Biomarkers of IL7R modulator activity

Abstract

The present invention relates to a method for evaluating or predicting a therapeutic response to treatment with an IL7R modulator, such as an IL7R antagonist or agonist, in a patient, and more particularly to the identification of biomarkers for evaluating or predicting whether an IL7R modulator is effective in treating a patient. The present invention also relates to a method for screening for a compound that is effective in treating a patient. The biomarkers are BCL2, CISH, SOCS2, FLT3LG, PTGER2, and DPP4.

Description

Technical Field

[0001] (Field of the Invention) The present invention relates to a method for evaluating or predicting a therapeutic response to treatment with an IL7R modulator, such as an IL7R antagonist or agonist, in a patient, and more particularly to the identification of biomarkers for evaluating or predicting whether an IL7R modulator is effective in treating a patient. The present invention also relates to a method for screening for a compound effective in treating a patient.

Background Art

[0002] (Background of the Invention) IL-7 is one of the most important cytokines for the homeostasis, proliferation, differentiation, and survival of both naive and memory T cells. IL-7 transmits signals through the IL-7 receptor (IL-7R), which is composed of an alpha chain (IL-7Rα), also called CD127, and a common cytokine receptor gamma chain (IL-2Rγ, CD132). Activation of IL-7R mainly induces proliferative and anti-apoptotic signals by activating the JAK-STAT pathway. Some studies have reported that the IL-7 canal also activates the PI3K or MAPK / ERK pathway, suggesting that IL-7 can use different signaling pathways depending on both cell type and the physiological state of the cell.

[0003] IL-7 and IL-7R are involved in the etiology of different disease states mediated by lymphoid dysfunction, such as autoimmune diseases, for example, diabetes and multiple sclerosis (Lee, L.F. et al., Proc Natl Acad Sci U S A 109, 12674-12679 (2012)), chronic inflammatory diseases, for example, rheumatoid arthritis, ankylosing spondylitis and inflammatory bowel disease (Churchman, S.M. & Ponchel, F., Rheumatology (Oxford) 47, 753-759 (2008); Krzystek-Korpacka, M. et al., Cancer Immunol Immunother 66, 171-179 (2017); Anderson, C.A. et al., Nat Genet 43, 246-252 (2011); Gracey, E. et al., Ann Rheum Dis 75, 2124-2132 (2016)) and blood cancers (Oliveira, M.L. et al., Adv Biol Regul 71, 88-96 (2018)).

[0004] CD127 is highly expressed on naive T cells and memory T cells, but is poorly expressed in FoxP3+ regulatory T cells (Tregs). This represents a unique opportunity to selectively target pathogenic effectors while maintaining natural regulators (Liu W, et al., J. Exp. Med. 2006; 203:1701-1711; Seddiki N, et al., J. Exp. Med. 2006; 203:1693-1700; Michel L, et al., J. Clin. Invest. 2008;118:3411-3419). Many antagonists of IL7R are currently being developed. Several studies have been conducted to analyze gene expression profiles in cancer patients and identify biomarkers, either in vitro or in vivo, to predict or monitor treatment responses such as anti-PD1 antibody. To the applicant's knowledge, gene signatures associated with IL7R antagonist or agonist therapy are not known or publicly available. The IL7 signaling pathway is complex and involves several genes. Indeed, 481 genes were differentially expressed in human PBMCs incubated with anti-human IL-7Rα mAb compared to control conditions (Belarif L., et al., Nat Commun. 2018 Oct 26;9(1):4483).

[0005] Therefore, it is necessary to identify drug activity biomarkers to accurately monitor patients' responses to such drugs in clinical trials. Such markers would facilitate the individualization of treatment regimens for each patient.

Summary of the Invention

Problems to be Solved by the Invention

[0006] (Summary of the Invention) By a method of analyzing the gene expression profile in a subject treated with an anti-IL7R antagonist antibody, 41 differentially expressed genes were identified. On the other hand, by a method of analyzing the gene expression profile in human cells treated in vitro with the IL7 cytokine, 56 differentially expressed genes were identified. Therefore, there remains a need to identify a limited number of drug-related biomarkers in order to easily evaluate the effectiveness of an anti-IL7R antagonist or agonist.

[0007] Furthermore, the present applicant has previously shown that only a small proportion of IL7-induced transcriptional modifications are common to mice and humans, and thus, when examining the IL7-IL7R signaling pathway, it is emphasized that the possibility of translating mouse studies to humans is low. Therefore, it is not possible to identify drug-related biomarkers for use in humans by performing in vitro gene signatures in mouse models.

Means for Solving the Problems

[0008] To solve these problems, the inventors have developed, in the present application, a new and powerful method performed in human peripheral blood mononuclear cells, and identified highly relevant drug-related biomarkers (also referred to herein as gene signatures) for monitoring IL7R agonists or antagonists such as anti-IL7R antagonist antibody activity. Comparative screening was used to identify a very limited number of genes that can be used as highly effective drug-related biomarkers for accurately monitoring the effectiveness of IL7R antagonist or agonist treatment. By measuring the expression level of at least one of these 4 to 6 target genes, the efficiency of IL7R antagonist or agonist compounds can be evaluated much more easily and at a lower cost in order to avoid risks in clinical trials. The use of these biomarkers can be extended to evaluate the potential for a therapeutic response to the IL7R modulator. Therefore, the selection of therapeutic compounds can be made much more selectively.

Advantages of the Invention

[0009] The present invention relates to an in vitro method for evaluating the therapeutic response of an IL7R modulator, such as treatment with an IL7R antagonist (e.g., an anti-IL7R antagonist antibody or an antigen-binding fragment thereof) or an IL7R agonist (e.g., IL7) in a human patient, comprising determining the gene expression profile of at least 1, 2, 3, or 4 genes selected from the group consisting of the BCL2, CISH, PTGER2, and DPP4 genes, preferably at least 1, 2, 3, 4, 5, or 6 genes selected from the group consisting of the BCL2, CISH, PTGER2, DPP4, FLT3LG, and SOCS2 genes, in a sample from a patient administered with at least one dose of an IL7R antagonist (e.g., an anti-IL7R antagonist antibody or an antigen-binding fragment thereof) or an IL7R agonist (e.g., IL7), wherein a lower or higher gene expression profile of said gene in said patient sample compared to a control value indicates that the patient is likely to respond to the antagonist or agonist treatment, respectively. This set of genes corresponds to a unique signature for monitoring IL7R modulator activity.

[0010] In another aspect, the present invention relates to an IL7R antagonist (e.g., an anti-IL7R antagonist antibody or an antigen-binding fragment thereof) for use in the treatment of a disease associated with enhanced IL7R signaling pathway induced by IL7 in a human patient in need thereof, wherein after the patient has been administered at least one dose of the antagonist (e.g., an anti-IL7R antagonist antibody or an antigen-binding fragment thereof), when the patient is evaluated to be highly likely to respond to the treatment with the antagonist (e.g., an anti-IL7R antagonist antibody or an antigen-binding fragment thereof) by the above method, the antagonist is administered to the patient. Preferably, the disease associated with enhanced IL7R signaling pathway induced by IL7 is an autoimmune disease, an inflammatory disease, an allergic disease, a cancer disease, an infectious disease, a respiratory disease, and a disease related to transplantation, which is accompanied by activation or proliferation of CD127-positive diseased cells, particularly an autoimmune disease or an inflammatory disease accompanied by activation or proliferation of CD127-positive cells, more preferably selected from the group consisting of chronic inflammatory diseases such as Sjögren's syndrome or inflammatory bowel diseases such as ulcerative colitis. The present invention relates to the above IL7R antagonist.

[0011] In another aspect, the present invention relates to an IL7R agonist for use in the treatment of a disease associated with impairment of normal T cell activity in a human patient in need thereof, preferably a disorder (s) that requires immune response stimulation, more preferably a cancer disease or an infectious disease whose onset itself does not depend on the IL7R signaling pathway. Here, after the patient has been administered at least one dose of the agonist, when it is evaluated that the patient is highly likely to respond to the treatment with the agonist by the above method, the agonist is administered to the patient. The present invention relates to the above IL7R agonist.

[0012] The present invention also relates to an in vitro method for evaluating the potential for a therapeutic response to an IL7R modulator such as an IL7 receptor antagonist (e.g., an anti-IL7R antagonist antibody or an antigen-binding fragment thereof) or an agonist treatment in a human patient. a) culturing a sample containing cells previously collected from a patient prior to said IL7R modulator treatment, in the presence of said IL7R modulator such as an IL7R antagonist (e.g., an anti-IL7R antagonist antibody or an antigen-binding fragment thereof) or an IL7R agonist; b) determining the gene expression profile of at least 1, 2, 3, or 4 genes selected from the group consisting of the BCL2, CISH, PTGER2, and DPP4 genes in the cultured sample, preferably at least 1, 2, 3, 4, 5, or 6 genes selected from the group consisting of the BCL2, CISH, PTGER2, DPP4, FLT3LG, and SOCS2 genes; comprising, wherein a lower or higher gene expression profile of said gene(s) in the cultured sample as compared to a control value indicates that the patient is likely to respond to said antagonist (e.g., an anti-IL7R antagonist antibody or an antigen-binding fragment thereof) or agonist treatment, regarding said in vitro method.

[0013] In another aspect, the present invention relates to an IL7R antagonist (e.g., an anti-IL7R antagonist antibody or an antigen-binding fragment thereof) for use in the treatment of a disease associated with enhanced IL7R signaling pathway induced by IL7 in a human patient in need thereof, wherein the patient has been previously evaluated as likely to respond to said antagonist (e.g., an anti-IL7R antagonist antibody or an antigen-binding fragment thereof) treatment by the method defined above, preferably, said disease associated with enhanced IL7R signaling pathway induced by IL7 is an autoimmune disease, an inflammatory disease, an allergic disease, a cancer disease, an infectious disease, a respiratory disease, and a disease associated with transplantation, involving activation or proliferation of CD127-positive diseased cells, particularly an autoimmune disease or an inflammatory disease involving activation or proliferation of CD127-positive cells, more preferably, selected from the group consisting of chronic inflammatory diseases such as Sjögren's syndrome or inflammatory bowel diseases such as ulcerative colitis, regarding said IL7R antagonist.

[0014] In another aspect, the present invention relates to an IL7R agonist for use in the treatment of a disease associated with impairment of normal T cell activity, preferably a cancer disease or an infectious disease in a human patient whose onset itself does not depend on the IL7R pathway, wherein the patient has been previously evaluated as likely to respond to the IL7R agonist treatment by the method defined above, and wherein the patient has been previously evaluated as likely to respond to the treatment with the agonist (e.g., an anti-IL7R antagonist antibody or an antigen-binding fragment thereof) by the method defined above, with respect to the IL7R agonist.

[0015] The present invention also relates to an in vitro method for selecting an IL7R modulator, such as an IL7R antagonist or an IL7R agonist compound, effective for the treatment of a disease associated with enhancement of the IL7R signaling pathway induced by IL7, or a disease associated with impairment of normal T cell activity, a) culturing human cells in the presence of the compound, b) determining the gene expression profile of at least 1, 2, 3, or 4 genes selected from the group consisting of the BCL2, CISH, PTGER2, and DPP4 genes in the cells, preferably at least 1, 2, 3, 4, 5, or 6 genes selected from the group consisting of the BCL2, CISH, PTGER2, DPP4, SOCS2, and FLT3LG genes, and c) selecting an antagonist or agonist compound that induces a lower or higher gene expression profile of the gene(s) in the cells, respectively, compared to a control value comprising the in vitro method.

[0016] In another aspect, the present invention relates to a kit comprising a set of reagents for specifically detecting the gene expression profile of at least 1, 2, 3 or 4 genes selected from the group consisting of the BCL2, CISH, PTGER2 and DPP4 genes, preferably at least 1, 2, 3, 4, 5 or 6 genes selected from the group consisting of the BCL2, CISH, PTGER2, DPP4, SOCS2 and FLT3LG genes, preferably wherein said reagents are primer pairs and / or probes specific for each gene.

BEST MODE FOR CARRYING OUT THE INVENTION

[0017] (Detailed Description of the Invention) The term "biomarker" refers to a characteristic biological or biogenic indicator (i.e., cellular, biochemical, molecular, genetic, protein, metabolite, specific post-translational modification, or physiological or physical sign) of a process, event, or condition. According to the present disclosure, the biomarker may refer to a gene signature. A gene signature is a single or combined group of genes within a cell having a unique and characteristic gene expression profile that results from a modified or unmodified biological process or a diseased medical condition.

[0018] The terms "subject" and "patient" are used interchangeably herein and refer to both human and non-human animals. As used herein, the term "patient" refers to mammals such as rodents, cats, dogs, and primates. Preferably, the patient according to the present invention is a human.

[0019] The term "patient sample" means any biological sample derived from a patient. Examples of such samples include tissue samples, cell samples, organs, biopsies, preferably tumor tissue containing infiltrating immune cells from cancer patients or blood patient samples. A preferred biological sample is a cell sample, preferably a blood cell sample such as isolated human peripheral blood mononuclear cells.

[0020] The term "likely responder" or "likely to respond to treatment" refers to a subject in which the onset of at least one symptom of the condition to be treated is delayed or prevented during or after treatment, or in which the symptom or at least one of the symptoms is stabilized, reduced or eliminated.

[0021] "Treatment response" refers to the result of a medical treatment in a patient that is determined to be useful or favorable. For example, a treatment response can be the delay or prevention of at least one symptom during or after treatment, or the stabilization, reduction or elimination of a symptom or at least one of the symptoms in a patient.

[0022] According to the present disclosure, "treatment response in a patient treated with an IL7R modulator" or "IL7R modulator treatment response" means that when an administration step of an IL7R modulator such as an IL7R antagonist (for example, an anti-IL7R antagonist antibody or an antigen-binding fragment thereof) or an IL7R agonist (for example, IL7) is carried out, the clinical condition of an animal or human patient in need thereof is improved, and the patient suffers from a disorder (s) associated with an enhancement of a signal transduction pathway induced by IL7, i.e., an IL7-IL7R signal transduction pathway (also referred to herein as the IL7-IL7R pathway), that is, an IL7-IL7 receptor interaction accompanied by activation and / or proliferation of CD127-positive cells, or a disorder associated with a disorder of normal T cell activity. Such treatment aims to improve the clinical condition of an animal or human patient by eliminating or reducing symptoms associated with a disorder (s) associated with IL-7 or TSLP (thymic stromal lymphopoietin), and in a preferred embodiment, by eliminating or reducing symptoms associated with a disorder (s) associated with an IL7R signal transduction pathway induced by IL7 or a disorder associated with a disorder of normal T cell activity.

[0023] In certain embodiments, the therapeutic response in a patient treated with an IL7R antagonist (e.g., an anti-IL7R antagonist antibody or an antigen-binding fragment thereof) refers to the administration of an IL7R antagonist that improves the clinical condition of a disorder associated with enhanced IL7R signaling pathway induced by IL-7, such as an autoimmune disorder, an inflammatory disorder, an allergic disorder, a cancer disorder, an infectious disease, a respiratory disease, and a disease related to transplantation, particularly an autoimmune disorder or an inflammatory disorder accompanied by activation or proliferation of CD127-positive diseased cells, more preferably a chronic inflammatory disorder such as Sjögren's syndrome or an inflammatory bowel disease such as ulcerative colitis.

[0024] In another certain embodiment, the therapeutic response in a patient treated with an IL7R agonist (e.g., IL7) refers to the administration of an IL7R agonist that improves the clinical condition of a disorder(s) or disease(s) associated with impaired normal T cell activity, preferably a disorder(s) that requires immune response stimulation, more preferably a cancer disorder or an infectious disease whose onset itself does not depend on the IL7R pathway.

[0025] Typically, an IL7R modulator can be a compound having antagonist or agonist properties for the IL7-IL7 receptor interaction and regulating (increasing or decreasing) IL7 / IL7R-mediated signaling (the IL7R signaling pathway induced by IL7).

[0026] In a preferred embodiment, the IL7R modulator (e.g., an IL7R antagonist or an IL7R agonist) can be selected from the group consisting of antibodies and antigen-binding antibody mimetics against the components of the IL7-IL7R signaling pathway, such as proteins, polypeptides or peptide modulators (e.g., IL7), mutants or derivatives thereof (fragments, fusion proteins, soluble proteins, dominant negative mutants), fragments thereof, and expressible derivatives.

[0027] The binding of IL-7 to IL-7R induces the activation of several signaling pathways, including Janus kinase (JAK)-1 and Janus kinase (JAK)-3, signal transducer and activator of transcription 5 (STAT5), and phosphatidylinositol 3-kinase (PI3-k). Although the activation of STAT1 and STAT3 pathways has been reported, they do not appear to be the major pathways. The activation of the STAT5 pathway is required for the induction of the anti-apoptotic protein Bcl-2 and the prevention of the apoptosis-promoting protein Bax from entering the mitochondria, and thus is required for the survival of T cell progenitor cells generated in the thymus. The activation of the PI3-k pathway results in the phosphorylation and cytoplasmic retention of the apoptosis-promoting protein Bad.

[0028] In certain embodiments, the IL7-R signaling pathway induced by IL7 can be identified by measuring STAT5 phosphorylation, as described in the examples of WO2018 / 104483. The IL-7-induced phosphorylation of STAT5 is a marker of IL7-R activation. Compounds that antagonize the IL7-IL7-R interaction are expected to reduce the IL-7-induced phosphorylation of STAT5, and compounds that stimulate the IL7-IL7-R interaction are expected to increase the IL-7-induced phosphorylation of STAT5.

[0029] In certain embodiments, an antagonist of IL7-R according to the present disclosure inhibits the IL-7-induced phosphorylation of STAT5. In preferred embodiments, the inhibition of STAT5 phosphorylation is greater than 50% at an IL7R antagonist concentration as low as about 55 ng / ml, and / or the inhibition of STAT5 phosphorylation is greater than 80% at an antibody concentration as low as about 100 ng / ml. The inhibition of STAT5 phosphorylation can be evaluated by methods known to those skilled in the art, in particular, by the methods described in Example 5 of WO2018 / 104483, and / or paragraphs

[69] and

[70] on page 21, and Example 3 of WO2015 / 189302. An agonist of IL7-R according to the present disclosure increases the IL-7-induced phosphorylation of STAT5.

[0030] Modulators of IL7R, such as the IL7R antagonists (especially antibodies, antigen-binding fragments thereof) or agonists of the present disclosure, can be identified by measuring the activation of the PI3-K and / or ERK (extracellular signal-regulated kinase) signaling pathways. In certain embodiments, the IL7R antagonist inhibits and / or does not activate or enhance the activation of the PI3-k and / or ERK (extracellular signal-regulated kinase) signaling pathways, particularly inhibiting and / or not inducing or enhancing the phosphorylation of PI3-k and / or ERK1 and / or ERK2. In particular, the IL7R antagonists provided herein do not induce the activation of the PI3-k and / or ERK pathways (preferably, the PI3-k and ERK pathways), particularly do not induce the phosphorylation of PI3-k and / or ERK 1 and / or ERK 2, and more particularly do not induce the phosphorylation of PI3-k and ERK 1 and ERK 2. In particular, the IL7R antagonist inhibits the activation of the PI3-k and / or ERK pathways, particularly inhibits the phosphorylation of PI3-k and / or ERK 1 and / or ERK 2, and more particularly inhibits the phosphorylation of PI3-k and ERK 1 and ERK 2. The activation of the protein pathway and / or phosphorylation can be tested by methods known to those skilled in the art, particularly by Western blotting as shown in Figure 7 and Example 8 of WO2018 / 104483, preferably by inhibiting the activation of PI3-k and / or ERK.

[0031] In another particular embodiment, the IL7R agonist induces or enhances the activation of the PI3-k and / or ERK (extracellular signal-regulated kinase) signaling pathways, particularly induces or enhances the phosphorylation of PI3-k and / or ERK 1 and / or ERK 2.

[0032] The IL7R antagonists (particularly, antibodies, their antigen-binding fragments) or agonists of the present disclosure can also be identified by measuring the internalization of CD127. In certain embodiments, the IL7R antagonists provided herein inhibit the IL-7-induced internalization of CD127, and the IL7R agonists increase the IL-7-induced internalization of CD127. The internalization of CD127 can be measured by comparing the cell surface expression level of CD127 in cells incubated with an IL7R modulator (e.g., an IL7R antagonist or agonist) to a control value such as cells not incubated with the modulator.

[0033] In certain embodiments, when incubated with the IL7R antagonist, the presence of IL7 does not induce, or induces a less strong decrease in, the cell surface expression of CD127 than in cells incubated without the antagonist. In certain embodiments, when incubated with the antagonist, the level of CD127 cell surface expression when the cells are incubated with 5 ng / mL of IL7 at 37 °C for 15 minutes is at least 80%, preferably at least 90%, of the cell surface expression level in cells incubated without IL7. In vitro, cell surface expression is preferably measured after a limited time as shown above. Moreover, since most cell internalization processes are inhibited at low temperature, the effect is usually best observed at physiological temperature, particularly 37 °C. However, incubation of cells at low temperature, particularly 4 °C, is also contemplated.

[0034] In another preferred embodiment, the IL7R antagonists provided herein do not induce internalization of CD127. Thus, the cell surface expression of CD127 in cells incubated in the presence of the IL7R antagonist is not reduced, or otherwise is not significantly reduced compared to the cell surface expression in cells incubated under the same conditions but in the absence of the antagonist. In certain embodiments, in the presence of 50 ng / mL of the antagonist, when incubated at 37 °C for 30-45 minutes, the level of CD127 cell surface expression is at least 80%, preferably at least 90% of that level in cells incubated in the absence of the antagonist. This antagonist effect can be observed in the absence of IL7 (in both antibody-treated and untreated cells), in the presence of IL7, and / or in both.

[0035] The above two CD127 internalization-related features (i.e., inhibition of IL7-induced internalization or non-induction of internalization) can be further defined and / or tested as described in WO2015 / 189302, particularly in paragraphs

[59] -

[63] on pages 19-20 and Figures 16 and Example 5.

[0036] For example, an IL7R modulator according to the present disclosure can be a) providing a plurality of cells expressing CD127 on the surface, b) incubating the cells with a candidate compound, c) determining whether the candidate compound binds to CD127 and decreases or increases the biological activity of CD127 (e.g., STAT5 phosphorylation, activation of the phosphatidylinositol 3-kinase and / or ERK signaling pathways, CD127 internalization) as described above, and d) selecting a candidate compound that decreases or increases the biological activity of CD127 (e.g., STAT5 phosphorylation, activation of the phosphatidylinositol 3-kinase and / or ERK signaling pathways, CD127 internalization) and can be identified by a method comprising the steps of.

[0037] In a preferred embodiment, the IL7R modulator according to the present disclosure is an IL7R antagonist.

[0038] As used herein, "IL7 receptor antagonist (e.g., anti-IL7R antagonist antibody or antigen-binding fragment thereof)" refers to a compound (e.g., anti-IL7R antagonist antibody or antigen-binding fragment thereof) that has antagonist properties for the IL7-IL7 receptor interaction and impairs the function of IL7 / IL7R-mediated signaling cells. According to certain embodiments, the IL7R antagonist according to the present disclosure further has antagonist properties for interleukin 7 (IL7), thereby antagonizing the access of IL7 to CD127 on CD127-positive cells, i.e., the binding of IL7 to CD127.

[0039] "Antagonist properties for the IL7-IL7-R interaction" means that an IL7R antagonist (e.g., an IL7R antagonist antibody or antigen-binding fragment thereof) targeting IL7-R alpha has the effect of preventing the accessibility of its binding partner IL7, particularly human IL7, to the IL7 receptor expressed on CD127 cells, particularly human effector T cells, particularly human memory T cells. As a result of antagonizing the binding of IL7, the IL7R antagonist results in lymphopenia by preventing the generation of IL7-dependent thymic T cells. This antagonist property can be, in particular, an antagonistic action against IL7-induced IL7-R signaling.

[0040] In a preferred embodiment, the IL7R antagonist provided herein does not bind to CD127 in the TSLP (thymic stromal lymphopoietin) receptor (i.e., does not bind to CD127 when complexed with CRLF2 that forms the TSLP receptor). Thus, the antagonist provided herein does not interfere with TSLP-induced and / or TSLP receptor-mediated signaling. In another preferred embodiment, the IL7R antagonist cannot bind to non-T cells in whole blood; ((d) cannot bind to CD127IL-7Ra expressed on non-T cells (e.g., monocytes) in whole blood, such as human whole blood).

[0041] In certain embodiments, the IL7R antagonist can be an anti-IL7R antagonist antibody or an antigen-binding fragment thereof, preferably a monoclonal antibody or an antigen-binding fragment thereof that specifically binds to the alpha chain of the receptor for IL-7, preferably the alpha chain of the receptor for human IL-7. The alpha chain of the receptor for interleukin 7 (IL-7) is named CD127 or p90 IL-7R, IL-7R alpha or IL-7Rα (NCBI accession number: XP_005248356.1, updated November 22, 2021). According to the present disclosure, the IL7R antagonist is preferably an IL7Rα antagonist, more preferably an anti-IL7Rα antagonist antibody or an antigen-binding fragment thereof.

[0042] As used herein, the terms "specifically binds to" or "specifically binding" mean that the antigen receptor has at least about 1×10 -6 M, 1×10 -7 M, 1×10 -8 M, 1×10 -9 M, 1×10 -10 M, 1×10 -11 M, 1×10 -12Refers to the ability to bind to an antigen with an affinity of M or more, and / or the ability to bind to a target with an affinity that is at least two-fold greater than the affinity for a non-specific antigen. Affinity can be determined by various methods well-known to those skilled in the art. These methods include, but are not limited to, Biacore analysis, Blitz analysis, and Scatchard plots.

[0043] In an embodiment, the antibody or antigen-binding fragment thereof according to the present invention can be determined by biosensor analysis, particularly by Biacore analysis, to have a KD value of 10 -8 M or less, preferably 10 -9 M or less, more preferably 1.10 -10 M or less for CD127, particularly human CD127.

[0044] As used herein, the term "antibody" refers to an immunoglobulin molecule and an immunologically active portion of an immunoglobulin molecule, i.e., a molecule that contains an antigen-binding site that immunospecifically binds to an antigen. Thus, the term "antibody" encompasses not only the entire antibody molecule, but also antibody fragments and variants (including derivatives) of the antibody.

[0045] In the natural antibodies of rodents and primates, two heavy chains are linked to each other by disulfide bonds, and each heavy chain is linked to a light chain by a disulfide bond. There are two types of light chains: lambda (λ) and kappa (κ). There are five major heavy chain classes (or isotypes) that determine the functional activities of the antibody molecules: IgM, IgD, IgG, IgA, and IgE. Each chain contains separate sequence domains. In a typical IgG antibody, the light chain contains two domains: a variable domain (VL) and a constant domain (CL). The heavy chain contains four domains: a variable domain (VH) and three constant domains (CH1, CH2, and CH3, collectively called CH). The variable regions of both the light chain (VL) and the heavy chain (VH) determine the binding recognition and specificity for the antigen. The constant region domains of the light chain (CL) and the heavy chain (CH) confer important biological properties such as antibody chain association, secretion, transplacental mobility, complement binding, and binding to Fc receptors (FcR).

[0046] The Fv fragment is the N-terminal portion of the Fab fragment of an immunoglobulin and consists of the variable portions of one light chain and one heavy chain. The specificity of an antibody lies in the structural complementarity between the antibody's antigen-binding site and the antigen determinant. The antigen-binding site of an antibody is mainly composed of residues derived from the hypervariable regions or complementarity-determining regions (CDRs). In some cases, residues from non-hypervariable regions or framework regions (FRs) may be involved in the antibody binding site or may affect the overall domain structure and thus the binding site to the antigen. The complementarity-determining region, i.e., CDR, refers to the amino acid sequence that defines both the binding affinity and specificity of the native Fv region of the native immunoglobulin binding site. The light and heavy chains of an immunoglobulin each have three CDRs named L-CDR1, L-CDR2, L-CDR3 and H-CDR1, H-CDR2, H-CDR3. Thus, the antigen-binding site typically contains six CDRs, including a set of CDRs from each of the heavy and light chain V regions. The framework region (FR) refers to the amino acid sequence sandwiched between the CDRs. Thus, the variable regions of the light and heavy chains typically contain four framework regions and three CDRs in the following sequence: FR1-CDR1-FR2-CDR2-FR3-CDR3-FR4.

[0047] Residues in the antibody variable domains are conventionally numbered according to the system devised by Kabat et al. This system is described in Kabat et al., 1987, Sequences of Proteins of Immunological Interest, US Department of Health and Human Services, NIH, USA (Kabat et al., 1992, hereinafter "Kabat et al."). This numbering system is used herein. The Kabat residue designations do not necessarily directly correspond to the numbering of the linear amino acid residues in the sequences of SEQ ID NOs. The actual linear amino acid sequence may contain fewer or additional amino acids compared to the strict Kabat numbering corresponding to deletions or insertions into the framework of the basic variable domain structure or into the complementarity determining regions (CDRs). The correct Kabat numbering of residues can be determined for a given antibody by aligning homologous residues in the antibody sequence with the "canonical" Kabat numbering sequence.

[0048] As used herein, the term "monoclonal antibody" refers to a preparation of antibody molecules of single specificity. Monoclonal antibodies exhibit a single binding specificity and affinity for a particular epitope. Thus, the term "human monoclonal antibody" refers to an antibody having a single binding specificity and having variable and constant regions derived from or based on human germline immunoglobulin sequences or from fully synthetic sequences. The method of preparing the monoclonal antibody is not relevant to the binding specificity.

[0049] As used herein, the term "antigen-binding fragment" of an antibody (or simply "antibody fragment") refers to one or more fragments of an antibody that retain the ability to specifically bind to an antigen (e.g., IL7R), and in particular, has antagonist properties against the IL7-IL7 receptor interaction, impairs the function of IL7 / IL7R-mediated signal transduction such as the PI3K / Akt / mTOR and JAK / STAT pathways, and can exhibit cytotoxic activity against CD127-positive cells. It has been shown that the antigen-binding function of an antibody can be performed by fragments of a full-length antibody.

[0050] Examples of binding fragments included within the term "antigen-binding fragment" of an antibody include: a monovalent fragment consisting of the Fab fragment, VL, VH, CL, and CH1 domains; a bivalent fragment comprising the F(ab)2 fragment, two Fab fragments linked by a disulfide bridge in the hinge region; the Fd fragment consisting of the VH domain and the CH1 domain; the Fv fragment consisting of the VL domain and the VH domain of a single arm of an antibody; the dAb fragment consisting of the VH domain (Ward et al., 1989 Nature 341:544-546), or any fusion protein comprising such an antigen-binding fragment. Furthermore, although the two domains VL and VH of the Fv fragment are encoded by separate genes, they can be joined by a synthetic linker using recombinant methods, and by this synthetic linker, they can be made as a single-chain protein, in which the VL region and the VH region pair to form a monovalent molecule (known as single-chain Fv (scFv)) (see, for example, Bird et al., 1988 Science 242:423-426; and Huston et al., 1988 Proc. Natl. Acad. Sci. 85:5879-5883). Such single-chain antibodies are also intended to be included within the term "antigen-binding fragment" of an antibody. These antibody fragments are obtained using conventional techniques known to those skilled in the art, and the fragments are screened for utility in the same manner as intact antibodies.

[0051] In certain embodiments, the anti-IL7R antagonist antibody or antigen-binding fragment thereof according to the present disclosure is an antibody or antigen-binding fragment thereof of N13B2h1, N13B2h2 or N13B2h3 described in WO2015 / 189302, or an antibody or antigen-binding fragment thereof of N13B2-hVL3, N13B2-hVL4, N13B2-hVL5 or N13B2-hVL6 described in WO2018 / 104483, and comprises: (a) a variable heavy chain comprising three CDRs wherein VHCDR1 comprises or consists of the amino acid sequence of SEQ ID NO: 1, VHCDR2 comprises or consists of the amino acid sequence of SEQ ID NO: 2, and VHCDR3 comprises or consists of the amino acid sequence of SEQ ID NO: 3 or 11; and (b) a variable light chain comprising three CDRs wherein VLCDR1 comprises or consists of the amino acid sequence of SEQ ID NO: 4 or 12, VLCDR2 comprises or consists of the amino acid sequence of SEQ ID NO: 5 or 13, and VLCDR3 comprises or consists of the amino acid sequence of SEQ ID NO: 6.

[0052] In more particular embodiments, the anti-IL7R antagonist antibody or antigen-binding fragment thereof according to the present disclosure is an antibody or antigen-binding fragment thereof of N13B2h3 described in WO2015 / 189302, or an antibody or antigen-binding fragment thereof of N13B2-hVL3, N13B2-hVL4, N13B2-hVL5 or N13B2-hVL6 described in WO2018 / 104483, preferably an antibody or antigen-binding fragment thereof of N13B2-hVL6 described in WO2018 / 104483, and comprises: (a) a variable heavy chain comprising three CDRs wherein VHCDR1 comprises or consists of the amino acid sequence of SEQ ID NO: 1, VHCDR2 comprises or consists of the amino acid sequence of SEQ ID NO: 2, and VHCDR3 comprises or consists of the amino acid sequence of SEQ ID NO: 3; and (b) a variable light chain comprising three CDRs wherein VLCDR1 comprises or consists of the amino acid sequence of SEQ ID NO: 4, VLCDR2 comprises or consists of the amino acid sequence of SEQ ID NO: 5, and VLCDR3 comprises or consists of the amino acid sequence of SEQ ID NO: 6.

[0053] It is further contemplated that the antibodies or antigen-binding fragments thereof can be further screened or optimized for their affinity binding and / or antagonist activity. In particular, the antibody or antigen-binding fragment thereof may have one, two, three, four, five, or six modifications in the amino acid sequences of one, two, three, four, five, or six CDRs of the monoclonal antibodies provided herein. It is contemplated that the amino acids at positions 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 of CDR1, CDR2, CDR3, CDR4, CDR5, or CDR6 of the VJ or VDJ region of the light or heavy chain variable region of the antibody may have insertions, deletions, or substitutions with conservative or non-conservative amino acids. Such amino acids that can be substituted or that can constitute a substitution are disclosed above.

[0054] In some embodiments, the amino acid differences are conservative substitutions, i.e., the substitution of one amino acid with another amino acid having similar chemical or physical properties (size, charge or polarity), which generally does not adversely affect the biochemical, biophysical and / or biological properties of the antibody. In particular, the substitution does not inhibit the interaction between the antibody and the IL7R antigen and the antagonistic properties. The conservative substitution(s) are preferably selected from one of the following five groups: Group 1 - small aliphatic, non-polar or slightly polar residues (A, S, T, P, G); Group 2 - polar, negatively charged residues and their amides (D, N, E, Q); Group 3 - polar, positively charged residues (H, R, K); Group 4 - large aliphatic, non-polar residues (M, L, I, V, C); and Group 5 - large aromatic residues (F, Y, W).

[0055] In certain embodiments, the anti-IL7R antagonist is an antibody or antigen-binding fragment thereof of N13B2h1, N13B2h2 or N13B2h3 described in WO2015 / 189302, and comprises a heavy chain variable domain comprising or consisting of an amino acid selected from the group consisting of SEQ ID NOs: 14-17, and a light chain variable domain comprising or consisting of an amino acid sequence selected from the group consisting of SEQ ID NOs: 18-21. Preferably, the anti-IL7R antagonist is an antibody or antigen-binding fragment thereof of N13B2h3 described in WO2015 / 189302, and comprises a heavy chain variable domain comprising or consisting of the amino acid sequence of SEQ ID NO: 17, and a light chain variable domain comprising or consisting of the amino acid sequence of SEQ ID NO: 21. More preferably, the antibody or antigen-binding fragment thereof comprises a complete heavy chain comprising or consisting of SEQ ID NO: 22 and a complete light chain comprising or consisting of SEQ ID NO: 23.

[0056] In another particular embodiment, the anti-IL7R antagonist is an antibody or antigen-binding fragment thereof of N13B2-hVL3, N13B2-hVL4, N13B2-hVL5 or N13B2-hVL6 described in WO2018 / 104483, and comprises a heavy chain variable domain comprising or consisting of the amino acid sequence of SEQ ID NO: 7, and a light chain variable domain comprising or consisting of an amino acid sequence selected from the group consisting of SEQ ID NOs: 8, 24-26.

[0057] In a more preferred embodiment, the anti-IL7R antagonist antibody or antigen-binding fragment thereof is an antibody or antigen-binding fragment thereof of N13B2-hVL6 described in WO2018 / 104483, and comprises a heavy chain variable domain comprising or consisting of the amino acid sequence of SEQ ID NO: 7 and a light chain variable domain comprising or consisting of the amino acid sequence of SEQ ID NO: 8. Preferably, the antibody or antigen-binding fragment thereof comprises a complete heavy chain comprising or consisting of SEQ ID NO: 9 and a complete light chain comprising or consisting of SEQ ID NO: 10.

[0058] An anti-IL7R antagonist antibody or an antigen-binding fragment thereof having an amino acid sequence with at least 90%, for example, at least 95%, 96%, 97%, 98%, or 99% identity to any one of the amino acid sequences defined above is also part of the present disclosure. Typically, the anti-IL7R antagonist antibody or an antigen-binding fragment thereof has at least equivalent or greater antagonist activity for the above IL7-IL7R interaction than the above IL7R antagonist antibody or an antigen-binding fragment thereof consisting of VH and VL of the heavy and light chains of the N13B2-hVL6 antibody.

[0059] As used herein, percent identity between two sequences is a function of the number of identical positions shared by the sequences, taking into account the number of gaps that need to be introduced for optimal alignment of the two sequences and the length of each gap (i.e., % identity = number of identical positions / total number of positions × 100). Comparison of sequences and determination of percent identity between two sequences can be accomplished using the following mathematical algorithms.

[0060] The percent identity between two amino acid sequences or nucleotide sequences can be determined using the algorithm of E. Meyers and W. Miller (Comput. Appl. Biosci., 4:11-17, 1988) incorporated into the ALIGN program (version 2.0), using a PAM120 weight residue table, a 12 gap length penalty, and a 4 gap penalty. Alternatively, the percent identity between two amino acid sequences or nucleotide sequences can be determined using the algorithm of Needleman and Wunsch (J. Mol, Biol. 48:444-453, 1970) incorporated into the GAP program (available at http: / / www.gcg.com) within the GCG software package, using either a Blossom 62 matrix or a PAM250 matrix, and a gap weight of 16, 14, 12, 10, 8, 6, or 4 and a length weight of 1, 2, 3, 4, 5, or 6. The percent identity between two nucleotide sequences or amino acid sequences can also be determined, for example, using an algorithm such as the BLASTN program for nucleic acid or amino acid sequences, with a default of an 11 word length (W), an expectation value (E) of 10, M = 5, N = 4, and a comparison of both strands.

[0061] In another specific embodiment of the present disclosure, the IL7R antagonist (e.g., an IL7R antagonist antibody or an antigen-binding fragment thereof) further has cytotoxic properties against human cells expressing the receptor, particularly human T cells.

[0062] According to certain embodiments of the present disclosure, the cytotoxic IL7R antagonist further has the ability to enhance the antibody-dependent cell phagocytosis (ADCP) activity of CD127-positive tumor cells by macrophages, and in particular is an antibody that does not have antibody-dependent cell cytotoxicity (ADCC) activity in immune cells. In certain embodiments of the present invention, the anti-IL-7R antagonist antibody or antigen-binding fragment thereof used in the methods of the present invention or for use in accordance with the present invention, in particular, the anti-CD127 antibody or fragment thereof, has the ability to enhance the antibody-dependent cell phagocytosis (ADCP) activity of CD127-positive tumor cells by macrophages, and preferably, in particular, does not have antibody-dependent cell cytotoxicity (ADCC) activity in immune cells.

[0063] In another embodiment, the IL7R modulator (i.e., an IL7R antagonist or an IL7R agonist) is an antigen-binding antibody mimetic. As used herein, the term "antigen-binding antibody mimetic" refers to artificial proteins, peptides, and any compounds having antigen-binding ability that mimics the ability of an antibody. Such mimics include affitins and anticalins as well as aptamers (peptide aptamers and oligonucleotide aptamers).

[0064] In one embodiment, the IL7R modulator (i.e., antagonist or IL7R agonist) is an aptamer. An aptamer is a class of molecules that represents an alternative to antibodies from the perspective of molecular recognition. An aptamer is an oligonucleotide or oligopeptide sequence that has the ability to recognize substantially any class of target molecule with high affinity and specificity. Such ligands can be isolated by Systematic Evolution of Ligands by EXponential enrichment (SELEX) of a random sequence library. The random sequence library is obtained by combinatorial chemical synthesis of DNA. In this library, each member is a finally chemically modified linear oligomer of a unique sequence. A peptide aptamer consists of a structurally constrained antibody variable region presented by a platform protein such as Escherichia coli thioredoxin A and is selected from a combinatorial library by the two-hybrid method.

[0065] In some embodiments, the IL7R modulator (i.e., IL7R antagonist or IL7R agonist) is a polypeptide. The term "polypeptide" as used herein means a polymer of amino acids having no specific length. Thus, peptides, oligopeptides and proteins are included in the definition of "polypeptide", and these terms are used interchangeably throughout this specification and in the claims. The term "polypeptide" does not exclude post-translational modifications including, but not limited to, phosphorylation, acetylation, glycosylation, etc.

[0066] In one embodiment, the polypeptide of the present disclosure has a length comprising 2 to 200 amino acids. In one embodiment, the polypeptide of the present disclosure has a length comprising 2 to 190, particularly 10 to 180, 10 to 170, 10 to 160, 10 to 150, 10 to 140, 10 to 130, 10 to 120, 10 to 110 amino acids. In one embodiment, the polypeptide of the present invention has a length comprising 10 to 100 amino acids. In one embodiment, the polypeptide of the present invention has a length comprising 50 to 100 amino acids. In one embodiment, the polypeptide of the present invention has a length of 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99 or 100 amino acids.

[0067] In certain embodiments, the polypeptide is a functional equivalent of IL7R. As used herein, an “functional equivalent” of IL7R is a compound that can bind to at least one IL7 ligand and thereby prevent interaction with IL7R. The term “functional equivalent” includes fragments, mutants, and muteins of IL7R. The term “functionally equivalent” thus includes any equivalent of IL7R obtained by modifying the amino acid sequence so that the protein analog retains its ability to bind to its ligand, for example, by deletion, substitution or addition of one or more amino acids. Amino acid substitutions may be made, for example, by point mutation of the DNA encoding the amino acid sequence.

[0068] Functional equivalents include, but are not limited to, molecules that include all or part of the extracellular domain of IL7R and that form a soluble receptor that can bind to and capture the ligand of IL7R. Thus, functional equivalents include the soluble form of IL7R. Suitable soluble forms of these proteins, or their functional equivalents, may include, for example, cleaved forms of the proteins in which the transmembrane domain has been removed by chemical, proteolytic or recombinant methods. In particular, over the entire length of the corresponding protein, at least 80% identity, more particularly at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, and even more particularly at least 99% identity with the corresponding protein. As used herein, the term "corresponding protein" refers to a protein to which the functional equivalent of the invention has a similar function. The percentage of identity referred to in the present disclosure is determined based on the global alignment of the sequences to be compared, i.e., the alignment of the sequences over their entire length, using, for example, the algorithm of Needleman and Wunsch 1970. This sequence comparison can be performed using the needle software, for example, by using parameters "Gap open" equal to 10.0, parameter "Gap Extend" equal to 0.5, and matrix "BLOSUM 62". Software such as needle is available on the world wide web site ebi.ac.uk under the name "needle". As used herein, the term "functionally equivalent fragment" can also mean any fragment or assembly of fragments of IL7R that binds to IL7.Accordingly, the present disclosure provides a polypeptide capable of inhibiting the binding of IL7R to IL7, in particular a functional equivalent, comprising a continuous amino acid having a sequence corresponding to at least a part of the extracellular domain sequence of IL7R, which is the part binding to IL7. In some embodiments, the polypeptide, in particular the functional equivalent, corresponds to the extracellular domain of IL7R.

[0069] In some embodiments, the functional equivalent of IL7R is fused to a heterologous polypeptide to form a fusion protein. As used herein, a "fusion protein" includes all or (typically, a bioactive) part of a functional equivalent of the present disclosure operably linked to a heterologous polypeptide (i.e., a polypeptide other than the polypeptide). In a fusion protein, the term "operably linked" is intended to indicate that the functional equivalent of the present disclosure and the heterologous polypeptide are fused in-frame with each other. The heterologous polypeptide can be fused to the N-terminus or C-terminus of the functional equivalent of the present disclosure.

[0070] In some embodiments, a functional equivalent of IL7R is fused to an immunoglobulin constant domain (Fc region) to form an immunoadhesin. The immunoadhesin can possess many of the valuable chemical and biological properties of human antibodies. The immunoadhesin can be constructed from a human protein sequence having the desired specificity linked to an appropriate human immunoglobulin hinge and constant domain (Fc) sequence, so that the binding specificity of interest can be achieved using fully human components. Such immunoadhesins have minimal immunogenicity in patients and are safe for chronic or repeated use. In some embodiments, the Fc region is the native sequence Fc region. In some embodiments, the Fc region is a variant Fc region. In yet another embodiment, the Fc region is a functional Fc region. As used herein, the term "Fc region" is used to define the C-terminal region of an immunoglobulin heavy chain, including the native sequence Fc region and variant Fc regions. The boundaries of the Fc region of an immunoglobulin heavy chain can vary, but the human IgG heavy chain Fc region is typically defined as extending from the amino acid residue at position Cys226 or Pro230 to its carboxyl terminus. The adhesion portion of the immunoadhesin and the immunoglobulin sequence portion can be linked by a minimal linker. The immunoglobulin sequence is typically, but not necessarily, an immunoglobulin constant domain. The immunoglobulin portion in the chimeras of the invention can be obtained from IgG1, IgG2, IgG3 or IgG4 subtypes, IgA, IgE, IgD or IgM, but typically from IgG1 or IgG4. In some embodiments, the functional equivalent of IL7R and the immunoglobulin sequence portion of the immunoadhesin are linked by a minimal linker. As used herein, the term "linker" refers to a sequence of at least one amino acid that links the polypeptide of the invention to the immunoglobulin sequence portion. Such linkers can be useful in preventing steric hindrance.In some embodiments, the linker has 4; 5; 6; 7; 8; 9; 10; 11; 12; 13; 14; 15; 16; 17; 18; 19; 20; 21; 22; 23; 24; 25; 26; 27; 28; 29; 30 amino acid residues. However, the upper limit is not critical and is selected, for example, for reasons of convenience regarding the manufacture of such polypeptides as biopharmaceuticals. The linker sequence may be a naturally occurring sequence or a non-naturally occurring sequence. When used for therapeutic purposes, the linker is typically non-immunogenic in the subject to whom the immunoadhesin is administered. One useful group of linker sequences is the linker derived from the hinge region of a heavy chain antibody as described in WO 96 / 34103 and WO 94 / 04678. Another example is a polyalanine linker sequence.

[0071] According to the present disclosure, when the antagonist (e.g., an anti-IL7R antagonist antibody or an antigen-binding fragment thereof) has an IL7R antagonist effect that can be correlated with a therapeutic response to the antagonist treatment in the patient, the patient is likely to respond to the IL7R antagonist (e.g., an anti-IL7R antagonist antibody or an antigen-binding fragment thereof) treatment.

[0072] In another specific embodiment, according to the present disclosure, when the agonist has an IL7R agonist effect that can be correlated with a therapeutic response to the agonist treatment in the patient, the patient is likely to respond to the IL7R agonist (e.g., IL7) treatment.

[0073] According to different methods of the present disclosure, the analysis of the gene expression profiles of at least 1, 2, 3, or 4 genes selected from the group consisting of the BCL2, CISH, PTGER2, and DPP4 genes, preferably at least 1, 2, 3, 4, 5, or 6 genes selected from the group consisting of the BCL2, CISH, PTGER2, DPP4, SOCS2, and FLT3LG genes, in patient samples previously treated with an IL7R modulator such as an IL7R antagonist (e.g., an anti-IL7R antagonist antibody or an antigen-binding fragment thereof) or an IL7R agonist can determine the effectiveness of an IL7R modulator such as an IL7R antagonist (e.g., an anti-IL7R antagonist antibody or an antigen-binding fragment thereof) or an IL7R agonist in a patient, evaluate the treatment response of the patient to the antagonist (e.g., an anti-IL7R antagonist antibody or an antigen-binding fragment thereof) or agonist treatment, or assess the likelihood of a treatment response, or can be used as a biomarker for selecting an IL7R antagonist or agonist compound that is likely to be effective in the treatment.

[0074] Therefore, different methods according to the present disclosure include determining the expression levels of at least 1, 2, 3, or 4 genes selected from the group consisting of the BCL2, CISH, PTGER2, and DPP4 genes, preferably at least 1, 2, 3, 4, 5, and 6 genes selected from the group consisting of the BCL2, CISH, PTGER2, DPP4, SOCS2, and FLT3LG genes.

[0075] The term "determining the expression profile of at least 1, 2, 3, or 4 genes selected from the group consisting of the BCL2, CISH, PTGER2, and DPP4 genes" means that the expression levels of at least 1, 2, 3, or 4 of the said genes are evaluated. In certain embodiments, the expression level of each of all the said genes is evaluated.

[0076] The human BCL2 gene, also named BCL2 apoptosis regulator (Gene ID: 596, updated on June 20, 2021), encodes a complete mitochondrial outer membrane protein that blocks apoptotic death in some cells such as lymphocytes. The human BCL2 gene encodes a 6881bp BCL2 transcript variant alpha (NCBI reference sequence: NM_000633.3, updated on July 1, 2021) and a 1595bp BCL2 transcript variant beta (NCBI reference sequence: NM_000657.3, updated on July 1, 2021) that encode BCL2 isoform alpha protein (NCBI reference sequence: NP_000624.2, updated on July 1, 2021) and BCL2 isoform beta protein (NCBI reference sequence: NP_000648.2, updated on July 1, 2021), respectively, which are generated by alternative splicing.

[0077] The human CISH gene (cytokine-inducible SH2-containing protein, Gene ID: 1154, updated on June 20, 2021) encodes a protein containing an SH2 domain and a SOCS box domain, and belongs to the cytokine-inducible STAT inhibitor (CIS), also known as the suppressor of cytokine signaling (SOCS) or STAT-induced STAT inhibitor (SSI) protein family. The human CISH gene encodes a 2176 bp CISH transcript variant 1 (NCBI reference sequence: NM_013324.7, updated on July 1, 2021) and a 2019 bp CISH transcript variant 2 (NCBI reference sequence: NM_145071.4, updated on July 1, 2021) that encode CISH isoform 1 protein (NCBI reference sequence: NP_037456.5, updated on July 1, 2021) and CISH isoform 2 protein (NCBI reference sequence: NP_659508.1, updated on July 1, 2021), respectively.

[0078] The human PTGER2 gene (prostaglandin E receptor 2, Gene ID: 5732, updated on June 20, 2021) encodes a receptor for prostaglandin E2, a metabolite of arachidonic acid that has different biological activities in a wide range of tissues. The human PTGER2 gene encodes a 2548 bp (NCBI reference sequence: NM_000956.4, updated on June 24, 2021) transcript that encodes the PTGER2 protein (NCBI reference sequence: NP_000947.2, updated on June 24, 2021).

[0079] The human DPP4 gene (dipeptidyl peptidase 4, Gene ID: 1803, updated on June 28, 2021), also known as ADABP, ADCP2, CD26, DPPIV, and TP103, encodes dipeptidyl peptidase 4, which is identical to adenosine deaminase complexing protein-2 and T cell activation antigen CD26. The human DPP4 gene encodes four transcripts: 3573 bp DPP4 transcript variant 1 (NCBI reference sequence: NM_001935.4, updated on June 29, 2021), 3570 bp DPP4 transcript variant 2 (NCBI reference sequence: NM_001379604.1, updated on July 1, 2021), 3567 bp DPP4 transcript variant 3 (NCBI reference sequence: NM_001379605.1, updated on June 30, 2021), and 3519 bp DPP4 transcript variant 4 (NCBI reference sequence: NM_001379606.1, updated on July 1, 2021), which encode four protein isoforms: DPP4 isoform 1 (NCBI reference sequence: NP_001926.2, updated on June 29, 2021), DPP4 isoform 2 (NCBI reference sequence: NP_001366533.1, updated on July 1, 2021), DPP4 isoform 3 (NCBI reference sequence: NP_001366534.1, updated on June 30, 2021), and DPP4 isoform 4 (NCBI reference sequence: NP_001366535.1, updated on July 1, 2021).

[0080] The human SOCS2 gene (Gene ID: 8835, updated on June 11, 2021), also known as CIS2, Cish2, SOCS-2, SSI-2, SSI2, STATI2, encodes a member of the suppressor of cytokine signaling (SOCS) family. The human SOCS2 gene encodes five transcripts: the 2606 bp SOCS2 transcript variant 1 (NCBI reference sequence: NM_003877.5, updated on June 27, 2021), the 2653 bp SOCS2 transcript variant 2 (NCBI reference sequence: NM_001270467.2, updated on June 27, 2021), the 2581 bp SOCS2 transcript variant 3 (NCBI reference sequence: NM_001270468.2, updated on June 27, 2021), the 2431 bp SOCS2 transcript variant 4 (NCBI reference sequence: NM_001270469.2, updated on June 27, 2021), the 2888 bp SOCS2 transcript variant 5 (NCBI reference sequence: NM_001270470.1, updated on June 23, 2021), and the 2386 bp SOCS2 transcript variant 6 (NCBI reference sequence: NM_001270471.2, updated on June 26, 2021). These transcripts encode a 198-amino acid SOCS2 protein (NCBI reference sequence: NP_003868.1, updated on June 27, 2021).

[0081] The human FLT3LG gene (fms-related receptor tyrosine kinase 3 ligand, Gene ID: 2323, updated on June 11, 2021), also known as FL, FLG3L, and FLT3L, encodes a protein that controls the development of dendritic cells. The human FLT3LG gene encodes five transcripts: FLT3LG transcript variant 1 of 1101 bp (NCBI reference sequence: NM_001204502.2, updated on June 27, 2021), FLT3LG transcript variant 2 of 1051 bp (NCBI reference sequence: NM_001204503.2, updated on June 27, 2021), FLT3LG transcript variant 3 of 1050 bp (NCBI reference sequence: NM_001459.4, updated on June 26, 2021), FLT3LG transcript variant 4 of 1078 bp (NCBI reference sequence: NM_001278637.2, updated on May 3, 2021), and FLT3LG transcript variant 5 of 1067 bp (NCBI reference sequence: NM_001278638.2, updated on May 3, 2021). These transcripts encode two protein isoforms, the FLT3LG isoform 1 precursor protein (NCBI reference sequence: NP_001450.2, updated on June 26, 2021) and FLT3LG isoform 2 (NCBI reference sequence: NP_001265566.1, updated on May 3, 2021).

[0082] The gene expression profile of the said gene can be determined by any suitable method known to those skilled in the art.

[0083] The term "gene expression profile" or "gene signature" refers to the pattern of expression of at least one gene of the above gene set within a cell, which is a biological process and in this specification, results from the intracellular IL7R antagonist or agonist effect. According to the present disclosure, the gene expression profile of the biomarker can be determined by measuring the relative amounts of gene products expressed in cells treated with the above ILR7 antagonist (for example, an anti-IL7R antagonist antibody or an antigen-binding fragment thereof) or IL7R agonist, preferably, by calculating the gene set enrichment score per sample as the difference normalized by the empirical cumulative distribution function of the gene expression ranks inside and outside the gene set, by performing single-sample GSEA (ssGSEA) with the GSVA algorithm.

[0084] Typically, these methods involve measuring the amount of the above mRNA or protein. Methods for determining the amount of mRNA are well known in the art. For example, mRNA contained in a sample is first extracted according to standard methods, for example, using lysozyme or a chemical solution, or extracted by a nucleic acid binding resin according to the manufacturer's instructions. The extracted mRNA is then detected by hybridization (for example, Northern blot analysis) and / or amplification (for example, RT-PCR) using the primer pairs and probes specific for the genes described in the examples of the present disclosure. Quantitative or semi-quantitative RT-PCR is preferred. In another specific embodiment, the mRNA expression level is measured by the RNA seq method.

[0085] In some embodiments, for example, the expression level of the gene measured by quantitative RT-PCR is normalized by subtracting the expression level of a housekeeping gene determined in the same experiment from each gene, and the gene expression profile preferably corresponds to the normalized gene expression level of one gene selected from the group consisting of the BCL2, CISH, PTGER2, and DPP4 genes or at least two, three genes, preferably four genes, more preferably the sum of the normalized gene expression levels of each of 4, 5, or 6 genes selected from the group consisting of BCL2, CISH, PTGER2, DPP4, SOCS2, and FLT3LG.

[0086] In another embodiment, determining the gene expression profile specifically includes determining the expression level of the gene(s) by RNA-seq or DNA microarray.

[0087] (In Vitro Method for Evaluating the Therapeutic Response of IL7 Receptor Modulator Therapy during Treatment) In certain embodiments, the expression profile of at least one, preferably 2, 3, 4, 5 of these genes, more preferably all combinations thereof, is useful for monitoring the antagonist or agonist effect of a compound (IL7R antagonist (e.g., anti-IL7R antagonist antibody or its antigen-binding fragment) or IL7R agonist) and thus the effectiveness of the treatment in a patient during the treatment process.

[0088] The biomarkers according to the present disclosure enable classifying a patient as a responder to the treatment by determining the gene expression profile of at least 1, 2, 3, or 4 genes selected from the group consisting of the BCL2, CISH, PTGER2, and DPP4 genes in a patient sample, preferably at least 1, 2, 3, 4, 5, or 6 genes selected from the group consisting of the BCL2, CISH, PTGER2, DPP4, SOCS2, and FLT3LG genes, and by determining whether the gene expression profile of said genes is high or low compared to a control value.

[0089] The present disclosure relates to an in vitro method for evaluating the treatment response of a human patient to treatment with an IL7R antagonist (e.g., an anti-IL7 receptor antagonist antibody or an antigen-binding fragment thereof) by determining the gene expression profile of 1, 2, 3, or 4 genes selected from the group consisting of the human BCL2, CISH, PTGER2, and DPP4 genes in a sample from a patient administered at least one dose of the IL7R antagonist (e.g., an anti-IL7 receptor antagonist antibody or an antigen-binding fragment thereof), preferably at least 1, 2, 3, 4, 5, or 6 genes selected from the group consisting of the BCL2, CISH, PTGER2, DPP4, SOCS, and FLT3LG genes, wherein a lower gene expression profile of said gene(s) in said patient sample compared to a control value indicates that the patient is likely to respond to the antagonist treatment.

[0090] In another specific embodiment, the present disclosure provides an in vitro method for evaluating the therapeutic response of a human patient to treatment with an IL7R agonist (e.g., IL7) by determining the gene expression profile of 1, 2, 3, or 4 genes selected from the group consisting of the human BCL2, CISH, PTGER2, and DPP4 genes, preferably at least 1, 2, 3, 4, 5, or 6 genes selected from the group consisting of the BCL2, CISH, PTGER2, DPP4, SOCS, and FLT3LG genes, in a sample from a patient administered at least one dose of the IL7R agonist, wherein a higher gene expression profile of the gene(s) in the patient sample compared to a control value indicates that the patient is likely to respond to the agonist treatment.

[0091] As used herein, the term "evaluating the therapeutic response to treatment" refers to the ability to assess whether a patient's treatment is likely to be effective (e.g., provide a measurable benefit or a positive medical response) in a patient after a period of treatment. In other words, according to the present disclosure, evaluating the response to treatment refers to the ability to assess the effect of an IL7R antagonist (e.g., an anti-IL7R antagonist antibody or an antigen-binding fragment thereof) or an IL7R agonist in patient cells, which may correlate with the likely responsiveness of the patient to the treatment.

[0092] In particular, the antagonist or agonist effect of a compound in a patient, and thus the likelihood that the patient will respond to the antagonist or agonist treatment, is evaluated by comparing the gene expression profile of at least 1, 2, 3, or 4 genes selected from the group consisting of the BCL2, CISH, PTGER2, and DPP4 genes, preferably at least 1, 2, 3, 4, 5, or 6 genes selected from the group consisting of the BCL2, CISH, PTGER2, DPP4, SOCS2, and FLT3LG genes, in a patient administered at least one dose of the antagonist to a control value.

[0093] According to this method for evaluating a treatment response, the sample has been previously collected from a patient administered with at least one dose of an IL7R antagonist (e.g., an anti-IL7R antagonist antibody or an antigen-binding fragment thereof) or an IL7R agonist.

[0094] The term "patient sample" means any biological sample derived from a patient. Examples of such samples include tissues, cell samples, organs, biopsies, tumor samples, and the like.

[0095] Preferred biological samples are peripheral blood mononuclear cells, tissues or tumors containing infiltrating immune cells.

[0096] In certain embodiments, the sample comprises immune cells. Immune cells include lymphocytes such as B cells, T cells and natural killer cells. The immune cells can be obtained from a number of non-limiting sources including peripheral blood mononuclear cells, bone marrow, lymph node tissue, cord blood, thymus tissue, tissue from an infected site, ascites, pleural effusion, spleen tissue, and tumors, using any number of techniques known to those skilled in the art to obtain units of blood taken from a subject. Immune cells can be extracted from blood or can be stem cell-derived. In a preferred embodiment, the sample is a tissue sample, preferably a tumor tissue containing infiltrating immune cells from a cancer patient or a blood patient sample, more preferably isolated human peripheral blood mononuclear cells.

[0097] According to the present disclosure, a patient who has received IL7R antagonist or agonist treatment refers to a patient administered with at least one dose of an IL7R antagonist (e.g., an anti-IL7R antagonist antibody or an antigen-binding fragment thereof) or an IL7R agonist. The treatment response can be evaluated according to this method after each administration of a dose of an IL7R antagonist or an IL7R agonist over the course of the treatment to monitor the treatment response over time.

[0098] As used herein, the term "control value" preferably refers to a biological sample obtained from the patient at different times before the administration of the IL7R antagonist (e.g., anti-IL7R antagonist antibody or antigen-binding fragment thereof) dose, or before any administration of the antagonist dose, or before any treatment of the condition or disease, or a gene signature in a biological sample obtained from patient cells cultured in vitro without the antagonist or agonist, or patient cells cultured in vitro with a compound (e.g., an antibody) known not to affect the IL7 / IL7R pathway (i.e., at least 1, 2, 3, or 4 genes selected from the group consisting of the BCL2, CISH, PTGER2, and DPP4 genes, preferably at least 1, 2, 3, 4, 5, or 6 genes selected from the group consisting of the BCL2, CISH, PTGER2, DPP4, SOCS2, and FLT3LG genes). The control value can alternatively be a predetermined value such as a threshold, standard value, or range obtained from a source other than the patient's data. The predetermined value of the control can be based on a comparative measurement between the patient before the administration of the antagonist (e.g., anti-IL7R antagonist antibody or antigen-binding fragment thereof) dose and the patient administered the antagonist dose, or based on a comparative measurement between patient cells cultured in vitro with the antagonist or agonist, or patient cells cultured in vitro without the antagonist or agonist, or patient cells cultured in vitro with a compound (e.g., an antibody) known not to affect the IL7 / IL7R signaling pathway.

[0099] In certain embodiments, according to the method of evaluating the therapeutic response to IL7 receptor modulator therapy in the treatment process of the present disclosure, the control value is preferably the gene signature (i.e., at least 1, 2, 3, or 4 genes selected from the group consisting of BCL2, CISH, PTGER2, and DPP4 genes, preferably at least 1, 2, 3, 4, 5, or 6 genes selected from the group consisting of BCL2, CISH, PTGER2, DPP4, SOCS2, and FLT3LG genes) in a biological sample obtained from the patient at different time points before the administration of the IL7R antagonist (e.g., anti-IL7R antagonist antibody or antigen-binding fragment thereof) dose, or before any administration of the antagonist dose, or before any treatment of the condition or disease, or the control value refers to a predetermined value of a control established based on a comparative measurement between the patient before the administration of the antagonist (e.g., anti-IL7R antagonist antibody or antigen-binding fragment thereof) dose and the patient administered the antagonist dose.

[0100] In a preferred embodiment, the gene expression profile of the biomarker is evaluated in a sample collected from the patient at least 1, 2, 3, or 7, preferably 10, 14, 21, 28 days after the administration of the antibody dose, more particularly at least 30, 35, 40, 45, 50, 55, 60, 70, 80, 90, 100, 120 days after the administration of the antibody dose.

[0101] A lower gene expression profile of the biomarker compared to the control value in a sample of a patient administered at least one dose of an IL7R antagonist (e.g., anti-IL7R antagonist antibody or antigen-binding fragment thereof) correlates with the antagonist effect of the antagonist (e.g., anti-IL7R antagonist antibody or antigen-binding fragment thereof) in the patient, and thus correlates with the therapeutic response of the patient to the IL7R antagonist (e.g., anti-IL7R antagonist antibody or antigen-binding fragment thereof) treatment.

[0102] Typically, when the log2 fold change of the gene expression profile of at least 1, 2, 3, or 4 genes selected from the group consisting of the BCL2, CISH, PTGER2, and DPP4 genes, preferably at least 1, 2, 3, 4, 5, or 6 genes selected from the group consisting of the BCL2, CISH, PTGER2, DPP4, SOCS2, and FLT3LG genes, in the patient relative to that of the control value is at least 0.1, preferably 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, more preferably 1, 2, 3, 4, and even more preferably lower than 5, the gene expression profile of the biomarker in the patient sample is considered to be lower than the control value.

[0103] In particular, when the gene expression level is determined by quantitative RT-PCR, the log2 fold change of the gene expression profile of at least 1, 2, 3, or 4 genes selected from the group consisting of the BCL2, CISH, PTGER2, and DPP4 genes, preferably at least 1, 2, 3, 4, 5, or 6 genes selected from the group consisting of the BCL2, CISH, PTGER2, DPP4, SOCS2, and FLT3LG genes, in the patient relative to that of the control value is lower than at least 0.4, the gene expression profile of the biomarker in the patient sample is considered to be lower than the control value.

[0104] In another specific embodiment, when the gene expression level is determined by RNA-seq, the log2 fold change of the gene expression profile of at least 1, 2, 3, or 4 genes selected from the group consisting of the BCL2, CISH, PTGER2, and DPP4 genes, preferably at least 1, 2, 3, 4, 5, or 6 genes selected from the group consisting of the BCL2, CISH, PTGER2, DPP4, SOCS2, and FLT3LG genes, in the patient relative to that of the control value is lower than at least 0.1, the gene expression profile of the biomarker in the patient sample is considered to be lower than the control value.

[0105] After treatment with an IL7R antagonist (e.g., an anti-IL7R antagonist antibody or an antigen-binding fragment thereof), if the gene expression profile of the biomarker in a sample from a patient administered at least one dose of the antagonist is not lower than the control value, for example, by changing the IL7R antagonist compound or by changing the concentration of the IL7R antagonist, the treatment should be interrupted or changed. The method for evaluating the treatment response according to the present disclosure can indicate the success or failure of treating a patient with an IL7R antagonist (e.g., an anti-IL7R antagonist antibody or an antigen-binding fragment).

[0106] In another specific embodiment, a higher gene expression profile of the biomarker in a sample from a patient administered at least one dose of an IL7R agonist, compared to the control value, correlates with the agonist effect of the agonist in the patient and, thus, with the treatment response of the patient to the IL7R agonist treatment.

[0107] Typically, when the log2 fold change of the gene expression profile of at least 1, 2, 3, or 4 genes selected from the group consisting of the BCL2, CISH, PTGER2, and DPP4 genes, preferably at least 1, 2, 3, 4, 5, or 6 genes selected from the group consisting of the BCL2, CISH, PTGER2, DPP4, SOCS2, and FLT3LG genes, in the patient, relative to that of the control value, is higher than at least 0.1, preferably 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, more preferably 1, 2, 3, again more preferably 4, and even more preferably 5, the gene expression profile of the biomarker in the patient sample is considered to be higher than the control value.

[0108] In particular, when the gene expression level is determined by quantitative RT-PCR, if the log2 fold change of the gene expression profile of at least 1, 2, 3 or 4 genes selected from the group consisting of the BCL2, CISH, PTGER2 and DPP4 genes, preferably at least 1, 2, 3, 4, 5 or 6 genes selected from the group consisting of the BCL2, CISH, PTGER2, DPP4, SOCS2 and FLT3LG genes, in the patient, relative to that of the control value, is higher than at least 0.4, the gene expression profile of the biomarker in the patient sample is considered to be higher than the control value.

[0109] In another specific embodiment, when the gene expression level is determined by RNA-seq, if the log2 fold change of the gene expression profile of at least 1, 2, 3 or 4 genes selected from the group consisting of the BCL2, CISH, PTGER2 and DPP4 genes, preferably at least 1, 2, 3, 4, 5 or 6 genes selected from the group consisting of the BCL2, CISH, PTGER2, DPP4, SOCS2 and FLT3LG genes, in the patient, relative to that of the control value, is higher than at least 0.1, the gene expression profile of the biomarker in the patient sample is considered to be higher than the control value.

[0110] If, after treatment with an IL7R agonist, the gene expression profile of the biomarker in a sample of a patient administered at least 1 dose of the agonist is not higher than the control value, the treatment should be interrupted or changed, for example, by changing the IL7R antagonist compound or by changing the concentration of the IL7R antagonist. The method for evaluating the treatment response according to the present disclosure can indicate the success or failure of the treatment of a patient with an IL7R agonist.

[0111] According to the present disclosure, the patient treated with an IL7R antagonist (e.g., an anti-IL7R antagonist antibody or antigen-binding fragment) suffers from a disorder or disease associated with enhanced IL7R signaling pathway induced by IL7 (IL7 / IL7R pathway).

[0112] Various pathological conditions are associated with abnormally high levels of IL-7 and / or IL-7R. The disorders associated with the enhancement of the IL7 signaling pathway induced by IL7 are autoimmune diseases, inflammatory diseases, allergic diseases, cancer diseases, infectious diseases, respiratory diseases, and diseases associated with transplantation, particularly autoimmune diseases or inflammatory diseases accompanied by the activation or proliferation of CD127-positive diseased cells, more preferably, chronic inflammatory diseases such as Sjögren's syndrome or inflammatory bowel diseases such as ulcerative colitis, which can be selected from the group consisting of.

[0113] Sjögren's disease is an autoimmune disease that mainly affects exocrine glands and causes their dysfunction. Sjögren's disease is an autoimmune disease characterized by signs of inflammation and can be classified as an inflammatory disease. The symptoms of Sjögren's syndrome include, for example, dry eyes and dry mouth. Sjögren's syndrome is often accompanied by other immune system disorders such as rheumatoid arthritis, lupus, erythematosus, or progressive systemic sclerosis.

[0114] In certain embodiments, the disorder associated with the enhancement of the IL7R signaling pathway induced by IL7 is preferably an infectious disease selected from the group consisting of chronic infectious diseases, primary infectious diseases, and secondary infectious diseases.

[0115] In certain embodiments, the disorder associated with the enhancement of the IL7R signaling pathway induced by IL7 is preferably an autoimmune disease selected from the group consisting of diabetes, multiple sclerosis, Sjögren's syndrome, lupus, psoriasis, and atopic dermatitis.

[0116] In another specific embodiment, the disease associated with the enhancement of the IL7R signaling pathway induced by IL7 is preferably a chronic inflammatory disease selected from the group consisting of rheumatoid arthritis, ankylosing spondylitis, and inflammatory bowel disease.

[0117] In another specific embodiment, the disease associated with the enhancement of the IL7R signaling pathway induced by IL7 is an allergic disease.

[0118] The disease associated with the enhancement of the IL7R signaling pathway induced by IL7 may also preferably be a cancer such as a blood cancer selected from the group consisting of T-cell acute lymphoblastic leukemia, B-cell acute lymphoblastic leukemia, chronic lymphocytic leukemia, T-cell prolymphocytic leukemia, and Hodgkin lymphoma.

[0119] In a preferred embodiment, the disorder associated with the enhancement of the IL7R signaling pathway induced by IL7 is an inflammatory bowel disease such as Sjögren's syndrome or ulcerative colitis or Crohn's disease.

[0120] According to the present disclosure, the patient treated with an IL7R agonist has a disorder associated with impaired normal T-cell activity, preferably a disorder or disease that can be rescued through the IL7R signaling pathway induced by IL7, preferably a cancer disease or an infectious disease whose onset itself does not depend on the IL7R pathway. In a specific embodiment, the patient treated with an IL7R agonist has a disorder or disease that requires enhancement of the immune response such as immunotherapy in cancer diseases or infectious diseases.

[0121] In another specific embodiment, the present disclosure relates to the therapeutic use of an IL7R antagonist (e.g., an anti-IL7 receptor antagonist antibody or an antigen-binding fragment thereof) or an agonist in a human patient in need thereof, wherein after the patient has been administered at least one dose of the IL7R antagonist (e.g., an anti-IL7 receptor antagonist antibody or an antigen-binding fragment thereof) or the IL7R agonist, when the patient is evaluated as being likely to respond to the antagonist or agonist treatment in the above method, the antagonist, particularly the antibody or agonist, is administered to the patient.

[0122] The present invention also relates to a method for treating a disease associated with enhanced IL7R signaling pathway induced by IL7 in a human patient in need thereof, or a disease associated with the impairment of the above-described normal T cell activity, comprising: (a) providing a sample previously collected from said patient administered with at least one dose of an IL7R antagonist (e.g., an anti-IL7 receptor antagonist antibody or an antigen-binding fragment thereof) or an IL7R agonist; (b) determining whether the gene expression profile of at least 1, 2, 3, or 4 genes selected from the group consisting of the BCL2, CISH, PTGER2, and DPP4 genes, preferably at least 1, 2, 3, 4, 5, or 6 genes selected from the group consisting of BCL2, CISH, PTGER2, DPP4, FLT3LG, and SOCS2, is lower or higher compared to a control value, wherein a lower or higher gene expression profile of said gene indicates that said patient is likely to respond to said IL7R antagonist (e.g., an anti-IL7 receptor antagonist antibody or an antigen-binding fragment thereof) or IL7R agonist treatment, respectively, determining whether it is lower or higher compared to said control value; (c) re-administering a therapeutically effective amount of an IL7R antagonist (e.g., an anti-IL7 receptor antagonist antibody or an antigen-binding fragment thereof) or an IL7R agonist to said patient evaluated as likely to respond to said antagonist or agonist treatment. The method as described above.

[0123] In other words, the present disclosure relates to a method for evaluating the therapeutic response of IL7R antagonist treatment in a human patient in need thereof suffering from a disease associated with enhanced IL7R signaling pathway induced by IL7 as described above, comprising: (a) providing a sample collected from said patient at a first time point or earlier; (b) providing a sample collected from said patient at a second time point or later; (b) Determining whether the gene expression profile of at least 1, 2, 3, or 4 genes selected from the group consisting of the BCL2, CISH, PTGER2, and DPP4 genes, preferably at least 1, 2, 3, 4, 5, or 6 genes selected from the group consisting of the BCL2, CISH, PTGER2, DPP4, FLT3LG, and SOCS2 genes, in the patient sample collected at the second time point or a later time point is lower compared to the patient sample collected at the first time point or an earlier time point, wherein a lower gene expression profile indicates that the patient is likely to respond to the treatment, and determining whether it is lower compared to the above, (c) Readministering a therapeutically effective amount of an IL7R antagonist (e.g., an anti-IL7 receptor antagonist antibody or an antigen-binding fragment thereof) to the patient who has been evaluated as likely to respond to the antagonist treatment relating to the above method.

[0124] The present disclosure also relates to a method for evaluating the therapeutic response of an IL7R agonist treatment in a human patient in need thereof who is suffering from a disorder or disease associated with impaired healthy T cell activity and can preferably be rescued via the IL7R signaling pathway induced by IL7, (a) Providing a sample collected from the patient at a first time point or an earlier time point, (b) Providing a sample collected from the patient at a second time point or a later time point, (b) Determining whether the gene expression profile of at least 1, 2, 3, or 4 genes selected from the group consisting of BCL2, CISH, PTGER2, and DPP4 genes, preferably at least 1, 2, 3, 4, 5, or 6 genes selected from the group consisting of BCL2, CISH, PTGER2, DPP4, FLT3LG, and SOCS2 genes, in the patient sample collected at the second time point or a later time point is higher compared to the patient sample collected at the first time point or an earlier time point, and a higher gene expression profile indicates that the patient is likely to respond to the treatment, determining whether it is higher compared to said comparison, (c) Re-administering a therapeutically effective amount of an IL7R agonist to the patient evaluated to be likely to respond to the agonist treatment relates to said method, comprising

[0125] The present disclosure relates to the use of an IL7R antagonist (e.g., the anti-IL7 receptor antagonist antibody or antigen-binding fragment thereof described above) or an IL7R agonist in the manufacture of a medicament for the treatment of a disease associated with the IL7-induced IL7R signaling pathway in a human patient in need thereof or a disease associated with a disorder of normal T cell activity, wherein after the patient has been administered at least one dose of said IL7R antagonist (e.g., the anti-IL7 receptor antagonist antibody or antigen-binding fragment thereof) or IL7R agonist, when the patient is evaluated to be likely to respond to the antagonist or agonist treatment by the method described above, said antagonist or agonist is administered to the patient, relating to said use.

[0126] In certain embodiments, the disorder or disease associated with enhanced IL7 - induced IL7 signaling pathway and treated with an IL7R antagonist is an autoimmune disease, inflammatory disease, allergic disease, cancer disease, infectious disease, respiratory disease, and a disease related to transplantation, accompanied by activation or proliferation of CD127 - positive diseased cells, in particular, an autoimmune disease or inflammatory disease accompanied by activation or proliferation of CD127 - positive cells, more preferably, a chronic inflammatory disease such as Sjögren's syndrome or an inflammatory bowel disease such as ulcerative colitis, preferably, diabetes, sarcoidosis, cancer, preferably, an autoimmune disease selected from the group consisting of multiple sclerosis, Sjögren's syndrome, lupus, psoriasis and atopic dermatitis, or preferably, a chronic inflammatory disease selected from the group consisting of rheumatoid arthritis, ankylosing spondylitis and inflammatory bowel disease, preferably, selected from the group consisting of ulcerative colitis or Crohn's disease.

[0127] In certain embodiments, the IL7R antagonist is an anti - IL7R antagonist antibody or an antigen - binding fragment thereof according to the present disclosure, comprising: (a) a variable heavy chain comprising three CDRs, wherein VHCDR1 comprises or consists of the amino acid sequence of SEQ ID NO: 1, VHCDR2 comprises or consists of the amino acid sequence of SEQ ID NO: 2, and VHCDR3 comprises or consists of the amino acid sequence of SEQ ID NO: 3 or 11; and (b) a variable light chain comprising three CDRs, wherein VLCDR1 comprises or consists of the amino acid sequence of SEQ ID NO: 4 or 12, VLCDR2 comprises or consists of the amino acid sequence of SEQ ID NO: 5 or 13, and VLCDR3 comprises or consists of the amino acid sequence of SEQ ID NO: 6.

[0128] In a preferred embodiment, the anti-IL7 receptor antagonist antibody or antigen-binding fragment thereof comprises: (a) a variable heavy chain comprising three CDRs, wherein VHCDR1 comprises or consists of the amino acid sequence of SEQ ID NO: 1, VHCDR2 comprises or consists of the amino acid sequence of SEQ ID NO: 2, and VHCDR3 comprises or consists of the amino acid sequence of SEQ ID NO: 3; and (b) a variable light chain comprising three CDRs, wherein VLCDR1 comprises or consists of the amino acid sequence of SEQ ID NO: 4, VLCDR2 comprises or consists of the amino acid sequence of SEQ ID NO: 5, and VLCDR3 comprises or consists of the amino acid sequence of SEQ ID NO: 6.

[0129] In certain embodiments, the anti-IL7R antagonist antibody or antigen-binding fragment thereof comprises a heavy chain variable domain comprising or consisting of an amino acid sequence selected from the group consisting of SEQ ID NOs: 14-17, and a light chain variable domain comprising or consisting of an amino acid sequence selected from the group consisting of SEQ ID NOs: 18-21. Preferably, the anti-IL7R antagonist is the N13B2h3 antibody or antigen-binding fragment thereof described in WO2015 / 189302, which comprises a heavy chain variable domain comprising or consisting of the amino acid sequence of SEQ ID NO: 17 and a light chain variable domain comprising or consisting of the amino acid sequence of SEQ ID NO: 21. More preferably, the antibody or antigen-binding fragment thereof comprises a complete heavy chain comprising or consisting of SEQ ID NO: 22 and a complete light chain comprising or consisting of SEQ ID NO: 23.

[0130] In another particular embodiment, the anti-IL7R antagonist antibody or antigen-binding fragment thereof comprises a heavy chain variable domain comprising or consisting of the amino acid sequence of SEQ ID NO: 7, and a light chain variable domain comprising or consisting of an amino acid sequence selected from the group consisting of SEQ ID NOs: 8, 24-26.

[0131] In a more preferred embodiment, the anti-IL7R antagonist antibody or antigen-binding fragment thereof comprises a heavy chain variable domain comprising or consisting of the amino acid sequence of SEQ ID NO: 7 and a light chain variable domain comprising or consisting of the amino acid sequence of SEQ ID NO: 8, and preferably, the antibody or antigen-binding fragment thereof comprises a complete heavy chain comprising or consisting of SEQ ID NO: 9 and a complete light chain comprising or consisting of SEQ ID NO: 10.

[0132] In the context of the present disclosure, the terms "treat" or "treatment" mean, as used herein, the reversal, alleviation, inhibition of progression, or prevention of a disorder or condition to which such terms apply, or the reversal, alleviation, inhibition of progression, or prevention of one or more symptoms of a disorder or condition to which such terms apply.

[0133] As used herein, the term "therapeutically effective amount" or "effective amount" means an amount of a composition that is sufficient to produce a therapeutic effect when administered to a subject for treating a condition, disorder or disease. The therapeutically effective amount will vary depending on the compound, formulation or composition, the disease and its severity, and the age, weight, physical condition and responsiveness of the subject being treated.

[0134] The IL7R antagonists (e.g., anti-IL7R antagonist antibodies or antigen-binding fragments thereof) or IL7R agonists described herein can be administered by any means known to those skilled in the art including, but not limited to, intravenous, oral, intratumoral, intralesional, intradermal, topical, intraperitoneal, intramuscular, parenteral, subcutaneous, and local administration. Thus, the composition can be formulated as an injectable, topical, or ingestible preparation. Administration of the compounds or therapeutic agents according to the present disclosure to a subject can exhibit a dose-dependent beneficial effect. Thus, within a wide range, administration of a greater amount of the composition is expected to achieve an enhancement of the beneficial biological effect over administration of a lesser amount. Furthermore, effectiveness is also contemplated at dosages below the level at which toxicity is observed.

[0135] As is well known to those skilled in the art, the specific dosage of the IL7R antagonist or agonist administered in any given case is adjusted according to the administered composition, the amount of the composition that can be effectively delivered to the administration site, the disease to be treated or inhibited, the condition of the subject, and other relevant medical factors that can change the activity of the composition or the response of the subject.

[0136] For example, the specific dosage of the IL7R antagonist or agonist for a particular subject depends on age, weight, general health, diet, timing and mode of administration, rate of excretion, drugs used in combination, and the severity of the particular disorder to which the therapy is applied. The dosage for a given patient can be determined using conventional considerations, such as by the usual comparison of the different activities of the compositions described herein and known drugs according to appropriate conventional pharmacological protocols. The composition can be administered on a single-dose schedule or on a multiple-dose schedule.

[0137] The appropriate dosage range of the IL7R antagonist (e.g., anti-IL7R antagonist antibody or its antigen-binding fragment) or IL7R agonist can be on the order of several hundred micrograms of a drug in the range of about 0.001 to 100 mg / kg, preferably about 0.1 to 20 mg / kg, more preferably about 1 to 12 mg / kg, and still more preferably 8 to 12 mg / kg. For example, a fixed amount such as 400, 450, 500, 600, 700, 850, 900 mg can be used.

[0138] (Method for evaluating the likelihood of a therapeutic response to IL7R modulator therapy in a human patient prior to said treatment) The combination of the expression profiles of the biomarkers can also be classified as likely to respond prior to any treatment, and can inform the condition of patients for whom treatment with the antagonists (e.g., anti-IL7 receptor antagonist antibodies or their antigen-binding fragments) or IL7R agonists described herein is recommended.

[0139] For example, the assay can include culturing a sample of the patient prior to treatment in the presence of an IL7R antagonist or agonist to evaluate its biological response.

[0140] Therefore, in another specific embodiment, the present disclosure provides a method for evaluating the likelihood of a therapeutic response in a human patient to treatment with an IL7R antagonist (e.g., an anti-IL7 receptor antagonist antibody or an antigen-binding fragment thereof) or agonist prior to said antagonist or agonist treatment, a) culturing a sample previously collected from the patient prior to said antagonist or agonist treatment in the presence of an IL7 receptor antagonist and / or an IL7R agonist, b) determining the gene expression profile of at least 1, 2, 3, or 4 genes selected from the group consisting of the BCL2, CISH, PTGER2, and DPP4 genes, preferably at least 1, 2, 3, 4, 5, or 6 genes selected from the group consisting of the BCL2, CISH, PTGER2, DPP4, SOCS2, and FLT3LG genes, wherein a lower or higher gene expression profile of said gene in the cultured sample, as compared to a control value, indicates that the patient is likely to respond to the antagonist or agonist treatment respectively, the step of determining the gene expression profile relating to said method, comprising.

[0141] As used herein, the term "evaluating the likelihood of a therapeutic response" refers to the ability to assess whether a patient will respond to treatment with an IL7R antagonist (e.g., an anti-IL7 receptor antagonist antibody or an antigen-binding fragment thereof) or an IL7R agonist, in other words, whether said antagonist or agonist treatment is effective in providing a measurable benefit or a positive medical response to the patient prior to the therapy. Such an ability to assess whether a patient will respond to treatment with said antagonist or agonist is typically exercised prior to the initiation of the antagonist or agonist treatment in the patient.

[0142] According to this method for evaluating the possibility of treatment response, the sample is collected in advance from the patient before the start of antagonist or agonist treatment. Preferably, the sample is immune cells collected from the patient before treatment with the antagonist or agonist as described above, more preferably peripheral blood mononuclear cells.

[0143] In certain embodiments, the cells are immune cells. Immune cells include lymphocytes such as B cells and T cells, natural killer cells. The immune cells can be obtained from a number of non-limiting sources including peripheral blood mononuclear cells, bone marrow, lymph node tissue, cord blood, thymus tissue, tissue from an infected site, ascites, pleural effusion, spleen tissue, and tumors, using any number of techniques known to those skilled in the art to obtain a unit of blood taken from a subject. Immune cells can be extracted from blood or can be stem cell-derived. In a preferred embodiment, the sample is a tissue sample, preferably tumor tissue containing infiltrating immune cells from a cancer patient or a blood patient sample, more preferably isolated human peripheral blood mononuclear cells.

[0144] The sample is then cultured in vitro in the presence of an IL7R antagonist (e.g., an anti-IL7 receptor antagonist antibody or an antigen-binding fragment thereof) and / or an IL7R agonist.

[0145] The sample may be cultured and grown in a medium under culture conditions well known in the art before being collected for gene expression level analysis. In a preferred embodiment, the sample is cultured at 37°C for at least 3 hours, preferably 2, 4, 5, or 6 hours before being collected for further analysis.

[0146] Many media are commercially available and well-known to those skilled in the art. This medium can be, in particular, a minimal medium containing mineral salts, amino acids, vitamins, a carbon source essential for cells, and a buffer system for adjusting pH. Basic media that can be used in the method according to the present invention include, for example, but are not limited to, DMEM / F12 medium, DMEM medium, RPMI medium, Ham's F12 medium, IMDM medium, and KnockOut™ DMEM medium (Life Technologies). The medium may contain interleukin-7, preferably human recombinant interleukin-7.

[0147] In certain embodiments, in the method of assessing the potential for a therapeutic response in a human patient to an IL7R antagonist, the sample pre-collected from the patient prior to the antagonist treatment is cultured in step a) to allow cell proliferation in the presence of an IL7 receptor antagonist (e.g., an anti-IL7 receptor antagonist antibody or an antigen-binding fragment thereof) and an IL7 cytokine, preferably human recombinant interleukin-7.

[0148] The medium more preferably contains an IL7R antagonist, preferably an anti-IL7 receptor antagonist antibody or an antigen-binding fragment thereof, at a concentration of 1 to 10,000 ng / mL, preferably 500 to 10,000 ng / mL, more preferably 1,000 to 10,000 ng / mL.

[0149] The medium preferably contains an IL7R agonist at a concentration of 1 to 10,000 ng / mL, preferably 500 to 10,000 ng / mL, more preferably 1,000 to 10,000 ng / mL.

[0150] After a cell culture step with an IL7R antagonist (e.g., an anti-IL7 receptor antagonist antibody or an antigen-binding fragment thereof) and / or an IL7R agonist, the cells are collected, and the gene expression profile of at least 1, 2, 3, or 4 genes selected from the group consisting of the BCL2, CISH, PTGER2, and DPP4 genes in the cultured cells, preferably at least 1, 2, 3, 4, 5, or 6 genes selected from the group consisting of the BCL2, CISH, PTGER2, DPP4, SOCS2, and FLT3LG genes, is determined as described above.

[0151] The method further includes determining whether the gene expression profile of the gene is higher or lower compared to a control value. For example, a lower or higher gene expression profile of at least one of the genes compared to the control value may indicate a patient likely to respond to antagonist or agonist treatment, respectively.

[0152] In certain embodiments, according to the method for evaluating the likelihood of a treatment response in a human patient of the present disclosure, the control value may be the gene expression profile of the gene(s) in patient cells cultured in vitro without the antagonist or agonist, or in patient cells cultured in vitro with a compound (e.g., an antibody) known not to affect the IL7 / IL7R pathway, or may be a given value of a control established based on a comparative measurement between patient cells cultured in vitro with the antagonist or agonist, or patient cells cultured in vitro without the antagonist or agonist, or patient cells cultured in vitro with a compound (e.g., an antibody) known not to affect the IL7 / IL7R signaling pathway.

[0153] A lower gene expression profile of at least one of the biomarkers in patient cells cultured in vitro with an IL7R antagonist (e.g., an anti-IL7 receptor antagonist antibody or an antigen-binding fragment thereof), compared to a control value, indicates an IL7R antagonist effect in the patient cells and, thus, correlates with the likelihood of a therapeutic response to IL7R antagonist treatment in the patient.

[0154] Typically, when the log2 fold change of the gene expression profile of the gene(s) in the patient relative to that of the control value is at least 0.1, preferably 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, more preferably 1, 2, 3, 4, and even more preferably lower than 5, the gene expression profile of the gene(s) in the patient sample cultured in vitro with an IL7R antagonist (e.g., an anti-IL7R antagonist antibody or an antigen-binding fragment thereof) is considered to be lower than the control value.

[0155] In particular, when the gene expression level is determined by quantitative RT-PCR, the log2 fold change of the gene expression profile of at least 1, 2, 3, or 4 genes selected from the group consisting of the BCL2, CISH, PTGER2, and DPP4 genes in the patient, preferably at least 1, 2, 3, 4, 5, or 6 genes selected from the group consisting of the BCL2, CISH, PTGER2, DPP4, SOCS2, and FLT3LG genes relative to that of the control value, is lower than at least 0.4, the gene expression profile of the biomarker in the patient sample is considered to be lower than the control value.

[0156] In another specific embodiment, when the gene expression level is determined by RNA-seq, the gene expression profile of at least 1, 2, 3, or 4 genes selected from the group consisting of the BCL2, CISH, PTGER2, and DPP4 genes in the patient, preferably at least 1, 2, 3, 4, 5, or 6 genes selected from the group consisting of the BCL2, CISH, PTGER2, DPP4, SOCS2, and FLT3LG genes, if the log2 fold change of the gene expression profile relative to that of the control value is lower than at least 0.1, the gene expression profile of the biomarker in the patient sample is considered to be lower than the control value.

[0157] In another specific embodiment, a gene expression profile of at least one of the biomarkers in patient cells cultured in vitro with an IL7R agonist that is higher compared to the control value indicates an IL7R agonist effect in the patient cells and, thus, correlates with the likelihood of a therapeutic response to IL7R agonist treatment in the patient.

[0158] Typically, if the log2 fold change of the gene expression profile of the gene(s) in the patient relative to that of the control value is at least 0.1, preferably 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, more preferably 1, 2, 3, 4, and even more preferably higher than 5, the gene expression profile of the gene(s) in the patient sample cultured in vitro with an IL7R agonist is considered to be higher than the control value.

[0159] In particular, when the gene expression level is determined by quantitative RT-PCR, if the log2 fold change of the gene expression profile of at least 1, 2, 3, or 4 genes selected from the group consisting of the BCL2, CISH, PTGER2, and DPP4 genes in the patient, preferably at least 1, 2, 3, 4, 5, or 6 genes selected from the group consisting of the BCL2, CISH, PTGER2, DPP4, SOCS2, and FLT3LG genes, relative to that of the control value, is higher than at least 0.4, then the gene expression profile of the biomarker in the patient sample is considered to be higher than the control value.

[0160] In another specific embodiment, when the gene expression level is determined by RNA-seq, if the log2 fold change of the gene expression profile of at least 1, 2, 3, or 4 genes selected from the group consisting of the BCL2, CISH, PTGER2, and DPP4 genes in the patient, preferably at least 1, 2, 3, 4, 5, or 6 genes selected from the group consisting of the BCL2, CISH, PTGER2, DPP4, SOCS2, and FLT3LG genes, relative to that of the control value, is higher than at least 0.1, then the gene expression profile of the biomarker in the patient sample is considered to be higher than the control value.

[0161] According to the present disclosure, the patient treated with an IL7R antagonist (e.g., an anti-IL7 receptor antagonist antibody or an antigen-binding fragment thereof) suffers from a disorder or disease associated with the enhancement of the IL7R signaling pathway induced by IL7, preferably an autoimmune disease, an inflammatory disease, an allergic disease, a cancer disease, an infectious disease, a respiratory disease, and a disease related to transplantation, particularly an autoimmune disease or an inflammatory disease accompanied by the activation or proliferation of CD127-positive diseased cells, more preferably a chronic inflammatory disease such as Sjögren's syndrome, or an inflammatory bowel disease such as ulcerative colitis, and is selected from the group consisting of disorders or diseases.

[0162] According to the present disclosure, the patient to be treated with an IL7R agonist suffers from a disorder or disease associated with the above-described impairment of normal T cell activity, preferably a cancer disease or an infectious disease whose onset itself does not depend on the IL7R pathway.

[0163] The present disclosure also relates to the therapeutic use of an IL7R antagonist (e.g., an anti-IL7 receptor antagonist antibody or an antigen-binding fragment thereof) or an IL7R agonist in a human patient who has been previously evaluated as likely to respond to the antagonist or agonist treatment by the above method prior to said treatment.

[0164] The present disclosure also provides a method of treating a disease associated with upregulation of the IL7R pathway induced by IL7 in a human patient in need thereof with an IL7R antagonist (e.g., an anti-IL7 receptor antagonist antibody or an antigen-binding fragment thereof), (a) culturing a sample previously collected from a patient prior to said treatment in the presence of an IL7R antagonist (e.g., an anti-IL7 receptor antagonist antibody or an antigen-binding fragment thereof), and (b) determining whether the gene expression profile of at least 1, 2, 3, or 4 genes selected from the group consisting of the BCL2, CISH, PTGER2, and DPP4 genes, preferably at least 1, 2, 3, 4, 5, or 6 genes selected from the group consisting of the BCL2, CISH, PTGER2, DPP4, FLT3LG, and SOCS2 genes, in said sample is lower compared to a control value, wherein a gene expression profile lower compared to the control value in said patient sample indicates that the patient is likely to respond to said treatment, determining whether it is lower compared to said control value, (c) administering a therapeutically effective amount of an IL7R antagonist (e.g., an anti-IL7 receptor antagonist antibody or an antigen-binding fragment thereof) to said patient evaluated as likely to respond to said treatment comprising the above method.

[0165] The present disclosure also relates to a method of treating a disease associated with the impairment of the above-described normal T cell activity in a human patient in need thereof with an IL7R agonist, comprising: (a) culturing a sample previously collected from the patient with an IL7R agonist prior to said treatment; and (b) determining whether the gene expression profile of at least 1, 2, 3, or 4 genes selected from the group consisting of the BCL2, CISH, PTGER2, and DPP4 genes in said sample, preferably at least 1, 2, 3, 4, 5, or 6 genes selected from the group consisting of the BCL2, CISH, PTGER2, DPP4, FLT3LG, and SOCS2 genes, is higher compared to a control value, wherein a gene expression profile higher compared to the control value in said patient sample indicates that the patient is likely to respond to said treatment; (c) administering a therapeutically effective amount of an IL7R agonist to said patient determined to be likely to respond to said treatment. The present disclosure also relates to the use of an IL7R antagonist (e.g., an anti-IL7 receptor antagonist antibody or an antigen-binding fragment thereof) or an IL7R agonist in the manufacture of a medicament for the treatment of a disease associated with the above-described IL7 / IL7R pathway or a disease associated with the impairment of normal T cell activity in a human patient in need thereof, who has been previously evaluated to be likely to respond to said antagonist or agonist treatment by the above method.

[0166]

[0167] ​In certain embodiments, the anti-IL7R antagonist is an antibody or antigen-binding fragment thereof of N13B2h1, N13B2h2, or N13B2h3 described in WO2015 / 189302, or an antibody or antigen-binding fragment thereof of N13B2-hVL3, N13B2-hVL4, N13B2-hVL5, or N13B2-hVL6 described in WO2018 / 104483, comprising: (a) a variable heavy chain comprising three CDRs wherein VHCDR1 comprises or consists of the amino acid sequence of SEQ ID NO: 1, VHCDR2 comprises or consists of the amino acid sequence of SEQ ID NO: 2, and VHCDR3 comprises or consists of the amino acid sequence of SEQ ID NO: 3 or 11; and (b) a variable light chain comprising three CDRs wherein VLCDR1 comprises or consists of the amino acid sequence of SEQ ID NO: 4 or 12, VLCDR2 comprises or consists of the amino acid sequence of SEQ ID NO: 5 or 13, and VLCDR3 comprises or consists of the amino acid sequence of SEQ ID NO: 6.

[0168] In a preferred embodiment, the anti-IL7 receptor antagonist antibody or antigen-binding fragment thereof is an antibody or antigen-binding fragment thereof of N13B2h3 described in WO2015 / 189302, or an antibody or antigen-binding fragment thereof of N13B2-hVL3, N13B2-hVL4, N13B2-hVL5, or N13B2-hVL6 described in WO2018 / 104483, preferably an N13B2-hVL6 antibody or antigen-binding fragment thereof, comprising: (a) a variable heavy chain comprising three CDRs wherein VHCDR1 comprises or consists of the amino acid sequence of SEQ ID NO: 1, VHCDR2 comprises or consists of the amino acid sequence of SEQ ID NO: 2, and VHCDR3 comprises or consists of the amino acid sequence of SEQ ID NO: 3; and (b) a variable light chain comprising three CDRs wherein VLCDR1 comprises or consists of the amino acid sequence of SEQ ID NO: 4, VLCDR2 comprises or consists of the amino acid sequence of SEQ ID NO: 5, and VLCDR3 comprises or consists of the amino acid sequence of SEQ ID NO: 6.

[0169] In certain embodiments, the anti-IL7R antagonist antibody or antigen-binding fragment thereof comprises or consists of a heavy chain variable domain comprising an amino acid selected from the group consisting of SEQ ID NOs: 14-17, and a light chain variable domain comprising or consisting of an amino acid sequence selected from the group consisting of SEQ ID NOs: 18-21. Preferably, the anti-IL7R antagonist is the N13B2h3 antibody or antigen-binding fragment thereof described in WO2015 / 189302, comprising or consisting of a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO: 17, and a light chain variable domain comprising or consisting of the amino acid sequence of SEQ ID NO: 21. More preferably, the antibody or antigen-binding fragment thereof comprises or consists of a complete heavy chain comprising SEQ ID NO: 22 and a complete light chain comprising or consisting of SEQ ID NO: 23.

[0170] In another specific embodiment, the anti-IL7R antagonist is an antibody or antigen-binding fragment thereof of N13B2-hVL3, N13B2-hVL4, N13B2-hVL5 or N13B2-hVL6 described in WO2018 / 104483, comprising or consisting of a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO: 7, and a light chain variable domain comprising or consisting of an amino acid sequence selected from the group consisting of SEQ ID NO: 8 and SEQ ID NOs: 24-26.

[0171] In a more preferred embodiment, the anti-IL7R antagonist antibody or antigen-binding fragment thereof comprises or consists of a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO: 7 and a light chain variable domain comprising or consisting of the amino acid sequence of SEQ ID NO: 8. Preferably, the antibody or antigen-binding fragment thereof comprises or consists of a complete heavy chain comprising SEQ ID NO: 9 and a complete light chain comprising or consisting of SEQ ID NO: 10.

[0172] The antagonists described herein, preferably antibodies or antigen-binding fragments or agonists thereof, can be administered by any means known to those skilled in the art as described above.

[0173] The appropriate dosage range of an IL7R antagonist, preferably an anti-IL7R antagonist antibody or its antigen-binding fragment, or an IL7R agonist, can be on the order of several hundred micrograms of a drug in the range of about 0.001 to 100 mg / kg, preferably about 0.1 to 20 mg / kg, more preferably about 1 to 12 mg / kg, and still more preferably in the range of 8 to 12 mg / kg. For example, a fixed amount such as 400, 450, 500, 600, 700, 850, 900 mg can be used.

[0174] (Method for Selecting IL7R Modulator Compounds) In another specific embodiment, the present disclosure is an in vitro method for selecting an IL7R antagonist compound, preferably an IL7Rα antagonist compound, that is likely to be effective in the treatment of diseases associated with enhanced IL7R signaling pathways induced by the above-mentioned IL7, a) culturing the compound with human cells, b) determining the gene expression profile of at least 1, 2, 3, or 4 genes selected from the group consisting of the BCL2, CISH, PTGER2, and DPP4 genes in the cells, preferably at least 1, 2, 3, 4, 5, or 6 genes selected from the group consisting of the BCL2, CISH, PTGER2, DPP4, SOCS2, and FLT3LG genes, and c) selecting a compound that induces a lower gene expression profile of the gene(s) in the cells compared to a control value relates to the above in vitro method.

[0175] In another specific embodiment, the present disclosure is an in vitro method for selecting an IL7R agonist compound that is likely to be effective in the treatment of diseases associated with the impairment of the above-mentioned normal T cell activity, a) culturing the agonist compound with human cells, b) determining the gene expression profile of at least 1, 2, 3, or 4 genes selected from the group consisting of the BCL2, CISH, PTGER2, and DPP4 genes in the cell, preferably at least 1, 2, 3, 4, 5, or 6 genes selected from the group consisting of the BCL2, CISH, PTGER2, DPP4, SOCS2, and FLT3LG genes, and c) selecting a compound that induces a higher gene expression profile of the gene(s) in the cell as compared to a control value relates to the in vitro method comprising the same.

[0176] According to the method of selecting an IL7R antagonist or agonist compound, the control value can be the gene expression profile of the gene(s) in cells cultured without including a compound known not to affect the IL7 / IL7R signaling pathway.

[0177] In a preferred embodiment, the compound is an anti-IL7R antagonist antibody or an antigen-binding fragment thereof.

[0178] The cells that can be used to select the compound can be immortalized cell lines expressing CD127 such as T cell lines, for example, Jurkat cells, or cells collected from donors, preferably healthy human donors.

[0179] In a particular embodiment, the cells are lymphocytes, particularly immune cells including T cells and natural killer cells. The immune cells can be obtained from a number of non-limiting sources including peripheral blood mononuclear cells, bone marrow, lymph node tissue, cord blood, thymus tissue, tissue from an infected site, ascites, pleural effusion, spleen tissue, and tumors, and units of blood collected from a subject using any number of techniques known to those skilled in the art. The immune cells can be extracted from blood or can be stem cell-derived. In a preferred embodiment, the cells are peripheral blood mononuclear cells.

[0180] The antagonist is a suitable IL7-R antagonist, preferably an anti-IL7R antagonist antibody or an antigen-binding fragment thereof, and at least 1, 2, 3, or 4 genes selected from the group consisting of the BCL2, CISH, PTGER2, and DPP4 genes in cells cultured with the compound, preferably at least 1, 2, 3, 4, 5, or 6 genes selected from the group consisting of the BCL2, CISH, PTGER2, DPP4, SOCS2, and FLT3LG genes, when inducing a lower gene expression profile compared to the control value, preferably at least 1, 2, 3, or 4 genes selected from the group consisting of BCL2, CISH, PTGER2, and DPP4 in the cells cultured with the compound, preferably 1, 2, 3, 4, 5, or 6 genes selected from the group consisting of the BCL2, CISH, PTGER2, DPP4, SOCS2, and FLT3LG genes, and the log2 fold change of the gene expression profile of these genes relative to that of the control value is at least 0.1, preferably 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, more preferably 1, 2, 3, 4, even more preferably lower than 5, it is likely to be effective in the treatment of diseases associated with the enhancement of the IL7R signaling pathway induced by IL7.

[0181] In particular, when the gene expression level is determined by quantitative RT-PCR, at least 1, 2, 3, or 4 genes selected from the group consisting of the BCL2, CISH, PTGER2, and DPP4 genes in the patient, preferably at least 1, 2, 3, 4, 5, or 6 genes selected from the group consisting of the BCL2, CISH, PTGER2, DPP4, SOCS2, and FLT3LG genes, and the log2 fold change of the gene expression profile of these genes relative to that of the control value is lower than at least 0.4, then the gene expression profile of the biomarker in the patient sample is considered to be lower than the control value.

[0182] In another specific embodiment, when the gene expression level is determined by RNA-seq, if the log2 fold change of the gene expression profile of at least 1, 2, 3 or 4 genes selected from the group consisting of the BCL2, CISH, PTGER2 and DPP4 genes, preferably at least 1, 2, 3, 4, 5 or 6 genes selected from the group consisting of the BCL2, CISH, PTGER2, DPP4, SOCS2 and FLT3LG genes in the patient, relative to that of the control value, is lower than at least 0.1, then the gene expression profile of the biomarker in the patient sample is considered to be lower than the control value.

[0183] Otherwise, the agonist is a suitable IL7R agonist, and when it induces a higher gene expression profile of at least 1, 2, 3 or 4 genes selected from the group consisting of the BCL2, CISH, PTGER2 and DPP4 genes, preferably at least 1, 2, 3, 4, 5 or 6 genes selected from the group consisting of the BCL2, CISH, PTGER2, DPP4, SOCS2 and FLT3LG genes in the cells cultured with the compound, preferably the log2 fold change of the gene expression profile of 1, 2, 3, 4, 5 or 6 genes selected from the group consisting of the BCL2, CISH, PTGER2, DPP4 genes, preferably at least 1, 2, 3, 4, 5 or 6 genes selected from the group consisting of the BCL2, CISH, PTGER2, DPP4, SOCS2 and FLT3LG genes in the cells cultured with the compound, relative to that of the control value, is higher than 0.1, preferably 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, more preferably 1, 2, 3, 4, even more preferably 5, it may be considered likely to be effective in the treatment of diseases associated with the impairment of the above-mentioned healthy T cell activity.

[0184] In particular, when the gene expression level is determined by quantitative RT-PCR, if the log2 fold change of the gene expression profile of at least 1, 2, 3, or 4 genes selected from the group consisting of the BCL2, CISH, PTGER2, and DPP4 genes in the patient, preferably at least 1, 2, 3, 4, 5, or 6 genes selected from the group consisting of the BCL2, CISH, PTGER2, DPP4, SOCS2, and FLT3LG genes, relative to that of the control value, is higher than at least 0.4, the gene expression profile of the biomarker in the patient sample is considered to be higher than the control value.

[0185] In another specific embodiment, when the gene expression level is determined by RNA-seq, if the log2 fold change of the gene expression profile of at least 1, 2, 3, or 4 genes selected from the group consisting of the BCL2, CISH, PTGER2, and DPP4 genes in the patient, preferably at least 1, 2, 3, 4, 5, or 6 genes selected from the group consisting of the BCL2, CISH, PTGER2, DPP4, SOCS2, and FLT3LG genes, relative to that of the control value, is higher than at least 0.1, the gene expression profile of the biomarker in the patient sample is considered to be higher than the control value.

[0186] (Kit) In another aspect, the present disclosure relates to a kit for use in an in vitro method for evaluating the therapeutic response of an IL7R antagonist or agonist, or an in vitro method for evaluating the potential of a therapeutic response in a human patient to an IL7R antagonist or agonist, preferably comprising or consisting of a set of reagents for specifically detecting the gene expression profile of at least 1, 2, 3, or 4 genes selected from the group consisting of the BCL2, CISH, PTGER2, and DPP4 genes, preferably at least 1, 2, 3, 4, 5, or 6 genes selected from the group consisting of the BCL2, CISH, PTGER2, DPP4, SOCS2, and FLT3LG genes.

[0187] Preferably, the kit comprises a container containing one or more compounds, each at a concentration or amount that facilitates the reconstruction and / or use of a set of reagents for specifically detecting the gene expression profiles of at least 1, 2, 3, or 4 genes selected from the group consisting of the BCL2, CISH, PTGER2, and DPP4 genes, preferably at least 1, 2, 3, 4, 5, or 6 genes selected from the group consisting of the BCL2, CISH, PTGER2, DPP4, SOCS2, and FLT3LG genes, and / or the implementation of the methods according to the present disclosure.

[0188] In particular, the kit comprises primer pairs and / or probes specific for at least 1, 2, 3, or 4 genes selected from the group consisting of the BCL2, CISH, PTGER2, and DPP4 genes, preferably at least 1, 2, 3, 4, 5, or 6 genes selected from the group consisting of the BCL2, CISH, PTGER2, DPP4, SOCS2, and FLT3LG genes.

[0189] The kit may also include instructions for preparing and / or using reagents for determining the expression levels of the genes according to the methods of the present disclosure.

[0190] The present invention also relates to the use of a kit for the methods according to the present disclosure.

[0191] The present invention will now be illustrated, with reference to the accompanying drawings, by the following non-limiting examples.

Brief Description of the Drawings

[0192] (Brief Description of the Drawings)

Figure 1

Figure 2

Figure 3

Figure 4

Figure 5

Figure 6

Figure 7

Example

[0193] (Example) RNA of the anti-IL7R antagonist antibody with rRNA and globulin removed from paxgen blood samples in a healthy volunteer population was sequenced from both ends of 150 base pairs by Genewiz using Illumina HiSeq.

[0194] The quality of the sequencing by the fastQC bioinformatics tool was evaluated for the raw fastq files. Using the index creation option -genecode and the salmon bioinformatics tool, the reads were pseudo-aligned to CRGh38 / hg38, and using the option -geneMap and the salmon bioinformatics tool, a raw count table and a normalized TPM table for the read counts of each gene were created. The pseudo mapping quality reads were evaluated using the fastQC bioinformatics tool, and the average value of the reads for all samples was 20 - 30M, and more than 90% of the bases were over 30.

[0195] This cohort of healthy volunteers included peripheral blood mononuclear cell (PBMC) samples collected on day 15 after injection for the entire cohort of 48 samples, and peripheral blood mononuclear cell (PBMC) samples collected on day 57 after injection for 4 of them, paired with the anti-human IL7R antagonist antibody (N13B2-hVL6) (10 mg / kg) or 22 peripheral blood mononuclear cell (PBMC) samples before placebo injection. Only one sample did not pass different quality controls and was removed from the study.

[0196] The inventors have identified six genes that are actually involved in the IL7 / IL7R signaling pathway: BCL2, CISH, SOCS2, FLT3LG, PTGER2, and DPP4.

[0197] To evaluate the performance of gene signatures containing these genes, a single-sample GSEA signature was calculated (ssGSEA).

[0198] For samples from healthy volunteers treated with N13B2-hVL6, a significantly high decrease in the ssGSEA signature of six genes was observed at D15 during treatment, but not for samples from healthy volunteers treated with placebo (Figure 1A). This decrease was also observed at D57 after N13B2-hVL6 injection, which was consistent with the long-term detection of antibodies (over 3 months) in the sera of healthy volunteers treated with N13B2-hVL6 (Figure 1B).

[0199] For samples from healthy volunteers treated with N13B2-hVL6, a significantly high decrease in the ssGSEA signature of only four genes was also observed at D15 during treatment, but not for samples from healthy volunteers treated with placebo (Figure 2A). Similar to the signature of six genes, this decrease was also observed at D57 after N13B2-hVL6 injection (Figure 2B).

[0200] As shown in Table 1 below, higher sensitivity was observed with ssGSEA signatures of four and six genes compared to ssGSEA analysis of only one gene after short-term and long-term treatments.

[0201] [Table 1] Table 1: ssGSEA analysis of four and six genes performed on subject samples treated with 10 mg / kg of N13B2-hVL6

[0202] This new signature strongly correlates with the biological anti-IL7R antibody antagonist effect and could be used as a tool to track the biological effects of anti-IL7R antagonist antibodies in different new clinical trials.

[0203] RNA seq technology is very sensitive but expensive. The gene signature was also verified by RT-qPCR, another cheaper technology, but this is less sensitive. The FLT3G and SOCS2 genes were not analyzed by RT-qPCR.

[0204] The inventors performed RT-qPCR verification of these four genes as a companion test for the anti-IL7R antagonist antibody.

[0205] Real-time quantitative analysis (RT-qPCR) of these four genes was performed on five paired samples of D1 and D15 from placebo and five paired samples of D1 and D15 from volunteers (10 mg / kg) treated in vivo with N13B2-hVL6, which had already been analyzed by RNAseq sequencing by Genewiz.

[0206] For this purpose, reverse transcription (RT) was performed using EzDNASE #11766050 Thermo and SuperScript IV VILO Master Mix to obtain complementary DNA (cDNA), which was treated with Dnase to remove genomic DNA. TaqMan probes and TaqMan® Fast Advanced Master Mix #4444557 (5 mL) Thermo were used to quantify each gene and the housekeeping gene on a real-time analyzer ViiA7 (Table 2).

[0207]

Table 2

[0208] The sum of -DeltaCt normalized to the TBP housekeeping gene for the four genes (BCL2, CISH, PTGER2, DPP4) detected by RT-qPCR was calculated and used as a signature.

[0209] This box plot (Figure 3) of the signature shows that the decrease observed by RNA sequencing in the D15 samples from healthy volunteers treated with N13B2-hVL6 is also observable by RT-qPCR analysis.

[0210] To verify the RT-qPCR signature of these four genes, RT-qPCR analysis of these four genes was performed in the subject samples treated with 1 mg / kg, 4 mg / kg, and 6 mg / kg of N13B2-hVL6 using TBP as the housekeeping gene (Table 3), and the log2 fold change (log2FC) was calculated between D15 and D1 in the paired samples.

[0211]

Table 3

[0212] A significant decrease in the RT-qPCR signature of the four genes was observed at all doses of N13B2-hVL6 treatment.

[0213] This signature was verified using other antibodies. For this purpose, RNAseq sequencing of human PBMCs pre-incubated with three anti-human IL7Rα mAbs (mAb1: N13B2-hVL6, mAb2: MD707-13, and mAb13: GSKWO2011) and stimulated with recombinant IL7 for 3 hours was performed.

[0214] A significantly high decrease in the ssGSEA signature related to the IL7 / IL7R pathway was observed for three antibodies using 4 and 6 genes (Figures 4 and 5).

[0215] To verify that this new signature can be used as a tool to track the biological effects of IL7R agonists, correlating with the IL7R agonist effect, RNA sequencing of PBMCs treated with IL7 cytokine was performed. Freshly isolated human PBMCs from healthy donors (obtained from the Etablissement Français du Sang) were incubated with 5 ng / ml of recombinant human IL-7 (AbDSerotec) at 37°C for 3 hours. The reaction was stopped on ice and the cell pellet was resuspended in RLT buffer (Qiagen) containing 1% β-mercaptoethanol in RNase / Dnase-free water and stored at -80°C.

[0216] RNA was extracted using an RNA mini extraction kit according to the manufacturer's (Qiagen) instructions. The quality and quantity of RNA were evaluated by infrared spectroscopy measurement (Nanodrop) and Agilent Bioanalyzer (Agilent RNA 6000 Pico Kit).

[0217] Smart-Seq2 libraries were sequenced by the Broad Genomics Platform according to a SmartSeq2 protocol adapted and partially modified by the Broad Technology Lab (Trombetta, J. J. et al., Curr. Protoc. Mol. Biol. 107, 4.22.1-4.22.17 (2014)). Briefly, total RNA was purified using RNA-SPRI beads, polyA+ mRNA was reverse transcribed into cDNA, the amplified cDNA was subjected to transposon-based fragmentation using dual indices, and each fragment of each converted transcript was barcoded using a combination of barcodes specific to each sample.

[0218] Eight cycles were added for each index, and sequencing was performed as paired-end 2×25 bp. The data were separated by barcodes and aligned using Tophat version 2.0.10 with default settings. Transcripts were quantified using Cuffquant version 2.2.167 by the computational pipeline of Broad Technology Labs. Briefly, the data were processed by CuffNorm when 50% of the reads were aligned and at least 100,000 pairs were aligned for each sample. Normalization used default settings including "geometric" normalization, and the expression level information as log2-transformed FPKM values (fragments per kilobase of transcript per million mapped fragments) was used for subsequent analysis. The results of RNA-seq are accessible in the GEO database under accession code GSE103643 (Belarif et al., Nature communications, (2018) 9:4483).

[0219] Single-sample GSEA (ssGSEA) signatures were calculated based on the genes of interest BCL2, CISH, SOCS2, FLT3LG, PTGER2, and DPP4.

[0220] As shown in Figures 6 and 7, significant and high increases in the ssGSEA signatures of 4 and 6 genes related to the IL7 / IL7R pathway were observed for the samples stimulated with IL7.

[0221] As shown in Table 4, higher sensitivity was observed for the ssGSEA signatures of 4 and 6 genes compared to the ssGSEA analysis of only one gene.

[0222]

Table 4

[0223] (Conclusion) This study enabled the discovery that the gene expression profiles of six genes, particularly four genes, reflect IL7 / IL7R pathway activity. The gene expression profile of at least one of these four genes, which can be easily quantified by RT-qPCR, can be used to evaluate IL7R antagonist or agonist activity and can serve as a companion test for monitoring the biological effects of IL7R antagonists or agonists.

Claims

1. An in vitro method for evaluating the therapeutic response to IL7R antagonist or IL7R agonist treatment in human patients, The process includes determining the gene expression profiles of each of the following genes in a sample from a patient who has been administered at least one dose of an IL7R antagonist or IL7R agonist: BCL2, CISH, PTGER2, and DPP4 genes, and optionally further FLT3LG and / or SOCS2 genes, wherein, The in vitro method, wherein a lower or higher gene expression profile of the gene in the patient sample compared to a control value indicates that the patient is more likely to respond to the IL7R antagonist or IL7R agonist treatment, respectively.

2. The method according to claim 1, wherein the sample is collected in advance from the patient at least 15 days after the administration of at least one dose of the IL7R antagonist or IL7R agonist to the patient.

3. An in vitro method for evaluating the potential therapeutic response in human patients to an IL7R antagonist or agonist before treatment with said IL7R antagonist or agonist, a) A step of culturing a sample collected in advance from the patient before the treatment in the presence of the IL7R antagonist or IL7R agonist, b) A step of determining the gene expression profile of each of the following genes in the cultured sample: BCL2, CISH, PTGER2, and DPP4 genes, and optionally further FLT3LG and / or SOCS2 genes, wherein a lower or higher gene expression profile of the gene in the cultured sample compared to a control value indicates that the patient is more likely to respond to treatment with the IL7R antagonist or IL7R agonist, respectively. The in vitro method, including the aforementioned method.

4. The aforementioned IL7R antagonist, a) VHCDR1 contains or consists of the amino acid sequence of SEQ ID NO: 1, - VHCDR2 contains or consists of the amino acid sequence of SEQ ID NO: 2, and - VHCDR3 contains or consists of the amino acid sequence of SEQ ID NO: 3 or 11. Variable heavy chains containing three CDRs; and (b)- VLCDR1 contains or consists of the amino acid sequence of SEQ ID NO: 4 or 12, - VLCDR2 contains or consists of the amino acid sequence of SEQ ID NO: 5 or 13, and - VLCDR3 contains or consists of the amino acid sequence of SEQ ID NO: 6 Variable light chain containing 3 CDRs The method according to claim 1 or 3, wherein the anti-IL7R antibody or antigen-binding fragment thereof contains (for example, the anti-IL7R antagonist antibody or antigen-binding fragment thereof contains or comprises a heavy chain variable domain containing or comprising the amino acid sequence of SEQ ID NO: 7 and a light chain variable domain containing or comprising the amino acid sequence of SEQ ID NO: 8, or contains a full-length heavy chain containing or comprising the amino acid sequence of SEQ ID NO: 9 and a full-length light chain containing or comprising the amino acid sequence of SEQ ID NO:

10.

5. A method for evaluating the therapeutic response to the IL7R antagonist according to claim 1 when the patient suffers from a disease associated with enhanced IL7R signaling pathways induced by IL7 (for example, a disease selected from the group consisting of autoimmune diseases, inflammatory diseases, allergic diseases, cancers, infectious diseases, respiratory diseases, and transplant-related diseases, involving activation or proliferation of CD127-positive pathogenic cells), or a method for evaluating the potential therapeutic response to the IL7R antagonist in a human patient according to claim 3.

6. A method for evaluating the therapeutic response to an IL7R agonist according to claim 1, wherein the patient suffers from a disease associated with impaired healthy T-cell activity (for example, a cancer or infection whose onset is not dependent on the IL7R pathway itself), or a method for evaluating the potential therapeutic response to an IL7R agonist in a human patient according to claim 3.

7. The method according to claim 1 or 3, wherein the patient sample is a tissue sample (for example, tumor tissue containing infiltrating immune cells from a cancer patient, or a blood sample from a patient, or isolated human peripheral blood mononuclear cells).

8. The method according to claim 1 or 3, wherein the gene expression profile is determined by detecting the mRNA expression of the gene.

9. An IL7R antagonist for use in treating a disease associated with the IL7-induced IL7R signaling pathway in a human patient requiring such treatment (for example, a disease selected from the group consisting of autoimmune diseases, inflammatory diseases, allergic diseases, cancerous diseases, infectious diseases, respiratory diseases, and transplant-related diseases, involving activation or proliferation of CD127-positive pathogenic cells), the IL7R antagonist being administered to the patient when the patient is evaluated to be likely to respond to the treatment in the manner described in claim 1 or 3, after being administered at least one dose of the IL7R antagonist.

10. An IL7R antagonist for use in treating a disease associated with the IL7-induced IL7R signaling pathway in a human patient requiring such treatment (for example, a disease selected from the group consisting of autoimmune diseases, inflammatory diseases, allergic diseases, cancerous diseases, infectious diseases, respiratory diseases, and transplant-related diseases, involving activation or proliferation of CD127-positive pathogenic cells), wherein the patient is pre-evaluated to be likely to respond to the IL7R antagonist treatment in the manner described in claim 3.

11. The aforementioned IL7R antagonist, a) VHCDR1 contains or consists of the amino acid sequence of SEQ ID NO: 1, - VHCDR2 contains or consists of the amino acid sequence of SEQ ID NO: 2, and - VHCDR3 contains or consists of the amino acid sequence of SEQ ID NO: 3 or 11. Variable heavy chains containing 3 CDRs; and (b)- VLCDR1 contains or consists of the amino acid sequence of SEQ ID NO: 4 or 12, - VLCDR2 contains or consists of the amino acid sequence of SEQ ID NO: 5 or 13, and - VLCDR3 contains or consists of the amino acid sequence of SEQ ID NO: 6 Variable light chain containing 3 CDRs An IL7R antagonist for use according to claim 9, which is an anti-IL7R antagonist antibody or an antigen-binding fragment thereof (for example, the antibody or the antigen-binding fragment thereof comprises a heavy chain variable domain comprising or consisting of the amino acid sequence of SEQ ID NO: 7 and a light chain variable domain comprising or consisting of the amino acid sequence of SEQ ID NO: 8, or comprises a full-length heavy chain comprising or consisting of the amino acid sequence of SEQ ID NO: 9 and a full-length light chain comprising or consisting of the amino acid sequence of SEQ ID NO: 10).

12. An IL7R agonist for use in treating a disease associated with impaired healthy T-cell activity in a human patient requiring such treatment (e.g., a cancer or infection whose onset is not itself dependent on the IL7R pathway), the IL7R agonist being administered to the patient when the patient is evaluated to be likely to respond to the treatment in the manner described in claim 1 or 3, after being administered at least one dose of the IL7R agonist.

13. An IL7R agonist for use in treating a disease associated with impaired healthy T-cell activity in a human patient requiring such treatment (e.g., a cancer or infection whose onset is not itself dependent on the IL7R pathway), wherein the patient is pre-evaluated to be likely to respond to the IL7R antagonist treatment in the manner described in claim 3.

14. An in vitro method for selecting compounds that are likely to be effective in treating diseases associated with the IL7R signaling pathway induced by IL7 or diseases associated with impaired healthy T cell activity, a) A step of culturing human cells (e.g., immune cells or peripheral blood mononuclear cells) in the presence of the compound. b) A step of determining the gene expression profile of each of the following genes in the cell: BCL2, CISH, PTGER2, and DPP4 genes, and optionally further FLT3LG and / or SOCS2 genes, and c) The step of selecting a compound that induces a lower or higher gene expression profile of the gene in the cell compared to a control value. The in vitro method, which optionally includes a gene expression profile determined by detecting the mRNA expression of the gene.

15. A kit for use in the method according to any one of claims 1, 3, and 14, comprising a set of reagents for specifically detecting the gene expression profiles of each of the following genes: BCL2, CISH, PTGER2, and DPP4 genes, and optionally further FLT3LG and / or SOCS2 genes, wherein the reagents are optionally specific primer pairs and / or probes for each gene.

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