Treatment for Prurigo Nodularis

Abstract

Described herein are treatments and preventions for prurigo nodularis (PN), antibodies and pharmaceutical compositions for use in the treatment or prevention of PN, and the use of anti-IL-31RA antibodies (e.g., nemolizumab) in the manufacture of a medicament for the treatment or prevention of PN. Also described herein are biomarkers for PN and methods of altering or ameliorating these biomarkers through treatment with an antibody that binds IL-31RA (e.g., nemolizumab).

Description

[Technical field]

[0001] cross reference This application claims priority under 35 U.S.C. §119(e) to U.S. Provisional Application No. 63 / 172,020, filed April 7, 2021, and U.S. Provisional Application No. 63 / 323,409, filed March 24, 2022, the entire contents of which are incorporated herein by reference.

[0002] Statement of government support This invention was made with Government support under Grants R01-AR069071, R01-AR073196, P30-AR075043, and K01-AR072129 awarded by the National Institutes of Health (NIH). The U.S. Government has certain rights in this invention.

[0003] Field Described herein are treatments and preventions of prurigo nodularis (PN), antibodies and pharmaceutical compositions for use in the treatment or prevention of PN, and the use of anti-IL-31RA antibodies (e.g., nemolizumab) in the manufacture of a medicament for the treatment or prevention of PN. Also described herein are biomarkers of PN and methods of altering or ameliorating these biomarkers through treatment with an antibody that binds IL-31RA (e.g., nemolizumab). [Background technology]

[0004] background The following discussion is provided merely to aid the reader in understanding the present disclosure and is not admitted to describe or constitute prior art to the present disclosure.

[0005] Prurigo chronica (CP) is a skin disorder caused by sensitization of nerve cells to itch and an extension of the itch-scratch cycle. Prurigo nodularis (PN), a subtype of CP, is a skin disorder that causes hard itchy nodules (nodules) to form on the skin. The itching (scratching) can be intense and cause bleeding or painful scratching. Scratching can cause more skin lesions to appear. The itch is worsened by heat, sweating, or irritation from clothing. In some cases, people with PN have a history of other diseases, including eczema (atopic dermatitis), diabetes, lymphoma, HIV infection, severe anemia, or kidney disease.

[0006] The exact cause of PN is unknown, and diagnosis of the disease is based on observing symptoms such as extremely itchy skin with the formation of nodules. In some cases, a skin biopsy is used to confirm the diagnosis. Currently, treatment may include corticosteroid creams, oral medications, cryotherapy, or photochemotherapy.

[0007] Treatment for PN remains necessary, and patients are identified as likely to respond to, or are responding to, such treatment. Summary of the Invention

[0008] overview Described herein are treatments and preventions for prurigo nodularis (PN) that achieve specific therapeutic results. In general, the treatments and preventions include administering an anti-IL-31RA antibody (e.g., nemolizumab) to a subject with PN. Also described herein are biomarkers for PN and methods of using the biomarkers of the present disclosure to determine whether a subject responds to treatment.

[0009] In a first aspect, the disclosure provides a method of treating or preventing prurigo nodularis (PN) in a subject, the method comprising administering an anti-IL-31RA antibody to a subject having PN, wherein the subject differentially expresses at least one gene selected from the genes disclosed in Table 1 as compared to a reference expression level of the at least one gene.

[0010] In a second aspect, the present disclosure provides a method of normalizing differentially expressed genes (DEGs) in a subject with PN, comprising administering an anti-IL-31RA antibody to a subject with PN, wherein the subject differentially expresses at least one gene selected from the genes disclosed in Table 1 compared to a reference expression level of the at least one gene, and wherein administration of the anti-IL-31RA antibody normalizes the expression level of the at least one gene.

[0011] In some embodiments of these aspects, normalization can be determined about 4 weeks, about 8 weeks, or about 12 weeks after administration of the anti-IL-31RA antibody.

[0012] In some embodiments of these aspects, the differential expression of at least one gene can be determined by RT-qPCR, RT-PCR, RNA-seq, Northern blotting, serial analysis of gene expression (SAGE), or DNA or RNA microarrays, or the differential expression of at least one gene can be determined at the protein level by Western blotting, ELISA, surface plasmon resonance, or mass spectrometry.

[0013] In some embodiments of these aspects, at least two, at least three, at least four, or at least five of the genes disclosed in Table 1 may be differentially expressed.

[0014] In some embodiments of these aspects, the differentially expressed genes are KRT6C (keratin 6C, NCBI Entrez Gene: 286887, UniProt: P48668), DEFB4A (defensin beta 4A, NCBI Entrez Gene: 1673, UniProt: O15263), KRT16 (keratin 16, NCBI Entrez Gene: 3868, UniProt: P08779), LCE5A (late cornified envelope 5A, NCBI Entrez Gene: 254910, UniProt: Q5TCM9), AQP7 (aquaporin 7, NCBI Entrez Gene: 364, UniProt: O14520), IL36A (interleukin 36 alpha, NCBI Entrez Gene: 1673, UniProt: O15263), IL40A (interleukin 40 alpha, NCBI Entrez Gene: 1673, UniProt: O15263), IL40B (interleukin 40 alpha, NCBI Entrez Gene: 1673, UniProt: O15263), IL40C (interleukin 40 alpha, NCBI Entrez Gene: 1673, UniProt: O15263), IL40D (interleukin 40 alpha, NCBI Entrez Gene: 1673, UniProt: O15263), IL40E (interleukin 40 alpha, NCBI Entrez Gene: 1673, UniProt: O15263), IL40F ... IL36G (Interleukin 36 gamma, NCBI Entrez Gene: 56300, UniProt: Q9NZH8), IL19 (Interleukin 19, NCBI Entrez Gene: 29949, UniProt: Q9UHD0), IL20 (Interleukin 20, NCBI Entrez Gene: 50604, UniProt: Q9NYY1), IL22 (Interleukin 22, NCBI Entrez Gene: 50616, UniProt: Q9GZX6), IL24 (Interleukin 24, NCBI Entrez Gene: 11009, UniProt: Q13007), and IL26 (Interleukin 26, NCBI Entrez Gene: 55801, UniProt: Q9NPH9).

[0015] In some embodiments of these aspects, KRT6C, DEFB4A, and / or KRT16 are overexpressed compared to a reference expression level. In some embodiments of these aspects, LCE5A and / or AQP7 are underexpressed compared to a reference expression level. In some embodiments of these aspects, IL1A, IL1B, and / or IL4R are overexpressed compared to a reference expression level. In some embodiments of these aspects, Ki67 (MKI67), CDKN1A, and / or IL-1 and IL-36 are overexpressed in the skin of the subject compared to a reference expression level. In some embodiments of these aspects, the reference level is an expression level in a human without PN.

[0016] In some embodiments of these aspects, the subject may have a gene ontology as disclosed in Table 2.

[0017] In some embodiments of these aspects, the anti-IL-31RA antibody may be administered subcutaneously.

[0018] In some embodiments of these aspects, the anti-IL-31RA antibody may be administered once per week, once per two weeks, once per three weeks, once per four weeks, once per five weeks, once per six weeks, once per seven weeks, or once per eight weeks.

[0019] In some embodiments of these aspects, the anti-IL-31RA antibody may be administered at a dose of about 0.01 mg / kg to about 0.1 mg / kg, about 0.1 mg / kg to about 0.5 mg / kg, about 0.5 mg / kg to about 1.5 mg / kg, about 1.5 mg / kg to about 2.5 mg / kg, or about 2.5 mg / kg to about 10 mg / kg.

[0020] In some embodiments of these aspects, the anti-IL-31RA antibody may be administered at a dose of about 10 mg, about 15 mg, about 20 mg, about 25 mg, about 30 mg, about 35 mg, about 40 mg, about 45 mg, about 50 mg, about 55 mg, about 60 mg, about 65 mg, about 70 mg, about 75 mg, about 80 mg, about 85 mg, about 90 mg.

[0021] In some embodiments of these aspects, the anti-IL-31RA antibody is administered according to a flat dosing regimen. In some embodiments of these aspects, the anti-IL-31RA antibody is administered according to a loading dose regimen.

[0022] In some embodiments of these aspects, itch scoring is improved in the subject.

[0023] In some embodiments of these aspects, the anti-IL-31RA antibody comprises a heavy chain variable region comprising an HCDR1 comprising SEQ ID NO:8, an HCDR2 comprising SEQ ID NO:9, and an HCDR3 comprising SEQ ID NO:10, and a light chain variable region comprising an LCDR1 comprising SEQ ID NO:12, an LCDR2 comprising SEQ ID NO:13, and an LCDR3 comprising SEQ ID NO:14. In some embodiments of these aspects, the anti-IL-31RA antibody is nemolizumab or a fragment or variant thereof. In some embodiments of these aspects, the anti-IL-31RA antibody is nemolizumab.

[0024] In a third aspect, the disclosure provides a method of normalizing epidermal hyperproliferation and / or normalizing epidermal differentiation in a subject with prurigo nodularis (PN), comprising administering an anti-IL-31RA antibody to a subject with PN, thereby normalizing epidermal hyperproliferation and / or normalizing epidermal differentiation compared to a human without PN. In some embodiments, normalization can be determined about 4 weeks, about 8 weeks, or about 12 weeks after administration of the anti-IL-31RA antibody.

[0025] In a fourth aspect, the disclosure provides a method of reducing an inflammatory response in the skin of a subject with prurigo nodularis (PN), comprising administering an anti-IL-31RA antibody to a subject with PN, thereby reducing the inflammatory response in the skin. In some embodiments, the inflammatory response can be a Th2 response, optionally including overexpression of IL-13, IL-17A, IL-1, and / or IL-36. In some embodiments, the inflammatory response can be a Th1 response.

[0026] In some embodiments of the third and fourth aspects, the subject may differentially express at least one gene selected from the genes disclosed in Table 1 compared to a reference expression level of the at least one gene. In some embodiments, at least two, at least three, at least four, or at least five genes disclosed in Table 1 are differentially expressed. In some embodiments, the differentially expressed genes are selected from KRT6C, DEFB4A, KRT16, LCE5A, AQP7, IL36A, IL36G, IL19, IL20, IL22, IL24, and IL26. In some embodiments, KRT6C, DEFB4A, and / or KRT16 are overexpressed compared to a reference expression level. In some embodiments, LCE5A and / or AQP7 are underexpressed compared to a reference expression level. In some embodiments, IL1A, IL1B, and / or IL4R are overexpressed compared to a reference expression level. In some embodiments, Ki67 (MKI67), CDKN1A, and / or IL-1 and IL-36 are overexpressed in the skin of a subject relative to a reference expression level. In some embodiments, the reference level is an expression level in a human without PN.

[0027] In some embodiments of the third and fourth aspects, the subject may have a gene ontology as disclosed in Table 2.

[0028] In some embodiments of the third and fourth aspects, the anti-IL-31RA antibody is administered subcutaneously.

[0029] In some embodiments of the third and fourth aspects, the anti-IL-31RA antibody is administered once per week, once per two weeks, once per three weeks, once per four weeks, once per five weeks, once per six weeks, once per seven weeks, or once per eight weeks.

[0030] In some embodiments of the third and fourth aspects, the anti-IL-31RA antibody is administered at a dose of about 0.01 mg / kg to about 0.1 mg / kg, about 0.1 mg / kg to about 0.5 mg / kg, about 0.5 mg / kg to about 1.5 mg / kg, about 1.5 mg / kg to about 2.5 mg / kg, or about 2.5 mg / kg to about 10 mg / kg.

[0031] In some embodiments of the third and fourth aspects, the anti-IL-31RA antibody is administered at a dose of about 10 mg, about 15 mg, about 20 mg, about 25 mg, about 30 mg, about 35 mg, about 40 mg, about 45 mg, about 50 mg, about 55 mg, about 60 mg, about 65 mg, about 70 mg, about 75 mg, about 80 mg, about 85 mg, or about 90 mg.

[0032] In some embodiments of the third and fourth aspects, the anti-IL-31RA antibody is administered according to a flat dosing regimen. In some embodiments of the third and fourth aspects, the anti-IL-31RA antibody is administered according to a loading dose regimen.

[0033] In some embodiments of the third and fourth aspects, itch scoring is improved in the subject.

[0034] In some embodiments of the third and fourth aspects, the anti-IL-31RA antibody comprises a heavy chain variable region comprising an HCDR1 comprising SEQ ID NO:8, an HCDR2 comprising SEQ ID NO:9, and an HCDR3 comprising SEQ ID NO:10, and a light chain variable region comprising an LCDR1 comprising SEQ ID NO:12, an LCDR2 comprising SEQ ID NO:13, and an LCDR3 comprising SEQ ID NO:14. In some embodiments, the anti-IL-31RA antibody is nemolizumab or a fragment or variant thereof. In some embodiments, the anti-IL-31RA antibody is nemolizumab.

[0035] In a fifth aspect, the present disclosure provides a method for diagnosing prurigo nodularis (PN), comprising detecting expression levels of at least one, at least two, at least three, at least four, or at least five of the differentially expressed genes (DEGs) of Table 1 in a sample obtained from a subject suspected of having PN, and comparing the expression levels of the DEGs to a reference level, where the reference level is a corresponding gene expression level of each DEG in a sample from an individual not having PN.

[0036] In some embodiments of the fifth aspect, the sample obtained from a subject suspected of having PN may be a skin sample, optionally containing a lesion or nodule.

[0037] In some embodiments of the fifth aspect, the expression levels of DEGs are determined by RT-qPCR, RT-PCR, RNA-seq, Northern blotting, serial analysis of gene expression (SAGE), or DNA or RNA microarrays.

[0038] In some embodiments of the fifth aspect, the DEGs are selected from KRT6C, DEFB4A, KRT16, LCE5A, AQP7, IL36A, IL36G, IL19, IL20, IL22, IL24, and IL26. In some embodiments, the DEGs include overexpression of KRT6C, DEFB4A, and / or KRT16 compared to a reference expression level, which is indicative of PN. In some embodiments, underexpression of LCE5A and / or AQP7 compared to a reference expression level is indicative of PN. In some embodiments, the DEGs include overexpression of IL1A, IL1B, and / or IL4R compared to a reference expression level, which is indicative of PN. In some embodiments, the DEGs include overexpression of Ki67 (MKI67), CDKN1A, and / or IL-1 and IL-36 compared to a reference expression level, which is indicative of PN.

[0039] In some embodiments of the fifth aspect, the subject has a gene ontology disclosed in Table 2.

[0040] In a sixth aspect, the present disclosure provides a method of determining whether a subject with prurigo nodularis (PN) will respond to treatment with an anti-IL-31RA antibody, comprising detecting expression levels of at least one, at least two, at least three, at least four, or at least five of the differentially expressed genes (DEGs) of Table 1 in a sample obtained from a subject with PN, and comparing the expression levels of the DEGs to a reference level, where the reference level is a corresponding gene expression level for each DEG in a sample from an individual without PN.

[0041] In some embodiments of the sixth aspect, the sample obtained from the subject with PN is a skin sample, optionally containing a lesion or nodule.

[0042] In some embodiments of the fifth aspect, the expression levels of DEGs are determined by RT-qPCR, RT-PCR, RNA-seq, Northern blotting, serial analysis of gene expression (SAGE), or DNA or RNA microarrays.

[0043] In some embodiments of the sixth aspect, the DEGs are selected from KRT6C, DEFB4A, KRT16, LCE5A, AQP7, IL36A, IL36G, IL19, IL20, IL22, IL24, and IL26. In some embodiments, the DEGs include overexpression of KRT6C, DEFB4A, and / or KRT16 compared to a reference expression level, which is indicative of PN. In some embodiments, underexpression of LCE5A and / or AQP7 compared to a reference expression level is indicative of PN. In some embodiments, the DEGs include overexpression of IL1A, IL1B, and / or IL4R compared to a reference expression level, which is indicative of PN. In some embodiments, the DEGs include overexpression of Ki67 (MKI67), CDKN1A, and / or IL-1 and IL-36 compared to a reference expression level, which is indicative of PN.

[0044] In some embodiments of the sixth aspect, the anti-IL-31RA antibody comprises a heavy chain variable region comprising HCDR1 comprising SEQ ID NO:8, HCDR2 comprising SEQ ID NO:9, and HCDR3 comprising SEQ ID NO:10, and a light chain variable region comprising LCDR1 comprising SEQ ID NO:12, LCDR2 comprising SEQ ID NO:13, and LCDR3 comprising SEQ ID NO:14. In some embodiments, the anti-IL-31RA antibody is nemolizumab or a fragment or variant thereof. In some embodiments, the anti-IL-31RA antibody is nemolizumab.

[0045] In a seventh aspect, the present disclosure provides a method of determining whether a subject with prurigo nodularis (PN) will respond to treatment with an anti-IL-31RA antibody, comprising detecting expression levels of at least one, at least two, at least three, at least four, or at least five of the differentially expressed genes DEGs of Table 1 in a post-treatment sample obtained from a subject with PN who was administered at least one dose of an anti-IL-31RA antibody, and comparing the expression levels of the DEGs to a baseline level of expression, wherein the baseline level of gene expression is from a sample from the same individual before treatment was initiated.

[0046] In some embodiments of the seventh aspect, the sample obtained from the subject with PN is a skin sample, optionally containing a lesion or nodule.

[0047] In some embodiments of the seventh aspect, the expression levels of DEGs are determined by RT-qPCR, RT-PCR, RNA-seq, Northern blotting, serial analysis of gene expression (SAGE), or DNA or RNA microarrays.

[0048] In some embodiments of the seventh aspect, the DEGs are selected from KRT6C, DEFB4A, KRT16, LCE5A, AQP7, IL36A, IL36G, IL19, IL20, IL22, IL24, and IL26. In some embodiments, the DEGs include overexpression of KRT6C, DEFB4A, and / or KRT16 compared to a reference expression level, which is indicative of PN. In some embodiments, underexpression of LCE5A and / or AQP7 compared to a reference expression level is indicative of PN. In some embodiments, the DEGs include overexpression of IL1A, IL1B, and / or IL4R compared to a reference expression level, which is indicative of PN. In some embodiments, the DEGs include overexpression of Ki67 (MKI67), CDKN1A, and / or IL-1 and IL-36 compared to a reference expression level, which is indicative of PN.

[0049] In some embodiments of the seventh aspect, the post-treatment sample is obtained about 4 weeks, about 8 weeks, or about 12 weeks after administration of the anti-IL-31RA antibody.

[0050] In some embodiments of the seventh aspect, the anti-IL-31RA antibody comprises a heavy chain variable region comprising HCDR1 comprising SEQ ID NO:8, HCDR2 comprising SEQ ID NO:9, and HCDR3 comprising SEQ ID NO:10, and a light chain variable region comprising LCDR1 comprising SEQ ID NO:12, LCDR2 comprising SEQ ID NO:13, and LCDR3 comprising SEQ ID NO:14. In some embodiments, the anti-IL-31RA antibody is nemolizumab or a fragment or variant thereof. In some embodiments, the anti-IL-31RA antibody is nemolizumab.

[0051] In an eighth aspect, the disclosure provides a method of treating or preventing prurigo nodularis (PN) in a subject, comprising administering an anti-IL-31RA antibody to a subject having PN, wherein treatment with the anti-IL-31RA antibody results in (a) reduced leukocyte migration or leukocyte cell migration, (b) inhibition of the STAT3 pathway, (c) inhibition of the STAT5b pathway, (d) downregulation of IL-6 or the IL-6 pathway, (e) downregulation of VEGF or the VEGF pathway, (f) reduced TGFB1 pathway activation, or (g) a combination thereof.

[0052] In a ninth aspect, the disclosure provides a method of altering an immune response in a subject having PN, comprising administering an anti-IL-31RA antibody to the subject having PN, wherein administration of the anti-IL-31RA antibody results in (a) reduced leukocyte migration or leukocyte cell migration, (b) inhibition of the STAT3 pathway, (c) inhibition of the STAT5b pathway, (d) downregulation of IL-6 or the IL-6 pathway, (e) downregulation of VEGF or the VEGF pathway, (f) reduced TGFB1 pathway activation, or (g) a combination thereof.

[0053] In some embodiments of the eighth and ninth aspects, (a) reduced leukocyte migration or leukocyte cell movement, (b) inhibition of the STAT3 pathway, (c) inhibition of the STAT5b pathway, (d) downregulation of IL-6 or the IL-6 pathway, (e) downregulation of VEGF or the VEGF pathway, (f) reduced TGFB1 pathway activation, or (g) a combination thereof is determined with respect to (i) a control sample obtained from one or more individuals not having PN, or (ii) a biological sample obtained from the subject prior to administration of an anti-IL-31RA antibody.

[0054] In some embodiments of the eighth and ninth aspects, a) reduced leukocyte migration or leukocyte cell movement, (b) inhibition of the STAT3 pathway, (c) inhibition of the STAT5b pathway, (d) downregulation of IL-6 or the IL-6 pathway, (e) downregulation of VEGF or the VEGF pathway, (f) reduced TGFB1 pathway activation, or (g) a combination thereof is assessed about 2 weeks, about 4 weeks, about 6 weeks, about 8 weeks, about 10 weeks, or about 12 weeks after administration of the anti-IL-31RA antibody.

[0055] In some embodiments of the eighth and ninth aspects, (a) reduced leukocyte migration or leukocyte cell movement, (b) inhibition of the STAT3 pathway, (c) inhibition of the STAT5b pathway, (d) downregulation of IL-6 or the IL-6 pathway, (e) downregulation of VEGF or the VEGF pathway, (f) reduced TGFB1 pathway activation, or (g) a combination thereof is determined by mass spectrometry performed on one or more biological samples obtained from the subject. In some embodiments, the biological sample is a plasma sample.

[0056] In some embodiments of the eighth and ninth aspects, the subject exhibits at least two, at least three, at least four, at least five, or all six of: (a) decreased leukocyte migration or leukocyte cell movement; (b) inhibition of the STAT3 pathway; (c) inhibition of the STAT5b pathway; (d) downregulation of IL-6 or the IL-6 pathway; (e) downregulation of VEGF or the VEGF pathway; and (f) decreased TGFB1 pathway activation.

[0057] In a tenth aspect, the disclosure provides a method of reducing an inflammatory response in plasma of a subject with prurigo nodularis (PN), comprising administering an anti-IL-31RA antibody to the subject with PN, thereby reducing the inflammatory response in the skin. In some embodiments, reducing the inflammatory response comprises reducing the plasma concentration of IL-6, VEGF, or a combination thereof, compared to a reference level. In some embodiments, the reference level is determined from (i) a control sample obtained from one or more individuals without PN, or (ii) a biological sample obtained from the subject prior to administration of the anti-IL-31RA antibody. In some embodiments, after administration of the anti-IL-31RA antibody, the subject further exhibits (a) reduced leukocyte migration or leukocyte cell migration, (b) inhibition of the STAT3 pathway, (c) inhibition of the STAT5b pathway, (d) reduced TGFB1 pathway activation, or (e) a combination thereof. In some embodiments, at least 4 weeks, at least 8 weeks, or at least 12 weeks after administration of the anti-IL-31RA antibody, the subject further exhibits at least two, at least three, or all four of: (a) decreased leukocyte migration or leukocyte cell migration; (b) inhibition of the STAT3 pathway; (c) inhibition of the STAT5b pathway; and (d) decreased TGFB1 pathway activation.

[0058] In some embodiments of the eighth, ninth, or tenth aspects, the subject may also exhibit downregulation in a neuron ontology selected from CREB signaling in neurons, synaptogenesis signaling pathways, glial cell death and glial apoptosis, and combinations thereof, following administration of an anti-IL-31RA antibody.

[0059] In some embodiments of the eighth, ninth, or tenth aspect, the anti-IL-31RA antibody is administered subcutaneously. In some embodiments of the eighth, ninth, or tenth aspect, the anti-IL-31RA antibody is administered once per week, once per two weeks, once per three weeks, once per four weeks, once per five weeks, once per six weeks, once per seven weeks, or once per eight weeks.

[0060] In some embodiments of the eighth, ninth, or tenth aspect, the anti-IL-31RA antibody is administered at a dose of about 0.01 mg / kg to about 0.1 mg / kg, about 0.1 mg / kg to about 0.5 mg / kg, about 0.5 mg / kg to about 1.5 mg / kg, about 1.5 mg / kg to about 2.5 mg / kg, or about 2.5 mg / kg to about 10 mg / kg. In some embodiments of the eighth, ninth, or tenth aspect, the anti-IL-31RA antibody is administered at a dose of about 10 mg, about 15 mg, about 20 mg, about 25 mg, about 30 mg, about 35 mg, about 40 mg, about 45 mg, about 50 mg, about 55 mg, about 60 mg, about 65 mg, about 70 mg, about 75 mg, about 80 mg, about 85 mg, about 90 mg.

[0061] In some embodiments of the eighth, ninth, or tenth aspect, the anti-IL-31RA antibody is administered according to a flat dosing regimen. In some embodiments of the eighth, ninth, or tenth aspect, the anti-IL-31RA antibody is administered according to a loading dose regimen.

[0062] In some embodiments of the eighth, ninth, or tenth aspect, itch scoring is improved in the subject.

[0063] In some embodiments of the eighth, ninth, or tenth aspects, the anti-IL-31RA antibody comprises a heavy chain variable region comprising an HCDR1 comprising SEQ ID NO:8, an HCDR2 comprising SEQ ID NO:9, and an HCDR3 comprising SEQ ID NO:10, and a light chain variable region comprising an LCDR1 comprising SEQ ID NO:12, an LCDR2 comprising SEQ ID NO:13, and an LCDR3 comprising SEQ ID NO:14. In some embodiments, the anti-IL-31RA antibody is nemolizumab or a fragment or variant thereof. In some embodiments, the anti-IL-31RA antibody is nemolizumab.

[0064] In an eleventh aspect, the disclosure provides a method of determining whether a subject with prurigo nodularis (PN) responds to treatment with an anti-IL-31RA antibody, comprising detecting at least one, at least two, at least three, at least four, or at least five of the following in a post-treatment plasma sample obtained from a subject with PN administered at least one dose of an anti-IL-31RA antibody: (a) a decrease in leukocyte migration or leukocyte cell movement; (b) inhibition of the STAT3 pathway; (c) inhibition of the STAT5b pathway; (d) downregulation of IL-6 or the IL-6 pathway; (e) downregulation of VEGF or the VEGF pathway; (f) a decrease in TGFB1 pathway activation; or (g) a combination thereof, wherein the decrease or inhibition is measured relative to a baseline amount, and the decrease or inhibition is indicative of responsiveness to treatment, the baseline amount being determined from a plasma sample obtained from the same subject before treatment was initiated. In some embodiments, the decrease or inhibition is determined by mass spectrometry. In some embodiments, the subject may also exhibit downregulation in a neuronal ontology selected from CREB signaling in neurons, synaptogenesis signaling pathways, glial cell death and glial apoptosis, and combinations thereof, following administration of an anti-IL-31RA antibody.

[0065] In some embodiments of the eleventh aspect, the post-treatment sample is obtained about 2 weeks, about 4 weeks, about 6 weeks, about 8 weeks, about 10 weeks, or about 12 weeks after administration of the anti-IL-31RA antibody.

[0066] In some embodiments of the eleventh aspect, the anti-IL-31RA antibody comprises a heavy chain variable region comprising an HCDR1 comprising SEQ ID NO:8, an HCDR2 comprising SEQ ID NO:9, and an HCDR3 comprising SEQ ID NO:10, and a light chain variable region comprising an LCDR1 comprising SEQ ID NO:12, an LCDR2 comprising SEQ ID NO:13, and an LCDR3 comprising SEQ ID NO:14. In some embodiments, the anti-IL-31RA antibody is nemolizumab or a fragment or variant thereof. In some embodiments, the anti-IL-31RA antibody is nemolizumab.

[0067] The foregoing summary and the following detailed description are exemplary and explanatory and are intended to provide further explanation of the present disclosure as claimed. Other objects, advantages, and novel features will become readily apparent to those skilled in the art from the following brief description of the drawings and detailed description of the present disclosure. [Brief description of the drawings]

[0068] [Figure 1A] Figures 1A-1E show that prurigo nodularis (PN) is characterized by immune activation and abnormal keratinocyte differentiation. Number of differentially expressed genes (DEGs) in PN lesional vs. non-lesional skin (n=62, FC>=2 or FC<=-2, FDR<=0.1) (A). Enriched GO categories in PN lesional skin (B). Literature-based gene network derived from the top 1,000 DEGs in PN skin generated using Genomatix Pathway System (GePS, Genomatix.de). Pictures show the top 50 most connected genes co-cited in PubMed abstracts in the same sentence linked to function terms (most relevant genes / interactions). Orange represents upregulated genes and green represents downregulated genes in PN lesional vs. non-lesional skin. Significant nodes included the proliferation marker Ki67 (MKI67), IL-1 family members IL36G and IL1A, as well as CXCL8 and CDKN1A (C). Number of genes and intracluster correlation in modules identified from weighted co-expression gene network (WCGNA) analysis of non-lesional and lesional PN skin (D). Functions enriched in major co-expression modules from PN skin (modules #5, #6, #8) (E). [Figure 1B]Figures 1A-1E show that prurigo nodularis (PN) is characterized by immune activation and abnormal keratinocyte differentiation. Number of differentially expressed genes (DEGs) in PN lesional vs. non-lesional skin (n=62, FC>=2 or FC<=-2, FDR<=0.1) (A). Enriched GO categories in PN lesional skin (B). Literature-based gene network derived from the top 1,000 DEGs in PN skin generated using Genomatix Pathway System (GePS, Genomatix.de). Pictures show the top 50 most connected genes co-cited in PubMed abstracts in the same sentence linked to function terms (most relevant genes / interactions). Orange represents upregulated genes and green represents downregulated genes in PN lesional vs. non-lesional skin. Significant nodes included the proliferation marker Ki67 (MKI67), IL-1 family members IL36G and IL1A, as well as CXCL8 and CDKN1A (C). Number of genes and intracluster correlation in modules identified from weighted co-expression gene network (WCGNA) analysis of non-lesional and lesional PN skin (D). Functions enriched in major co-expression modules from PN skin (modules #5, #6, #8) (E). [Figure 1C]Figures 1A-1E show that prurigo nodularis (PN) is characterized by immune activation and abnormal keratinocyte differentiation. Number of differentially expressed genes (DEGs) in PN lesional vs. non-lesional skin (n=62, FC>=2 or FC<=-2, FDR<=0.1) (A). Enriched GO categories in PN lesional skin (B). Literature-based gene network derived from the top 1,000 DEGs in PN skin generated using Genomatix Pathway System (GePS, Genomatix.de). Pictures show the top 50 most connected genes co-cited in PubMed abstracts in the same sentence linked to function terms (most relevant genes / interactions). Orange represents upregulated genes and green represents downregulated genes in PN lesional vs. non-lesional skin. Significant nodes included the proliferation marker Ki67 (MKI67), IL-1 family members IL36G and IL1A, as well as CXCL8 and CDKN1A (C). Number of genes and intracluster correlation in modules identified from weighted co-expression gene network (WCGNA) analysis of non-lesional and lesional PN skin (D). Functions enriched in major co-expression modules from PN skin (modules #5, #6, #8) (E). [Figure 1D]Figures 1A-1E show that prurigo nodularis (PN) is characterized by immune activation and abnormal keratinocyte differentiation. Number of differentially expressed genes (DEGs) in PN lesional vs. non-lesional skin (n=62, FC>=2 or FC<=-2, FDR<=0.1) (A). Enriched GO categories in PN lesional skin (B). Literature-based gene network derived from the top 1,000 DEGs in PN skin generated using Genomatix Pathway System (GePS, Genomatix.de). Pictures show the top 50 most connected genes co-cited in PubMed abstracts in the same sentence linked to function terms (most relevant genes / interactions). Orange represents upregulated genes and green represents downregulated genes in PN lesional vs. non-lesional skin. Significant nodes included the proliferation marker Ki67 (MKI67), IL-1 family members IL36G and IL1A, as well as CXCL8 and CDKN1A (C). Number of genes and intracluster correlation in modules identified from weighted co-expression gene network (WCGNA) analysis of non-lesional and lesional PN skin (D). Functions enriched in major co-expression modules from PN skin (modules #5, #6, #8) (E). [Figure 1E]Figures 1A-1E show that prurigo nodularis (PN) is characterized by immune activation and abnormal keratinocyte differentiation. Number of differentially expressed genes (DEGs) in PN lesional vs. non-lesional skin (n=62, FC>=2 or FC<=-2, FDR<=0.1) (A). Enriched GO categories in PN lesional skin (B). Literature-based gene network derived from the top 1,000 DEGs in PN skin generated using Genomatix Pathway System (GePS, Genomatix.de). Pictures show the top 50 most connected genes co-cited in PubMed abstracts in the same sentence linked to function terms (most relevant genes / interactions). Orange represents upregulated genes and green represents downregulated genes in PN lesional vs. non-lesional skin. Significant nodes included the proliferation marker Ki67 (MKI67), IL-1 family members IL36G and IL1A, as well as CXCL8 and CDKN1A (C). Number of genes and intracluster correlation in modules identified from weighted co-expression gene network (WCGNA) analysis of non-lesional and lesional PN skin (D). Functions enriched in major co-expression modules from PN skin (modules #5, #6, #8) (E). [Figure 2A] Figures 2A-2C show enriched transcriptomic cell signatures and overlap with psoriasis and AD. Cell type inference analysis for non-lesional (NL) and lesional (L) PN skin samples using xCell. Enriched cell signatures are shown in red, whereas underrepresented cell signatures are shown in blue. Bars on the left side show statistical differences in enrichment between lesional and non-lesional PN skin, with colors representing different p-value thresholds (A). Comparison of PN-associated DEGs to DEGs in psoriasis (Pso) and atopic dermatitis (AD) for increased and decreased DEGs (B). Correlation analysis between effect sizes in PN lesions and effect sizes in psoriasis (Pso) and atopic dermatitis (AD). Spearman's rank correlation was included. Genes significant in the x-axis, y-axis, and both axes are colored red / blue / purple, respectively (C). [Figure 2B]Figures 2A-2C show enriched transcriptomic cell signatures and overlap with psoriasis and AD. Cell type inference analysis for non-lesional (NL) and lesional (L) PN skin samples using xCell. Enriched cell signatures are shown in red, whereas underrepresented cell signatures are shown in blue. Bars on the left side show statistical differences in enrichment between lesional and non-lesional PN skin, with colors representing different p-value thresholds (A). Comparison of PN-associated DEGs to DEGs in psoriasis (Pso) and atopic dermatitis (AD) for increased and decreased DEGs (B). Correlation analysis between effect sizes in PN lesions and effect sizes in psoriasis (Pso) and atopic dermatitis (AD). Spearman's rank correlation was included. Genes significant in the x-axis, y-axis, and both axes are colored red / blue / purple, respectively (C). [Figure 2C] Figures 2A-2C show enriched transcriptomic cell signatures and overlap with psoriasis and AD. Cell type inference analysis for non-lesional (NL) and lesional (L) PN skin samples using xCell. Enriched cell signatures are shown in red, whereas underrepresented cell signatures are shown in blue. Bars on the left side show statistical differences in enrichment between lesional and non-lesional PN skin, with colors representing different p-value thresholds (A). Comparison of PN-associated DEGs to DEGs in psoriasis (Pso) and atopic dermatitis (AD) for increased and decreased DEGs (B). Correlation analysis between effect sizes in PN lesions and effect sizes in psoriasis (Pso) and atopic dermatitis (AD). Spearman's rank correlation was included. Genes significant in the x-axis, y-axis, and both axes are colored red / blue / purple, respectively (C). [Figure 3A]Figures 3A-3B show transcriptomic changes associated with the anti-IL31R inhibitor nemolizumab. Principal component analysis (PCA) of transcriptomic data from PN biopsies before and after 12 weeks of a prospective, placebo-controlled, double-blind clinical trial with the anti-IL-31R inhibitor nemolizumab. Different colors represent different treatment arms, with lesional samples shown as triangles and non-lesional skin shown as circles (A). Heatmap showing 2-way clustering (using genes that were differentially expressed between non-lesional and lesional skin at baseline) for all samples (B). [Figure 3B] Figures 3A-3B show transcriptomic changes associated with the anti-IL31R inhibitor nemolizumab. Principal component analysis (PCA) of transcriptomic data from PN biopsies before and after 12 weeks of a prospective, placebo-controlled, double-blind clinical trial with the anti-IL-31R inhibitor nemolizumab. Different colors represent different treatment arms, with lesional samples shown as triangles and non-lesional skin shown as circles (A). Heatmap showing 2-way clustering (using genes that were differentially expressed between non-lesional and lesional skin at baseline) for all samples (B). [Figure 4A] Figures 4A-4C show the effect of nemolizumab on PN-related transcriptome changes. Three-way Bend plot of increases and decreases in DEGs in PN skin and their overlap with DEGs in nemolizumab and placebo cohorts (compared to baseline) (A). Correlation analysis between different groups (PN baseline vs. placebo and nemolizumab DEGs) (Spearman's rank correlation) (B). [Figure 4B] Figures 4A-4C show the effect of nemolizumab on PN-related transcriptome changes. Three-way Bend plot of increases and decreases in DEGs in PN skin and their overlap with DEGs in nemolizumab and placebo cohorts (compared to baseline) (A). Correlation analysis between different groups (PN baseline vs. placebo and nemolizumab DEGs) (Spearman's rank correlation) (B). [Figure 4C]Figures 4A-4C show the effect of nemolizumab on PN-related transcriptome changes. Three-way Bend plot of increases and decreases in DEGs in PN skin and their overlap with DEGs in nemolizumab and placebo cohorts (compared to baseline) (A). Correlation analysis between different groups (PN baseline vs. placebo and nemolizumab DEGs) (Spearman's rank correlation) (B). [Figure 5A] 5A-5D show that nemolizumab treatment results in normalized epidermal differentiation and reduced IL-31 / Th2 responses in PN skin. Nemolizumab treatment resulted in reduced IL-31 and IL-13 responses in PN skin compared to placebo, along with reduced expression of IL-17A-responsive genes (A). Nemolizumab treatment was accompanied by reduced transcriptomic signatures of Th1, Th17, and Th2 cells (B). Cross-comparison of transcriptomic responses in PN skin against cellular signatures derived from single-cell data of healthy epidermis demonstrated that nemolizumab treatment resulted in normalization of epidermal gene expression associated with the differentiation layer of the epidermis (KRT10+), corresponding to normalization of epidermal differentiation. The different nomenclature corresponds to different layers of the epidermis with "basal" corresponding to KRT5+ basal cells, KRT10+ "differentiated" corresponding to the epidermal spinous layer, and "keratinized" corresponding to the granular layer (FLG+) (C). Transcription factor binding sites (TFBS) of PN-associated DEGs with greater normalization between nemolizumab downregulated genes compared to placebo (5D). [Figure 5B]5A-5D show that nemolizumab treatment results in normalized epidermal differentiation and reduced IL-31 / Th2 responses in PN skin. Nemolizumab treatment resulted in reduced IL-31 and IL-13 responses in PN skin compared to placebo, along with reduced expression of IL-17A-responsive genes (A). Nemolizumab treatment was accompanied by reduced transcriptomic signatures of Th1, Th17, and Th2 cells (B). Cross-comparison of transcriptomic responses in PN skin against cellular signatures derived from single-cell data of healthy epidermis demonstrated that nemolizumab treatment resulted in normalization of epidermal gene expression associated with the differentiation layer of the epidermis (KRT10+), corresponding to normalization of epidermal differentiation. The different nomenclature corresponds to different layers of the epidermis with "basal" corresponding to KRT5+ basal cells, KRT10+ "differentiated" corresponding to the epidermal spinous layer, and "keratinized" corresponding to the granular layer (FLG+) (C). Transcription factor binding sites (TFBS) of PN-associated DEGs with greater normalization between nemolizumab downregulated genes compared to placebo (5D). [Figure 5C] 5A-5D show that nemolizumab treatment results in normalized epidermal differentiation and reduced IL-31 / Th2 responses in PN skin. Nemolizumab treatment resulted in reduced IL-31 and IL-13 responses in PN skin compared to placebo, along with reduced expression of IL-17A-responsive genes (A). Nemolizumab treatment was accompanied by reduced transcriptomic signatures of Th1, Th17, and Th2 cells (B). Cross-comparison of transcriptomic responses in PN skin against cellular signatures derived from single-cell data of healthy epidermis demonstrated that nemolizumab treatment resulted in normalization of epidermal gene expression associated with the differentiation layer of the epidermis (KRT10+), corresponding to normalization of epidermal differentiation. The different nomenclature corresponds to different layers of the epidermis with "basal" corresponding to KRT5+ basal cells, KRT10+ "differentiated" corresponding to the epidermal spinous layer, and "keratinized" corresponding to the granular layer (FLG+) (C). Transcription factor binding sites (TFBS) of PN-associated DEGs with greater normalization between nemolizumab downregulated genes compared to placebo (5D). [Figure 5D] 5A-5D show that nemolizumab treatment results in normalized epidermal differentiation and reduced IL-31 / Th2 responses in PN skin. Nemolizumab treatment resulted in reduced IL-31 and IL-13 responses in PN skin compared to placebo, along with reduced expression of IL-17A-responsive genes (A). Nemolizumab treatment was accompanied by reduced transcriptomic signatures of Th1, Th17, and Th2 cells (B). Cross-comparison of transcriptomic responses in PN skin against cellular signatures derived from single-cell data of healthy epidermis demonstrated that nemolizumab treatment resulted in normalization of epidermal gene expression associated with the differentiation layer of the epidermis (KRT10+), corresponding to normalization of epidermal differentiation. The different nomenclature corresponds to different layers of the epidermis with "basal" corresponding to KRT5+ basal cells, KRT10+ "differentiated" corresponding to the epidermal spinous layer, and "keratinized" corresponding to the granular layer (FLG+) (C). Transcription factor binding sites (TFBS) of PN-associated DEGs with greater normalization between nemolizumab downregulated genes compared to placebo (5D). [Figure 6A] Figures 6A-6B show nemolizumab-driven reductions in pruritus scoring, accompanied by tighter clustering of PN samples in PCA analysis, after 12 weeks of treatment. Peak pruritus numerical rating scale (PP-NRS) was overlaid onto transcriptomic data from PN lesional skin and assessed using PCA analysis. The nemolizumab group is shown as large dots and the placebo group as small dots. Baseline (top) and week 12 (bottom) of treatment are shown (A). Nemolizumab treatment resulted in tighter clustering of PN samples in PCA analysis compared to biopsy samples from the placebo-treated cohort (B). [Figure 6B]Figures 6A-6B show nemolizumab-driven reductions in pruritus scoring, accompanied by tighter clustering of PN samples in PCA analysis, after 12 weeks of treatment. Peak pruritus numerical rating scale (PP-NRS) was overlaid onto transcriptomic data from PN lesional skin and assessed using PCA analysis. The nemolizumab group is shown as large dots and the placebo group as small dots. Baseline (top) and week 12 (bottom) of treatment are shown (A). Nemolizumab treatment resulted in tighter clustering of PN samples in PCA analysis compared to biopsy samples from the placebo-treated cohort (B). [Figure 7] Figure 1 shows the cytokine signature in PN in placebo vs. nemolizumab-treated skin. A literature-based network generated using the Genomatix Pathway System (GePS, genomatix.de), with a function term co-citation filter, shows key cytokines as key nodes in PN skin. [Figure 8] 1 shows the expression of selected TFs in PN skin before and after nemolizumab treatment. [Figure 9] PN, distances between PCA components before and after nemolizumab treatment are shown. [Figure 10] Clinical scores of patients selected for mass spectrometry are shown. Left panel: PNR score at baseline. Right panel: NRS change at 12 weeks. [Figure 11] Enriched canonical pathways from mass spectrometry are shown. Enriched canonical pathways were sorted according to (right) z-score only, (left) z-score and p-value (p<0.05). [Figure 12] Upstream regulator analysis from mass spectrometry. Top: Upstream regulators sorted according to right) z-score only, left) z-score and p-value (p<0.05). [Figure 13] The enriched biological functions, as identified by mass spectrometry, are shown. The enriched biological function ontologies were sorted according to z-score and p-value (p<0.05). DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0069] Detailed Description Described herein is the treatment and prevention of prurigo nodularis (PN) using anti-IL-31RA antibodies (e.g., nemolizumab) and previously unknown biomarkers and gene signatures associated with PN. The disclosed biomarkers, including differentially expressed genes (DEGs), PN-specific gene ontologies (GOs), and other inflammatory and hyperproliferative markers, can be used to identify subjects with PN, determine whether the subject is likely to respond to treatment (e.g., with anti-IL-31RA antibodies), and track the subject's responsiveness to treatment. The disclosed treatments and preventions achieve therapeutic endpoints (e.g., normalizing DEGs, normalizing epidermal hyperproliferation, normalizing epidermal differentiation, and / or reducing inflammatory responses in the skin) that were previously unknown or unobtainable with conventional treatments for PN.

[0070] I. Definition It is to be understood that the terminology used herein is for the purpose of describing particular embodiments only, and is not intended to be limiting.

[0071] Unless otherwise defined, technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art. Unless otherwise specified, materials and / or methodologies known to those of ordinary skill in the art may be utilized in carrying out the methods described herein, based on the guidance provided herein.

[0072] As used herein, the singular terms "a," "an," and "the" include plural referents unless the context clearly dictates otherwise. Reference to an object in the singular is not intended to mean "one and only one," unless expressly so stated, but rather "one or more."

[0073] As used herein, when used in conjunction with a numerical value, "about" means the numerical value set forth, as well as plus or minus 10% of the numerical value. For example, "about 10" should be understood as both "10" and "from 9 to 11."

[0074] As used herein, "and / or" refers to and includes any and all possible combinations with one or more of the associated listed items, as well as the lack of combinations when interpreted in the alternative ("or").

[0075] As used herein, a phrase in the form "A / B" or in the form "A and / or B" means (A), (B), or (A and B), and a phrase in the form "at least one of A, B, and C" means (A), (B), (C), (A and B), (A and C), (B and C), or (A, B and C).

[0076] As used herein, the phrase "therapeutically effective amount" with respect to an anti-IL31R antibody (e.g., nemolizumab) refers to a dose of an antibody that provides the specific pharmacological effect for which the drug is administered to a subject in need of such treatment. A therapeutically effective amount may be effective to reduce, improve, or eliminate itching, scratching, and / or the formation of lesions or nodules, and / or improve the quality of life in a subject with PN. It is emphasized that in certain cases, a therapeutically effective amount of an anti-IL31R (e.g., nemolizumab) may not always be effective in treating PN in an individual subject, even if it is considered a therapeutically effective amount by those skilled in the art. Those skilled in the art can adjust what is considered a therapeutically effective amount according to standard practices as necessary to treat a particular subject. A therapeutically effective amount may vary based on, for example, the age and weight of the subject, and / or the general health of the subject, and / or the severity of the subject's PN.

[0077] The terms "treat," "treatment," or "treating" as used herein with respect to PN refer to reducing, ameliorating, or eliminating itching, scratching, and / or the formation of lesions or nodules, and / or improving quality of life in a subject with PN.

[0078] The terms "prevent," "preventing," or "prevention" as used herein with respect to PN refer to eliminating or reducing the risk of developing a lesion or nodule, or preventing the development of a biomarker signature of the present disclosure associated with PN. Prevention can also refer to the prevention of PN flares or recurrences after an initial flare has been treated or cured.

[0079] As used herein, the terms "individual," "subject," and "patient" are used interchangeably and refer to any individual mammalian subject, e.g., a bovine, canine, feline, equine, or human. In specific embodiments, a subject, individual, or patient is a human.

[0080] II. Prurigo Nodularis (PN) and Biomarkers Prurigo nodularis (or "PN") is a skin disorder that causes hard, itchy lumps (nodules) to form on the skin. The itching (pruritus) can be intense and cause bleeding or painful scratching. Scratching can cause more skin lesions to appear. The itching is made worse by heat, sweating, or irritation from clothing. In some cases, people with PN have a history of other diseases, including eczema (atopic dermatitis), diabetes, lymphoma, HIV infection, severe anemia, or kidney disease. The exact cause of PN was previously not well understood. It was thought that nodules were likely to form if the skin was injured or irritated in some way. Thus, the act of a person scratching their skin could cause nodules to form. However, originally, the cause of the skin becoming so itchy was unknown.

[0081] Approximately 50% of people with PN have a history of atopy. The main symptom of PN is the formation of hard, very itchy lumps (nodules) on the skin. The size of the nodules can range from very small to about half an inch in diameter. The nodules often have a rough, dry top and can range from a few to hundreds. The nodules most commonly form on the outer arms, shoulders, and legs. They can also form on the neck and torso, and rarely on the face and palms. They may be lighter or darker in color than the surrounding skin. Scarring may occur after the nodules begin to heal. Symptoms of PN can begin at any age, but are most common in adults after age 50. People with PN can be very concerned about the appearance of the nodules, and the intensely itchy skin can interfere with sleep or daily activities. This can cause people with PN to develop stress and depression.

[0082] Pruritus refers to the itching and / or itchy sensation of the skin. Pruritus may be caused by other diseases or conditions such as PN or dry skin. In some cases, pruritus involves generalized skin itching over the entire body. In some cases, pruritus is localized to a specific area of ​​the body, such as the arms or legs. Pruritus may be chronic or acute. Symptoms of pruritus include, but are not limited to, peeling, redness, bumps, spots, blisters, dry skin, cracked skin, and a leathery or scaly texture to the skin. In some cases, pruritus does not result in detectable changes to the skin. Behavioral responses to pruritus include, but are not limited to, scratching the skin and / or massaging the skin. In some cases, scratching the skin can result in peeling ranging from mild to severe. In some cases, patients with pruritus refrain from scratching and / or massaging the skin. Conventional treatments for PN include, but are not limited to, skin moisturizers, topical emollients, antihistamines such as diphenhydramine, topical corticosteroids, topical calcineurin inhibitors, and phototherapy, narrow-band UVB, and systemic immunosuppressants such as cyclosporine or methotrexate.

[0083] The present disclosure illuminates for the first time underlying gene expression patterns that are associated with PN and can be used to diagnose PN, identify subjects likely to respond to treatment (e.g., treatment with an anti-IL-31RA antibody), and determine whether a subject is responding appropriately to treatment.

[0084] In particular, the present disclosure shows that subjects with PN may differentially express at least 5,934 genes (known as differentially expressed genes or DEGs), as shown in FIG. 1A and Table 1 below. This differential gene expression may be observed in the skin of the subject, particularly in skin samples that contain or consist of nodules or lesions. In some embodiments, the differential gene expression may be at least 1, at least 2, at least 3, at least 4, at least 5, at least 10, at least 20, at least 30, at least 40, at least 50, at least 60, at least 70, at least 80, at least 90, at least 100, at least 150, at least 200, at least 250, at least 300, at least 350, at least 400, at least 450, at least 500, at least 550, at least 600, at least 650, at least 700, at least 750, at least 800, at least 800, at least 900, at least 1000, at least 1500, at least 2000, at least 2500, at least 3000, at least 3500, at least 4000, at least 4500, at least 5000, at least 5500, at least 6000, at least 6500, at least 7000, at least 7500, at least 8000, at least 9000, at least 10 ... DEGs of the present disclosure may be differentially expressed in subjects with PN, including 50, at least 900, at least 950, at least 1000, at least 1100, at least 1200, at least 1300, at least 1400, at least 1500, at least 1600, at least 1700, at least 1800, at least 1900, at least 2000, at least 2100, at least 2200, at least 2300, at least 2400, or at least 2500, and up to 2500, 3000, 3500, 4000, 4500, 5000, 5500, or about 6000. Of these DEGs, 2,060 may be increased (i.e., overexpressed) and 3,874 may be decreased (i.e., not expressed). Genes that may be most increased include: KRT6C, which may be increased by at least 100-fold, at least 150-fold, at least 200-fold, at least 250-fold, at least 300-fold, at least 350-fold, at least 400-fold, at least 450-fold, at least 500-fold, at least 550-fold, or 588-fold compared to expression levels in a sample (e.g., a skin sample) from an individual without PN; DEFB4A, which may be increased by at least 25-fold, at least 50-fold, at least 75-fold, at least 100-fold, at least 125-fold, or at least 150-fold compared to expression levels in a sample (e.g., a skin sample) from an individual without PN; and KRT16, which may be increased by at least 10-fold, at least 20-fold, at least 30-fold, at least 40-fold, at least 50-fold, at least 60-fold, at least 70-fold, at least 80-fold, or at least 90-fold compared to expression levels in a sample (e.g., a skin sample) from an individual without PN.

[0085] The genes that were decreased include: LCE5A, which may be decreased by at least 2-fold, at least 3-fold, at least 4-fold, at least 5-fold, at least 6-fold, at least 7-fold, at least 8-fold, at least 9-fold, at least 10-fold, or at least 11-fold compared to expression levels in a sample (e.g., a skin sample) from an individual without PN; and AQP7, which may be reduced by at least 2-fold, at least 3-fold, at least 4-fold, at least 5-fold, at least 6-fold, at least 7-fold, or at least 7.9-fold compared to expression levels in a sample from an individual without PN (e.g. a skin sample).

[0086] Genes encoding cytokines may also be overexpressed in subjects suffering from PN. For cytokine genes, the most prominent upregulated genes are IL-36 family members and IL-20 family members. These upregulated or overexpressed genes may include: ●IL36A (for example, about 6.8 times, FDR = 1.8 × 10-4 ), ●IL36G (e.g., about 8.4 times, FDR = 3.9 × 10 -25 ), ●IL19 (e.g., about 5.1 times, FDR = 7.4 × 10 -4 ), ●IL20 (for example, about 3.5 times, FDR = 1.7 × 10 -3 ), ●IL22 (e.g., about 2.7 times, FDR = 2.9 × 10 -2 ), ●IL24 (e.g., about 5.8 times, FDR = 3.8 × 10 -10 ), and ●IL26 (e.g., about 4.9 times, FDR = 3.3 × 10 -3 ). Each of these IL-36 and IL-20 family member cytokine genes can be overexpressed by at least about 2-fold, at least about 2.5-fold, at least about 3-fold, at least about 3.5-fold, at least about 4-fold, at least about 4.5-fold, at least about 5-fold, at least about 5.5-fold, at least about 6-fold, at least about 6.5-fold, at least about 7-fold, at least about 7.5-fold, at least about 8-fold, or at least about 8-5-fold compared to expression levels in a sample (e.g., a skin sample) from an individual without PN.

[0087] Other factors that may be upregulated or overexpressed include IL1A (e.g., about 4.7-fold, FDR=1.0×10 -12 ) and IL1B (e.g., about 4.1-fold, FDR = 3.7 × 10 -6 In addition, the IL4R gene has a gene expression level of about 2.6 times (FDR=6.3×10 -19 ) may be increased. Table 1 at the end of the Examples section of this specification provides a more comprehensive list of DEGs.

[0088] In addition to the aforementioned genes, the present disclosure also shows that certain plasma markers or signatures may be altered as a result of successful treatment of PN with an anti-IL-31RA antibody such as nemolizumab. Such markers or signatures may be detectable, for example, by mass spectrometry and other protein evaluation methods (e.g., ELISA, Western blot, etc.). Circulating plasma protein markers or signatures that may be modulated as a result of treatment with an anti-IL-31RA antibody such as nemolizumab may include leukocyte migration and cell migration, IL-6 pathway, vascular endothelial growth factor (VEGF) pathway, STAT3 (signal transducer and activator of transcription 3) pathway, STAT5b (signal transducer and activator of transcription 5b) pathway, TGFB1 (transforming growth factor beta-1) pathway, and neuroontology.

[0089] The STAT3 pathway, a direct target of IL-31 signaling, is also inhibited in the nemolizumab responder signature of the present disclosure, suggesting target engagement. STAT3 activity and expression may be relatively high in subjects with PN or in subjects prior to initiating treatment with an anti-IL-31RA antibody, such as nemolizumab, compared to individuals or populations without PN.

[0090] The amount of circulating proinflammatory cytokines may be relatively high in subjects with PN, or in subjects prior to initiating treatment with an anti-IL-31RA antibody, such as nemolizumab, compared to individuals or populations without PN. Such proinflammatory cytokine signatures include, but are not limited to, IL-6 and VEGF. In PN subjects receiving treatment with an anti-IL-31RA antibody, such as nemolizumab, the treatment may result in a decrease in one or both of IL-6 and VEGF, or a decrease or inhibition of the signaling pathways of IL-6, VEGF, or both, compared to baseline levels. The baseline levels may be determined with respect to (i) a control sample obtained from one or more individuals (i.e., a population) without PN, or (ii) a biological sample obtained from the subject prior to administration of the anti-IL-31RA antibody.

[0091] TGFB1 activity is also inhibited in the nemolizumab responder signature of the present disclosure. TGFB1 activity and expression may be relatively high in subjects with PN or in subjects prior to initiating treatment with an anti-IL-31RA antibody, such as nemolizumab, compared to individuals or populations without PN.

[0092] For purposes of plasma protein markers of the present disclosure, the amount of a protein marker of the present disclosure in the plasma of a subject with PN may be at least 2-fold, at least 3-fold, at least 4-fold, at least 5-fold, at least 6-fold, at least 7-fold, at least 8-fold, at least 9-fold, at least 10-fold, at least 15-fold, at least 20-fold, at least 25-fold, at least 30-fold, at least 35-fold, at least 40-fold, at least 45-fold, or at least 50-fold higher than the baseline level. The baseline level may be determined with respect to (i) a control sample obtained from one or more individuals (i.e., a population) without PN, or (ii) a biological sample obtained from the subject prior to administration of an anti-IL-31RA antibody. Similarly, after a subject with PN has been treated with an anti-IL-31RA antibody, such as nemolizumab (e.g., 2, 4, 6, 8, 10, or 12 weeks after administration of the antibody), the amount of a plasma protein marker of the present disclosure in the subject may be decreased by at least 2-fold, at least 3-fold, at least 4-fold, at least 5-fold, at least 6-fold, at least 7-fold, at least 8-fold, at least 9-fold, at least 10-fold, at least 15-fold, at least 20-fold, at least 25-fold, at least 30-fold, at least 35-fold, at least 40-fold, at least 45-fold, or at least 50-fold compared to baseline levels. The baseline levels may be determined with respect to (i) a control sample obtained from one or more individuals (i.e., a population) without PN, or (ii) a biological sample obtained from the subject prior to administration of the anti-IL-31RA antibody.

[0093] In addition, neural ontologies (e.g., CREB signaling in neurons, synaptogenesis signaling pathways, glial cell death, and glial apoptosis) can be upregulated in subjects with PN and subsequently downregulated in nemolizumab responder subjects, thus highlighting the impact of IL-31 as a neuroinflammatory cytokine in PN.

[0094] Thus, the present disclosure provides a method for diagnosing PN, comprising detecting the expression levels of at least one, at least two, at least three, at least four, or at least five of the differentially expressed genes (DEGs) in Table 1 (e.g., KRT6C, DEFB4A, KRT16, LCE5A, AQP7, IL-36 family members, IL-20 family members, etc.) in a sample obtained from a subject suspected of having PN, and comparing the expression levels of the DEGs to a reference level based on the gene expression levels in a sample (e.g., a skin sample) from an individual not having PN. In some embodiments, the sample obtained from a subject suspected of having PN is a skin sample that may include a lesion or nodule. If certain genes are upregulated or overexpressed (e.g., KRT6C, DEFB4A, KRT16) and / or other genes are downregulated or underexpressed (e.g., LCE5A, AQP7), the subject may be diagnosed with PN.

[0095] The present disclosure also provides a method of determining the likelihood of a positive response to a treatment (e.g., treatment with an anti-IL-31RA antibody such as nemolizumab) in a subject with PN, comprising detecting the expression levels of at least one, at least two, at least three, at least four, or at least five of the differentially expressed genes (DEGs) of Table 1 (e.g., KRT6C, DEFB4A, KRT16, LCE5A, AQP7, IL-36 family members, IL-20 family members, etc.) in a sample obtained from a subject with PN, and comparing the expression levels of the DEGs to a reference level based on the gene expression levels in a sample (e.g., a skin sample) from an individual without PN. In some embodiments, the sample obtained from a subject with PN is a skin sample that may include a lesion or nodule. If certain genes are upregulated or overexpressed (e.g., KRT6C, DEFB4A, KRT16) and / or other genes are downregulated or underexpressed (e.g., LCE5A, AQP7), the subject is more likely to respond to the treatment.

[0096] The disclosure also provides a method of determining whether a subject with PN is responding to a treatment (e.g., treatment with an anti-IL-31RA antibody such as nemolizumab), comprising detecting the expression levels of at least one, at least two, at least three, at least four, or at least five of the differentially expressed genes (DEGs) of Table 1 (e.g., KRT6C, DEFB4A, KRT16, LCE5A, AQP7, IL-36 family members, IL-20 family members, etc.) in a sample obtained from a subject with PN before treatment is initiated, and comparing the expression levels of the DEGs to a baseline level of expression, where the baseline level of gene expression is from a sample (e.g., a skin sample) from the same individual. In some embodiments, the sample may be a skin sample that may contain a lesion or nodule, and if the expression levels of certain genes (e.g., KRT6C, DEFB4A, KRT16) decrease and / or the expression levels of other genes (e.g., LCE5A, AQP7) increase, the subject is responding to the treatment.

[0097] The disclosure also provides a method of determining whether a subject with PN is responding to treatment (e.g., treatment with an anti-IL-31RA antibody such as nemolizumab), comprising detecting one or more biomarkers selected from leukocyte migration and cell movement, IL-6 pathway, VEGF pathway, STAT3 pathway, STAT5b pathway, TGFB1 pathway, and a neuroontology of the disclosure in a post-treatment plasma sample obtained from a subject with PN administered at least one dose of an anti-IL-31RA antibody, wherein a decrease in leukocyte migration and cell movement, a decrease in IL-6 or a decrease in IL-6 pathway signaling, a decrease in VEGF or a decrease in VEGF pathway signaling, a decrease in STAT3 or a decrease in STAT3 pathway signaling, a decrease in STAT5b or a decrease in STAT5b pathway signaling, a decrease in TGFB1 or a decrease in TGFB1 pathway signaling, or an increase in a neuroontology of the disclosure indicates responsiveness to treatment relative to a baseline amount, the baseline amount being determined from a plasma sample obtained from the same subject before treatment was initiated.

[0098] This disclosure also defines for the first time the biological processes enriched in the PN skin gene ontology (GO). The most prominent GO categories related to PN are: ● "Cornified envelope" (FDR=1.5×10 -12 ), ●“Epithelial cell differentiation” (FDR=6.4×10 -10 ), - Keratinization (FDR=1.6×10 -12 ), ● "Peptidase regulatory factor activity" (FDR = 1.1 x 10 -4 ), ● "Interleukin-4 and 13 signaling" (FDR = 6.8 × 10 -7 ), "Interferon alpha / beta signaling" and "response to interferon gamma" (FDR=4.1×10 -7 and FDR = 4.1 × 10 -6 ), ● "IL23 pathway" (FDR = 2 × 10 -5 ), and ● "Metaphase and anaphase of mitosis" (FDR = 3.8 × 10 -10 ).

[0099] These GO categories are defined in more detail by Figure 1B and Table 2 at the end of the Examples section of this specification. These categories reflect the hyperproliferative nature of PN, which was found to be associated with alterations in epidermal differentiation and inflammatory components. For example, an inflammatory network including the proliferation marker Ki67 (MKI67), the cell cycle gene CDKN1A, and IL-1 and IL-36 was found to be potentially relevant to the pathogenesis of PN, as shown in Figure 1C.

[0100] The present disclosure also discloses 20 co-expression modules identified in non-lesional skin and 10 clusters in lesional PN skin, as shown in FIG. 1D and Table 3 at the end of the Examples section of this specification. "Immune response" (FDR=1.8×10 -47 ), “protective response” (FDR = 1.2 × 10 -39 ), the most prominent involved immunological processes (module #8), including "cell cycle" (FDR=2.9×10 -94 ), "DNA metabolic process" (FDR = 8.7 × 10 -67 ), and “Epidermal Development” (FDR 3.5 × 10 -10 ), “keratinization” (FDR=1.7×10 -6 Different functions could be defined for these co-expressed gene modules, especially for those co-expressed modules in PN lesional skin, including epidermal processes such as “extracellular matrix” (module #5) and “metastasis” (module #6). See also Figure 1E. Another notable finding is the low expression of “extracellular matrix” (FDR=1.16×10 -59 , module #2), including genes such as MMP14, MMP16, COL1A1, COL1A2, and COL3A1, which were modestly elevated in lesional skin (FC ≥ 1.4, FDR ≤ 6 × 10), consistent with an association between PN and dermal fibrosis. -2 ).

[0101] The present disclosure also provides specific cell type signatures for non-lesional and lesional PN skin samples. As shown in Figure 2A, enrichment of transcriptomic signatures related to epithelial cells and keratinocytes was observed (p<0.001 and p<0.0001, respectively). Subjects with PN may show a Th2-related signature (p<0.0001) that coincides with the enriched GO category of IL-4 / IL-13 (as described above and shown in Figure 1B). Other inflammatory signatures, such as macrophages (p<0.01), may be more diverse.

[0102] As shown in Figure 2B, a three-way comparison of the PN transcriptomes of atopic dermatitis (AD) and psoriasis showed that all three diseases shared significant overlap for both up- and down-regulated genes. However, the effect size correlation in lesional skin was more pronounced between PN and psoriasis (Spearman correlation ρ = 0.64) than between PN and AD (ρ = 0.55). Commonly up-regulated genes in both psoriasis and PN include those involved in cytokine activity (CCL3, CXCL10, IFNG, IL12B, IL19, IL1B, IL20, etc.) and keratinization (KRT16, KRT17, LCE3A, LCE3E, etc.) (Table 4).

[0103] Thus, the present disclosure provides a method for identifying or diagnosing PN based on gene ontology, co-expression module, and / or gene signature. For example, a subject may present the PN skin gene ontology (GO) shown in FIG. 1B or Table 2. Additionally or alternatively, the skin sample may also show upregulation or overexpression of Ki67 (MKI67), CDKN1A, and / or inflammatory network including IL-1 and IL-36. Additionally or alternatively, the subject may present the co-expression module as shown in FIG. 1D or Table 3. Additionally or alternatively, a subject with PN may present (e.g., in a skin sample) the transcriptome or Th2 signature disclosed in FIG. 2A.

[0104] The expression levels of the genes and markers disclosed herein may be determined by any suitable method known in the art, including, but not limited to, RT-qPCR, RT-PCR, RNA-seq, Northern blotting, serial analysis of gene expression (SAGE), DNA or RNA microarrays, and in situ hybridization. At the protein level, the biomarkers of the present disclosure may be detected or measured using, for example, Western blotting, ELISA (enzyme-linked immunosorbent assay), surface plasmon resonance, and mass spectrometry.

[0105] Subjects with or suspected of having PN that display any of the DEGs, gene ontology, co-expression modules, or gene signatures of the present disclosure are suitable for treatment or prevention with an anti-IL-31RA antibody, such as nemolizumab, as described in further detail herein.

[0106] The present disclosure provides a combination of: (1) a pharmaceutical composition for use in the treatment or prevention of prurigo nodularis (PN), comprising an anti-IL31RA antibody (e.g., nemolizumab or a fragment or variant thereof) as an active ingredient; and (2) a diagnostic agent for detecting the expression level of at least one gene selected from the genes disclosed in Table 1 in a subject suspected of having PN, compared to a reference expression level of the at least one gene.

[0107] III. Therapeutic Antibodies and Interleukin-31 Receptor Subunit Alpha (IL-31RA) Interleukin-31 (IL-31) is a neuroinflammatory cytokine that can activate both structural and immune cells and peripheral nerves. IL-31 has been implicated in a number of chronic inflammatory diseases, including atopic dermatitis. IL-31 is produced by a variety of cells, including type 2 helper (Th2) T cells. IL-31 signals through a receptor complex consisting of IL-interleukin 31 receptor subunit alpha ("IL-31RA", also known as NR10, glm-r, and GPL) and oncostatin M receptor beta (OSMRβ), which are expressed in immune and epithelial cells, as well as a subset of neurons.

[0108] When IL-31RA functions as an IL-31 receptor, it forms a heterodimer with oncostatin M receptor (OSMR). There are several known splicing variants of human-derived IL-31RA (WO00 / 075314): NR10.1 is composed of 662 amino acids and contains a transmembrane domain. NR10.2 is a soluble receptor-like protein composed of 252 amino acids without a transmembrane domain. Further known IL-31RA splicing variants that function as transmembrane receptor proteins include NR10.3 and IL-31RAv3. Preferred IL-31RA variants include NR10.3 (ILRAv4 (also referred to as Nat Immunol 5, 752-60, 2004) and IL-31RAv3. NR10.3 (IL31RAv4) consists of 662 amino acids (WO00 / 075314, Nat Immunol 5, 752-60, 2004), and IL31RAv3 consists of 732 amino acids (GenBank accession number: NM-139017).

[0109] The amino acid sequence of IL31RAv4 is as follows: (Sequence number 1).

[0110] The amino acid sequence of IL31RAv3 is as follows: (Sequence number 2).

[0111] Mouse-derived IL-31RA contains the following amino acid sequence: (Sequence number 3).

[0112] Cynomolgus monkey-derived IL-31RA contains the following amino acid sequence: (Sequence number 4).

[0113] For purposes of this disclosure, an anti-IL-31RA antibody (ie, a therapeutic antibody) such as nemolizumab must bind at least human IL-31RA or a splice variant thereof.

[0114] As used herein, the term "antibody" refers collectively to immunoglobulin or immunoglobulin-like molecules, including IgA, IgD, IgE, IgG, and IgM, combinations thereof, or fragments thereof. Antibody fragments may include, for example, Fab fragments and single-chain variable fragments (scFv). Antibodies generally comprise a heavy (H) chain and a light (L) chain interconnected by disulfide bonds. There are two types of light chains: lambda (λ) and kappa (κ). There are five main heavy chain classes (or isotypes) that determine the functional activity of antibody molecules: IgG, IgA, IgM, IgD, and IgE. Each heavy and light chain contains a constant region and a variable region (also known as a "domain"). In combination, the heavy and light chain variable regions, also referred to as "Fab regions", specifically bind to a given antigen. The light and heavy chain variable regions contain a "framework" region interrupted by three hypervariable regions, also referred to as "complementarity determining regions" or "CDRs". The extent of the framework regions and CDRs are defined (see Kabat et al., Sequences of Proteins of Immunological Interest, USDepartment of Health and Human Services, 1991). The Kabat database is currently maintained online. The sequences of the framework regions of different light or heavy chains are relatively conserved within a species, and the framework regions act to form a scaffold for orienting the CDRs through non-covalent interactions between the chains.

[0115] CDRs are primarily responsible for binding to an epitope on an antigen. The CDRs of each chain are typically referred to as CDR1, CDR2, and CDR3, are numbered sequentially starting from the N-terminus, and are typically identified by the chain in which the particular CDR is located. Thus, HCDR3 is located in the variable domain of the heavy chain of the antibody in which it is found, while LCDR1 is the CDR1 from the variable domain of the light chain of the antibody in which it is found. Antibodies that bind to IL-31RA have a specific VDR1. H Area and V LEach CDR has a specific CDR sequence and therefore a specific CDR sequence. Antibodies with different specificities generally have different CDRs. It is the CDRs that differ from antibody to antibody, but only a limited number of amino acid positions in the CDR are directly involved in antigen binding. These positions in the CDR are called specificity determining residues (SDRs).

[0116] The Fc fragment region (Fc) of an antibody plays a role in regulating immune cell activity. The Fc region functions to ensure that each antibody generates an appropriate immune response to a given antigen by binding to a specific class of proteins found on specific cells, such as B lymphocytes, follicular dendritic cells, natural killer cells, macrophages, and neutrophils, called "Fc receptors." The class of heavy chains in antibodies determines their class effect, since the constant domains of the heavy chains constitute the Fc region of an antibody. Heavy chains in antibodies include alpha, gamma, delta, epsilon, and mu, which correlate with the antibody's isotypes IgA, IgG, IgD, IgE, and IgM, respectively. Thus, different isotypes of antibodies have different class effects, since their different Fc regions bind and activate different types of receptors.

[0117] IgG, the most abundant antibody isotype in human serum, has four subclasses. The four highly conserved subclasses are IgG1, IgG2, IgG3, and IgG4. The amino acid sequences of the constant regions of these peptides are known in the art, see, for example, Rutishauser, U. et al. (1968) "Amino acid sequence of the Fc region of a human gamma G-immunoglobulin" PNAS61(4):1414-1421, Shinoda et al. (1981) "Complete amino acid sequence of the Fc region of a human delta chain" PNAS78(2):785-789, and Robinson et al. (1980) "Complete amino acid sequence of a mouse immunoglobulin alpha chain (MOPC511)" PNAS77(8):4909-4913.

[0118] All therapeutic antibodies for purposes of the methods and pharmaceutical uses of the present disclosure are antibodies or fragments thereof that bind to IL-31RA, but the specific anti-IL-31RA antibody is not limited. Nemolizumab is a preferred anti-IL-31RA antibody, but other anti-IL-31RA antibodies can be used as well. Therapeutic antibodies suitable for use in the methods and pharmaceutical uses of the present disclosure can be human, humanized, or chimeric, and can be IgA, IgG (i.e., IgG1, IgG2, IgG3, and IgG4), IgD, IgE, or IgM.

[0119] Nemolizumab is a humanized monoclonal antibody that binds to IL-31RA. Nemolizumab is annotated as follows: Immunoglobulin G2-kappa, anti-[Homo sapiens IL31RA (Interleukin 31 receptor subunit alpha)], humanized monoclonal antibody; gamma 2 heavy chain (1-445) [humanized VH(Homo sapiens IGHV1-2*02 (83.70%)-(IGHD)-IGHJ5*01)[8.8.14](1-121)-Homo sapiens IGHG2*01 (CH1 C10>S(135), R12>K(137), E16>G(141), S17>G(142)(122-219), hinge C4>S(223)(220-231), CH2 H30>Q(268)(232-340), CH3 R11>Q(355), Q98>E(419)(341-445))(122-445)], (224-214')-disulfide containing kappa light chain, (1'-214') [humanized V-kappa (Homo sapiens IGKV1-39*01(82.10%)-IGKJ4*01)[6.3.9](1'-107')-Homo sapiens IGKC*01(108'-214')], dimeric (227-227":230-230")-bisdisulfide. Nemolizumab has disulfide bridges at the following positions: Intra-H(C23-C104) 22-96 148-204 261-321 367-425 22''-96'' 148''-204'' 261''-321'' 367''-425'', Intra-L(C23-C104) 23'-88' 134'-194' 23'''-88''' 134'''-194'''', Inter-HL(h5-CL126) 224-214' 224''-214'''', Inter-HH(h8,h11) 227-227'' 230-230''. Nemolizumab has N-glycosylation sites at the following positions: H CH2 N84.4: 297,297''. Nemolizumab lacks the C-terminal glycine and lysine of the H chain (CHS G1>del, K2>del).

[0120] Nemolizumab contains the following heavy chain amino acid sequence: QVQLVQSGAEVKKPGASVKVSCKASGYTFTGYIMNWVRQAPGQGLEWMGLINPYNGGTDYNPQFQDRVTITADKSTSTAYMELSSLRSEDTAVYYCARDGYDDGPYTLETWGQ GTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSNFGTQTYTCNVDHKPSNTKVDKTVERKSCVE CPPCPAPPVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTFRVVSVLTVVHQDWLNGKEYKCKVSNKGLPAPIEKTISKTKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPMLDSDGSFFLYSKLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSP (SEQ ID NO: 5).

[0121] Nemolizumab contains the following light chain amino acid sequence: DIQMTQSPSSLSASVGDRVTITCQASEDIYSFVAWYQQKPGKAPKLLIYNAQTEAQGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQHHYDSPLTFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC (SEQ ID NO: 6).

[0122] The heavy chain variable region of nemolizumab comprises the following amino acid sequence: QVQLVQSGAEVKKPGASVKVSCKASGYTFTGYIMNWVRQAPGQGLEWMGLINPYNGGTDYNPQFQDRVTITADKSTSTAYMELSSLRSEDTAVYYCARDGYDDGPYTLETWGQGTLVTVSS (SEQ ID NO: 7).

[0123] HCDR1 of nemolizumab comprises the amino acid sequence GYIMN (SEQ ID NO: 8), HCDR2 comprises the amino acid sequence LINPYNGGTDYNPQFQD (SEQ ID NO: 9), and HCDR3 comprises the amino acid sequence DGYDDGPYTLET (SEQ ID NO: 10).

[0124] The light chain variable region of nemolizumab comprises the following amino acid sequence: DIQMTQSPSSLSASVGDRVTITCQASEDIYSFVAWYQQKPGKAPKLLIYNAQTEAQGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQHHYDSPLTFGGGTKVEIKR (SEQ ID NO: 11).

[0125] LCDR1 of nemolizumab comprises the amino acid sequence QASEDIYSFVA (SEQ ID NO: 12), LCDR2 comprises the amino acid sequence NAQTEAQ (SEQ ID NO: 13), and LCDR3 comprises the amino acid sequence QHHYDSPLT (SEQ ID NO: 14).

[0126] For purposes of this disclosure, a "variant antibody" or "variant" of nemolizumab includes (i) an antibody having a heavy chain that comprises at least 55%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 97%, at least 98%, at least 99%, or 100% amino acid sequence identity to the heavy chain sequence of nemolizumab; (ii) an antibody having a light chain that comprises at least 55%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 97%, at least 98%, at least 99%, or 100% amino acid sequence identity to the light chain sequence of nemolizumab. (iii) an antibody having a variable region that contains at least 55%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 97%, at least 98%, at least 99%, or 100% amino acid sequence identity to the variable region sequence of nemolizumab, (iv) an antibody having a CDR that contains at least 55%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 97%, at least 98%, at least 99%, or 100% amino acid sequence identity to the CDR sequence of nemolizumab, and combinations thereof. For example, suitable variants include immunoglobulins or immunoglobulin-like molecules that have heavy and light chain amino acid sequences identical or substantially similar to those of nemolizumab. Other suitable therapeutic antibodies may bind to the same isoform of IL-31RA as nemolizumab (e.g., IL31-RAv3), optionally to the same epitope of IL-31RA, block or neutralize IL-31RA, or a combination thereof. Additional exemplary therapeutic antibodies are described, for example, in WO2010 / 064697.

[0127] The variants of nemolizumab and suitable therapeutic antibodies may be monoclonal or polyclonal antibodies. Such monoclonal antibodies having IL31-RA binding activity and / or neutralizing activity can be obtained, for example, by the following procedure: anti-IL31-RA monoclonal antibodies are prepared by using IL31-RA or a fragment thereof derived from a mammal such as a human or mouse as an antigen by known methods, and then antibodies having IL31-RA binding activity and / or neutralizing activity are selected from the anti-IL31-RA monoclonal antibodies thus obtained. Specifically, a desired antigen or cells expressing a desired antigen are used as a sensitizing antigen for immunization by a conventional immunization method. Anti-IL31-RA monoclonal antibodies can be prepared by fusing the obtained immune cells with known parent cells using a conventional cell fusion method and screening for monoclonal antibody-producing cells (hybridomas) by conventional screening methods. Animals to be immunized include, for example, mammals such as mice, rats, rabbits, sheep, monkeys, goats, donkeys, cows, horses, and pigs. Antigens can be prepared using known IL31-RA gene sequences according to known methods, for example, by baculovirus-based methods (e.g., WO98 / 46777). Variants of nemolizumab and suitable therapeutic antibodies include intrabodies, peptibodies, nanobodies, single domain antibodies, multispecific antibodies (e.g., bispecific antibodies, diabodies, triabodies, tetrabodies, tandem di-scFv, tandem tri-scFv), darpins, heavy chain monomers, heavy chain dimers, or single domain antibodies (i.e., V, VL ... H H-fragment or "lactam-like" antibodies).

[0128] Hybridomas can be prepared, for example, according to the method of Milstein et al. (Kohler, G. and Milstein, C., Methods Enzymol. (1981) 73:3-46). When the antigen is lowly immunogenic, the antigen may be bound to a macromolecule having immunogenicity such as albumin before immunization. The antigen used to prepare a monoclonal antibody having binding activity and / or neutralizing activity against human IL31-RA is not particularly limited, so long as it allows the preparation of an antibody having binding activity and / or neutralizing activity against human IL31-RA. For example, it is known that there are many variants of human IL31-RA, and any variant can be used as an immunogen, so long as it allows the preparation of an antibody having binding activity and / or neutralizing activity against human IL31-RA. Alternatively, under the same conditions, a peptide fragment of IL31-RA or a protein in which an artificial mutation has been introduced into the natural IL31-RA sequence may be used as an immunogen. Human IL31-RA.3 is one of the preferred immunogens for preparing antibodies having IL31-RA binding and / or neutralizing activity in the present disclosure.

[0129] The IL31-RA binding activity of therapeutic antibodies can also be determined by methods known to those skilled in the art. Methods for determining the antigen-binding activity of antibodies include, for example, ELISA (enzyme-linked immunosorbent assay), EIA (enzyme immunoassay), RIA (radioimmunoassay), and fluorescent antibody technique. For example, when using an enzyme immunoassay, an antibody-containing sample, such as purified antibody and culture supernatant of antibody-producing cells, is added to an antigen-coated plate. A secondary antibody labeled with an enzyme such as alkaline phosphatase is added, and the plate is incubated. After washing, an enzyme substrate such as p-nitrophenyl phosphate is added, and the absorbance is measured to evaluate the antigen-binding activity. The binding activity and / or neutralizing activity of therapeutic antibodies against IL31-RA can be measured, for example, by observing the effect of suppressing the proliferation of IL-31-dependent cell lines. For example, the activity of purified mouse IL-31 antibodies can be assayed by evaluating the IL-31-dependent growth of Ba / F3 cells transfected with mouse IL-31 receptor alpha and mouse OSMR genes.

[0130] Any of the anti-IL31RA antibodies (i.e., "therapeutic antibodies") disclosed herein, including nemolizumab and fragments or variants thereof, can be used to treat and / or prevent PN and achieve the therapeutic endpoints of the disclosure. Optimal dosages and routes of administration may vary.

[0131] IV. Pharmaceutical Compositions Provided herein is a pharmaceutical composition for use in the treatment or prevention of prurigo nodularis (PN), including skin lesions, nodules, or pruritus caused by PN. The pharmaceutical composition comprises, as an active ingredient, an anti-IL31RA antibody (i.e., a "therapeutic antibody"), such as nemolizumab, or a fragment or variant thereof.

[0132] The phrase "comprising nemolizumab or a fragment or variant thereof as an active ingredient" means that nemolizumab or a fragment or variant thereof is included as at least one of the active ingredients, and does not limit the percentage of the antibody. Furthermore, the therapeutic agent for PN of the present disclosure may include other ingredients that enhance the treatment or prevention of PN in combination with nemolizumab or a fragment or variant thereof. For example, the composition may include one or more of a topical corticosteroid cream or injection, an ointment containing menthol or phenol for cooling and soothing itchy skin, a capsaicin cream, an oral corticosteroid, a selective serotonin reuptake inhibitor (SSRI), and an oral antihistamine.

[0133] Pharmaceutical compositions of nemolizumab or fragments or variants thereof can be prepared as formulations according to standard methods (see, for example, Remington's Pharmaceutical Science, Mark Publishing Company, Easton, USA). Pharmaceutical compositions generally include carriers and / or excipients in addition to the antibody. For example, in some embodiments, the pharmaceutical composition comprises one or more surfactants (e.g., PEG and Tween), excipients, antioxidants (e.g., ascorbic acid), colorants, flavoring agents, preservatives, stabilizers, buffers (e.g., phosphoric acid, citric acid, and other organic acids), chelating agents (e.g., EDTA), suspending agents, isotonizing agents, binders, disintegrants, lubricants, flow enhancers, Corrigent, light anhydrous silicic acid, lactose, crystalline cellulose, mannitol, starch, carmellose calcium, carmellose sodium, hydroxypropyl cellulose, hydroxypropyl methylcellulose, polyvinyl acetal diethylaminoacetate, polyvinylpyrrolidone, gelatin, medium chain triglycerides, polyoxyethylene hydrogenated castor oil 60, sucrose, carboxymethyl cellulose, corn starch, and inorganic salts. In some embodiments, the pharmaceutical composition comprises one or more other low molecular weight polypeptides, proteins such as serum albumin, gelatin, and immunoglobulins, and amino acids such as glycine, glutamine, asparagine, arginine, and lysine.

[0134] Nemolizumab or a fragment or variant thereof may be prepared as an aqueous solution for injection, and nemolizumab or a fragment or variant thereof may be dissolved in an isotonic solution containing, for example, saline, dextrose, or other excipients or isotonic agents (i.e., tonicity agents). Isotonic agents may include, for example, D-sorbitol, D-mannose, D-mannitol, and sodium chloride. In addition, suitable solubilizers, such as alcohols (e.g., ethanol), polyalcohols (e.g., propylene glycol and PEG), and non-ionic detergents (polysorbate 80 and HCO-50), may be used simultaneously.

[0135] In some embodiments, nemolizumab or a fragment or variant thereof may be encapsulated in microcapsules (microcapsules made of hydroxymethylcellulose, gelatin, polymethylmethacrylate, etc.) and made into components of colloid drug delivery systems (liposomes, albumin microparticles, microemulsions, nanoparticles, and nanocapsules) (see, e.g., "Remington's Pharmaceutical Science 16th edition" &, Oslo Ed. (1980)). Furthermore, methods for making sustained release drugs are known and can be applied to nemolizumab or a fragment or variant thereof (Langer et al., J. Biomed. Mater. Res. (1981) 15, 167-277; Langer, Chem. Tech. (1982) 12, 98-105; U.S. Pat. No. 3,773,919; European Patent Application (EP) No. 58,481; Sidman et al., Biopolymers (1983) 22, 547-56; EP 133,988).

[0136] The pharmaceutical composition of the present disclosure may be administered orally or parenterally, but is preferably administered parenterally. Specifically, the pharmaceutical composition is administered to the patient by injection or transdermal administration. Injection includes, for example, intravenous injection, intramuscular injection, and subcutaneous injection for systemic or local administration. The pharmaceutical composition may be given to the site where inflammation and / or itch is suppressed, or the area surrounding the site by local injection or intramuscular or subcutaneous injection. In some embodiments, the pharmaceutical composition is administered at the site of one or more skin peels, lesions, or nodules, or proximal to the site of one or more skin peels, lesions, or nodules.

[0137] The administration method can be appropriately selected depending on the age, body weight, and condition of the patient. The single administration dose can be selected, for example, within the range of 0.0001 to 100 mg of antibody (e.g., nemolizumab or a fragment or variant thereof) per kg of body weight. Alternatively, for example, when the antibody is administered to a human patient, the dose of the antibody can be selected within the range of 0.001 to 1,000 mg / kg of body weight. In some embodiments, the composition may be administered in a concentration of, for example, about 0.01 to 50 mg / kg, about 0.01 mg / kg to about 0.1 mg / kg, about 0.05 mg / kg to 0.15 mg / kg, about 0.1 mg / kg to about 0.6 mg / kg, about 0.1 mg / kg to about 1 mg / kg, about 0.25 mg / kg to about 0.75 mg / kg, about 0.4 mg / kg to about 0.8 mg / kg, about 0.4 mg / kg to about 1.8 mg / kg, about 0.5 to about 2.5 mg / kg, about 0.8 mg / kg to about 2.2 mg / kg, about 1 mg / kg to about 2.5 mg / kg, about 1 mg / kg to about 3.5 mg / kg, The formulation is formulated to administer a dose containing about 1 mg / kg to about 5 mg / kg, about 2 mg / kg to about 4 mg / kg, about 2.5 mg / kg to about 10 mg / kg, about 5 mg / kg to about 10 mg / kg, about 10 mg / kg to about 20 mg / kg, about 10 mg / kg to about 40 mg / kg, about 20 mg / kg to about 50 mg / kg, about 25 mg / kg to about 75 mg / kg, about 50 mg / kg to about 100 mg / kg, or about 100 mg / kg to about 500 mg / kg, or about 100 mg / kg to about 1000 mg / kg of body weight of nemolizumab or a fragment or variant thereof. In preferred embodiments, the dosage is in the range of about 0.01 mg / kg to about 0.1 mg / kg, about 0.1 mg / kg to about 0.5 mg / kg, about 0.5 mg / kg to about 1.5 mg / kg, about 1.5 mg / kg to about 2.5 mg / kg, or about 2.5 mg / kg to about 10 mg / kg.In some embodiments, the dosage is about 0.01 mg / kg, about 0.02 mg / kg, about 0.03 mg / kg, about 0.04 mg / kg, about 0.05 mg / kg, about 0.06 mg / kg, about 0.07 mg / kg, about 0.08 mg / kg, about 0.09 mg / kg, about 0.1 mg / kg, about 0.2 mg / kg, about 0.3 mg / kg, about 0.4 mg / kg, about 0.5 mg / kg, about 0.6 mg / kg, about 0.7 mg / kg, about 0.8 mg / kg, about 0.9 mg / kg, about 1 mg / kg, about 1.1 mg / kg, about 1.2 mg / kg, about 1.3 mg / kg, about 1.4 mg / kg, about 1.5 mg / kg, about 1.6 mg / kg, about 1.7 mg / kg, about 1.8 mg / kg, about 1.9 mg / kg, about 2.0 mg / kg, about 2.1 mg / kg, about 2.2 mg / kg, about 2.3 mg / kg, about 2.4 mg / kg, about 2.5 mg / kg, about 2.6 mg / kg, about 2.7 mg / kg, about 2.8 mg / kg, about 2.9 mg / kg, about 3.0 mg / kg, about 3.1 mg / kg, about 3.2 mg / kg, about 3.3 mg / kg, about 3.4 mg / kg, about 3.5 mg / kg, about 3.6 mg / kg, about 3.7 mg / kg, about 3.8 mg / kg, about 3.9 mg / kg, about 3.1 mg / kg, about 3.2 mg / kg, about 3.3 mg / kg, about 3.4 mg / kg, about 3.5 mg / kg, about 3.6 mg / kg, about 3.7 mg / kg, about mg / kg, about 1.8 mg / kg, about 1.9 mg / kg, about 2 mg / kg, about 2.1 mg / kg, about 2.2 mg / kg, about 2.3 mg / kg, about 2.4 mg / kg, about 2.5 mg / kg, about 2.6 mg / kg, about 2.7 mg / kg, about 2.8 mg / kg, about 2.9 mg / kg, about 3 mg / kg, about 3.5 mg / kg, about 4 mg / kg, about 4.5 mg / kg, about 5 mg / kg, about 6 mg / kg, about 7 mg / kg, about 8 mg / kg, about 9 mg / kg, about 10 mg / kg, about 15 mg / kg, about 25 mg / kg, about 50 mg / kg, about 75 mg / kg, about 100 mg / kg, about 500 mg / kg or about 1,000 mg / kg. In certain embodiments, the effective amount of nemolizumab or a fragment or variant thereof is about 0.1 mg / kg, about 0.5 mg / kg, about 1 mg / kg, about 1.5 mg / kg, about 2 mg / kg, or about 2.5 mg / kg. In a preferred embodiment, the dosage is about 0.5 mg / kg.

[0138] The present disclosure provides a pharmaceutical composition comprising an anti-IL31RA antibody (e.g., nemolizumab or a fragment or variant thereof) as an active ingredient for use in treating or preventing prurigo nodularis (PN) in a subject, wherein the subject differentially expresses at least one gene selected from the genes disclosed in Table 1 compared to a reference expression level of the at least one gene.

[0139] The present disclosure provides a pharmaceutical composition for use in treating or preventing prurigo nodularis (PN) in a subject, comprising an anti-IL31RA antibody (e.g., nemolizumab or a fragment or variant thereof) as an active ingredient, wherein the subject differentially expresses at least one gene selected from the genes disclosed in Table 1 compared to a reference expression level of at least one gene, and the subject is diagnosed as having PN by detecting expression levels of at least one, at least two, at least three, at least four, or at least five of the differentially expressed genes (DEGs) in Table 1 in a sample obtained from a subject suspected of having PN, and comparing the expression levels of the DEGs to a reference level, wherein the reference level is the corresponding gene expression level of each DEG in a sample from an individual not having PN.

[0140] Any of the pharmaceutical compositions disclosed herein comprising nemolizumab and fragments or variants thereof can be used to treat and / or prevent PN and achieve the therapeutic endpoints of the disclosure. Optimal dosages and routes of administration may vary.

[0141] V. Methods for Treating / Preventing PN and Compositions for Treating or Preventing PN The present disclosure provides a method of treating or preventing pruritus in a subject with prurigo nodularis (PN), the method comprising, consisting of, or consisting essentially of administering to the subject an anti-IL-31RA antibody, such as nemolizumab or a fragment or variant thereof (i.e., a "therapeutic antibody"). The disclosed methods may be carried out to achieve certain therapeutic endpoints, which are described in more detail below. Also disclosed herein is the use of an anti-IL-31RA antibody, such as nemolizumab or a fragment or variant thereof (i.e., a "therapeutic antibody"), to a subject to treat or prevent PN and / or achieve the therapeutic endpoints of the disclosure. Also disclosed herein is the use of an anti-IL-31RA antibody, such as nemolizumab or a fragment or variant thereof (i.e., a "therapeutic antibody"), to a subject to treat or prevent PN and / or achieve the therapeutic endpoints of the disclosure. In addition, certain subgroups of subjects with PN may be particularly suitable for treatment with the methods and uses of the present disclosure (eg, patients exhibiting any of the DEGs disclosed in Table 1).

[0142] The present disclosure is the first to report a transcriptomic signature of PN that can not only identify and positively diagnose PN, but also identify subjects with PN who will likely respond to treatment with an anti-IL-31RA antibody (e.g., nemolizumab) and track the response to treatment with an anti-IL-31RA antibody (e.g., nemolizumab).

[0143] This disclosure is also the first to report a plasma proteome signature of PN subjects successfully treated with an anti-IL-31RA antibody such as nemolizumab. This "responder signature" can be used not only as a marker for positive clinical endpoints, but also to identify subjects with PN who are likely to respond to treatment with an anti-IL-31RA antibody (e.g., nemolizumab) and to track the response of treatment with an anti-IL-31RA antibody (e.g., nemolizumab).

[0144] A. Subjects to be Treated A subject treated for PN according to the methods and uses of the present disclosure may exemplify one or more of the underlying gene expression patterns disclosed herein. In particular, a subject with PN treated according to the methods and uses of the present disclosure may differentially express up to 5,943 genes (known as differentially expressed genes or DEGs) as shown in FIG. 1A and Table 1 below. This differential gene expression may be observed in the subject's skin, particularly in skin samples that contain or consist of nodules or lesions. Of these DEGs, 2,060 may be increased (i.e., overexpressed) and 3,874 may be decreased (i.e., not expressed). Genes that may be most increased include: KRT6C, which may be increased by at least 100-fold, at least 150-fold, at least 200-fold, at least 250-fold, at least 300-fold, at least 350-fold, at least 400-fold, at least 450-fold, at least 500-fold, at least 550-fold, or 588-fold compared to expression levels in a sample (e.g., a skin sample) from an individual without PN; DEFB4A, which may be increased by at least 25-fold, at least 50-fold, at least 75-fold, at least 100-fold, at least 125-fold, or at least 150-fold compared to expression levels in a sample (e.g., a skin sample) from an individual without PN; and KRT16, which may be increased by at least 10-fold, at least 20-fold, at least 30-fold, at least 40-fold, at least 50-fold, at least 60-fold, at least 70-fold, at least 80-fold, or at least 90-fold compared to expression levels in a sample (e.g., a skin sample) from an individual without PN.

[0145] The genes that were decreased include: LCE5A, which may be decreased by at least 2-fold, at least 3-fold, at least 4-fold, at least 5-fold, at least 6-fold, at least 7-fold, at least 8-fold, at least 9-fold, at least 10-fold, or at least 11-fold compared to expression levels in a sample (e.g., a skin sample) from an individual without PN; and AQP7, which may be reduced by at least 2-fold, at least 3-fold, at least 4-fold, at least 5-fold, at least 6-fold, at least 7-fold, or at least 7.9-fold compared to expression levels in a sample from an individual without PN (e.g. a skin sample).

[0146] Genes encoding cytokines may also be overexpressed in subjects suffering from PN, which can be treated or prevented according to the methods and uses of the present disclosure. For cytokine genes, the most prominent upregulated genes are IL-36 family members and IL-20 family members. These upregulated or overexpressed genes may include: ●IL36A (for example, about 6.8 times, FDR = 1.8 × 10 -4 ), ●IL36G (e.g., about 8.4 times, FDR = 3.9 × 10 -25 ), ●IL19 (e.g., about 5.1 times, FDR = 7.4 × 10 -4 ), ●IL20 (for example, about 3.5 times, FDR = 1.7 × 10 -3 ), ●IL22 (e.g., about 2.7 times, FDR = 2.9 × 10 -2 ), ●IL24 (e.g., about 5.8 times, FDR = 3.8 × 10 -10 ), and ●IL26 (e.g., about 4.9 times, FDR = 3.3 × 10 -3 ).

[0147] Each of these IL-36 and IL-20 family member cytokine genes can be overexpressed by at least about 2-fold, at least about 2.5-fold, at least about 3-fold, at least about 3.5-fold, at least about 4-fold, at least about 4.5-fold, at least about 5-fold, at least about 5.5-fold, at least about 6-fold, at least about 6.5-fold, at least about 7-fold, at least about 7.5-fold, at least about 8-fold, or at least about 8-5-fold compared to expression levels in a sample (e.g., a skin sample) from an individual without PN.

[0148] Other factors that may be upregulated or overexpressed in samples (e.g., skin samples) obtained from subjects with PN being treated include IL1A (e.g., about 4.7-fold, FDR=1.0×10 -12 ), IL1B (e.g., about 4.1-fold, FDR = 3.7 × 10 -6 ), and IL4R (e.g., about 2.6-fold (FDR = 6.3 × 10 -19 ) can be mentioned. Table 1 at the end of the Examples section of this specification provides a more comprehensive list of DEGs.

[0149] Certain Gene Ontology (GO) categories may also be evident in the skin of subjects with PN treated according to the methods and uses of the present disclosure. These GO categories are as follows: ● "Cornified envelope" (FDR=1.5×10 -12 ), ●“Epithelial cell differentiation” (FDR=6.4×10 -10 ), - Keratinization (FDR=1.6×10 -12 ), ● "Peptidase regulatory factor activity" (FDR = 1.1 x 10 -4 ), ● "Interleukin-4 and 13 signaling" (FDR = 6.8 × 10 -7 ), "Interferon alpha / beta signaling" and "response to interferon gamma" (FDR=4.1×10 -7 and FDR = 4.1 × 10 -6 ), ● "IL23 pathway" (FDR = 2 × 10 -5 ), and ● "Metaphase and anaphase of mitosis" (FDR = 3.8 × 10 -10 ).

[0150] These categories reflect the hyperproliferative nature of PN, which has been found to be associated with alterations in epidermal differentiation and inflammatory components. In some embodiments, subjects with PN treated according to the methods and uses of the present disclosure may overexpress the proliferation marker Ki67 (MKI67), the cell cycle gene CDKN1A, and / or the inflammatory network, including IL-1 and IL-36.

[0151] A subject with PN treated according to the methods or uses of the present disclosure may additionally or alternatively present one or more of the co-expression modules or clusters shown in Figure 1D and Table 3 at the end of the Examples section herein. Different functions are defined for these co-expressed gene modules as described herein.

[0152] The present disclosure also provides specific cell type signatures for non-lesional and lesional PN skin that may be present in subjects with PN treated according to the methods and uses of the present disclosure.For example, subjects with PN may have the transcriptome signatures associated with epithelial cells and keratinocytes observed in Figure 2A, as described herein, and / or may have a signature associated with Th2.Other inflammatory signatures, such as macrophages, may also be prominent in PN lesions and the skin of subjects with PN.

[0153] In general, the methods and uses of the present disclosure can treat or prevent PN in subjects suffering from mild, moderate, or severe pruritus. In some embodiments, PN may be classified as moderate to severe. In some embodiments, PN may be classified as moderate, and in other embodiments, PN may be classified as severe. In some embodiments, pruritus may be scored as none, mild, moderate, or severe. "None", "mild", "moderate", and "severe" are terms of the art in describing the presence, extent, and / or intensity of ablation. Those skilled in the art know the boundaries and borders of these terms. For example, pruritus can be characterized according to one or more of the following methods known to those skilled in the art: Intensity can be quickly measured on a unidimensional scale routinely used in clinical care. See Pereira et al., Allergology International (2017) 66:3-78. Additionally or alternatively, patients can be asked to rate their itch intensity using a numeric rating scale (NRS) from 0 ("no itch") to 10 ("worst itch imaginable"). Another unidimensional scale, the visual analog scale (VAS), offers patients the opportunity to indicate their itch intensity by marking on a ruler-like scale 10 cm long. Both endpoints are marked with a number corresponding to the intensity, with 0 representing "no itch" and 10 representing "worst itch imaginable". Scores below 3.0 VAS / NRS points are generally associated with mild itch, while scores above 6.9 indicate severe itch. Scores above 9.0 represent very severe itch. The verbal rating scale (VRS) is a further unidimensional scale that allows patients to describe their itch intensity by ascending adjectives (0-no itch, 4-worst itch imaginable). The NRS, VAS and VRS have been validated in large studies of chronic pruritus patients with pruritic skin diseases of various origins or pruritus. These instruments were highly reproducible and there was a high correlation between scales 6, 7 and 8. Chronic pruritus can significantly reduce the quality of life of patients. For this reason, the Dermatology Life Quality Index (DLQI) has been widely used and validated.DLQI scores range from 0 to 30, with higher scores indicating poorer quality of life. Investigator Global Assessment (IGA) scores range from 0 (clear) to 5 (very severe disease) and are presented as a percentage of patients in the indicated population. In this study, IGA scores range from 0 to 4.

[0154] A subject treated for PN according to the disclosed methods and uses may exemplify one or more of the disclosed plasma protein signature alterations. In particular, a subject with PN treated according to the disclosed methods and uses may exhibit (a) a decrease in leukocyte migration and cell migration, (b) inhibition of the STAT3 and STAT5b pathways, (c) downregulation of the IL-6 and VEGF pathways, (d) a decrease in the TGFB1 pathway, or (e) a combination thereof. The aforementioned decrease or inhibition may be observed at a particular time point after the start of treatment, for example, 2 weeks, 4 weeks, 6 weeks, 8 weeks, 10 weeks, or 12 weeks. The decrease or inhibition may be determined with respect to (i) a control sample obtained from one or more individuals without PN, or (ii) a biological sample obtained from the subject before administration of an anti-IL-31RA antibody. Additionally or alternatively, the subject may exhibit upregulation in neuronal terms, including CREB signaling in neurons, synaptogenesis signaling pathways, glial cell death and glial apoptosis, and combinations thereof, following administration of an anti-IL-31RA antibody.

[0155] A subject with PN suitable for treatment with the methods and uses of the present disclosure may exhibit a high level of leukocyte migration or leukocyte cell migration relative to individuals or populations without PN or in a subject prior to initiating treatment with an anti-IL-31RA antibody such as nemolizumab. Additionally or alternatively, a suitable subject may exhibit a relatively high STAT3 pathway activity or expression compared to individuals or populations without PN or in a subject prior to initiating treatment with an anti-IL-31RA antibody such as nemolizumab. Additionally or alternatively, a suitable subject may exhibit a relatively high TGFB1 pathway activity or expression compared to individuals or populations without PN or in a subject prior to initiating treatment with an anti-IL-31RA antibody such as nemolizumab. Additionally or alternatively, a suitable subject may exhibit a relatively high amount of a circulating cytokine signature compared to individuals or populations without PN or in a subject prior to initiating treatment with an anti-IL-31RA antibody such as nemolizumab. Such cytokine pathways include, but are not limited to, the IL-6 and VEGF pathways.

[0156] Prior to treatment with an anti-IL-31Ra antibody, such as nemolizumab, the amount of a protein marker of the present disclosure in the plasma of a subject with PN may be at least 2-fold, at least 3-fold, at least 4-fold, at least 5-fold, at least 6-fold, at least 7-fold, at least 8-fold, at least 9-fold, at least 10-fold, at least 15-fold, at least 20-fold, at least 25-fold, at least 30-fold, at least 35-fold, at least 40-fold, at least 45-fold, or at least 50-fold higher than the baseline level. The baseline level may be determined with respect to (i) a control sample obtained from one or more individuals (i.e., a population) without PN, or (ii) a biological sample obtained from the subject prior to administration of the anti-IL-31RA antibody.

[0157] In some embodiments, the subject has been diagnosed with PN for at least about 6 months. In certain embodiments, the subject has at least about 20 nodules in the body with a bilateral distribution. In certain embodiments, the subject has pruritus lesions on the upper limbs with or without lesions on the trunk or lower limbs. In some embodiments, the pruritus is assigned a score of at least 7 on the Numeric Rating Scale (NRS). In some embodiments, the mean of the worst daily intensity of the NRS score is at least 7 compared to the previous 3 days. In some embodiments, the mean of the worst daily intensity of the NRS score is at least 7 compared to the previous week.

[0158] In some embodiments, the subject does not have atopic dermatitis (AD). In some embodiments, the subject does not have chronic pruritus due to conditions other than PN, such as scabies, bites, lichen simplex chronicus, psoriasis, acne, folliculitis, habitual picking, lymphomatoid papulosis, chronic actinic dermatitis, dermatitis herpetiformis, sporotrichosis, or bullous disease. In some embodiments, the subject does not have neurogenic or psychogenic pruritus, such as hypoesthesia, palmar pruritus, dilutional parasitosis, or pathological pruritus.

[0159] B. Therapeutic Endpoints for Treatment Based on the data provided herein, IL-31 signaling is believed to be upstream of IL-17 and IL-4 signaling, at least in subjects with PN, because treatment with the anti-IL-31RA antibody nemolizumab could alter the transcriptome expression signatures of IL-17 and IL-4. Thus, the present disclosure provides a method for normalizing differentially expressed genes (DEGs) in subjects with PN, comprising administering to the subject with PN an anti-IL-31RA antibody, such as nemolizumab or a fragment or variant thereof. As shown in the Examples section below, treatment or prevention with nemolizumab results in normalization of several PN-related DEGs, both for increased (e.g., about 969 genes) and decreased (e.g., about 1,268 genes). See FIG. 4A. In some embodiments, the concentration is about 5, about 10, about 15, about 20, about 25, about 50, about 75, about 100, about 150, about 200, about 250, about 300, about 350, about 400, about 450, about 500, about 550, about 600, about 650, about 700, about 750, about 800, about 850, about 900, about 950, about 1000, about 1100, about 1200, about 1300, about 1400, about 1500, about 1600, about 1700, about 1800, about 1900, about 2100, about 2200, about 2300, about 2400, about 2500, about 2600, about 2700, about 2800, about 2900, about 3000, about 3100, about 3200, about 3300, about 3400, about 3500, about 3600, about 3700, about 3800, about 3900, about 4000, about 4000, about 4500, about 5000, about 5500, about 6000, about 6500, about 7000, about 7500, about 8000, about 8500, about 9000, about 9500, about 1000, about 1100, about 1200, about 2500, about 3500, about 4000, about 4500, about 5000, about 5500, about 600 , about 1300, about 1400, about 1500, about 1600, about 1700, about 1800, about 1900, about 2000, about 2100, about 2200, about 2300, about 2400, or about 2500 DEGs may be normalized in subjects with PN after treatment with an anti-IL-31RA antibody, such as nemolizumab or a fragment or variant thereof.In some embodiments, the number of saturation saturation saturation saturation is at least 5, at least 10, at least 15, at least 20, at least 25, at least 50, at least 75, at least 100, at least 150, at least 200, at least 250, at least 300, at least 350, at least 400, at least 450, at least 500, at least 550, at least 600, at least 650, at least 700, at least 750, at least 800, at least 850, at least 900, at least 950, at least 1000, at least 1100, at least 1200, at least 1300, at least 1400, at least 1500, at least 1600, at least 1700, at least 1800, at least 1900, at least 2000, at least 2100, at least 2200, at least 2300, at least 2400, or at least 2500 DEGs, and up to 2500, 3000, 3500, 4000, 4500, 5000, 5500, or about 6000 DEGs may be normalized in subjects with PN after treatment with an anti-IL-31RA antibody, such as nemolizumab or a fragment or variant thereof. In some embodiments, the treatment or prevention may include administration of an anti-IL-31RA antibody (e.g., nemolizumab) once a week, once every two weeks, once every three weeks, once every four weeks, once every five weeks, once every six weeks, once every seven weeks, or once every eight weeks. Administration about once every 4 weeks may be preferred. Administration may be performed via injection, such as subcutaneous injection. The time frame for determining / comparing normalization of DEG may be 4 weeks, 5 weeks, 6 weeks, 7 weeks, 8 weeks, 9 weeks, 10 weeks, 11 weeks, or 12 weeks or more.For example, at baseline (i.e., at or before treatment initiation), a sample (e.g., a skin sample) may be obtained from the subject to detect expression levels of some or all of the putative DEGs disclosed in Table 1 (e.g., KRT6C, DEFB4A, KRT16, LCE5A, AQP7, IL-36 family members, IL-20 family members, etc.), and another sample may be taken and evaluated at 4, 5, 6, 7, 8, 9, 10, 11, or 12 weeks or more after treatment initiation, with a reference expression level of the DEGs associated with normal healthy skin (e.g., expression levels obtained from a skin sample from a subject without PN) and a more consistent (i.e., "normalized") expression level to determine whether the expression level of the DEGs has changed. Such methods can be used to track treatment and evaluate whether the subject is responsive to treatment with an anti-IL-31RA antibody (e.g., nemolizumab).

[0160] GO categories enriched among DEGs may also be decreased or altered as a result of treatment with an anti-IL-31RA antibody (e.g., nemolizumab). For example, after 12 weeks of treatment, the following GO categories: "cell cycle" (FDR=5.6×10 -14 ), "keratinocyte differentiation" (FDR = 1.8 × 10 -4 ), and “Interleukin-4 and 13 signaling” (FDR = 1.5 × 10 -2) was observed. Indeed, data provided in the Examples section demonstrate that nemolizumab treatment (and treatment with other anti-IL-31RA antibodies) normalizes epidermal hyperproliferation, normalizes differentiation, and reduces inflammatory responses, particularly those associated with Th2 responses. Thus, the present disclosure provides a method of normalizing epidermal hyperproliferation, normalizing epidermal differentiation, and / or reducing inflammatory responses in the skin, comprising administering an anti-IL-31RA antibody (e.g., nemolizumab) to a subject with PN. In some embodiments, the inflammatory response may be a Th2 response. In some embodiments, administration of the anti-IL-31RA antibody (e.g., nemolizumab) may be once a week, once every two weeks, once every three weeks, once every four weeks, once every five weeks, once every six weeks, once every seven weeks, or once every eight weeks. Administration about once every four weeks may be preferred. Administration may be performed via injection, such as subcutaneous injection.

[0161] With respect to the disclosed method of reducing inflammatory response in the skin of a subject with PN, in some embodiments, the cytokine response signature generated in human epidermal rafts of the skin of the subject may be altered. For example, the expression of IL17A mRNA was not significantly different in non-lesional versus lesional skin, and was not significantly different after 12 weeks of treatment with nemolizumab. A consistent reduction in IL-31 response, alone or in combination with other inflammatory cytokines, including the Th2 cytokines IL-13 or IL-17A, can be seen in FIG. 5A, thus providing evidence of the blockade of the IL-31 pathway by nemolizumab. However, IL-17A-responsive genes are shown herein to be enriched in PN skin and downregulated by nemolizumab. This suggests that IL-17A itself is not the predominant cytokine in PN, but is downstream of IL-31 signaling. Indeed, a consistent reduction in IL-31 responses, alone or in combination with other inflammatory cytokines (including the Th2 cytokines IL-13, or IL-17A), was observed upon treatment with nemolizumab (Figure 5A), with more robust reductions observed in Th1, Th17, and Th2 markers (Figure 5B). Thus, IL-31 signaling is likely upstream of IL-17 and IL-4 signaling.

[0162] In some embodiments, the basal keratinocyte (KRT14+) signature may be elevated in PN lesional skin, and this signature may be restored to normal (i.e., non-PN) when treated according to the methods and uses of the present disclosure. In some embodiments, the epidermal spinous (KRT10+) signature may be elevated in PN lesional skin, and this signature may be restored to normal (i.e., non-PN) when treated according to the methods and uses of the present disclosure (FIG. 5C).

[0163] In some embodiments, transcription factor binding sites (TFBS) enriched among genes upregulated in baseline lesional skin are more likely to be enriched among genes downregulated by nemolizumab after treatment with an anti-IL-31RA antibody such as nemolizumab (e.g., by 12 weeks post-treatment, as shown in FIG. 5D). In some embodiments, the transcription factors downregulated by treatment can be EGR4 (a member of the EGF family of zinc finger transcription factors), STAT3, and / or KLF16.

[0164] The present disclosure further provides plasma protein markers identified in subjects successfully treated with nemolizumab, and thus the resulting changes in biomarkers can be considered a "responder signature" to establish success and a means to track responsiveness. Thus, disclosed herein is a method of treating or preventing prurigo nodularis (PN) in a subject, comprising administering an anti-IL-31RA antibody to a subject with PN, wherein treatment with the anti-IL-31RA antibody results in (a) reduced leukocyte migration and cell migration, (b) reduced IL-6 or reduced IL-6 pathway signaling, (c) reduced VEGF or reduced VEGF pathway signaling, (d) reduced STAT3 or reduced STAT3 pathway signaling, (e) reduced STAT5b or reduced STAT5b pathway signaling, (f) reduced TGFB1 or reduced TGFB1 pathway signaling, or (g) a combination thereof. In some embodiments, the subject may also exhibit an increase in the neuron ontology of the present disclosure. Additionally, disclosed herein is a method of altering immune response in a subject with PN, comprising administering an anti-IL-31RA antibody to the subject with PN, wherein treatment with the anti-IL-31RA antibody results in (a) decreased leukocyte migration and cell migration, (b) decreased IL-6 or decreased IL-6 pathway signaling, (c) decreased VEGF or decreased VEGF pathway signaling, (d) decreased STAT3 or decreased STAT3 pathway signaling, (e) decreased STAT5b or decreased STAT5b pathway signaling, (f) decreased TGFB1 or decreased TGFB1 pathway signaling, or (g) a combination thereof. In some embodiments, the subject may also exhibit an increase in the neuron ontology of the present disclosure. The disclosed plasma protein markers are detectable, for example, by mass spectrometry and other protein evaluation methods (e.g., ELISA, Western blot, etc.).

[0165] In particular, when subjects with PN are treated with an anti-IL-31RA antibody, such as nemolizumab, immune cells (eg, white blood cells) may experience reduced migration or cell migration, or both.

[0166] STAT3 pathway activity may be decreased in subjects with PN treated with an anti-IL-31RA antibody, such as nemolizumab, compared to individuals or populations without PN, or compared to the subject before initiating treatment with an anti-IL-31RA antibody, such as nemolizumab.

[0167] The amount of cytokine activity may be relatively high in subjects with PN, or in subjects prior to initiating treatment with an anti-IL-31RA antibody, such as nemolizumab, compared to individuals or populations without PN. Such cytokine pathways include, but are not limited to, the IL-6 and VEGF pathways. In PN subjects treated with an anti-IL-31RA antibody, such as nemolizumab, the treatment may result in a decrease in the IL-6 or VEGF signature, or both, compared to baseline levels. The baseline level may be determined with respect to (i) a control sample obtained from one or more individuals (i.e., a population) without PN, or (ii) a biological sample obtained from the subject prior to administration of the anti-IL-31RA antibody.

[0168] TGFB1 pathway activity may be decreased in subjects with PN treated with an anti-IL-31RA antibody, such as nemolizumab, compared to individuals or populations without PN, or compared to the subject before initiating treatment with an anti-IL-31RA antibody, such as nemolizumab.

[0169] For purposes of plasma protein markers of the present disclosure, the amount of a protein marker of the present disclosure in the plasma of a subject with PN may be at least 2-fold, at least 3-fold, at least 4-fold, at least 5-fold, at least 6-fold, at least 7-fold, at least 8-fold, at least 9-fold, at least 10-fold, at least 15-fold, at least 20-fold, at least 25-fold, at least 30-fold, at least 35-fold, at least 40-fold, at least 45-fold, or at least 50-fold higher than the baseline level. The baseline level may be determined with respect to (i) a control sample obtained from one or more individuals (i.e., a population) without PN, or (ii) a biological sample obtained from the subject prior to administration of an anti-IL-31RA antibody. Similarly, after a subject with PN has been treated with an anti-IL-31RA antibody, such as nemolizumab (e.g., 2, 4, 6, 8, 10, or 12 weeks after administration of the antibody), the amount of a plasma protein marker of the present disclosure in the subject may be decreased by at least 2-fold, at least 3-fold, at least 4-fold, at least 5-fold, at least 6-fold, at least 7-fold, at least 8-fold, at least 9-fold, at least 10-fold, at least 15-fold, at least 20-fold, at least 25-fold, at least 30-fold, at least 35-fold, at least 40-fold, at least 45-fold, or at least 50-fold compared to baseline levels. The baseline levels may be determined with respect to (i) a control sample obtained from one or more individuals (i.e., a population) without PN, or (ii) a biological sample obtained from the subject prior to administration of the anti-IL-31RA antibody.

[0170] In addition, neural ontologies (e.g., CREB signaling in neurons, synaptogenesis signaling pathways, glial cell death, and glial apoptosis) can be upregulated in subjects with PN and subsequently downregulated in nemolizumab responder subjects.

[0171] In some embodiments of the methods and uses of the present disclosure, treatment or prevention with an anti-IL-31RA antibody, such as nemolizumab or a fragment or variant thereof, results in a reduction in pruritus scores. The reduction in scores may be measured, for example, by the Peak Pruritus Numeric Rating Scale (PP-NRS). See, for example, FIG. 6A. Indeed, the data provided herein shows that all subjects with PN treated with nemolizumab showed an improvement in pruritus scores. In some embodiments, the measured distance between principal component 1 and principal component 2 (i.e., PC1 / PC2 components) may be shorter than that of a placebo group that was not treated with nemolizumab or another anti-IL-31RA antibody. More specifically, at week 12 (i.e., 12 weeks after the start of treatment), the proportion of patients who achieved a weekly average reduction of 4 points in the PP NRS was significantly higher in the nemolizumab group compared to placebo (52.9% vs. 8.3%, p<0.001). At week 12, the proportion of subjects achieving IGA success (defined as IGA 0 [clear] or 1 [almost clear]) was significantly higher in the nemolizumab group compared to placebo (20.6% vs. 2.8%, p=0.02).

[0172] C. Dosage and Administration Regimens for the Disclosed Methods and Uses An effective amount of an anti-IL-31RA antibody, such as nemolizumab or a fragment or variant thereof, is an amount sufficient to produce a beneficial or desired result, such as alleviating at least one or more symptoms of PN. As used herein, an effective amount would also include an amount sufficient to delay or prevent the onset of pruritus, alter the course of PN symptoms, or reverse PN symptoms. Thus, an exact "effective amount" cannot be specified. However, the appropriate "effective amount" in any given case can be determined by one of ordinary skill in the art using only routine experimentation.

[0173] An effective amount can be administered in one or more administrations, applications, or dosages. Such delivery depends on several variables, including the period for which the individual dosage units are used, the bioavailability of the therapeutic agent, the route of administration, and the like. However, it is understood that the specific dosage level of the therapeutic agent of the present disclosure for any particular subject will depend on a variety of factors, including the activity of the particular compound used, the age, weight, general health, sex, and diet of the subject, the time of administration, the rate of excretion, the drug combination, and the severity and form of administration of the particular disorder being treated. Treatment and prophylactic dosages can generally be titrated to optimize safety and efficacy. Dosages are determined by the physician and can be adjusted, if necessary, to the observed effects of the treatment. Typically, dose-effect relationships from in vitro and / or in vivo tests can initially provide useful guidance regarding appropriate dosages for patient administration. In general, it is desired to administer an amount of the compound effective to achieve serum levels commensurate with the concentrations found to be effective in vitro. Determination of these parameters is within the skill of the art. These considerations, as well as effective formulation and administration procedures, are well known in the art and are described in standard textbooks.

[0174] Dosing regimens for treating or preventing PN may include flat dosing (i.e., administering the same dose repeatedly at predetermined intervals) or may include a loading dose (i.e., administering an initial dose that is higher or different than subsequent successive doses). For purposes of either type of dosing regimen, an effective dose may be administered topically, parenterally, subcutaneously, intradermally, or intramuscularly. In a preferred embodiment, administration involves subcutaneous injection.

[0175] In some embodiments, the loading dose and the subsequent consecutive doses may be administered via the same route (e.g., subcutaneously), while in some embodiments, the loading dose and the subsequent consecutive doses may be administered via different routes (e.g., parenteral and subcutaneous, respectively). In some embodiments, the loading dose may be about 5 mg, about 10 mg, about 15 mg, about 20 mg, about 25 mg, about 30 mg, about 35 mg, about 40 mg, about 45 mg, about 50 mg, about 55 mg, about 60 mg, about 65 mg, about 70 mg, about 75 mg, about 80 mg, about 85 mg, about 90 mg, about 95 mg, about 100 mg, about 105 mg, about 110 mg, about 115 mg, about 120 mg, or more. In some embodiments, the loading dose can be 5 mg, 10 mg, 15 mg, 20 mg, 25 mg, 30 mg, 35 mg, 40 mg, 45 mg, 50 mg, 55 mg, 60 mg, 65 mg, 70 mg, 75 mg, 80 mg, 85 mg, 90 mg, 95 mg, 100 mg, 105 mg, 110 mg, 115 mg, 120 mg or more.In some embodiments, the loading dose is about 0.01 mg / kg, about 0.02 mg / kg, about 0.03 mg / kg, about 0.04 mg / kg, about 0.05 mg / kg, about 0.06 mg / kg, about 0.07 mg / kg, about 0.08 mg / kg, about 0.09 mg / kg, about 0.1 mg / kg, about 0.2 mg / kg, about 0.3 mg / kg, about 0.4 mg / kg, about 0.5 mg / kg, about 0.6 mg / kg, about 0.7 mg / kg, about 0.8 mg / kg, about 0.9 mg / kg, about 1 mg / kg, about 1.1 mg / kg, about 1.2 mg / kg, about 1.3 mg / kg, about 1.4 mg / kg, about 1.5 mg / kg, about 1.6 mg / kg, about 1.7 mg / kg, about 1.8 mg / kg, about 1.9 mg / kg, about 2.0 mg / kg, about 2.1 mg / kg, about 2.2 mg / kg, about 2.3 mg / kg, about 2.4 mg / kg, about 2.5 mg / kg, about 2.6 mg / kg, about 2.7 mg / kg, about 2.8 mg / kg, about 2.9 mg / kg, about 3.0 mg / kg, about 3.1 mg / kg, about 3.2 mg / kg, about 3.3 mg / kg, about 3.4 mg / kg, about 3.5 mg / kg, about 3.6 mg / kg, about 3.7 mg / kg, about 3.8 mg / kg, about 3.9 mg / kg, about 3.1 mg / kg, about 3.2 mg / kg, about 3.3 mg / kg, about 3.4 mg / kg, about 3.5 mg / kg, about 3.6 mg / kg, about 3.7 mg / kg, about g / kg, about 1.8 mg / kg, about 1.9 mg / kg, about 2 mg / kg, about 2.1 mg / kg, about 2.2 mg / kg, about 2.3 mg / kg, about 2.4 mg / kg, about 2.5 mg / kg, about 2.6 mg / kg, about 2.7 mg / kg, about 2.8 mg / kg, about 2.9 mg / kg, about 3 mg / kg, about 3.5 mg / kg, about 4 mg / kg, about 4.5 mg / kg, about 5 mg / kg, about 6 mg / kg, about 7 mg / kg, about 8 mg / kg, about 9 mg / kg, about 10 mg / kg, about 15 mg / kg, about 25 mg / kg, about 50 mg / kg, about 75 mg / kg, about 100 mg / kg, about 500 mg / kg or about 1,000 mg / kg.In some embodiments, the loading dose is 0.01 mg / kg, 0.02 mg / kg, 0.03 mg / kg, 0.04 mg / kg, 0.05 mg / kg, 0.06 mg / kg, 0.07 mg / kg, 0.08 mg / kg, 0.09 mg / kg, 0.1 mg / kg, 0.2 mg / kg, 0.3 mg / kg, 0.4 mg / kg, 0.5 mg / kg, 0.6 mg / kg, 0.7 mg / kg, 0.8 mg / kg, 0.9 mg / kg, 1 mg / kg, 1.1 mg / kg, 1.2 mg / kg, 1.3 mg / kg, 1.4 mg / kg, 1.5 mg / kg, 1.6 mg / kg, 1. The loading dose may be 7 mg / kg, 1.8 mg / kg, 1.9 mg / kg, 2 mg / kg, 2.1 mg / kg, 2.2 mg / kg, 2.3 mg / kg, 2.4 mg / kg, 2.5 mg / kg, 2.6 mg / kg, 2.7 mg / kg, 2.8 mg / kg, 2.9 mg / kg, 3 mg / kg, 3.5 mg / kg, 4 mg / kg, 4.5 mg / kg, 5 mg / kg, 6 mg / kg, 7 mg / kg, 8 mg / kg, 9 mg / kg, 10 mg / kg, 15 mg / kg, 25 mg / kg, 50 mg / kg, 75 mg / kg, 100 mg / kg, 500 mg / kg, or 1,000 mg / kg. In some embodiments, the loading dose is administered as a single injection. In some embodiments, the loading dose is administered as multiple injections that may be administered simultaneously or spaced apart by defined intervals.

[0176] The subsequent successive doses of the loading dose regimen are generally lower than the loading dose. For example, in some embodiments, the dosing regimen may include a loading dose of 60 mg and a successive dose of 30 mg, which may be administered at a defined interval, for example, once every 4 weeks. In some embodiments, the successive doses of the dosing regimen may be about 5 mg, about 10 mg, about 15 mg, about 20 mg, about 25 mg, about 30 mg, about 35 mg, about 40 mg, about 45 mg, about 50 mg, about 55 mg, about 60 mg, about 65 mg, about 70 mg, about 75 mg, about 80 mg, about 85 mg, about 90 mg, about 95 mg, about 100 mg, about 105 mg, about 110 mg, about 115 mg, about 120 mg or more. In some embodiments, the sequential dose can be 5 mg, 10 mg, 15 mg, 20 mg, 25 mg, 30 mg, 35 mg, 40 mg, 45 mg, 50 mg, 55 mg, 60 mg, 65 mg, 70 mg, 75 mg, 80 mg, 85 mg, 90 mg, 95 mg, 100 mg, 105 mg, 110 mg, 115 mg, 120 mg or more.In some embodiments, the continuous dose is about 0.01 mg / kg, about 0.02 mg / kg, about 0.03 mg / kg, about 0.04 mg / kg, about 0.05 mg / kg, about 0.06 mg / kg, about 0.07 mg / kg, about 0.08 mg / kg, about 0.09 mg / kg, about 0.1 mg / kg, about 0.2 mg / kg, about 0.3 mg / kg, about 0.4 mg / kg, about 0.5 mg / kg, about 0.6 mg / kg, about 0.7 mg / kg, about 0.8 mg / kg, about 0.9 mg / kg, about 1 mg / kg, about 1.1 mg / kg, about 1.2 mg / kg, about 1.3 mg / kg, about 1.4 mg / kg, about 1.5 mg / kg, about 1.6 mg / kg, about 1.7 mg / kg, about 1.8 mg / kg, about 1.9 mg / kg, about 2.0 mg / kg, about 2.1 mg / kg, about 2.2 mg / kg, about 2.3 mg / kg, about 2.4 mg / kg, about 2.5 mg / kg, about 2.6 mg / kg, about 2.7 mg / kg, about 2.8 mg / kg, about 2.9 mg / kg, about 3.0 mg / kg, about 3.1 mg / kg, about 3.2 mg / kg, about 3.3 mg / kg, about 3.4 mg / kg, about 3.5 mg / kg, about 3.6 mg / kg, about 3.7 mg / kg, about 3.8 mg / kg, about 3.9 mg / kg, about 3.1 mg / kg, about 3.2 mg / kg, about 3.3 mg / kg, about 3.4 mg / kg, about 3.5 mg / kg, about 3.6 mg / kg, about 3.7 mg / kg, about g / kg, about 1.8 mg / kg, about 1.9 mg / kg, about 2 mg / kg, about 2.1 mg / kg, about 2.2 mg / kg, about 2.3 mg / kg, about 2.4 mg / kg, about 2.5 mg / kg, about 2.6 mg / kg, about 2.7 mg / kg, about 2.8 mg / kg, about 2.9 mg / kg, about 3 mg / kg, about 3.5 mg / kg, about 4 mg / kg, about 4.5 mg / kg, about 5 mg / kg, about 6 mg / kg, about 7 mg / kg, about 8 mg / kg, about 9 mg / kg, about 10 mg / kg, about 15 mg / kg, about 25 mg / kg, about 50 mg / kg, about 75 mg / kg, about 100 mg / kg, about 500 mg / kg or about 1,000 mg / kg.In some embodiments, the sequential doses are 0.01 mg / kg, 0.02 mg / kg, 0.03 mg / kg, 0.04 mg / kg, 0.05 mg / kg, 0.06 mg / kg, 0.07 mg / kg, 0.08 mg / kg, 0.09 mg / kg, 0.1 mg / kg, 0.2 mg / kg, 0.3 mg / kg, 0.4 mg / kg, 0.5 mg / kg, 0.6 mg / kg, 0.7 mg / kg, 0.8 mg / kg, 0.9 mg / kg, 1 mg / kg, 1.1 mg / kg, 1.2 mg / kg, 1.3 mg / kg, 1.4 mg / kg, 1.5 mg / kg, 1.6 mg / kg, 1.7 mg / kg, 1.8 mg / kg, 1.9 mg / kg, 2.0 mg / kg, 2.1 mg / kg, 2.2 mg / kg, 2.3 mg / kg, 2.4 mg / kg, 2.5 mg / kg, 2.6 mg / kg, 2.7 mg / kg, 2.8 mg / kg, 2.9 ...9 mg / kg, 2.1 mg / kg, 2.2 mg / kg, 2.3 mg / kg, 2.4 mg / kg, 2.5 mg / kg, 2.6 mg / kg, 2.7 mg / kg, 2.8 mg / kg, 2.9 mg / kg, 2.8 mg / kg The dose may be 7mg / kg, 1.8mg / kg, 1.9mg / kg, 2mg / kg, 2.1mg / kg, 2.2mg / kg, 2.3mg / kg, 2.4mg / kg, 2.5mg / kg, 2.6mg / kg, 2.7mg / kg, 2.8mg / kg, 2.9mg / kg, 3mg / kg, 3.5mg / kg, 4mg / kg, 4.5mg / kg, 5mg / kg, 6mg / kg, 7mg / kg, 8mg / kg, 9mg / kg, 10mg / kg, 15mg / kg, 25mg / kg, 50mg / kg, 75mg / kg, 100mg / kg, 500mg / kg or 1,000mg / kg.

[0177] For the purpose of the loading dose regimen, the first consecutive dose may be administered 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, 7 days, 8 days, 9 days, 10 days, 11 days, 12 days, 13 days, 1 week, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 7 weeks, 8 weeks, 9 weeks, or 10 weeks after the initial loading dose. In some embodiments, the first consecutive dose is administered 4 weeks after the initial loading dose. In some embodiments, the subsequent consecutive doses are administered once every 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, 7 days, 8 days, 9 days, 10 days, 11 days, 12 days, 13 days, 1 week, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 7 weeks, 8 weeks, 9 weeks, or 10 weeks. In some embodiments, the successive doses are spaced four weeks apart (i.e., nemolizumab, or a fragment or variant thereof, is administered once every four weeks).

[0178] In some embodiments, the dosage of nemolizumab or a fragment or variant thereof administered to a subject may be within the range of 0.001 to 1,000 mg / kg of the subject's body weight. In some embodiments, the dosage may be about 0.01 to 50 mg / kg, about 0.01 mg / kg to about 0.1 mg / kg, about 0.05 mg / kg to 0.15 mg / kg, about 0.1 mg / kg to about 0.6 mg / kg, about 0.1 mg / kg to about 1 mg / kg, about 0.25 mg / kg to about 0.75 mg / kg, about 0.4 mg / kg to about 0.8 mg / kg, about 0.4 mg / kg to about 1.8 mg / kg, about 0.5 to about 2.5 mg / kg, about 0.8 mg / kg to about 2.2 mg / kg, about 1 mg / kg to about 2.5 mg / kg, about 1 mg / kg to about 3.5 mg / kg, about 0.01 mg / kg to about 50 mg / kg, about 0.01 mg / kg to about 0.1 mg / kg, about 0.05 mg / kg to about 0.15 mg / kg, about 0.1 mg / kg to about 0.6 mg / kg, about 0.1 mg / kg to about 1 mg / kg, about 0.25 mg / kg to about 0.75 mg / kg, about 0.4 mg / kg to about 0.8 mg / kg, about 0.4 mg / kg to about 1.8 mg / kg, about 0.5 to about 2.5 mg / kg, about 0.8 mg / kg to about 2.2 mg / kg, about 1 mg / kg to about 2.5 mg / kg, about 1 mg / kg to about 3.5 mg / kg, about 0.01 mg / kg to about 50 mg / kg, about 0.01 mg / kg to about 0 The range of nemolizumab or a fragment or variant thereof is about 1 mg / kg to about 5 mg / kg, about 2 mg / kg to about 4 mg / kg, about 2.5 mg / kg to about 10 mg / kg, about 5 mg / kg to about 10 mg / kg, about 10 mg / kg to about 20 mg / kg, about 10 mg / kg to about 40 mg / kg, about 20 mg / kg to about 50 mg / kg, about 25 mg / kg to about 75 mg / kg, about 50 mg / kg to about 100 mg / kg, or about 100 mg / kg to about 500 mg / kg, or about 100 mg / kg to about 1000 mg / kg of body weight. In preferred embodiments, the dosage is in the range of about 0.01 mg / kg to about 0.1 mg / kg, about 0.1 mg / kg to about 0.5 mg / kg, about 0.5 mg / kg to about 1.5 mg / kg, about 1.5 mg / kg to about 2.5 mg / kg, or about 2.5 mg / kg to about 10 mg / kg.In some embodiments, the dosage is about 0.01 mg / kg, about 0.02 mg / kg, about 0.03 mg / kg, about 0.04 mg / kg, about 0.05 mg / kg, about 0.06 mg / kg, about 0.07 mg / kg, about 0.08 mg / kg, about 0.09 mg / kg, about 0.1 mg / kg, about 0.2 mg / kg, about 0.3 mg / kg, about 0.4 mg / kg, about 0.5 mg / kg, about 0.6 mg / kg, about 0.7 mg / kg, about 0.8 mg / kg, about 0.9 mg / kg, about 1 mg / kg, about 1.1 mg / kg, about 1.2 mg / kg, about 1.3 mg / kg, about 1.4 mg / kg, about 1.5 mg / kg, about 1.6 mg / kg, about 1.7 mg / kg, about 1.8 mg / kg, about 1.9 mg / kg, about 2.0 mg / kg, about 2.1 mg / kg, about 2.2 mg / kg, about 2.3 mg / kg, about 2.4 mg / kg, about 2.5 mg / kg, about 2.6 mg / kg, about 2.7 mg / kg, about 2.8 mg / kg, about 2.9 mg / kg, about 3.0 mg / kg, about 3.1 mg / kg, about 3.2 mg / kg, about 3.3 mg / kg, about 3.4 mg / kg, about 3.5 mg / kg, about 3.6 mg / kg, about 3.7 mg / kg, about 3.8 mg / kg, about 3.9 mg / kg, about 3.1 mg / kg, about 3.2 mg / kg, about 3.3 mg / kg, about 3.4 mg / kg, about 3.5 mg / kg, about 3.6 mg / kg, about 3.7 mg / kg, about mg / kg, about 1.8 mg / kg, about 1.9 mg / kg, about 2 mg / kg, about 2.1 mg / kg, about 2.2 mg / kg, about 2.3 mg / kg, about 2.4 mg / kg, about 2.5 mg / kg, about 2.6 mg / kg, about 2.7 mg / kg, about 2.8 mg / kg, about 2.9 mg / kg, about 3 mg / kg, about 3.5 mg / kg, about 4 mg / kg, about 4.5 mg / kg, about 5 mg / kg, about 6 mg / kg, about 7 mg / kg, about 8 mg / kg, about 9 mg / kg, about 10 mg / kg, about 15 mg / kg, about 25 mg / kg, about 50 mg / kg, about 75 mg / kg, about 100 mg / kg, about 500 mg / kg or about 1,000 mg / kg. In certain embodiments, the dosage of nemolizumab or a fragment or variant thereof is about 0.1 mg / kg, about 0.5 mg / kg, about 1 mg / kg, about 1.5 mg / kg, about 2 mg / kg, or about 2.5 mg / kg. In a preferred embodiment, the dosage is about 0.5 mg / kg.

[0179] In some embodiments, the dosage of nemolizumab or a fragment or variant thereof administered to a subject is in the range of 1-100 mg, 25-75 mg, 30-60 mg, 40-80 mg, 20-80 mg, 1-25 mg, 1-50 mg, 10-90 mg, 15-85 mg, or any range therebetween. In some embodiments, the dosage can be about 5 mg, about 10 mg, about 15 mg, about 20 mg, about 25 mg, about 30 mg, about 35 mg, about 40 mg, about 45 mg, about 50 mg, about 55 mg, about 60 mg, about 65 mg, about 70 mg, about 75 mg, about 80 mg, about 85 mg, about 90 mg, about 95 mg, about 100 mg, about 105 mg, about 110 mg, about 115 mg, about 120 mg, or more. In some embodiments, the dosage can be 5 mg, 10 mg, 15 mg, 20 mg, 25 mg, 30 mg, 35 mg, 40 mg, 45 mg, 50 mg, 55 mg, 60 mg, 65 mg, 70 mg, 75 mg, 80 mg, 85 mg, 90 mg, 95 mg, 100 mg, 105 mg, 110 mg, 115 mg, 120 mg or more.

[0180] In some embodiments of the method and use of the present disclosure, a loading dose of about 60 mg of nemolizumab or a fragment or variant thereof may be administered to a subject with PN, followed by a continuous dose of about 30 mg of nemolizumab fragment or variant thereof once every 4 weeks. In some embodiments of the method and use of the present disclosure, a first dose of about 60 mg of nemolizumab or a fragment or variant thereof may be administered to a subject with PN, followed by a continuous dose of about 60 mg of nemolizumab fragment or variant thereof once every 4 weeks (i.e., the dose remains constant or is a "flat" dosing regimen). In some embodiments of the method and use of the present disclosure, a first dose of about 30 mg of nemolizumab or a fragment or variant thereof may be administered to a subject with PN, followed by a continuous dose of about 30 mg of nemolizumab fragment or variant thereof once every 4 weeks.

[0181] In some embodiments of the methods and uses of the present disclosure, nemolizumab or a fragment or variant thereof is administered by a topical route or a parenteral route. In some embodiments, nemolizumab or a fragment or variant thereof is administered subcutaneously. In some embodiments, the dose is administered subcutaneously at or proximal to one or more nodules, lesions, or excisions.

[0182] In some embodiments of the methods and uses of the present disclosure, nemolizumab or a fragment or variant thereof is administered daily, every other day, twice a week, three times a week, four times a week, five times a week, six times a week, once a week, once every two weeks, once every three weeks, once every four weeks, once every five weeks, once every six weeks, once every seven weeks, once every eight weeks, once every nine weeks, once every ten weeks, once every eleven weeks, once every twelve weeks, twice a year, once a year, and / or as needed based on the appearance of symptoms of PN. In a preferred embodiment, nemolizumab or a fragment or variant thereof is administered once every four weeks or once every eight weeks.

[0183] In some embodiments of the disclosed methods and uses, the duration of treatment or prevention is about 1 day, about 1 week, about 2 weeks, about 3 weeks, about 4 weeks, about 5 weeks, about 6 weeks, about 7 weeks, about 8 weeks, about 9 weeks, about 10 weeks, about 11 weeks, about 12 weeks, about 13 weeks, about 14 weeks, about 15 weeks, about 16 weeks, about 17 weeks, about 18 weeks, about 19 weeks, about 20 weeks, about 24 weeks, about 30 weeks, about 36 weeks, about 40 weeks, about 48 weeks, about 50 weeks, about 1 year, about 2 years, about 3 years, about 4 years, about 5 years, or as needed based on the appearance of symptoms of PN. In preferred embodiments, the duration of treatment or prevention is about 12 weeks to about 24 weeks, about 12 weeks to about 36 weeks, about 12 weeks to about 48 weeks, or about 24 weeks to about 36 weeks.

[0184] The present disclosure provides the use of nemolizumab or a fragment or variant thereof in the manufacture of a medicament for treating or preventing PN, for normalizing differentially expressed genes (DEGs) in subjects with PN, for normalizing epidermal hyperproliferation, for normalizing epidermal differentiation, and / or for reducing inflammatory responses in the skin. All of the doses, administration regimens, administration routes, biomarkers, and therapeutic endpoints of the present disclosure are also applicable to these uses.

[0185] The following examples are provided to illustrate the present disclosure, it being understood that the invention should not be limited to the specific conditions or details described in these examples. EXAMPLES

[0186] Example 1 - Treatment of Prurigo Nodularis with Nemolizumab method Patient cohort The current study was conducted in patients with moderate to severe PN. Briefly, 70 patients were randomized 1:1 to placebo (36 patients) or nemolizumab (34 patients), the latter at a dose of 0.5 mg / kg body weight administered at baseline, week 4, and week 8. Peak pruritus scores on a numerical rating scale (PP-NRS) were recorded: pruritus severity on a numerical rating scale ranged from 0 (no itch) to 10 (worst imaginable itch), and peak pruritus was estimated using the worst score once every 24 hours for 7 days, with the highest score recorded as the peak score. The primary outcome of the study was the percentage change from baseline in the PP-NRS at week 4.

[0187] Reconstructed human epidermis (RHE) model A three-dimensional RHE model was generated. Briefly, normal human dermal fibroblasts (NHDF) and normal human epidermal keratinocytes (NHEK) were used to generate RHE cultures. The RHE cultures used were full thickness with dermis and epidermis and were composed of autologous fibroblasts and keratinocytes. They were scaffold-free (no exogenous matrix), allowing the cells to self-organize the different layers of the skin and avoiding some potential inhibitors contained in the collagen matrix for RNA extraction. The RHE was inserted into the insert (1.2 cm 2 ) for 42 days to obtain at least two layers of dermis and six layers of epidermis. RHE cultures from six different donors were left unstimulated or stimulated with seven different cytokines or cytokine combinations (three replicates per donor per condition): IL-31, IL-13, IL-17A, IFNg, IL-31+IL-13, IL-31+IL-17A, IL-31+IFNg and IL-13+IL-17A. The concentrations of each of the cytokines used were as follows: IL-31 (500 ng / mL), IL-13 (100 ng / mL), IL-17A (200 ng / mL), IFNg (50 ng / mL). RHE cultures were lysed 72 hours after stimulation and RNA extraction was performed using MagMAX mirVana Total RNA isolation from Tissue Kit (ThermoFisher Scientific). RNA was purified and concentrated using the RNA Clean&Concentrator-5 kit (Zymo Research) according to the protocol, and RNA concentrations ranged from 3 to 380 ng / μL. Total RNA was quantified using the QuantiFluor One RNA kit (Promega) on a GloMax-Multi+ detection system (Promega). Library preparation was performed using the Illumina® Stranded mRNA Prep Ligation Kit (Illumina) according to the manufacturer's recommendations.

[0188] Skin biopsy processing and RNA isolation Skin biopsies were taken from 16 placebo and 15 nemolizumab-treated subjects. Samples included lesional and non-lesional biopsies at baseline, and lesional samples (placebo or nemolizumab) after 12 weeks of treatment. RNA was extracted from the skin biopsies using Tripure Isolation Reagent (Sigma-Aldrich) according to the manufacturer's instructions. From these RNA samples, DNase treatment was applied using an RNase-Free DNase kit (catalog number 79254, Qiagen) followed by an RNeasy® MinElute® Cleanup kit (catalog number 74204, Qiagen).

[0189] Total RNA was quantified using the QuantiFluor One RNA kit (Promega). The resulting RNA concentrations range from 4 to 20 ng / μl. Total RNA was qualified using a Fragment Analyzer 5300 (Agilent) with the Agilent HS RNA Fragment Kit (Agilent). The resulting RNA quality numbers (RQNs) range from 1 to 6. Library preparation was performed using the SMARTer Stranded Total RNA-Seq Kit V2-Pico Input Mammalian Kit (TaKaRa).

[0190] RNA-seq Libraries were quantified using QuantiFluor One dsDNA kit (Promega) and library analysis was performed using the Agilent HS NGS Fragment kit (Agilent) on a Fragment Analyzer5300 (Agilent). Following size selection with AMPure XP beads, shotgun libraries were sequenced using NextSeq (Illumina) on an Illumina NextSeq500 sequencer at 2 × 75 bp (High Output Kit v2, 150 cycles).

[0191] RNA-seq data processing After adaptor trimming, sequence reads of 83 unique samples were aligned to the human genome (GRCh37) using STAR. Reads that uniquely mapped to one genomic location were then used to quantify gene (GENCODE v29) expression levels with HTSeq. Two RNA-seq samples were identified as outliers, and 81 samples were used for subsequent analysis. Only genes with an average of at least one read per sample were retained. DESeq2 was used for expression normalization, and a negative binomial distribution was used to model expression levels for differential expression analysis. For non-lesional vs. lesional and baseline vs. week 12 comparisons, individual effects were included as covariates, and for placebo vs. nemolizumab comparisons, age and sex were controlled. False discovery rate (FDR) ≤ 5% and |log2 fold change| >= 1 were used as criteria for declaring significant differentially expressed genes (DEGs).

[0192] Cytokines, cellular signatures, and functional inference analyses The most significant DEGs were compared to transcripts induced by cytokines in keratinocytes (defined by FDR<=10% and 1.5 fold change (FC)). A fair comparison was performed using the top 1,000 most significant DEGs from the PN data. For comparison with the epidermal compartment gene signature, scRNA-seq was performed on the epidermal layer of skin biopsies to identify the top 50 marker genes for the basal, differentiated, and keratinized layers, respectively. Effect sizes (log2FC) were then investigated for each marker gene in each differential expression comparison. For transcriptional analysis, promoter regions were defined as 5,000 base pairs upstream of the transcription start site, and enrichment statistics for transcription factor binging were calculated using meme suite.

[0193] Weighted Gene Correlation Network Analysis (WGCNA) Genes expressed in at least 20% of the samples were used in the WGCNA dataset. The "softPower" parameter was chosen as the minimum value that achieved at least r2>=0.75. Spearman correlation was used to calculate correlations and the minimum module size was set to 100. When modules were confluent, a high cut of 0.2 was used.

[0194] result Prurigo nodularis is characterized by abnormal keratinocyte differentiation and immune activation After quality control, we performed RNA sequencing data analysis of biopsy tissues from patients with prurigo nodularis (PN). There were 31 PN patients with transcriptome data in both lesional and non-lesional skin samples. Using a false discovery rate (FDR) of ≤10% and |log2| ≥ 1 as criteria, 5,943 differentially expressed genes (DEGs) were identified when comparing uninvolved and lesional skin at baseline, of which 2,060 genes were increased and 3,874 genes were decreased (Figure 1A). Genes that showed the most robust increase included KRT6C (588-fold, FDR = 8.2 × 10 -80 ), DEFB4A (150x, FDR=1.1×10 -12 ), and KRT16 (90-fold, FDR = 1.9 × 10 -52 ). Reduced genes included LCE5A (11-fold reduction, FDR = 8.1 × 10 -18 ) and AQP7 (7.9-fold decrease, FDR = 2.6 × 10 -17 Regarding cytokines, the most prominent upregulated genes were IL36 family members: IL36A (6.8-fold, FDR = 1.8 × 10 -4 ) and IL36G (8.4-fold, FDR = 3.9 × 10 -25 ), IL-20 family member: IL19 (5.1-fold, FDR = 7.4 × 10 -4 ), IL20(3.5-fold, FDR=1.7×10 -3 ), IL22 (2.7-fold, FDR = 2.9 × 10 -2 ), IL24 (5.8-fold, FDR = 3.8 × 10 -10), and IL26 (4.9-fold, FDR = 3.3 × 10 -3 ) were included. Other contributing factors were IL1A (4.7-fold, FDR = 1.0 × 10 -12 ), and IL1B (4.1-fold, FDR = 3.7 × 10 -6 The Th2 cytokines IL4 and IL13 did not reach significance, but IL4R was increased 2.6-fold (FDR = 6.3 × 10 -19 ) (see Table 1 at the end of the Examples section herein).

[0195] Functional enrichment analysis was then performed on the DEGs to define biological processes related to PN skin. The most prominent gene ontology (GO) terms included "cornified envelope" (FDR = 1.5 × 10 -12 ), “Epidermal cell differentiation” (FDR=6.4×10 -10 ), “keratinization” (FDR=1.6×10 -12 ), "peptidase regulator activity" (FDR = 1.1 × 10 -4 ), "Interleukin-4 and 13 signaling" (FDR = 6.8 × 10 -7 ), “interferon alpha / beta signaling” and “response to interferon gamma” (FDR=4.1×10 -7 , and FDR = 4.1 × 10 -6 ), "IL23 pathway" (FDR = 2 × 10 -5 ), and “metaphase and anaphase” (FDR = 3.8 × 10 -10 ) (Figure 1B) (see also Table 2 at the end of the Examples section herein). These reflect the hyperproliferative nature of PN associated with altered epidermal differentiation and inflammatory components, and focusing on key expression modules in the lesional skin transcriptome revealed an inflammatory network that included the proliferation marker Ki67 (MKI67), the cell cycle gene CDKN1A, and IL-1 and IL-36 (Figure 1C).

[0196] To better understand the disease regulatory network involved in PN skin weighted gene co-expression network analysis (WGCNA) was performed. 20 co-expression modules were identified in non-lesional skin and 10 clusters were identified in lesional PN skin (Figure 1D) (see Table 3 at the end of the Examples section herein). This analysis allowed the inventors to identify the "immune response" (FDR = 1.8 x 10 -47 ), “protective response” (FDR = 1.2 × 10 -39 ), the most prominent involved immunological processes (module #8), including "cell cycle" (FDR=2.9×10 -94 ), "DNA metabolic process" (FDR = 8.7 × 10 -67 ), and “Epidermal Development” (FDR 3.5 × 10 -10 ), “keratinization” (FDR=1.7×10 -6 We were able to assign distinct functions to these co-expressed gene modules, especially those co-expressed in PN lesion skin, including epidermal processes such as “extracellular matrix” (module #5) (Figure 1E). Another notable finding was the low FDR of “extracellular matrix” (FDR=1.16×10 -59 , module #2), including genes such as MMP14, MMP16, COL1A1, COL1A2, and COL3A1, which were modestly elevated in lesional skin (FC ≥ 1.4, FDR ≤ 6 × 10), consistent with an association between PN and dermal fibrosis. -2 ).

[0197] Transcriptomic alterations in PN lesions are enriched for keratinocyte and T cell signatures An in silico approach (xCell) was used to estimate specific cell type signatures for each non-lesional and lesional PN skin sample. Enrichment of transcriptomic signatures related to epithelial cells and keratinocytes was observed (p<0.001 and p<0.0001, respectively) (Figure 2A). There was also an increased prominence of Th2-related signatures (p<0.0001), consistent with the enriched GO category of IL-4 / IL-13 (Figure 1B). Other inflammatory signatures, such as macrophages (p<0.01), were more variable (Figure 2A). To address the relationship of PN to other hyperproliferative skin diseases with strong inflammatory signatures, the PN transcriptome was also compared to those of atopic dermatitis (AD) and psoriasis. In a three-way comparison, there was a large number of genes shared between all three diseases, both for up- and down-regulated genes (Figure 2B). The correlation of effect sizes in lesional skin was more pronounced between PN and psoriasis (Spearman correlation ρ=0.64) than between PN and AD (ρ=0.55). Genes commonly upregulated in both psoriasis and PN include those involved in cytokine activity (CCL3, CXCL10, IFNG, IL12B, IL19, IL1B, IL20, etc.) and keratinization (KRT16, KRT17, LCE3A, LCE3E, etc.). See Table 4 at the end of the Examples section of the specification.

[0198] Transcriptome changes in PN skin induced by IL31 receptor inhibitor nemolizumab Clinical results in patients with PN showed that nemolizumab resulted in a higher percentage of improvement in pruritus and skin lesions with an overall favorable safety profile. At week 12, the proportion of patients who achieved a 4-point reduction in the weekly average of PP-NRS was significantly higher in the nemolizumab group compared to placebo (52.9% vs. 8.3%, p<0.001). At week 12, the proportion of subjects who achieved IGA success (defined as IGA 0 [clear] or 1 [almost clear]) was significantly higher in the nemolizumab group compared to placebo (20.6% vs. 2.8%, p=0.02).

[0199] To address the therapeutic effect of the IL-31 receptor (IL-31R) inhibitor nemolizumab, we performed RNA-seq data from PN biopsies before and after 12 weeks of treatment and used a placebo control in a double-blind study. At baseline, there were 16 and 15 individuals in the placebo and nemolizumab groups, respectively, and lesion samples were obtained from 18 patients (11 placebo and 7 nemolizumab) at week 12. Using principal component analysis (PCA), we observed a mixture of samples from patients with PN at baseline between the two treatment groups (placebo vs. nemolizumab). After 12 weeks of treatment, there was a tendency to group the nemolizumab cohorts, but not the placebo cohorts (Figure 3A). This involved sample clustering using genes identified as differentially expressed in non-lesional vs. lesional skin comparisons, grouping six of seven nemolizumab samples (86%) at week 12 with baseline non-lesional skin samples, compared to 57% (4 of seven) in the placebo group (Figure 3B). Notably, nemolizumab treatment led to normalization of more PN-associated DEGs compared to placebo for both genes upregulated in PN lesional skin (969 nemolizumab vs. 211 placebo) and genes downregulated in PN lesional skin (1,268 genes nemolizumab vs. 166 placebo) (Figure 4A). This was also reflected in the correlation between placebo and nemolizumab-treated DEGs, where there was much greater overlap between nemolizumab-treated groups and PN compared to placebo vs. PN for both increased and decreased DEGs (Figure 4B). GO categories enriched among DEGs by week 12 were only decreased in the nemolizumab treatment group, with “cell cycle” (FDR=5.6×10 -14 ), "keratinocyte differentiation" (FDR = 1.8 × 10 -4 ), and “Interleukin-4 and 13 signaling” (FDR = 1.5 × 10 -2 ), whereas these GO categories were not found in the placebo control group (see Table 2 at the end of the Examples section herein). These data demonstrate that nemolizumab treatment normalized both epidermal hyperproliferation and differentiation in addition to reducing inflammatory responses, particularly those associated with Th2 responses.

[0200] Nemolizumab response is associated with a decrease in IL-31 / Th2 responses in PN skin To address the effect of nemolizumab treatment on inflammatory responses in PN skin, nemolizumab and placebo treatment responses were examined against the cytokine response signatures generated in RHE cultures as well as human epidermal rafts. A consistent reduction in IL-31 responses was observed alone or in combination with other inflammatory cytokines, including the Th2 cytokines IL-13 or IL-17A (Figure 5A), providing clear evidence of IL-31 pathway blockade by nemolizumab. In particular, IL-17A-responsive genes were enriched in PN skin (Table 4), likely corresponding to the overlap of specific downstream immunological cascades between psoriasis and PN. IL17A mRNA expression itself was not significantly different in non-lesional versus lesional skin or with nemolizumab treatment up to week 12 (see Table 1 at the end of the Examples section herein), suggesting that IL-17A is not the dominant cytokine in the PN IL-17A signature, but is downstream of IL-31 signaling. In terms of changes in the cellular transcriptome, a more robust reduction was observed in the nemolizumab group, including Th1 and Th17 (Figure 5B).

[0201] To determine the tissue compartment with the greatest contribution to the cellular response to anti-IL-31R blockade, transcriptome data from the placebo and nemolizumab groups were compared with the gene signature of the epidermal compartment obtained from single-cell RNA-seq data. Results show that while the basal keratinocyte (KRT14+) signature was elevated in PN lesional skin, the latter was restored to a similar extent in both placebo and treatment groups, induction of the epidermal spinous layer (KRT10+) signature in PN lesional skin was only restored by treatment, but not in the placebo group (Figure 5C). Transcription factor binding site (TFBS) analysis was then performed to further understand the transcriptional regulators of PN and transcriptome changes following placebo or nemolizumab treatment. Results demonstrated that binding sites enriched among genes upregulated in baseline lesional skin were more likely to be enriched among genes downregulated by nemolizumab by week 12 (Figure 5D). The most significantly regulated transcription factors included EGR4, a member of the EGF family of zinc finger transcription factors (enrichment in the promoters of upregulated genes and nemolizumab downregulated genes, respectively, p = 4.5 × 10 -6 and p = 1.2 × 10 -8 ), STAT3 (p = 2.2 × 10 -4 and p = 2.5 × 10 -5 ), and KLF16 (p=4.5×10 -5 and p = 2 × 10 -5 ) were included (see Table 5 at the end of the Examples section herein).

[0202] Nemolizumab results in a reduction in pruritus scores Peak Pruritus Numeric Rating Scale (PP-NRS), PP-NRS, was associated with the transcriptomic data (Figure 6A). PP-NRS was similar at baseline in both groups, with consistent reductions in pruritus only in the nemolizumab group, whereas a broad range of responses was seen in the placebo group. Also, all patients in the nemolizumab group showed improvement, whereas only a subset of patients in the placebo group showed significant changes. Notably, the measured distance between the principal component 1 and principal component 2 (PC1 / PC2) components in the nemolizumab treatment group was much shorter than that in the placebo group (Figure 6B), consistent with a treatment response.

[0203] Consideration The data presented herein are the first to provide a comprehensive view of global transcriptome changes in PN skin and reveal novel and important insights into the mechanism of action and efficacy of the anti-IL-31 receptor inhibitor nemolizumab. Notably, these data reflect at the transcriptome level many of the characteristic histological changes observed in PN, including epidermal changes, inflammatory responses (Figures 2A-C), fibrosis, and pruritus (Figures 6A-B), and how these are normalized with nemolizumab treatment.

[0204] Many of the gene expression changes in PN skin are associated with abnormal keratinocyte proliferation and differentiation (Figure 1A-1E). These epidermal changes account for the majority of PN's overlap with both AD and psoriasis (Figure 2A-2C, Table 4), both of which are also characterized by marked epidermal proliferation and altered epidermal differentiation. Notably, these changes showed significant improvement in the nemolizumab-treated group by week 12 of treatment, but not in the placebo group (Table 2). Furthermore, consistent with the therapeutic effect of nemolizumab, the greatest effect of normalization was observed in the differentiation layer of the epidermis (Figure 5C), which may reflect a decrease in keratinocyte proliferation and restoration of normal epidermal differentiation.

[0205] Consistent with PN being an inflammation-driven disease process, immune responses such as Th2 (IL-4 / IL-13) responses and type I and type II IFN responses were prominent (Figure 1A-Figure 1E). Th2 responses are closely correlated with pruritus in diseases such as atopic dermatitis, a common predisposing disease to PN development. Interestingly, anti-IL-31 receptor inhibition resulted in a reduction in Th2 responses, as well as a reduction in Th17 (Figure 5B), corresponding to a reduction in IL-31 responses in keratinocytes of PN skin (Figure 5A), as well as a reduction in Th17 (Figure 5B), corresponding to a reduction in IL-13 and IL-17 responses in keratinocytes (Figure 5A). The enriched IL-17 response in PN skin, and the contribution from IL-36, likely explain the greater overlap of PN skin with plaque psoriasis, in contrast to AD (Figure 2B and Figure 2C). No significant changes in IL17A mRNA expression in the PN data were observed, suggesting that IL-17A is not the dominant cytokine in PN. In addition, the alteration of both Th2 and Th17 responses, and to a lesser extent type II IFN responses, with nemolizumab treatment suggests that these cytokines act downstream of IL-31 in PN.

[0206] Fibrosis is a hallmark of PN, most prominent in the papillary dermis and less common in the reticular dermis. Fibrosis in PN is characterized by the deposition of vertically oriented collagen fibrils. A module of genes involved in extracellular matrix biology was found to be enriched in PN skin, including both collagen 1 and collagen 3 genes (Table 3). COL1A1, COL1A2, and COL1A3 mRNA were increased in PN skin at baseline (1.7-, 1.44-, and 1.52-fold, respectively, see Table 1) but did not show significant changes with nemolizumab treatment at week 12.

[0207] The data also show how nemolizumab induced changes in the transcriptome of PN correlate with improvements in pruritus. Chronic pruritus is a debilitating symptom of PN and severely impacts quality of life. Although the pathogenesis of pruritus in PN remains unclear, possible contributing factors include alterations in Th2 cytokines, IL-4 and IL-13, the major prurigens in atopic dermatitis, versus cutaneous nerve alterations, which have been shown to reduce the density of intraepidermal nerve fibers in both lesional and non-lesional skin. Data from nemolizumab treatment are consistent with both of these scenarios contributing to pruritus. Thus, nemolizumab treatment results in suppression of Th2 and IL-4 / IL-13 responses in PN skin, as well as reduced expression of factors such as KLF16, which have been shown to inhibit neurite outgrowth. In addition, nerve growth factor (NGF), identified as increased in PN skin, is also normalized to a greater extent with nemolizumab treatment at week 12 compared to the placebo group (Table 1). No changes were seen in CGRP or substance P (TAC1) expression. These data are highly suggestive of a broad effect of nemolizumab on pruritus and may explain the prolonged period of improvement in pruritus seen beyond the last dose of nemolizumab (>2 months).

[0208] In summary, PN is a debilitating and difficult to treat condition, with no currently approved therapies for its treatment. This study provides a detailed characterization of transcriptomic changes in PN skin and demonstrates a broad mechanism of action of the anti-IL-31 receptor inhibitor nemolizumab. These data demonstrate the broad therapeutic effects of anti-IL-31 receptor inhibition with nemolizumab on multiple aspects of PN pathogenesis, including epidermal differentiation, inflammatory responses, pruritus, and extracellular remodeling, confirming an upstream role for IL-31 in PN pathogenesis.

[0209] Example 2 - Plasma proteomic analysis of nemolizumab-treated patients with moderate to severe prurigo nodularis This example details a randomized, double-blind, phase 2 study of the anti-human IL31Ra blocking antibody nemolizumab at a dose of 0.5 mg / kg body weight administered subcutaneously at baseline, week 4, and week 8 compared with placebo in patients with moderate to severe prurigo nodularis. The aim of this study was to characterize the effect of nemolizumab on the entire plasma proteome using mass spectrometry.

[0210] Materials and Methods Patient Data Set Nineteen placebo non-responders and 19 nemolizumab responders were selected based on the change in PP-NRS (Peak Pruritus Score Numeric Rating Scale) clinical score at week 12 (19 placebo patients with a PP-NRS change ≥ -1.4 and 19 nemolizumab patients with a PP-NRS change < -5.4). The PP-NRS scores at baseline and after 12 weeks of nemolizumab treatment are shown in Figure 10.

[0211] protocol After plasma gY14 ultra-depletion, samples were analyzed using Proteome Science's TMT Calibrator MS2 workflow, using a pool of healthy skin biopsies 1) to allow batch correction between each plex and 2) to trigger the mass spectrometer to detect skin-associated peptides in blood.

[0212] statistics Only peptides corresponding to unique proteins were used for statistical analysis. Both baseline-corrected and baseline-uncorrected values ​​were used to calculate the differentially expressed proteins. Three statistical tests were used to calculate the following list of differentially expressed proteins: 1) least squares regression (with or without patient ID), 2) robust regression, and 3) general regression. 193 proteins were found to be differentially expressed (adjusted p-value < 0.05). No additional (e.g. logFC) filters were applied to this protein list. Enrichment analysis was performed using the 193 differentially expressed proteins using QIAGEN IPA (QIAGEN Inc., digitalinsights.qiagen.com / IPA).

[0213] result The list of 193 differentially expressed proteins was then analyzed using the enrichment software IPA. The p-value indicates the significance of the pathway affected in the dataset, while the z-score provides insight into the directionality, i.e., how pathways in the dataset are "activated" or "inhibited."

[0214] Classical pathways were sorted according to z-score only (Figure 11 right) or sorted by z-score and filtered by p-value (Figure 11 left, -log(p-value)>1.3 corresponding to p-value<0.05). IL-31 has been shown to induce activation of STAT3. Although STAT3 activation is not specific to IL-31, downregulation of this pathway in nemolizumab responders suggests target engagement by nemolizumab. Furthermore, compared to placebo non-responders, the signature of nemolizumab responders was characterized by downregulation of the IL-6 pathway. In addition, the vascular endothelial growth factor (VEGF) pathway, which has been shown to correlate with the severity of PN, was downregulated in nemolizumab responders, likely reflecting clinical improvement. Finally, two neuronal ontologies ("CREB signaling in neurons" and "synaptogenesis signaling pathway") were identified as downregulated in nemolizumab responder subjects, highlighting the influence of IL-31 as a neuroinflammatory cytokine in PNs.

[0215] The upstream regulator analysis allowed the identification of transcription factors and small molecules whose function is affected by the disease or treatment depending on the analyzed dataset. The proteins identified in this analysis may not be themselves differentially expressed in the studied dataset, but their activation / inhibition status can explain the expression changes in the dataset. The classical pathways were sorted according to z-score only (Figure 12 right) or sorted by z-score and filtered by p-value (Figure 12 left, p-value < 0.05). The profibrotic transforming growth factor beta 1 (TGFβ1) was found as a potential regulator of the nemolizumab responder signature and may indicate the resolution of fibrotic nodules. Another potential regulator was signal transduction and activator of transcription 5b (STAT5b), which may reflect target engagement by nemolizumab. These data clearly suggest that nemolizumab strongly influences inflammatory and tissue remodeling processes in PN.

[0216] Biological function analysis of nemolizumab responders revealed downregulated "leukocyte migration" and downregulated "leukocyte cell migration." In addition, two terms related to the neuraxis were downregulated: "glial cell death" and "glial cell apoptosis," possibly reflecting improvement in pruritus (FIG. 13).

[0217] Consideration This example aimed to perform a broad and exploratory analysis to identify plasma protein biomarkers that may also explain the mechanism of action of nemolizumab treatment.

[0218] To characterize the full range of plasma proteome changes, a preselected group of nemolizumab responders and placebo non-responders was evaluated. Due to this preselection of patients, the signature described in this example may reflect not only changes induced by nemolizumab, but also changes due to a decrease in PP-NRS.

[0219] The dataset analyzed in this study reveals that the Nemolizumab responder signature was characterized by amelioration of multiple aspects of PN pathophysiology, including inflammation, neuroimmune function, and tissue remodeling.

[0220] Enrichment analysis reveals that the nemolizumab responder signature was characterized by reduced leukocyte migration and cell migration. The STAT3 pathway, a direct target of IL-31 signaling, was also inhibited in nemolizumab responder subjects, suggesting target engagement. Nemolizumab responders also showed downregulation of other pro-inflammatory cytokine pathways, including the IL-6 and VEGF pathways.

[0221] Pathway analysis also revealed effects of nemolizumab treatment on neuron-related processes, including the CREB signaling in neurons and synaptogenesis signaling pathways, glial cell death, and glial apoptosis, consistent with the observed clinical impact of nemolizumab on pruritus.

[0222] Finally, the dataset showed a decrease in TGFB1 pathway activation suggesting an impact of nemolizumab treatment on tissue remodeling.

[0223] These observations strengthen the understanding that plasma proteomics can effectively capture, at least in part, the tissue-specific (i.e., skin) effects of nemolizumab in PN.

[0224] [Table 1] 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[0225]

Table 2

[0226]

Table 3

[0227]

Table 4

[0228]

Table 5

Claims

1. 1. A medicament comprising an anti-IL-31RA antibody for use in a method for treating or preventing prurigo nodularis (PN) in a subject, comprising: The method includes administering to a subject with PN an anti-IL-31RA antibody; and The subject differentially expresses at least one gene selected from the genes disclosed in Table 1 as compared to a reference expression level of the at least one gene. The said medicine.

2. 1. A medicament comprising an anti-IL-31RA antibody for use in a method for normalizing differentially expressed genes (DEGs) in a subject with PN, the method comprising: The method includes administering to a subject with PN an anti-IL-31RA antibody; the subject differentially expresses at least one gene selected from the genes disclosed in Table 1 as compared to a reference expression level of the at least one gene; and The normalization is determined about 4 weeks, about 8 weeks, or about 12 weeks after administration of the anti-IL-31RA antibody. The said medicine.

3. The pharmaceutical of claim 1 or 2, wherein the differential expression of the at least one gene is determined by RT-qPCR, RT-PCR, RNA-seq, Northern blotting, serial analysis of gene expression (SAGE), or DNA or RNA microarray, or the differential expression of the at least one gene is determined at the protein level by Western blotting, ELISA, surface plasmon resonance, or mass spectrometry.

4. 1. A medicament comprising an anti-IL-31RA antibody for use in a method for normalizing epidermal hyperproliferation and / or normalizing epidermal differentiation in a subject with prurigo nodularis (PN), comprising: The method includes administering to a subject with PN an anti-IL-31RA antibody; the subject differentially expresses at least one gene selected from the genes disclosed in Table 1 as compared to a reference expression level of the at least one gene; and The normalization is determined about 4 weeks, about 8 weeks, or about 12 weeks after administration of the anti-IL-31RA antibody. The said medicine.

5. 1. A medicament comprising an anti-IL-31RA antibody for use in a method for reducing an inflammatory response in the skin of a subject having prurigo nodularis (PN), The method includes administering to a subject with PN an anti-IL-31RA antibody; the subject differentially expresses at least one gene selected from the genes disclosed in Table 1 as compared to a reference expression level of the at least one gene; and The inflammatory response is a Th2 response. The said medicine.

6. The pharmaceutical composition of claim 5, wherein the inflammatory response comprises overexpression of IL-13, IL-17A, IL-1, and / or IL-36.

7. The method of claim 1 or 2, wherein at least two, at least three, at least four, or at least five genes disclosed in Table 1 are differentially expressed.

8. 3. The medicament of claim 1 or 2, wherein the differentially expressed genes are selected from KRT6C, DEFB4A, KRT16, LCE5A, AQP7, IL36A, IL36G, IL19, IL20, IL22, IL24, IL26, IL1A, IL1B, IL4R, Ki67 (MKI67), CDKN1A, IL-1, or IL-36.

9. The method of claim 1 or 2, wherein the subject has a gene ontology as disclosed in Table 2.

10. 1. A medicament comprising an anti-IL-31RA antibody for use in a method for treating or preventing prurigo nodularis (PN) in a subject, comprising: The method includes administering to a subject with PN an anti-IL-31RA antibody; The treatment with the anti-IL-31RA antibody, (a) a decrease in leukocyte migration or leukocyte cell movement; (b) inhibition of the STAT3 pathway; (c) inhibition of the STAT5b pathway; (d) downregulation of IL-6 or the IL-6 pathway; (e) downregulation of VEGF or the VEGF pathway; (f) a decrease in TGFB1 pathway activation, or (g) Combinations thereof Bringing about (a) the reduction in leukocyte migration or leukocyte cell movement, (b) the inhibition of the STAT3 pathway, (c) the inhibition of the STAT5b pathway, (d) the downregulation of IL-6 or the IL-6 pathway, (e) the downregulation of VEGF or the VEGF pathway, (f) the reduction in TGFB1 pathway activation, or (g) a combination thereof, is determined with respect to (i) a control sample obtained from one or more individuals without PN, or (ii) a biological sample obtained from the subject prior to administration of the anti-IL-31RA antibody. The said medicine.

11. 1. A medicament comprising an anti-IL-31RA antibody for use in a method of altering an immune response in a subject with PN, comprising: The method includes administering to a subject with PN an anti-IL-31RA antibody; administering the anti-IL-31RA antibody (a) a decrease in leukocyte migration or leukocyte cell movement; (b) inhibition of the STAT3 pathway; (c) inhibition of the STAT5b pathway; (d) downregulation of IL-6 or the IL-6 pathway; (e) downregulation of VEGF or the VEGF pathway; (f) a decrease in TGFB1 pathway activation, or (g) Combinations thereof Bringing about (a) the reduction in leukocyte migration or leukocyte cell movement, (b) the inhibition of the STAT3 pathway, (c) the inhibition of the STAT5b pathway, (d) the downregulation of IL-6 or the IL-6 pathway, (e) the downregulation of VEGF or the VEGF pathway, (f) the reduction in TGFB1 pathway activation, or (g) a combination thereof, is determined with respect to (i) a control sample obtained from one or more individuals without PN, or (ii) a biological sample obtained from the subject prior to administration of the anti-IL-31RA antibody. The said medicine.

12. A medicament for use in a method for reducing an inflammatory response in the plasma of a subject with prurigo nodularis (PN), comprising an anti-IL-31RA antibody, The method includes administering to a subject with PN an anti-IL-31RA antibody; Reducing the inflammatory response comprises decreasing a plasma concentration of IL-6, VEGF, or a combination thereof compared to a reference level; and The reference level is determined from (i) a control sample obtained from one or more individuals who do not have PN, or (ii) a biological sample obtained from the subject prior to administration of the anti-IL-31RA antibody. The said medicine.

13. The pharmaceutical agent of claim 12, wherein after administration of the anti-IL-31RA antibody, the subject further exhibits: (a) decreased leukocyte migration or leukocyte cell movement; (b) inhibition of the STAT3 pathway; (c) inhibition of the STAT5b pathway; (d) decreased TGFB1 pathway activation; or (e) a combination thereof.

14. The pharmaceutical agent according to any one of claims 10 to 13, wherein the subject exhibits downregulation in a neuron ontology selected from CREB signaling in neurons, synaptogenesis signaling pathways, glial cell death and glial cell apoptosis, and combinations thereof, after administration of the anti-IL-31RA antibody.

15. The pharmaceutical composition of claim 1 or 2, wherein, in the method, the anti-IL-31RA antibody is administered subcutaneously.

16. The pharmaceutical agent described in claim 1 or 2, wherein in the method, the anti-IL-31RA antibody is administered once a week, once every two weeks, once every three weeks, once every four weeks, once every five weeks, once every six weeks, once every seven weeks, or once every eight weeks.

17. The pharmaceutical described in claim 1 or 2, wherein in the method, the anti-IL-31RA antibody is administered at a dose of about 10 mg, about 15 mg, about 20 mg, about 25 mg, about 30 mg, about 35 mg, about 40 mg, about 45 mg, about 50 mg, about 55 mg, about 60 mg, about 65 mg, about 70 mg, about 75 mg, about 80 mg, about 85 mg, or about 90 mg.

18. The pharmaceutical composition of claim 1 or 2, wherein the anti-IL-31RA antibody is administered according to a flat dosing regimen.

19. The pharmaceutical composition of claim 1 or 2, wherein pruritus scoring is improved in the subject after treatment.

20. The pharmaceutical according to claim 1 or 2, wherein the anti-IL-31RA antibody comprises a heavy chain variable region comprising HCDR1 comprising SEQ ID NO: 8, HCDR2 comprising SEQ ID NO: 9, and HCDR3 comprising SEQ ID NO: 10, and a light chain variable region comprising LCDR1 comprising SEQ ID NO: 12, LCDR2 comprising SEQ ID NO: 13, and LCDR3 comprising SEQ ID NO:

14.

21. The pharmaceutical composition according to claim 1 or 2, wherein the anti-IL-31RA antibody is nemolizumab or a fragment or variant thereof.

22. 1. A method of testing whether a subject with prurigo nodularis (PN) responds to treatment with an anti-IL-31RA antibody, comprising: Detecting the expression levels of at least one, at least two, at least three, at least four, or at least five of the differentially expressed genes (DEGs) of Table 1 in a sample obtained from a subject suspected of having PN; and comparing the expression levels of the DEGs to a reference level; the reference level is the corresponding gene expression level of each DEG in a sample from an individual without PN; the sample obtained from the subject suspected of having PN is a skin sample; and the DEGs are selected from KRT6C, DEFB4A, KRT16, LCE5A, AQP7, IL36A, IL36G, IL19, IL20, IL22, IL24, IL26, IL1A, IL1B, IL4R, Ki67 (MKI67), CDKN1A, IL-1, or IL-36; The said medicine.

23. The method of claim 22, wherein the sample obtained from the subject suspected of having PN is a skin sample containing a lesion or nodule.

24. 1. A method for testing whether a subject with prurigo nodularis (PN) responds to treatment with an anti-IL-31RA antibody, comprising: detecting in a post-treatment plasma sample obtained from a subject with PN who has been administered at least one dose of an anti-IL-31RA antibody: (a) a decrease in leukocyte migration or leukocyte cell movement; (b) inhibition of the STAT3 pathway; (c) inhibition of the STAT5b pathway; (d) downregulation of IL-6 or the IL-6 pathway; (e) downregulation of VEGF or the VEGF pathway; (f) a decrease in TGFB1 pathway activation, or (g) Combinations thereof detecting at least one, at least two, at least three, at least four, or at least five of the amounts of said reduction or inhibition is measured relative to a baseline amount, and said reduction or inhibition indicates a responsiveness to treatment; The baseline amount is determined from a plasma sample obtained from the same subject before treatment was initiated. The method.