of suitable therapeutics that target the IFN-y-mediated response pathway include anti-IFN-y drugs or agents. Some nonlimiting examples of suitable anti- IFN-y drugs or agents include emapalumab, belimumab, fontolizumab, etc.

[0064] Some nonlimiting examples of suitable therapeutics that target the IL2 STAT 5 signaling pathway include JAK inhibitors. As discussed above, some non-limiting examples of suitable JAK inhibitors include barcitinib, tofacitinib, ruxolitinib, upadacitinib, fedratinib, abrocitinib, ritlecitinib, momelotinib, pacritinib, etc.

[0065] Some nonlimiting examples of suitable complement inhibitors include eculizumab, pegcetacoplan, ravulizumab, avacincaptadpegol, pozelimab, zilucoplan, crovalimab-akkz, iptacopan, danicopan, avacopan, complement Cl esterase inhibitors (e.g., CINRYZE® available from ViroPharma Biologies LLC (Massachusetts, USA), BERINERT® available from CSL Behring GmbH (Germany), RUCONEST® available from Pharming Group N.V. (Netherlands)), sutimlimab, etc. CINRYZE® is a registered trademark owned by ViroPharma Biologies LLC (Massachusetts, USA); BERINERT® is a registered trademark of CSL Behring GmbH (Germany); and RUCONEST® is a registered trademark of Pharming Intellectual Property B.V. (Netherlands).

[0066] Some nonlimiting examples of suitable leukotriene inhibitors include zileuton, zafirlukast, pranlukast, montelukast, etc.

[0067] Some nonlimiting examples of suitable anti-TNF agents and drugs include infliximab, etanercept, adalimumab, certolizumab pegol, golimumab, etc.

[0068] Some nonlimiting examples of suitable anti-MIF agents and drugs include imalumab, milatuzumab, ebselen, etc.

[0069] The checkpoint blockade therapy is also not particularly limited, and may be any suitable such therapy. Some nonlimiting examples of suitable checkpoint blockade therapies include: PD-1 inhibitors, some nonlimiting examples of which include pembrolizumab, nivolumab, cemiplimab, dostarlimab, retifanlimab-dlwr, tislelizumab, etc.; PD-L1 inhibitors, some nonlimiting examples of which include atezolizumab, avelumab, durvalumab, etc.; CTLA-4 inhibitors, some nonlimiting examples of which include ipilimumab, tremelimumab, etc.; and LAG-3 inhibitors, some nonlimiting examples of which relatlimab, etc.

[0070] Again drawing on the pathophysiological mechanisms discussed above, in aspects, the OHI index may also be used to identify patients or individuals likely to respond to or benefit from certain treatment regimens. For example, drawing on the correlation between OHI+ status and the presence of immune evasion, in aspects, a method of identifying cancer patients likely to respond to or benefit from checkpoint blockade therapy includes determining the OHI index status of the patient, and identifying the patient as a candidate for checkpoint blockade therapy based on the OHI index status. In aspects, an OHI+ status is indicative of patients that are candidates for checkpoint blockade therapy. The method may further include administering an effective amount of the checkpoint blockade therapy to those patients identified as candidates for that therapy.

[0071] Similarly, in some aspects, drawing on the correlation between OHI+ status and the enrichment of certain biological pathways, a method is directed to identifying cancer patients likely to respond to or benefit from therapies that target the IFN-y-mediated response pathway, the IL6 IAK STAT 3 signaling pathway, the complement activation pathway, the leukotriene activation pathway, the macrophage activation pathway, or the IL2 STAT 5 signaling pathway. For example, in some embodiments, the method may be directed to identifying cancer patientslikely to respond to or benefit from therapies that target the IFN-y-mediated response pathway, the IL6 JAK STAT 3 signaling pathway, the complement activation pathway, or the leukotriene activation pathway. In some aspects, for example, the method may be directed to identifying cancer patients likely to respond to or benefit from anti-inflammatory therapies, for example, anti- IFN-y agents, JAK inhibitors, complement inhibitors, leukotriene inhibitors, anti-IL-6 agents and antibodies, anti-TNF agents and antibodies, anti-MIF agents and antibodies, or combinations thereof. In some aspects, the method may be directed to identifying cancer patients likely to respond to anti-IFN-y agents, JAK inhibitors, or combinations thereof. These methods may include determining the OHI index status of the patient, and identifying the patient as a candidate for the therapy based on the OHI index status. In some aspects of such methods, an OHI+ status is indicative of patients that are candidates for the therapy. In aspects, the method may further include administering an effective amount of the therapy to those patients identified as candidates for that therapy.

[0072] In some aspects of the present disclosure, the OHI index may also be used to identify patients or individuals with cancer that are candidates for a clinical trial or that are likely to benefit from a treatment in a clinical trial. For example, according to embodiments of the present disclosure, a method of identifying cancer patients likely to respond to or benefit from a therapy in a clinical trial includes determining the OHI index status of the patient, and identifying the patient as a candidate for the clinical trial based on the OHI index status. In some aspects of such a method, an OHI+ status is indicative of patients that are candidates for the clinical trial. The method may further include enrolling the identified OHI+ patient(s) in the clinical trial. And the method may further include administering an effective amount of the clinical trial therapy to those patients identified as candidates for the clinical trial.

[0073] According to some aspects, the clinical trial may be for therapeutics or therapies for treating a hematologic malignancy, or for treating hyperinflammation associated with a hematologic malignancy. For example, in some embodiments, the clinical trial may be for therapeutics or therapies for treating a lymphoma, a leukemia, or a myeloma, or for treating hyperinflammation associated with a lymphoma, a leukemia, or a myeloma. In some embodiments, for example, clinical trial may be for treating a lymphoma, or for treating hyperinflammation associated with a lymphoma.EXPERIMENTAL

[0074] The following non-limiting examples are provided to further illustrate concepts and embodiments of this disclosure. It should be appreciated by those of skill in the art that the techniques disclosed in the examples that follow represent approaches that have been found to function well in the practice of this disclosure, and thus may be considered to constitute examples of modes for practice. However, those of skill in the art should, in light of the present disclosure, appreciate that many changes may be made in the specific embodiments that are disclosed and still obtain a like or similar result without departing from the spirit and scope of the present disclosure.EXAMPLEMethods

[0075] Study Participants. In these examples, OHI index is defined by a sCD25 threshold level of 3,900 U / mL and a ferritin threshold level of 1,000 ng / mL, where HM-0HI+ status is indicative of an sCD25 level above the 3,900 U / mL threshold as well as a ferritin level about the 1,000 ng / mL threshold. All HM-0HI+ patients in the study answered these criteria. The study population included a total of 14 HM-0HI+ patients, 14 HM-OHI- patients, 7 patients with FHL, and 7 healthy pediatric controls.

[0076] Proteomic Analysis: To elucidate the distinctive immune signature of HM-0HI+ patients, a SomaLogic proteomic study was conducted This proteomic analysis allowed comprehensive characterization of the immune activation profiles of HM-0HI+ patients in comparison to HM-OHI- patients, FHL patients, and healthy controls. The SomaLogic proteomic analysis assessed 7,596 serum proteins representing 6,412 unique genes.

[0077] Unsupervised Clustering: Unsupervised clustering techniques, including primary component analysis (PCA) and unbiased hierarchical clustering utilizing a heat map were applied to group patient results based on their proteomic profiles. Proteins were filtered based on the optimal standard deviation (S / Sma ) for the PCA and filter of Padjusted<0.05 for the heat map.

[0078] Gene Set Enrichment Analysis (GSEA): GSEA of the hallmark gene set was employed to identify enriched biological pathways in HM-0HI+ patients vs. HM-0HI+ and FHL vs. Pedctl.For the pairwise comparison analysis, only Hallmark pathways were utilized. For the direct deeper analyses of Hm-0HI+ vs. HM-OHI- the analysis was extended to Gene Ontogeny and Reactome pathways.

[0079] Differential Expression Analysis and Venn Analysis: Differential expression analysis was performed to identify differentially expressed proteins (DEPs) between HM-0HI+ patients and each of the comparison groups (HM-0HI-, FHL, and healthy controls). Subsequently, a Venn analysis was carried out to detect any overlapping DEPs between HM-0HI+ patients and FHL patients. These overlapping DEPs provide insights into shared immune activation signatures between these two groups.

[0080] Spearman’s Correlation Analysis and Logistic Regression: Similarly, spearman’s correlation analysis was conducted to explore the relationships between fold changes of proteins and normalized enrichment scores of hallmark GSEA comparisons within intersecting proteins and genes of HM-0HI+ patients and FHL patients. Logistic regression was then employed to create a quantitative representation of these correlations.

[0081] Classifier Development: Various machine learning algorithms were employed to develop pathway-specific classifiers. These included Supervised Learning utilizing algorithms such as Support Vector Machines (SVM), Random Forest (RF), and Gradient Boosting (XGBoost). These models were trained on labeled datasets where each sample was tagged with its corresponding pathway.

[0082] Orthogonal validation by ELISA: Enzyme-Linked Immunosorbent Assay (ELISA) was used to validate the results. A DLBCL external cohort was used including 80 patients - 40 OHI+ vs. 40 OHL patients.

[0083] Ethical Considerations: All data used in the study were obtained under Helsinki approved protocols, adhering to their respective data use agreements and ethical guidelines.ResultsFLH patients differ in their demographic, genetic, and outcome characteristics but have similar levels of inflammatory markers

[0084] FIGs. 1 A-E depict analysis of the proteomic cohort, highlighting distinctions between FHL and HM-0HI+, as well as their resemblances to their respective control groups.

[0085] In FIG. 1A, the age distribution of the FHL and Pediatric control (Pedctl) groups is presented. The means of the ages in these groups were 1.2 years (SD: 1.7 years) and 1.1 years (SD: 0.4 years), respectively. Statistical analysis produced a non-significant p-value of 0.3, indicating age equivalence.

[0086] FIG. IB demonstrates the age profiles of patients with malignancies and OHI+, which closely align with the OHI- group. The mean ages for these groups were 54.5 years (SD: 17.7 years) and 60.1 years (SD: 15 years), respectively, with a p-valye of 0.3, signifying comparable ages. However, there was a pronounced age disparity between OHI+ and FHL (p < 0.0001), highlighting their distinct demographic characteristics.

[0087] The most significant disparity between the two cohorts is observed in their disease outcomes, as depicted in FIG. IB. Specifically, there is a marked contrast in survival rates, with 57% of FHL patients surviving compared to only 14% survival amount OHI+ patients. This stark contrast reflects the distinct clinical trajectories of these conditions.

[0088] FIG. 1C presents the balanced OHI+ and OHI- cohort. The disease frequencies in our cohort align with their prevalence in the population, with Hemophagocytic Lymphohistiocytosis (HLH) / 0HI+ being most common in Diffuse Large B Cell Lymphoma (DLBCL), followed by T Cell Lymphoma (TCL), Hodgkin’s Lymphoma (HL), and Chronic Lymphocytic Leukemia (CLL).

[0089] FIG. ID underscores a notable genetic difference, with 71% of the FHL cohort manifesting germline mutations associated with HLH, while none of the OHI cohort exhibited underlying germline mutations. This genetic landscape includes instances of UNC13 positivity, homozygous and heterozygous cases, as well as PRF1 homozygosity and a RAB27A mutation.

[0090] However, despite these divergent genetic and demographic backgrounds, FIG. IE illustrates that the levels of inflammatory markers were comparable between the groups (p = 0.8). Particularly, median ferritin and sCD25 levels for FHL ad OHI+ stood at 5000 ng / mL (IQR: 1055- 11773 ng / mL) and 6664 U / mL (IQR: 3765-8221 U / mL), respectively. This consistency in marker levels underscores the presence of a shared inflammatory context despite genetic variations.FHL and 0H1+ patients exhibit noteworthy immunological parallels

[0091] The exclusion of outliers from the analysis unveiled distinct clustering patterns for FHL and HM-0HI+, setting them apart from their respective control groups, Pediatric Controls (Pedctl) and HM-OHI- individuals.

[0092] Utilizing an unsupervised primary component analysis, as depicted in FIG. 2A, and hierarchical clustering, as shown in FIG. 2B, a clear pattern of grouping emerged. OHI+ adult patients formed clusters that closely aligned with FHL infants, while healthy controls and OHL patients formed separate clusters.

[0093] These observations were further emphasized by a GSEA (FIG. 2C) revealing shared enriched biological pathways between OHI+ with High Inflammation (HM-0HI+) and FHL, as illustrated in FIG. 2C. These shared pathways encompassed the interferon-y response, IL6 JAK STAT3 signaling, !L2 STAT5 signaling, and oxidative phosphorylation. The normalized enrichment scores for these pathways were 1.82: 1.92, 1.76:2.23, 1.76: 1.76, and 1.68: 1.78, respectively, for HM-0HI+ and FHL.

[0094] These findings collectively indicate a substantial overlap of key proteins and biological pathways between FHL and OHI+, suggesting underlying mechanisms in these conditions.Overlap and significant correlation between OHI+ and FHL

[0095] Subsequent analysis investigated the extent of overlap between downregulated differentially expressed proteins (DEPs) in the dataset, where significance thresholds were set at p < 0.05 (FIG. 3A). 1,356 downregulated proteins in FHL vs. Pedctl were identified, and a higher statistical threshold was set to define DEGs in the OHI+ group (p=0.28) and 693 of these proteins, representing 51% of downregulated DEPs were found to overlap between the two groups. Similarly, for upregulated DEPs (p < 0.05 in FHL vs. Pedctl and p<0.046 in OHI+ vs. OHL, FIG. 3B), a total of 1,518 upregulated proteins were identified, with 55% of these upregulated DEPs showing overlap between the groups.

[0096] To directly assess the similarity between HM-0HI+ and FHL, a correlation analysis was conducted between the fold changes of proteins within the intersecting set (FIG. 3C). A highdegree of correlation (R = 0.96, p < 0.0001) was observed in the fold changes of these proteins. These proteins are known to be associated with FHL, including interleukin 18 (IL 18), IL 18 binding protein (IL18BP), Chemokine (C-X-C motif) ligand 9 (CXCL9), CXCL19, and LIL1B.

[0097] Moreover, the enriched pathways within this intersection also exhibited a high degree of correlation (FIG. 3D, q = 0.95). The pathways enriched in both FHL and OHI+ play pivotal roles in the pathophysiology of FHL. This includes the IFN-y response, which is a hallmark of FHL, as well as other enriched pathways such as IL6 JAK STAT3 signaling, IL2 STAT5 signaling, Inflammatory response, TNFa signaling via NFKB, and oxidative phosphorylation. Some pathways, such as the P53 pathway and myogenesis, were under-enriched in both FHL and OHI+ patients.OHI+ state is characterized by T cell activation, IFN-y response, IL-6 JAKSTAT3 and complement activation

[0098] Comparing protein profiles between the OHI+ and OHL groups (as depicted in FIGs. 4A-D), the presence in OHI+ of proteins of various biological processes was revealed. In particular, as shown in FIG. 4A, IL2RA, HLA-DRB3, and GZMB are present in OHI+ in the T cell activation pathway. As shown in FIG. 4B, GBP2, IL15, 1LI5R, and CXCL9 are present in OHI+ in the IFN-y response pathway. FIG. 4C shows that MIF, CSF1, CD 14, and IL6 are present in OHI+ in the IL6 JAK STAT3 signaling pathway. FIG. 4D shows that C3 and VSIG4 are present in OHI+ in the complement activation pathway.

[0099] An extensive GSEA was subsequently conducted, encompassing hallmark processes (FIG. 5A), Reactome processes (FIG. 5B), and Gene Ontogeny (GO) processes (FIG. 5C). Within the enriched pathways, several were pinpointed as targets for therapeutic intervention.

[0100] A T cell activation-driven response (FIG. 5C) was apparent within the enriched pathways. Specifically, pathways were identified that lend themselves to targeted therapeutic approaches, including those related to IFN-y signaling and response, IL6 JAK STAT3, complement activation, and leukotriene metabolism.

[0101] Specific classifiers were established to facilitate the detection of these activated pathways. These classifiers included a nine-protein ensemble for identifying IL6 JAK STAT3activation (FIG. 5D), IL 15 as a component for discerning the IFN-y response (FIG. 5D), GBP2 as a marker for IFN-y signaling (FIG. 5E), the inactivated form of C3b for detecting complement activation (FIG. 5D), and a four-protein classifier, which included LTA4H, for identifying leukotriene activation (FIG. 5F).OH1+ identifies immune evasion in lymphoma patients

[0102] To test a hypothesis that OHI+ associated hyperinflammation occurs when T and myeloid progenitor cells are perpetually and ineffectively “revving,” akin to the mechanism observed in FHL but in response to cancer, the enticement of immune evasion proteins in OHI+ was examined. Enrichment in immune evasion (FIG. 6A) was found in the majority of OHI+ patients (FIG. 6B). Checkpoint ligand and receptor pairs that exhibit strong correlations with OHI+ status were identified in patients with DLBCL (FIG. 6C) and CLL (FIG. 6D).

[0103] FIGs. 7A to 7F depict the ELISA orthogonal validation of the results discussed herein. Specifically, as noted above, a diffuse large B cell lymphoma (DLBCL) external cohort was used including 80 patients - 40 OHI+ vs. 40 OHI- patients. FIG. 7A shows the ELISA results for sCD8, 4-1BB, Granzyme B (GZMB), and Granzyme A (GZMA) in the T cell cytotoxicity and T cell receptor pathway. FIG. 7B shows the ELISA results for CXCL9, GBP2, CXCL10, and IL18BP in the IFN-y response and signaling pathway. FIG. 7C shows the ELISA results for sCD14 and HO-1 in the IL6 JAK STAT3 signaling pathway. FIG. 7D shows the ELISA results for MIF and sVSIG4 in the macrophage activation pathway. FIG. 7E shows the ELISA results for iC3b in the complement regulation activation and cytotoxicity activation pathway. FIG. 7F shows the ELISA results for the checkpoint proteins, sPDl, TIM3, sPDLl, GAL9, and HLA-G.

[0104] As shown in these studies, hematologic malignancy associated hyperinflammation and FHL share hyperinflammation pathways, with T cell activation being a player in pathway activation. These pathways are related to IFN-y and IL6 JAK STAT3 signaling. These studies have shown that the OHI index is a valuable tool for identifying and diagnosing hyperinflammation in patients with hematologic malignancies.

[0105] However, while cytotoxic T cell activation is harmful in FHL, adequate T cell function is important in cancer patients for tumor surveillance, elimination, and immunotherapy. Therefore,targeting T cells as a treatment of HM-OHI+ patients may be detrimental. Accordingly, aspects of the present disclosure are directed to treatment methods that target the shared pathways, e.g., the IL6 JAK STAT 3 signaling pathway, the IFN-y response pathway, and the IL2 STAT5 signaling pathway.

[0106] Additionally, as demonstrated in these studies, HM-0HI+ patients exhibit complement activation, leukotriene activation, macrophage activation, and elevated levels of checkpoint proteins and ligand / receptor pairs. Accordingly, aspects of the present disclosure are directed to treatment methods that target MIF, complement, or leukotriene inhibition. In some aspects, methods of treatment include checkpoint blockade therapy.

[0107] All percentages and ratios are calculated by weight unless otherwise indicated.

[0108] All percentages and ratios are calculated based on the total composition unless otherwise indicated.

[0109] It should be understood that every maximum numerical limitation given throughout this specification includes every lower numerical limitation, as if such lower numerical limitations were expressly written herein. Every minimum numerical limitation given throughout this specification will include every higher numerical limitation, as if such higher numerical limitations were expressly written herein. Every numerical range given throughout this specification will include every narrower numerical range that falls within such broader numerical range, as if such narrower numerical ranges were all expressly written herein.

[0110] The dimensions and values disclosed herein are not to be understood as being strictly limited to the exact numerical values recited. Instead, unless otherwise specified, each such dimension is intended to mean both the recited value and a functionally equivalent range surrounding that value. For example, a dimension disclosed as “20 mm” is intended to mean “about 20 mm.”

[0111] Every document cited herein, including any cross-referenced or related patent or application, is hereby incorporated herein by reference in its entirety unless expressly excluded or otherwise limited. All accessioned information (e.g., as identified by PUBMED, PUBCHEM, NCBI, UNTPROT, or EBI accession numbers) and publications in their entireties are incorporatedinto this disclosure by reference in order to more fully describe the state of the art as known to those skilled therein as of the date of this disclosure. The citation of any document is not an admission that it is prior art with respect to any invention disclosed or claimed herein or that it alone, or in any combination with any other reference or references, teaches, suggests or discloses any such invention. Further, to the extent that any meaning or definition of a term in this document conflicts with any meaning or definition of the same term in a document incorporated by reference, the meaning or definition assigned to that term in this document shall govern.

[0112] While particular embodiments of the present invention have been illustrated and described, it would be obvious to those skilled in the art that various other changes and modifications may be made without departing from the spirit and scope of the invention. It is therefore intended to cover in the appended claims all such changes and modifications that are within the scope of this invention.ILLUSTRATIVE COMBINATIONS

[0113] The following illustrative examples relate to various non-exhaustive ways in which the teachings herein may be combined or applied. It should be understood that the following examples are not intended to restrict the coverage of any claims that may be presented at any time in this application or in subsequent filings of this application. No disclaimer is intended. The following examples are being provided for nothing more than merely illustrative purposes. It is contemplated that the various teachings herein may be arranged and applied in numerous other ways. It is also contemplated that some variations may omit certain features referred to in the below examples. Therefore, none of the aspects or features referred to below should be deemed critical unless otherwise explicitly indicated as such at a later date by the inventors or by a successor in interest to the inventors. If any claims are presented in this application or in subsequent filings related to this application that include additional features beyond those referred to below, those additional features shall not be presumed to have been added for any reason relating to patentability.Example 1

[0114] A method of diagnosing and treating hematologic malignancy-associated hyperinflammation in an individual having a hematologic malignancy, the method comprising:determining an optimized Hemophagocytic lymphohistiocytosis (HLH) inflammatory (OHI) index status of the individual, the OHI index status being based on measured levels of soluble CD25 (sCD25) and ferritin in a biological sample of the individual; and based on the OHI index status of the individual, administering to the individual an effective amount of anti-inflammatory or checkpoint blockade therapy.Example 2

[0115] The method of Example 1, wherein the OHI index status is either:OHI+ in which the measured level of sCD25 is greater than a predetermined threshold sCD25 level, and the measured level of ferritin is greater than a predetermined threshold ferritin level; orOHI- in which the measured level of sCD25 is below the predetermined threshold sCD25 level, or the measured level of ferritin is below the predetermined threshold ferritin level.Example 3

[0116] The method of Example 1 or 2, wherein the predetermined threshold sCD25 level is 3,900 U / mL.Example 4

[0117] The method of any preceding Example, wherein the predetermined threshold ferritin level is 1,000 ng / mL.Example 5

[0118] The method of any preceding Example, wherein the OHI index status is either:OHI+ in which the measured level of sCD25 is greater than the predetermined threshold sCD25 level, and the measured level of ferritin is greater than 1,000 ng / mL; orOHI- in which the measured level of sCD25 is below the predetermined threshold sCD25 level, or the measured level of ferritin is below 1,000 ng / mL.Example 6

[0119] The method of any preceding Example, wherein the OHI index status is either:OHI+ in which the measured level of sCD25 is greater than 3,900 U / mL, and the measured level of ferritin is greater than the predetermined threshold ferritin level; orOHI- in which the measured level of sCD25 is below 3,900 U / mL, or the measured level of ferritin is below the predetermined threshold ferritin level.Example 7

[0120] The method of any preceding Example, wherein the OHI index status is either:OHI+ in which the measured level of sCD25 is greater than 3,900 U / mL, and the measured level of ferritin is greater than 1,000 ng / mL; orOHI- in which the measured level of sCD25 is below 3,900 U / mL, or the measured level of ferritin is below 1,000 ng / mL.Example 8

[0121] The method of any preceding Example, wherein the effective amount of antiinflammatory or checkpoint blockade therapy is administered to the individual when the OHI index status is determined to be OHI+.Example 9

[0122] The method of any preceding Example, wherein the anti-inflammatory therapy is selected from therapies that target the IFN-y-mediated response pathway, the IL6 JAK STAT 3 signaling pathway, the complement activation pathway, the leukotriene activation pathway, or the IL2 STAT 5 signaling pathway.Example 10

[0123] The method of any preceding Example, wherein the anti-inflammatory therapy is selected from therapies that target the IFN-y-mediated response pathway, or the IL6 JAK STAT 3 signaling pathway.Example 11

[0124] The method of any preceding Example, wherein the anti-inflammatory therapy is selected from: anti -interferon agents;JAK inhibitors; complement inhibitors; leukotriene inhibitors; anti-IL6 agents; anti-TNF agents; anti-MIF agents; and combinations thereof.Example 12

[0125] The method of any preceding Example, wherein the anti-inflammatory therapy is selected from anti-interferon agents, JAK inhibitors, complement inhibitors, and combinations thereof.Example 13

[0126] The method of any preceding Example, wherein the anti-inflammatory therapy is selected from anti-interferon agents, JAK inhibitors, and combinations thereof.Example 14

[0127] The method of any preceding Example, wherein the anti-inflammatory therapy is selected from: anti-IL-6 agents and antibodies such as tocilizumab, sarilumab, siltuximab, sirukumab, olokizumab, clazakizumab, vobarilizumba, and ziltivekimab;JAK inhibitors such as barcitinib, tofacitinib, ruxolitinib, upadacitinib, fedratinib, abrocitinib, ritlecitinib, momelotinib, and pacritinib ; anti-IFN-y drugs or agents such as emapalumab, belimumab, and fontolizumab; complement inhibitors such as eculizumab, pegcetacoplan, ravulizumab, avacincaptadpegol, pozelimab, zilucoplan, crovalimab-akkz, iptacopan, danicopan, avacopan, complement Cl esterase inhibitors, and sutimlimab; leukotriene inhibitors such as zileuton, zafirlukast, pranlukast, and montelukast ; anti-TNF agents and drugs such as infliximab, etanercept, adalimumab, certolizumab pegol, and golimumab; anti-MIF agents and drugs such as imalumab, milatuzumab, and ebselen; and combinations thereof.Example 15

[0128] The method of any of Examples 1 to 8, wherein the checkpoint blockade therapy is selected from PD-1 inhibitors, PD-L1 inhibitors, CTLA-4 inhibitors, LAG-3 inhibitors, and combinations thereof.Example 16

[0129] The method of any of Examples 1 to 8 and 15, wherein the checkpoint blockade therapy is selected from:PD-1 inhibitors such as pembrolizumab, nivolumab, cemiplimab, dostarlimab, retifanlimab-dlwr, and tislelizumab;PD-L1 inhibitors such as atezolizumab, avelumab, and durvalumab;CTLA-4 inhibitors such as ipilimumab, and tremelimumab;LAG-3 inhibitors such as relatlimab; and combinations thereof.Example 17

[0130] The method of any preceding Example, wherein the hematologic malignancy is selected from lymphomas, leukemias, and myelomas.Example 18

[0131] The method of any preceding Example, wherein the hematologic malignancy is a lymphoma.Example 19

[0132] The method of any preceding Example, wherein the hematologic malignancy is diffuse large B cell lymphoma (DLBCL), T cell lymphoma (TCL), Hodkin’s lymphoma, or Chronic lymphocytic Leukemia (CLL).Example 20

[0133] A method of diagnosing and treating lymphoma-associated hyperinflammation in an individual having a lymphoma, the method comprising: determining an optimized Hemophagocytic lymphohistiocytosis (HLH) inflammatory (OHI) index status of the individual, the OHI index status being based on measured levels of soluble CD25 (sCD25) and ferritin in a biological sample of the individual; and based on the OHI index status of the individual, administering to the individual an effective amount of anti-inflammatory or checkpoint blockade therapy.Example 21

[0134] The method of Example 20, wherein the OHT index status is either:OHI+ in which the measured level of sCD25 is greater than a predetermined threshold sCD25 level, and the measured level of ferritin is greater than a predetermined threshold ferritin level; orOHI- in which the measured level of sCD25 is below the predetermined threshold sCD25 level, or the measured level of ferritin is below the predetermined threshold ferritin level.Example 22

[0135] The method of Example 20 or 21, wherein the predetermined threshold sCD25 level is 3,900 U / mL.Example 23

[0136] The method of any of Examples 20 to 22, wherein the predetermined threshold ferritin level is 1,000 ng / mL.Example 24

[0137] The method of any of Examples 20 to 23, wherein the OHI index status is either:OHI+ in which the measured level of sCD25 is greater than the predetermined threshold sCD25 level, and the measured level of ferritin is greater than 1,000 ng / mL; orOHI- in which the measured level of sCD25 is below the predetermined threshold sCD25 level, or the measured level of ferritin is below 1,000 ng / mL.Example 25

[0138] The method of any of Examples 20 to 24, wherein the OHI index status is either:OHI+ in which the measured level of sCD25 is greater than 3,900 U / mL, and the measured level of ferritin is greater than the predetermined threshold ferritin level; orOHI- in which the measured level of sCD25 is below 3,900 U / mL, or the measured level of ferritin is below the predetermined threshold ferritin level.Example 26

[0139] The method of any of Examples 20 to 25, wherein the OHI index status is either:OHI+ in which the measured level of sCD25 is greater than 3,900 U / mL, and the measured level of ferritin is greater than 1,000 ng / mL; orOHI- in which the measured level of sCD25 is below 3,900 U / mL, or the measured level of ferritin is below 1,000 ng / mL.Example 27

[0140] The method of any of Examples 20 to 26, wherein the effective amount of antiinflammatory or checkpoint blockade therapy is administered to the individual when the OHI index status is determined to be OHI+.Example 28

[0141] The method of any of Examples 20 to 27, wherein the anti-inflammatory therapy is selected from therapies that target the IFN-y-mediated response pathway, the IL6 JAK STAT 3 signaling pathway, the complement activation pathway, the leukotriene activation pathway, or the IL2 STAT 5 signaling pathway.Example 29

[0142] The method of any of Examples 20 to 28, wherein the anti-inflammatory therapy is selected from therapies that target the IFN-y-mediated response pathway, or the IL6 JAK STAT 3 signaling pathway.Example 30

[0143] The method of any of Examples 20 to 29, wherein the anti-inflammatory therapy is selected from:anti-interferon agents;JAK inhibitors; complement inhibitors; leukotriene inhibitors; anti-IL6 agents; anti-TNF agents; anti-MIF agents; and combinations thereof.Example 31

[0144] The method of any of Examples 20 to 30, wherein the anti-inflammatory therapy is selected from anti-interferon agents, JAK inhibitors, complement inhibitors, and combinations thereof.Example 32

[0145] The method of any of Examples 20 to 31, wherein the anti-inflammatory therapy is selected from anti-interferon agents, JAK inhibitors, and combinations thereof.Example 33

[0146] The method of any of Examples 20 to 32, wherein the anti-inflammatory therapy is selected from: anti-IL-6 agents and antibodies such as tocilizumab, sarilumab, siltuximab, sirukumab, olokizumab, clazakizumab, vobarilizumba, and ziltivekimab;JAK inhibitors such as barcitinib, tofacitinib, ruxolitinib, upadacitinib, fedratinib, abrocitinib, ritlecitinib, momelotinib, and pacritinib ;anti-IFN-y drugs or agents such as emapalumab, belimumab, and fontolizumab; complement inhibitors such as eculizumab, pegcetacoplan, ravulizumab, avacincaptadpegol, pozelimab, zilucoplan, crovalimab-akkz, iptacopan, danicopan, avacopan, complement Cl esterase inhibitors, and sutimlimab ; leukotriene inhibitors such as zileuton, zafirlukast, pranlukast, and montelukast ; anti-TNF agents and drugs such as infliximab, etanercept, adalimumab, certolizumab pegol, and golimumab ; anti-MIF agents and drugs such as imalumab, milatuzumab, and ebselen; and combinations thereof.Example 34

[0147] The method of any of Examples 20 to 27, wherein the checkpoint blockade therapy is selected from PD-1 inhibitors, PD-L1 inhibitors, CTLA-4 inhibitors, LAG-3 inhibitors, and combinations thereof.Example 35

[0148] The method of any of Examples 20 to 27 and 34, wherein the checkpoint blockade therapy is selected from:PD-1 inhibitors such as pembrolizumab, nivolumab, cemiplimab, dostarlimab, retifanlimab-dlwr, and tislelizumab;PD-L1 inhibitors such as atezolizumab, avelumab, and durvalumab;CTLA-4 inhibitors such as ipilimumab, and tremelimumab;LAG-3 inhibitors such as relatlimab; and combinations thereof.Example 36

[0149] A method of identifying an individual with a hematologic malignancy that is likely to benefit from treatment with a checkpoint blockade therapy, the method comprising: determining an optimized Hemophagocytic lymphohistiocytosis (HLH) inflammatory (OHI) index status of the individual, the OHI index status being based on measured levels of soluble CD25 (sCD25) and ferritin in a biological sample of the individual; and based on the OHI index status of the individual, identifying the individual as likely to benefit from a checkpoint blockade therapy.Example 37

[0150] The method of Example 36, wherein the OHI index status is either:OHI+ in which the measured level of sCD25 is greater than a predetermined threshold sCD25 level, and the measured level of ferritin is greater than a predetermined threshold ferritin level; orOHI- in which the measured level of sCD25 is below the predetermined threshold sCD25 level, or the measured level of ferritin is below the predetermined threshold ferritin level.Example 38

[0151] The method of Example 36 or 37, wherein the predetermined threshold sCD25 level is 3,900 U / mL.Example 39

[0152] The method of any of Examples 36 to 38, wherein the predetermined threshold ferritin level is 1,000 ng / mL.Example 40

[0153] The method of any of Examples 36 to 39, wherein the OHI index status is either:OHI+ in which the measured level of sCD25 is greater than the predetermined threshold sCD25 level, and the measured level of ferritin is greater than 1,000 ng / mL; orOHI- in which the measured level of sCD25 is below the predetermined threshold sCD25 level, or the measured level of ferritin is below 1,000 ng / mL.Example 41

[0154] The method of any of Examples 36 to 40, wherein the OHI index status is either:OHI+ in which the measured level of sCD25 is greater than 3,900 U / mL, and the measured level of ferritin is greater than the predetermined threshold ferritin level; orOHI- in which the measured level of sCD25 is below 3,900 U / mL, or the measured level of ferritin is below the predetermined threshold ferritin level.Example 42

[0155] The method of any of Examples 36 to 41, wherein the OHI index status is either:OHI+ in which the measured level of sCD25 is greater than 3,900 U / mL, and the measured level of ferritin is greater than 1,000 ng / mL; orOHI- in which the measured level of sCD25 is below 3,900 U / mL, or the measured level of ferritin is below 1,000 ng / mL.Example 43

[0156] The method of any of Examples 36 to 42, wherein OHI+ status indicates that the individual is likely to benefit from the checkpoint blockade therapy.Example 44

[0157] The method of any of Examples 36 to 42, further comprising based on the OHI index status, administering to the individual the checkpoint blockade therapy.Example 45

[0158] The method of Example 44, wherein the checkpoint blockade therapy is selected from PD-1 inhibitors, PD-L1 inhibitors, CTLA-4 inhibitors, LAG-3 inhibitors, and combinations thereof.Example 46

[0159] The method of any of Examples 44 and 45, wherein the checkpoint blockade therapy is selected from:PD-1 inhibitors such as pembrolizumab, nivolumab, cemiplimab, dostarlimab, retifanlimab-dlwr, and tislelizumab;PD-L1 inhibitors such as atezolizumab, avelumab, and durvalumab;CTLA-4 inhibitors such as ipilimumab, and tremelimumab;LAG-3 inhibitors such as relatlimab; and combinations thereof.Example 47

[0160] A method for identifying an individual having a hematologic malignancy that is likely to benefit from a treatment in a clinical trial, the method comprising: determining an optimized Hemophagocytic lymphohistiocytosis (HLH) inflammatory (OHI) index status of the individual, the OHI index status being based on measured levels of soluble CD25 (sCD25) and ferritin in a biological sample of the individual; and based on the OHI index status of the individual, identifying the individual as a candidate for the clinical trial.Example 48

[0161] The method of Example 47, wherein the OHI index status is either:OHI+ in which the measured level of sCD25 is greater than a predetermined threshold sCD25 level, and the measured level of ferritin is greater than a predetermined threshold ferritin level; orOHI- in which the measured level of sCD25 is below the predetermined threshold sCD25 level, or the measured level of ferritin is below the predetermined threshold ferritin level.Example 49

[0162] The method of Example 47 or 48, wherein the predetermined threshold sCD25 level is 3,900 U / mL.Example 50

[0163] The method of any of Examples 47 to 49, wherein the predetermined threshold ferritin level is 1,000 ng / mL.Example 51

[0164] The method of any of Examples 47 to 50, wherein the OHI index status is either:OHI+ in which the measured level of sCD25 is greater than the predetermined threshold sCD25 level, and the measured level of ferritin is greater than 1,000 ng / mL; orOHI- in which the measured level of sCD25 is below the predetermined threshold sCD25 level, or the measured level of ferritin is below 1,000 ng / mL.Example 52

[0165] The method of any of Examples 47 to 51, wherein the OHI index status is either:OHI+ in which the measured level of sCD25 is greater than 3,900 U / mL, and the measured level of ferritin is greater than the predetermined threshold ferritin level; orOHI- in which the measured level of sCD25 is below 3,900 U / mL, or the measured level of ferritin is below the predetermined threshold ferritin level.Example 53

[0166] The method of any of Examples 47 to 52, wherein the OHI index status is either:OHI+ in which the measured level of sCD25 is greater than 3,900 U / mL, and the measured level of ferritin is greater than 1,000 ng / mL; orOHI- in which the measured level of sCD25 is below 3,900 U / mL, or the measured level of ferritin is below 1,000 ng / mL.Example 54

[0167] The method of any of Examples 47 to 53, wherein OHI+ status indicates that the individual is a candidate for the clinical trial.Example 55

[0168] The method of any of Examples 47 to 54, further comprising enrolling the individual in the clinical trial.Example 56

[0169] A method of identifying an individual with a hematologic malignancy that is likely to benefit from treatment with an anti-inflammatory therapy, the method comprising: determining an optimized Hemophagocytic lymphohistiocytosis (HLH) inflammatory (OHI) index status of the individual, the OHI index status being based on measured levels of soluble CD25 (sCD25) and ferritin in a biological sample of the individual; and based on the OHI index status of the individual, identifying the individual as likely to benefit from an anti-inflammatory therapy.Example 57

[0170] The method of Example 56, wherein the OHI index status is either:OHI+ in which the measured level of sCD25 is greater than a predetermined threshold sCD25 level, and the measured level of ferritin is greater than a predetermined threshold ferritin level; orOHI- in which the measured level of sCD25 is below the predetermined threshold sCD25 level, or the measured level of ferritin is below the predetermined threshold ferritin level.Example 58

[0171] The method of Example 56 or 57, wherein the predetermined threshold sCD25 level is 3,900 U / mL.Example 59

[0172] The method of any of Examples 56 to 58, wherein the predetermined threshold ferritin level is 1,000 ng / mL.Example 60

[0173] The method of any of Examples 56 to 59, wherein the OHI index status is either:OHI+ in which the measured level of sCD25 is greater than the predetermined threshold sCD25 level, and the measured level of ferritin is greater than 1,000 ng / mL; orOHI- in which the measured level of sCD25 is below the predetermined threshold sCD25 level, or the measured level of ferritin is below 1,000 ng / mL.Example 61

[0174] The method of any of Examples 56 to 60, wherein the OHI index status is either:OHI+ in which the measured level of sCD25 is greater than 3,900 U / mL, and the measured level of ferritin is greater than the predetermined threshold ferritin level; orOHI- in which the measured level of sCD25 is below 3,900 U / mL, or the measured level of ferritin is below the predetermined threshold ferritin level.Example 62

[0175] The method of any of Examples 56 to 61, wherein the OHI index status is either:OHI+ in which the measured level of sCD25 is greater than 3,900 U / mL, and the measured level of ferritin is greater than 1,000 ng / mL; orOHI- in which the measured level of sCD25 is below 3,900 U / mL, or the measured level of ferritin is below 1,000 ng / mL.Example 63

[0176] 63. The method of any of Examples 56 to 62, wherein OHI+ status indicates that the individual is likely to benefit from the anti-inflammatory therapy.Example 64

[0177] The method of any of Examples 56 to 63, further comprising administering to the individual the anti-inflammatory therapy.Example 65

[0178] The method of any of Examples 56 to 64, wherein the anti-inflammatory therapy is selected from therapies that target the IFN-y-mediated response pathway, the IL6 JAK STAT 3 signaling pathway, the complement activation pathway, the leukotriene activation pathway, or the IL2 STAT 5 signaling pathway.Example 66

[0179] The method of any of Examples 56 to 65, wherein the anti-inflammatory therapy is selected from therapies that target the IFN-y-mediated response pathway, or the IL6 JAK STAT 3 signaling pathway.Example 67

[0180] The method of any of Examples 56 to 66, wherein the anti-inflammatory therapy is selected from:anti-interferon agents;JAK inhibitors; complement inhibitors; leukotriene inhibitors; anti-IL6 agents; anti-TNF agents; anti-MIF agents; and combinations thereof.Example 68

[0181] The method of any of Examples 56 to 67, wherein the anti-inflammatory therapy is selected from anti-interferon agents, JAK inhibitors, complement inhibitors, and combinations thereof.Example 69

[0182] The method of any of Examples 56 to 68, wherein the anti-inflammatory therapy is selected from anti-interferon agents, JAK inhibitors, and combinations thereof.Example 70

[0183] The method of any of Examples 56 to 69, wherein the anti-inflammatory therapy is selected from: anti-IL-6 agents and antibodies such as tocilizumab, sarilumab, siltuximab, sirukumab, olokizumab, clazakizumab, vobarilizumba, and ziltivekimab;JAK inhibitors such as barcitinib, tofacitinib, ruxolitinib, upadacitinib, fedratinib, abrocitinib, ritlecitinib, momelotinib, and pacritinib;anti-IFN-y drugs or agents such as emapalumab, belimumab, and fontolizumab; complement inhibitors such as eculizumab, pegcetacoplan, ravulizumab, avacincaptadpegol, pozelimab, zilucoplan, crovalimab-akkz, iptacopan, danicopan, avacopan, complement Cl esterase inhibitors, and sutimlimab; leukotriene inhibitors such as zileuton, zafirlukast, pranlukast, and montelukast; anti-TNF agents and drugs such as infliximab, etanercept, adalimumab, certolizumab pegol, and golimumab; anti-MIF agents and drugs such as imalumab, milatuzumab, and ebselen; and combinations thereof.Supplemental FiguresSupplemental Table 1. The completeness of data in the cohort. sCD25- soluble CD25; Hb- hemoglobin; ANC- absolute neutrophils ; PLT- platelets; Tg- triglycerides; AST- alanine transferase; ALT- alanine aminotransferase; LDH- lactate dehydrogenase; NK- natural killer; NA- not applicable; ND- not described.Supplemental Table 2. The HLH-2004 parameters at initial presentation for patients examined as a routine investigation or due to HLH-suspicion. SD- standard deviation; sCD25- soluble CD25; Hb- hemoglobin; ANC- absolute neutrophils; PLT- platelets; Tg- triglycerides; The statistics of the quantitative parameters were analyzed Mann-Whitney U test. The statistics of the categorical parameters were analyzed with Fischer’s exact test.Supplemental Figure 1. Distribution of patient data reviewed for the study.A pie chart specifying the initial population, how many patients were excluded for each reason, and what fraction of the initial population is represented in the final study cohort. Malignancies that were not represented in the HLH group are plasma cell dyscrasias (n-3).Supplemental Figure 2. The distribution of HLH-defining features at peak / nadir values remains overlapping. Normal range and the diagnostic threshold level from the HLH-2004 study (used for FHL) are shown in patients examined as a routine at HM diagnosis (left) and patients tested due to HLH suspicion (right). (A) The distribution of the inflammatory markers. (B) The distribution of the blood lineages. (C) The distribution of other markers. (D) Categorical parameters. The statistics of the quantitative parameters were analyzed Mann-Whitney U test. The statistics of the categorical parameters were analyzed with Fischer’s exact test. Values shown were available for more than 90% of the patients. NK activity was available for 4% of the patients and therefore is not shown. HM: patients with uncomplicated hematologic malignancies; HM-HLH patients fulfilled 5 out of 8 HLH-2004 diagnostic criteria. *p<0.05, ***p<0.001 ****p<0.0001 . sCD25- soluble CD25; Hb- hemoglobin; ANC- absolute neutrophils count; PLT: platelets; Tg- triglycerides.Supplemental Figure 3. Classification and regression analysis of all markers (including peak / nadir) independently identifies ferritin and sCD25 as the individual features to best identify the entire syndrome of HLH. Classification and Regression Analysis (CART) with all available patient parameters, including initial and peak / nadir measurements of HLH associated features (24 parameters, see supplemental figure 1 ) is shown. The growth limits were defined with the Gini method with 4 cases in the child node and 5 cases in the parent node. The minimum change in improvement was defined as 0.0001 , and pruning was defined with the minimum difference in risk of 2 standard errors. The results were validated with 10% cross-validation. The performance of the CART analysis is specified in the table.Supplemental Figure 4. sCD25 and ferritin best predict mortality compared to other HLH-2004 defining parameters. Receiver operating characteristics (ROC) of HLH-2004 parameters in predicting mortality at 500 days are presented. The area under the curve (AUC), 95% Cl, and sensitivity and specificity for the 2004 thresholds and the optimized ones are presented for values recorded at their pathologic peak or nadir during the hospitalization in which HLH was identified. The dotted line indicates the pertinent point on the curve identified as the best balance between sensitivity and specificity (with the highest Youden index). sCD25- soluble CD25; Hb- hemoglobin; ANC- absolute neutrophils count; PLT: platelets; Tg- triglycerides.Supplemental Figure 5. sCD25 and ferritin best discriminate between patients that died and those that survived in the entire cohort. Receiver operating characteristics (ROC) of the HLH-2004 parameters with mortality at day 500. The blue arrow indicates the HLH-2004 cutoff; the red arrow represents the prognostic optimal cutoff (the point with the highest Youden index, the best balance between sensitivity and specificity). ; Hb- hemoglobin; ANC- absolute neutrophils count; PLT: platelets; Tg- triglycerides.Supplemental Figure 6. The Optimized HLH Inflammatory (OHI) index is highly predictive of mortality in T and B cell lymphoma patients. Receiver operating curve (ROC) analyses for sCD25 and ferritin for predicting HLH-2004 diagnosis (>5 diagnostic criteria) and mortality prediction in T / NK cell lymphoma patients (A) and B cell lymphoma patients (B). The black arrow indicates the pertinent point on the curve identified as the best balance between sensitivity and specificity (with the highest Youden index). The red arrow indicates the value of the OHI. C and D are showing Kaplan Meier curves of patients classified at their peak / nadir values (of the index hospitalization) by the HLH-2004 (> 5 HLH-2004 diagnostic criteria, blue) (C) in all patients with lymphoma T / NK cell lymphoma(n=58).(D) In all patients with B cell malignancies (including Hodgkin’s N=109). The number at risk is presented for each group. Statistics were calculated with the Long-Rank (Mantel-Cox) test. *p<0.05, ***p=0.001. All diagnostic indices in the lymphoma groups were highly distinct between patients that were + / - (p<0.0001).Su rvival of patients with d iscordant versus concordant indices HLH-2004(-sCD25) / OHI indexSupplemental Figure 7. sCD25 is essential for HLH diagnosis and prognostic estimation in patients with hematologic malignancies at initial presentation and peak / nadir values of the index hospitalization. To assess the added value of sCD25 in patients with diagnostic uncertainty at presentation and at determining the final diagnosis, we classified the patients by the OHI index 9sCD25>3,900 U / mL and ferritin >1 ,000 ng / mL) and the HLH-2004 without including sCD25 (patients still need to answer 5 / 7 criteria. Kaplan Meier curves for patients classified by discordant (HLH-2004(- sCD25) - OHI + / HLH-2004(-sCD25)+ OHI-) and concordant classification (HLH-2004(-sCD25)+ OHI+Z HLH-2004(-sCD25)- OHI-) at initial presentation (A) and peak / nadir values in the index hospitalization. (B) The number at risk is presented for each group. Statistics were calculated with the Long-Rank (Mantel-Cox) test. ****p<0.0001.Supplemental Figure 8. The Optimized HLH Inflammatory (OHI) index predicts the presence of other HLH features irrespective of the patient’s HLH-2004 status. Patients with hematologic malignancies were categorized by their OHI index (sCD25>3,900 U / rnL; and ferritin>1 ,000 ng / mL) and HLH-2004 (> 5 HLH-2004 diagnostic criteria) status. The normal range and the diagnostic threshold level from the HLH2004 study (used for FHL) are shown: (A) The inflammatory markers, (B) blood counts, (C) fibrinogen and triglycerides. Laboratory values shown are the initial assessments during the index encounter for malignancy. The statistics of the quantitative parameters were analyzed using Mann Whitney U-test. (D) The distribution of categorical parameters is shown. The statistics of the categorical parameters were analyzed with Fischer’s exact test. *p<0.05, ** p<0.01 , ***p<0.001 , ****p<0.0001.Survival of patients with lymphoid and myeloid neoplasms Based on the elevation of inflammatory markersSupplemental Figure 10. sCD25 and ferritin are highly predictive of mortality in all patients with malignancies, irrespective of the malignancy subtype. Kaplan Meier curves of patients classified by the OHI, OHI’s parameters or no inflammatory markers at all in in the peak / nadir of the index hospitalization; (A) only patients with lymphomas (n=167, cell lymphomas, T / NK cell lymphomas, and Hodgkin’s lymphoma) and (B) patients with myeloid malignancies (n=48, acute myeloid leukemia, myelodysplastic syndrome and myeloproliferative neoplasms). The number at risk is presented for each group. Statistics were calculated with the Long-Rank (Mantel-Cox) test. **p<0.01 , ****p<0.0001.Supplemental Figure 11. The distribution of malignant diagnoses in patients with a positive OHI index is consistent with patients answering > 5 HLH-2004 criteria. The distribution of malignancies in our cohort is shown in patients with HM (A) < 5 of the HLH-2004 diagnostic criteria; (B) >5 HLH-2004 diagnostic criteria; (C) OHI index negative; (D) OHI index positive; (E) patients with no elevation in inflammatory markers; (F) patients with isolated elevation of sCD25 >3,900 ll / rnl; or ferritin >1 ,000 U / rnL) (G); (H) patients that have > 5 of the HLH-2004 diagnostic criteria but are OHI index negative and (I) patients that have < 5 of the HLH-2004 diagnostic criteria but are OHI index positive.Supplemental Figure 12. Treatment decisions suggest that the OHI recognizes patients with aberrant inflammation by treating physicians. A and B are presenting the treatments the patients in the cohort received. The patients are classified by their OHI index or HLH-2004 criteria status in A, while in B, they have the discordant or concordant status of the OHI index and HLH-2004. *p<0.05, **p<0.01 , ****p<0.0001.Survival of patients with discordant versus concordant indices Other HM-HLH diagnostic indicesSupplemental Figure 13. The OHI index recognizes more accurately predict prognosis than the Tamamyan criteria and the sCD25 / ferritin ratio. (A) Kaplan Meier curve for patients classified by discordant indices (Tamamyan+ / OHI- / Tamamyan- / OHI+ and concordant indices (Tamamyan + OHI - / Tamamyan + OHI +); Data was available for 14 / 18 of the extended diagnostic parameters criteria. (B) Kaplan Meier curves for patients classified by discordant indices (Ratio>2 / / OHI- / Ratio<2 / OHI+ and concordant indices (Ratio<2 / OHI - / Ratio>2+ OHI +). Ratio = the ratio of sCD25 / ferritin. The number at risk is presented for each group. Statistics were calculated with the Long-Rank (Mantel-Cox) test. **p<0.01, ****p<0.0001.

Claims

CLAIMSWhat is claimed is:

1. A method of diagnosing and treating hematologic malignancy-associated hyperinflammation in an individual having a hematologic malignancy, the method comprising: determining an optimized Hemophagocytic lymphohistiocytosis (HLH) inflammatory (OHI) index status of the individual, the OHI index status being based on measured levels of soluble CD25 (sCD25) and ferritin in a biological sample of the individual; and based on the OHI index status of the individual, administering to the individual an effective amount of anti-inflammatory or checkpoint blockade therapy.

2. The method of claim 1, wherein the OHI index status is either:OHI+ in which the measured level of sCD25 is greater than a predetermined threshold sCD25 level, and the measured level of ferritin is greater than a predetermined threshold ferritin level; orOHI- in which the measured level of sCD25 is below the predetermined threshold sCD25 level, or the measured level of ferritin is below the predetermined threshold ferritin level.

3. The method of claim 1 or 2, wherein the predetermined threshold sCD25 level is 3,900 U / mL.

4. The method of any preceding claim, wherein the predetermined threshold ferritin level is 1,000 ng / mL.

5. The method of any preceding claim, wherein the OHI index status is either:OHI+ in which the measured level of sCD25 is greater than the predetermined threshold sCD25 level, and the measured level of ferritin is greater than 1,000 ng / mL; orOHI- in which the measured level of sCD25 is below the predetermined threshold sCD25 level, or the measured level of ferritin is below 1,000 ng / mL.

6. The method of any preceding claims, wherein the OHI index status is either:OHI+ in which the measured level of sCD25 is greater than 3,900 U / mL, and the measured level of ferritin is greater than the predetermined threshold ferritin level; orOHI- in which the measured level of sCD25 is below 3,900 U / mL, or the measured level of ferritin is below the predetermined threshold ferritin level.

7. The method of any preceding claim, wherein the OHI index status is either:OHI+ in which the measured level of sCD25 is greater than 3,900 U / mL, and the measured level of ferritin is greater than 1,000 ng / mL; orOHI- in which the measured level of sCD25 is below 3,900 U / mL, or the measured level of ferritin is below 1,000 ng / mL.

8. The method of any preceding claim, wherein the effective amount of anti-inflammatory or checkpoint blockade therapy is administered to the individual when the OHI index status is determined to be OHI+.

9. The method of any preceding claim, wherein the anti-inflammatory therapy is selected from therapies that target the IFN-y-mediated response pathway, the IL6 JAK STAT 3 signaling pathway, the complement activation pathway, the leukotriene activation pathway, or the IL2 STAT 5 signaling pathway.

10. The method of any preceding claim, wherein the anti-inflammatory therapy is selected from therapies that target the IFN-y-mediated response pathway, or the IL6 JAK STAT 3 signaling pathway.

11. The method of any preceding claim, wherein the anti-inflammatory therapy is selected from: anti -interferon agents;JAK inhibitors; complement inhibitors; leukotriene inhibitors; anti-IL6 agents;anti-TNF agents; anti-MIF agents; and combinations thereof.

12. The method of any preceding claim, wherein the anti-inflammatory therapy is selected from anti-interferon agents, JAK inhibitors, complement inhibitors, and combinations thereof.

13. The method of any preceding claim, wherein the anti-inflammatory therapy is selected from anti-interferon agents, JAK inhibitors, and combinations thereof.

14. The method of any preceding claim, wherein the anti-inflammatory therapy is selected from: anti-IL-6 agents and antibodies such as tocilizumab, sarilumab, siltuximab, sirukumab, olokizumab, clazakizumab, vobarilizumba, and ziltivekimab;JAK inhibitors such as barcitinib, tofacitinib, ruxolitinib, upadacitinib, fedratinib, abrocitinib, ritlecitinib, momelotinib, and pacritinib; anti-IFN-y drugs or agents such as emapalumab, belimumab, and fontolizumab; complement inhibitors such as eculizumab, pegcetacoplan, ravulizumab, avacincaptadpegol, pozelimab, zilucoplan, crovalimab-akkz, iptacopan, danicopan, avacopan, complement Cl esterase inhibitors, and sutimlimab; leukotriene inhibitors such as zileuton, zafirlukast, pranlukast, and montelukast; anti-TNF agents and drugs such as infliximab, etanercept, adalimumab, certolizumab pegol, and golimumab; anti-MIF agents and drugs such as imalumab, milatuzumab, and ebselen; and combinations thereof.

15. The method of any of claims 1 to 8, wherein the checkpoint blockade therapy is selected from PD-1 inhibitors, PD-L1 inhibitors, CTLA-4 inhibitors, LAG-3 inhibitors, and combinations thereof.

16. The method of any of claims 1 to 8 and 15, wherein the checkpoint blockade therapy is selected from:PD-1 inhibitors such as pembrolizumab, nivolumab, cemiplimab, dostarlimab, retifanlimab-dlwr, and tislelizumab;PD-L1 inhibitors such as atezolizumab, avelumab, and durvalumab;CTLA-4 inhibitors such as ipilimumab, and tremelimumab;LAG-3 inhibitors such as relatlimab; and combinations thereof.

17. The method of any preceding claim, wherein the hematologic malignancy is selected from lymphomas, leukemias, and myelomas.

18. The method of any preceding claim, wherein the hematologic malignancy is a lymphoma.

19. The method of any preceding claim, wherein the hematologic malignancy is diffuse large B cell lymphoma (DLBCL), T cell lymphoma (TCL), Hodkin’s lymphoma, or Chronic lymphocytic Leukemia (CLL).

20. A method of diagnosing and treating lymphoma-associated hyperinflammation in an individual having a lymphoma, the method comprising: determining an optimized Hemophagocytic lymphohistiocytosis (HLH) inflammatory (OHI) index status of the individual, the OHI index status being based on measured levels of soluble CD25 (sCD25) and ferritin in a biological sample of the individual; and based on the OHI index status of the individual, administering to the individual an effective amount of anti-inflammatory or checkpoint blockade therapy.

21. The method of claim 20, wherein the OHI index status is either:OHI+ in which the measured level of sCD25 is greater than a predetermined threshold sCD25 level, and the measured level of ferritin is greater than a predetermined threshold ferritin level; orOHI- in which the measured level of sCD25 is below the predetermined threshold sCD25 level, or the measured level of ferritin is below the predetermined threshold ferritin level.

22. The method of claim 20 or 21, wherein the predetermined threshold sCD25 level is 3,900 U / mL.

23. The method of any of claims 20 to 22, wherein the predetermined threshold ferritin level is 1,000 ng / mL.

24. The method of any of claims 20 to 23, wherein the OHI index status is either:OHI+ in which the measured level of sCD25 is greater than the predetermined threshold sCD25 level, and the measured level of ferritin is greater than 1,000 ng / mL; orOHI- in which the measured level of sCD25 is below the predetermined threshold sCD25 level, or the measured level of ferritin is below 1,000 ng / mL.

25. The method of any of claims 20 to 24, wherein the OHI index status is either:OHI+ in which the measured level of sCD25 is greater than 3,900 U / mL, and the measured level of ferritin is greater than the predetermined threshold ferritin level; orOHI- in which the measured level of sCD25 is below 3,900 U / mL, or the measured level of ferritin is below the predetermined threshold ferritin level.

26. The method of any of claims 20 to 25, wherein the OHI index status is either:OHI+ in which the measured level of sCD25 is greater than 3,900 U / mL, and the measured level of ferritin is greater than 1,000 ng / mL; orOHI- in which the measured level of sCD25 is below 3,900 U / mL, or the measured level of ferritin is below 1,000 ng / mL.

27. The method of any of claims 20 to 26, wherein the effective amount of anti-inflammatory or checkpoint blockade therapy is administered to the individual when the OHI index status is determined to be OHI+.

28. The method of any of claims 20 to 27, wherein the anti-inflammatory therapy is selected from therapies that target the IFN-y-mediated response pathway, the IL6 JAK STAT 3 signaling pathway, the complement activation pathway, the leukotriene activation pathway, or the IL2 STAT 5 signaling pathway.

29. The method of any of claims 20 to 28, wherein the anti-inflammatory therapy is selected from therapies that target the IFN-y-mediated response pathway, or the IL6 JAK STAT 3 signaling pathway.

30. The method of any of clams 20 to 29, wherein the anti-inflammatory therapy is selected from: anti -interferon agents;JAK inhibitors; complement inhibitors; leukotriene inhibitors; anti-IL6 agents; anti-TNF agents; anti-MIF agents; and combinations thereof.

31. The method of any of claims 20 to 30, wherein the anti-inflammatory therapy is selected from anti-interferon agents, JAK inhibitors, complement inhibitors, and combinations thereof.

32. The method of any of claims 20 to 31, wherein the anti-inflammatory therapy is selected from anti-interferon agents, JAK inhibitors, and combinations thereof.

33. The method of any claims 20 to 32, wherein the anti-inflammatory therapy is selected from: anti-IL-6 agents and antibodies such as tocilizumab, sarilumab, siltuximab, sirukumab, olokizumab, clazakizumab, vobarilizumba, and ziltivekimab;JAK inhibitors such as barcitinib, tofacitinib, ruxolitinib, upadacitinib, fedratinib, abrocitinib, ritlecitinib, momelotinib, and pacritinib;anti-IFN-y drugs or agents such as emapalumab, belimumab, and fontolizumab; complement inhibitors such as eculizumab, pegcetacoplan, ravulizumab, avacincaptadpegol, pozelimab, zilucoplan, crovalimab-akkz, iptacopan, danicopan, avacopan, complement C 1 esterase inhibitors, and sutimlimab; leukotriene inhibitors such as zileuton, zafirlukast, pranlukast, and montelukast; anti-TNF agents and drugs such as infliximab, etanercept, adalimumab, certolizumab pegol, and golimumab; anti-MIF agents and drugs such as imalumab, milatuzumab, and ebselen; and combinations thereof.

34. The method of any of claims 20 to 27, wherein the checkpoint blockade therapy is selected from PD-1 inhibitors, PD-L1 inhibitors, CTLA-4 inhibitors, LAG-3 inhibitors, and combinations thereof.

35. The method of any of claims 20 to 27 and 34, wherein the checkpoint blockade therapy is selected from:PD-1 inhibitors such as pembrolizumab, nivolumab, cemiplimab, dostarlimab, retifanlimab-dlwr, and tislelizumab;PD-L1 inhibitors such as atezolizumab, avelumab, and durvalumab;CTLA-4 inhibitors such as ipilimumab, and tremelimumab;LAG-3 inhibitors such as relatlimab; and combinations thereof.

36. A method of identifying an individual with a hematologic malignancy that is likely to benefit from treatment with a checkpoint blockade therapy, the method comprising: determining an optimized Hemophagocytic lymphohistiocytosis (HLH) inflammatory (OHI) index status of the individual, the OHI index status being based on measured levels of soluble CD25 (sCD25) and ferritin in a biological sample of the individual; and based on the OHI index status of the individual, identifying the individual as likely to benefit from a checkpoint blockade therapy.

37. The method of claim 36, wherein the OHI index status is either:OHI+ in which the measured level of sCD25 is greater than a predetermined threshold sCD25 level, and the measured level of ferritin is greater than a predetermined threshold ferritin level; orOHI- in which the measured level of sCD25 is below the predetermined threshold sCD25 level, or the measured level of ferritin is below the predetermined threshold ferritin level.

38. The method of claim 36 or 37, wherein the predetermined threshold sCD25 level is 3,900 U / mL.

39. The method of any of claims 36 to 38, wherein the predetermined threshold ferritin level is 1,000 ng / mL.

40. The method of any of claims 36 to 39, wherein the OHI index status is either:OHI+ in which the measured level of sCD25 is greater than the predetermined threshold sCD25 level, and the measured level of ferritin is greater than 1,000 ng / mL; orOHI- in which the measured level of sCD25 is below the predetermined threshold sCD25 level, or the measured level of ferritin is below 1,000 ng / mL.

41. The method of any of claims 36 to 40, wherein the OHI index status is either:OHI+ in which the measured level of sCD25 is greater than 3,900 U / mL, and the measured level of ferritin is greater than the predetermined threshold ferritin level; orOHI- in which the measured level of sCD25 is below 3,900 U / mL, or the measured level of ferritin is below the predetermined threshold ferritin level.

42. The method of any of claims 36 to 41, wherein the OHI index status is either:OHI+ in which the measured level of sCD25 is greater than 3,900 U / mL, and the measured level of ferritin is greater than 1,000 ng / mL; orOHI- in which the measured level of sCD25 is below 3,900 U / mL, or the measured level of ferritin is below 1,000 ng / mL.

43. The method of any of claims 36 to 42, wherein OHI+ status indicates that the individual is likely to benefit from the checkpoint blockade therapy.

44. The method of any of claims 36 to 42, further comprising based on the OHI index status, administering to the individual the checkpoint blockade therapy.

45. The method of claim 44, wherein the checkpoint blockade therapy is selected from PD-1 inhibitors, PD-L1 inhibitors, CTLA-4 inhibitors, LAG-3 inhibitors, and combinations thereof.

46. The method of any of claims 44 and 45, wherein the checkpoint blockade therapy is selected from:PD-1 inhibitors such as pembrolizumab, nivolumab, cemiplimab, dostarlimab, retifanlimab-dlwr, and tislelizumab;PD-L1 inhibitors such as atezolizumab, avelumab, and durvalumab;CTLA-4 inhibitors such as ipilimumab, and tremelimumab;LAG-3 inhibitors such as relatlimab; and combinations thereof.

47. A method for identifying an individual having a hematologic malignancy that is likely to benefit from a treatment in a clinical trial, the method comprising: determining an optimized Hemophagocytic lymphohistiocytosis (HLH) inflammatory (OHI) index status of the individual, the OHI index status being based on measured levels of soluble CD25 (sCD25) and ferritin in a biological sample of the individual; and based on the OHI index status of the individual, identifying the individual as a candidate for the clinical trial.

48. The method of claim 47, wherein the OHI index status is either:OHI+ in which the measured level of sCD25 is greater than a predetermined threshold sCD25 level, and the measured level of ferritin is greater than a predetermined threshold ferritin level; orOHI- in which the measured level of sCD25 is below the predetermined threshold sCD25 level, or the measured level of ferritin is below the predetermined threshold ferritin level.

49. The method of claim 47 or 48, wherein the predetermined threshold sCD25 level is 3,900 U / mL.

50. The method of any of claims 47 to 49, wherein the predetermined threshold ferritin level is 1,000 ng / mL.

51. The method of any of claims 47 to 50, wherein the OHI index status is either:OHI+ in which the measured level of sCD25 is greater than the predetermined threshold sCD25 level, and the measured level of ferritin is greater than 1,000 ng / mL; orOHI- in which the measured level of sCD25 is below the predetermined threshold sCD25 level, or the measured level of ferritin is below 1,000 ng / mL.

52. The method of any of claims 47 to 51, wherein the OHI index status is either:OHI+ in which the measured level of sCD25 is greater than 3,900 U / mL, and the measured level of ferritin is greater than the predetermined threshold ferritin level; orOHI- in which the measured level of sCD25 is below 3,900 U / mL, or the measured level of ferritin is below the predetermined threshold ferritin level.

53. The method of any of claims 47 to 52, wherein the OHI index status is either:OHI+ in which the measured level of sCD25 is greater than 3,900 U / mL, and the measured level of ferritin is greater than 1,000 ng / mL; orOHI- in which the measured level of sCD25 is below 3,900 U / mL, or the measured level of ferritin is below 1,000 ng / mL.

54. The method of any of claims 47 to 53, wherein OHI+ status indicates that the individual is a candidate for the clinical trial.

55. The method of any of claims 47 to 54, further comprising enrolling the individual in the clinical trial.

56. A method of identifying an individual with a hematologic malignancy that is likely to benefit from treatment with an anti-inflammatory therapy, the method comprising: determining an optimized Hemophagocytic lymphohistiocytosis (HLH) inflammatory (OHI) index status of the individual, the OHI index status being based on measured levels of soluble CD25 (sCD25) and ferritin in a biological sample of the individual; and based on the OHI index status of the individual, identifying the individual as likely to benefit from an anti-inflammatory therapy.

57. The method of claim 56, wherein the OHI index status is either:OHI+ in which the measured level of sCD25 is greater than a predetermined threshold sCD25 level, and the measured level of ferritin is greater than a predetermined threshold ferritin level; orOHI- in which the measured level of sCD25 is below the predetermined threshold sCD25 level, or the measured level of ferritin is below the predetermined threshold ferritin level.

58. The method of claim 56 or 57, wherein the predetermined threshold sCD25 level is 3,900 U / mL.

59. The method of any of claims 56 to 58, wherein the predetermined threshold ferritin level is 1,000 ng / mL.

60. The method of any of claims 56 to 59, wherein the OHI index status is either:OHI+ in which the measured level of sCD25 is greater than the predetermined threshold sCD25 level, and the measured level of ferritin is greater than 1,000 ng / mL; orOHI- in which the measured level of sCD25 is below the predetermined threshold sCD25 level, or the measured level of ferritin is below 1,000 ng / mL.

61. The method of any of claims 56 to 60, wherein the OHI index status is either:OHI+ in which the measured level of sCD25 is greater than 3,900 U / mL, and the measured level of ferritin is greater than the predetermined threshold ferritin level; orOHI- in which the measured level of sCD25 is below 3,900 U / mL, or the measured level of ferritin is below the predetermined threshold ferritin level.

62. The method of any of claims 56 to 61, wherein the OHI index status is either:OHI+ in which the measured level of sCD25 is greater than 3,900 U / mL, and the measured level of ferritin is greater than 1,000 ng / mL; orOHI- in which the measured level of sCD25 is below 3,900 U / mL, or the measured level of ferritin is below 1,000 ng / mL.

63. The method of any of claims 56 to 62, wherein OHI+ status indicates that the individual is likely to benefit from the anti-inflammatory therapy.

64. The method of any of claims 56 to 63, further comprising administering to the individual the anti-inflammatory therapy.

65. The method of any of claims 56 to 64, wherein the anti-inflammatory therapy is selected from therapies that target the IFN-y-mediated response pathway, the IL6 JAK STAT 3 signaling pathway, the complement activation pathway, the leukotriene activation pathway, or the IL2 STAT 5 signaling pathway.

66. The method of any of claims 56 to 65, wherein the anti-inflammatory therapy is selected from therapies that target the IFN-y-mediated response pathway, or the IL6 JAK STAT 3 signaling pathway.

67. The method of any of clams 56 to 66, wherein the anti-inflammatory therapy is selected from: anti -interferon agents;JAK inhibitors; complement inhibitors;leukotriene inhibitors; anti-IL6 agents; anti-TNF agents; anti-MIF agents; and combinations thereof.

68. The method of any of claims 56 to 67, wherein the anti-inflammatory therapy is selected from anti-interferon agents, JAK inhibitors, complement inhibitors, and combinations thereof.

69. The method of any of claims 56 to 68, wherein the anti-inflammatory therapy is selected from anti-interferon agents, JAK inhibitors, and combinations thereof.

70. The method of any of claims 56 to 69, wherein the anti-inflammatory therapy is selected from: anti-IL-6 agents and antibodies such as tocilizumab, sarilumab, siltuximab, sirukumab, olokizumab, clazakizumab, vobarilizumba, and ziltivekimab;JAK inhibitors such as barcitinib, tofacitinib, ruxolitinib, upadacitinib, fedratinib, abrocitinib, ritlecitinib, momelotinib, and pacritinib; anti-IFN-y drugs or agents such as emapalumab, belimumab, and fontolizumab; complement inhibitors such as eculizumab, pegcetacoplan, ravulizumab, avacincaptadpegol, pozelimab, zilucoplan, crovalimab-akkz, iptacopan, danicopan, avacopan, complement Cl esterase inhibitors, and sutimlimab; leukotriene inhibitors such as zileuton, zafirlukast, pranlukast, and montelukast; anti-TNF agents and drugs such as infliximab, etanercept, adalimumab, certolizumab pegol, and golimumab; anti-MIF agents and drugs such as imalumab, milatuzumab, and ebselen; and combinations thereof.

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