WO2023154953A1

WO2023154953A1 - Gdf15 polypeptides for treating and preventing autoimmune diseases

Info

Publication number: WO2023154953A1
Application number: PCT/US2023/062564
Authority: WO
Inventors: Zhiming Chen; Jer-Yuan HSU; Varun KAPOOR; Jonathan SITRIN; Jade BOJKOVIC; Igor MIKAELIAN
Original assignee: Ngm Biopharmaceuticals, Inc.
Priority date: 2022-02-14
Filing date: 2023-02-14
Publication date: 2023-08-17

Abstract

The present disclosure provides methods of treating and preventing autoimmune disorders (e.g., multiple sclerosis (MS)), said methods comprising administering to a subject (e.g., a human) in need thereof a therapeutically effective amount of a growth differentiation factor 15 (GDF15) polypeptide. The present disclosure also provides a method of reducing or blocking pathogenic immune cell infiltration into the central nervous system of a subject, said method comprising administering to the subject a therapeutically effective amount of a GDF15 polypeptide. The disclosure also provides related GDF15 polypeptides, methods of making same, and pharmaceutical compositions comprising the same.

Description

GDF15 POLYPEPTIDES FOR TREATING AND PREVENTING

AUTOIMMUNE DISEASES

CROSS-REFERENCE TO RELATED APPLICATION

[0001] This application claims the priority benefit of U.S. Provisional Application No. 63/267,979, filed February 14, 2022, the content of which is incorporated by reference in its entirety herein.

SEQUENCE LISTING

[0002] The instant application contains a Sequence Listing which has been submitted electronically in XML file format and is hereby incorporated by reference in its entirety. Said XML copy, created on February 9, 2022, is named 47702- OllOWOl.xml and is 26,118 bytes in size.

FIELD OF THE INVENTION

[0003] The present invention relates to, in part, methods of treating and preventing autoimmune disorders, said methods comprising administering to a subject (e.g., a human) in need thereof a therapeutically effective amount of a growth differentiation factor 15 (GDF15) polypeptide.

BACKGROUND

[0004] Autoimmune diseases are diseases in which a subject’s immune system mistakenly attacks the subject’s body. Normally, the immune system guards against foreign invaders such as bacteria and viruses. However, in autoimmune diseases the immune system mistakenly identifies the subject’s own cells as foreign invaders and mounts an immune response against the subject itself. Autoimmune diseases can be debilitating conditions, associated with pain, limitation of movement, and in some cases, loss of autonomy. Autoimmune diseases affect over 50 million Americans.

[0005] Currently, there are no cures for autoimmune diseases. Current treatments for autoimmune diseases include nonsteroidal anti-inflammatory drugs and immunosuppressive drugs. However, such treatments have serious side effects, including an increased risk of infection for immunosuppressive drugs. There is an unmet need for advances in treatments for autoimmune diseases.

[0006] Wild type GDF15, also known as MIC-1 (macrophage inhibitory cytokine- 1) has been linked to regulation of body weight (Tsai VW, et al., PLoS One 2013; 8 (2): e55174; US 8,192,735).

BRIEF SUMMARY

[0007] Autoimmune diseases are diseases in which a subject's immune system mistakenly attacks the subject's body. Normally, the immune system guards against foreign invaders such as bacteria and viruses. However, in autoimmune diseases the immune system mistakenly identifies the subject's own cells as foreign invaders and mounts an immune response against the subject itself. There is an unmet need for advances in treatments for autoimmune diseases.

[0008] The present disclosure is based on the surprising and unexpected finding in the working examples herein that a GDF15 polypeptide is capable of suppressing the onset and/or progression of autoimmune diseases. Both prophylactic and therapeutic administration of a GDF15 polypeptide attenuated autoimmune diseases, such as MS, inflammatory bowel disease, and type I diabetes in animal models. Administration of the GDF15 polypeptide blocked pathogenic immune cell infiltration in the central nervous system in an animal model of MS and modulated the immune cell profile.

The GDF15/glial cell derived neurotrophic factor family receptor alpha like (GFRAL) pathway mediated the role of GDF15 in attenuating autoimmune diseases. These data support targeting GDF15, e.g., via administration of a GDF15 polypeptide, in the treatment and prevention of autoimmune diseases (e.g., relapsing-remitting MS, primary progressive MS, secondary progressive MS).

[0009] Thus, provided herein is method for treating an autoimmune disease (e.g., MS) in a subject (e.g., a human) in need thereof, the method comprising: administering to the subject a therapeutically effective amount of a GDF15 polypeptide. In some instances, the GDF15 polypeptide binds GFRAL and activates GFRAL. In some instances, the autoimmune disease is MS. In some instances, the autoimmune disease is relapsing-remitting MS. In some instances, the autoimmune disease is primary progressive MS. In some instances, the autoimmune disease is secondary progressive MS. In some instances, the autoimmune disease is benign MS, primary progressive MS, secondary progressive MS, or relapsing-remitting MS. In some instances, the autoimmune disease is selected from the group consisting of MS, rheumatoid arthritis, psoriasis, psoriatic arthritis, systemic lupus erythematosus (SLE), cutaneous lupus erythematosus (CLE), discoid lupus erythematosus (DLE), inflammatory bowel disease (e.g., Crohn's disease, ulcerative colitis), polymyositis, dermatopolymyositis, Addison's disease, Graves' disease, Sjogren's syndrome, Hashimoto's thyroiditis, myasthenia gravis, autoimmune vasculitis, pernicious anemia, scleroderma, cytokine release syndrome, and celiac disease. In some instances, the autoimmune disease is not a glucose metabolism disorder. In some instances, the autoimmune disorder is not type 1 diabetes.

[0010] Also provided herein is method for preventing an autoimmune disease (e.g., MS) in a subject (e.g., a human) in need thereof, the method comprising: administering to the subject a therapeutically effective amount of a GDF15 polypeptide. In some instances, the GDF15 polypeptide binds GFRAL and activates GFRAL. In some instances, the autoimmune disease is MS. In some instances, the autoimmune disease is relapsing-remitting MS. In some instances, the autoimmune disease is primary progressive MS. In some instances, the autoimmune disease is secondary progressive MS. In some instances, the autoimmune disease is benign MS, primary progressive MS, secondary progressive MS, or relapsing-remitting MS. In some instances, the autoimmune disease is selected from the group consisting of MS, rheumatoid arthritis, psoriasis, psoriatic arthritis, systemic lupus erythematosus (SLE), cutaneous lupus erythematosus (CLE), discoid lupus erythematosus (DLE), inflammatory bowel disease (e.g., Crohn's disease, ulcerative colitis), polymyositis, dermatopolymyositis, Addison's disease, Graves' disease, Sjogren's syndrome, Hashimoto's thyroiditis, myasthenia gravis, autoimmune vasculitis, pernicious anemia, scleroderma, cytokine release syndrome, and celiac disease. In some instances, the autoimmune disease is not a glucose metabolism disorder. In some instances, the autoimmune disorder is not type 1 diabetes. [0011] Also provided herein is method for slowing down the onset of an autoimmune disease (e.g., MS) (e.g., slowing down the onset of one or more symptoms of an autoimmune disease) in a subject (e.g., a human) in need thereof, the method comprising: administering to the subject a therapeutically effective amount of a GDF15 polypeptide, wherein the subject is at risk of developing the autoimmune disease. In some instances, the GDF15 polypeptide binds GFRAL and activates GFRAL. In some instances, the autoimmune disease is MS. In some instances, the autoimmune disease is relapsing-remitting MS. In some instances, the autoimmune disease is primary progressive MS. In some instances, the autoimmune disease is secondary progressive MS. In some instances, the autoimmune disease is benign MS, primary progressive MS, secondary progressive MS, or relapsing-remitting MS. In some instances, the autoimmune disease is selected from the group consisting of MS, rheumatoid arthritis, psoriasis, psoriatic arthritis, systemic lupus erythematosus (SLE), cutaneous lupus erythematosus (CLE), discoid lupus erythematosus (DLE), inflammatory bowel disease (e.g., Crohn's disease, ulcerative colitis), polymyositis, dermatopolymyositis, Addison's disease, Graves' disease, Sjogren's syndrome, Hashimoto's thyroiditis, myasthenia gravis, autoimmune vasculitis, pernicious anemia, scleroderma, cytokine release syndrome, and celiac disease. In some instances, the autoimmune disease is not a glucose metabolism disorder. In some instances, the autoimmune disorder is not type 1 diabetes.

[0012] Also provided herein is a method of reducing or blocking pathogenic immune cell infiltration into the central nervous system of a subject (e.g., a subject, e.g., human, suffering from an autoimmune disease, e.g., relapsing remitting MS, primary progressive MS, secondary progressive MS), the method comprising administering to the subject a therapeutically effective amount of a GDF15 polypeptide. In some instances, the pathogenic immune cell is a pathogenic T cell. In some instances, the T cell is a CD4+ T cell. In some instances, the T cell is a Th- 17 cell. In some instances, the T cell is a Th-1 cell. In some instances, the T cell is a myelin oligodendrocyte glycoprotein (MOG)-specific CD4+ T cell.

[0013] In some instances, the GDF15 polypeptide comprises a complex comprising a first heterodimer and a second heterodimer; wherein each of the first heterodimer and the second heterodimer comprises a first polypeptide and a second polypeptide; wherein the first polypeptide comprises from the N-terminus to the C- terminus a first human IgGl Fc domain , a linker, and a GDF15 mutein, wherein the first human IgGl Fc domain comprises a first hinge region, a first CH2 domain, and a first CH3 domain; wherein the second polypeptide comprises from the N-terminus to the C-terminus a second human IgGl Fc domain, wherein the second human IgGl Fc domain comprises a second hinge region, a second CH2 domain, and a second CH3 domain; and wherein the GDF15 polypeptide binds to GFRAL. In some instances, the first human IgGl Fc domain comprises an engineered protuberance and the second human IgGl Fc domain comprises an engineered cavity. In some instances, the engineered protuberance comprises at least one substitution of the corresponding amino acid in a human IgGl Fc sequence selected from the group consisting of amino acid residues 347, 366 and 394, according to EU numbering; and the engineered cavity comprises at least one substitution of the corresponding amino acid in a human IgGl Fc sequence selected from the group consisting of consisting of amino acid residues 366, 368, 394, 405, and 407, according to EU numbering. In some instances, the engineered protuberance comprises at least one substitution of the corresponding amino acid in a human IgGl Fc sequence selected from the group consisting of Q347W/Y, T366W/Y. and T394W/Y. according to EU numbering; and the engineered cavity comprises at least one substitution of the corresponding amino acid in a human IgGl Fc sequence selected from the group consisting of T366S, L368A, T394S, F405I7V/A, and Y407I7V/A, according to EU numbering. In some instances, the engineered protuberance comprises the substitution T366W, according to EU numbering, and wherein the engineered cavity comprises the substitutions T366S, L368A, and Y407V, according to EU numbering. In some instances, the engineered protuberance comprises the substitution T366Y, according to EU numbering, and wherein the engineered cavity comprises the substitution Y407T, according to EU numbering. In some instances, the linker comprises the sequence Glycine-Glycine- Glycine-Glycine-Ser (G4S)n, wherein n=2-5 (SEQ ID NO: 17). In some instances, the GDF15 mutein comprises an amino acid sequence having at least 90%, at least 95%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to the amino acid sequence of any one of SEQ ID NOs:3-12. In some instances, the GDF15 mutein comprises an amino acid sequence having at least 90%, at least 95%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to the amino acid sequence of SEQ ID NO:3. In some instances, the GDF15 mutein comprises the amino acid sequence of SEQ ID NO:3. In some instances, the GDF15 mutein consists of the amino acid sequence of SEQ ID NO:3. In some instances, the first polypeptide comprises an amino acid sequence having at least 90%, at least 95%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to the amino acid sequence of SEQ ID NO: 13 or 14. In some instances, the first polypeptide comprises an amino acid sequence having at least 90%, at least 95%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to the amino acid sequence of SEQ ID NO: 13. In some instances, the first polypeptide comprises the amino acid sequence of SEQ ID NO: 13. In some instances, the first polypeptide consists of the amino acid sequence of SEQ ID NO: 13. In some instances, the second polypeptide comprises an amino acid sequence having at least 90%, at least 95%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to the amino acid sequence of SEQ ID NO: 15. In some instances, the first polypeptide comprises the amino acid sequence of SEQ ID NO: 13, and the second polypeptide comprises the amino acid sequence of SEQ ID NO: 15. In some instances, the first polypeptide consists of the amino acid sequence of SEQ ID NO: 13, and the second polypeptide consists of the amino acid sequence of SEQ ID NO: 15.

[0014] In some instances, the GDF15 polypeptide comprises a GDF15 mutein, wherein the GDF15 mutein comprises an amino acid sequence having at least 90%, at least 95%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to the amino acid sequence the amino acid sequence of any one of SEQ ID NOs:3-12. In some instances, the GDF15 mutein comprises the amino acid sequence of SEQ ID NO:7. In some instances, the GDF15 mutein consists of the amino acid sequence of SEQ ID NO:7.

[0015] In some instances, the GDF15 polypeptide comprises a GDF15 mutein comprising a contiguous amino acid sequence that is at least 90% identical to the amino acid sequence of wild type GDF15 (SEQ ID NO: 1). In some instances, the GDF15 mutein comprises at least one substitution of the corresponding amino acid in SEQ ID NO: 1 that creates the N-linked glycosylation consensus site. In some instances, the GDF15 mutein comprises at least one pair of substitutions of the corresponding amino acids in SEQ ID NO: 1 selected from the group consisting of: (i) K91N and D93T, (ii) K91N and D93S, (iii) D93N and G95T, or (iv) D93N and G95S. In some instances, the GDF15 mutein comprises an N-terminal deletion of one or more amino acids relative to the amino acid sequence of SEQ ID NO: 1.

[0016] In some instances, the GDF15 polypeptide comprises a dimer, wherein the dimer comprises a first polypeptide and a second polypeptide, wherein each of the first polypeptide and the second polypeptide comprises a GDF15 mutein.

[0017] In some instances, the GDF15 polypeptide comprises a dimer, wherein the dimer comprises a first polypeptide and a second polypeptide, wherein each of the first polypeptide and the second polypeptide comprises an amino acid sequence having at least 90%, at least 95%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to the amino acid sequence the amino acid sequence of any one of SEQ ID NOs:3-12, wherein the first polypeptide is identical to the second polypeptide.

[0018] In some instances, the GDF15 polypeptide comprises a dimer, wherein the dimer comprises a first polypeptide and a second polypeptide, wherein each of the first polypeptide and the second polypeptide comprises the amino acid sequence of SEQ ID NO:7.

[0019] In some instances, the GDF15 polypeptide comprises a dimer, wherein the dimer comprises a first polypeptide and a second polypeptide, wherein each of the first polypeptide and the second polypeptide consists of the amino acid sequence of SEQ ID NO:7.

[0020] In some instances of the foregoing methods, the subject is a human.

[0021] Also provided herein is a method of treating MS in a human subject in need thereof, the method comprising: administering to the human subject a therapeutically effective amount of a GDF15 polypeptide, wherein the GDF15 polypeptide consists of a complex consisting of a first heterodimer and a second heterodimer, wherein each of the first heterodimer and the second heterodimer consists of a first polypeptide and a second polypeptide, wherein the first polypeptide consists of the amino acid sequence of SEQ ID NO: 13 and the second polypeptide consists of the amino acid sequence of SEQ ID NO: 15, and wherein the first heterodimer dimerizes with the second heterodimer. In some cases, the MS is relapsing-remitting MS. In some instances, the MS is primary progressive MS. In some instances, the MS is secondary progressive MS.

[0022] Also provided herein is a method of preventing MS in a human subject in need thereof, the method comprising: administering to the human subject a therapeutically effective amount of a GDF15 polypeptide, wherein the GDF15 polypeptide consists of a complex consisting of a first heterodimer and a second heterodimer, wherein each of the first heterodimer and the second heterodimer consists of a first polypeptide and a second polypeptide, wherein the first polypeptide consists of the amino acid sequence of SEQ ID NO: 13 and the second polypeptide consists of the amino acid sequence of SEQ ID NO: 15; wherein the first heterodimer dimerizes with the second heterodimer. In some instances, the human subject is at risk of developing MS. In some cases, the MS is relapsing-remitting MS. In some instances, the MS is primary progressive MS. In some instances, the MS is secondary progressive MS.

[0023] Also provided herein is a method of treating MS in a human subject in need thereof, wherein the method comprises administering to the human subject a therapeutically effective amount of a GDF15 polypeptide, wherein the GDF15 polypeptide consists of a dimer, wherein the dimer consists of a first polypeptide and a second polypeptide, wherein each of the first polypeptide and the second polypeptide consists of the amino acid sequence of SEQ ID NO:7. In some cases, the MS is relapsing-remitting MS. In some instances, the MS is primary progressive MS. In some instances, the MS is secondary progressive MS.

[0024] Also provided herein is a method of preventing MS in a human subject in need thereof, wherein the method comprises administering to the human subject a therapeutically effective amount of a GDF15 polypeptide, wherein the GDF15 polypeptide consists of a dimer, wherein the dimer consists of a first polypeptide and a second polypeptide, wherein each of the first polypeptide and the second polypeptide consists of the amino acid sequence of SEQ ID NO:7. In some instances, the human subject is at risk of developing MS. In some cases, the MS is relapsingremitting MS. In some instances, the MS is primary progressive MS. In some instances, the MS is secondary progressive MS.

[0025] In some instances of the foregoing methods, the GDF15 polypeptide comprises AMG 171. In some instances of the foregoing methods, the GDF15 polypeptide comprises LY3463251. In some instances of the foregoing methods, the GDF15 polypeptide comprises NN9215. In some instances of the foregoing methods, the GDF15 polypeptide comprises CIN-109.

BRIEF DESCRIPTION OF THE FIGURES

[0026] FIG. 1 is a graph showing the experimental autoimmune encephalomyelitis (EAE) score (y-axis) at the indicated time induced by MOG35-55 immunization in C57BL/6 mice (n=10).

Key: (Group 1) EAE mice administered 0.3 mg/kg anti-KLH antibody at days -2, 6, and 13; (Group 2) EAE mice administered 0.3 mg/kg of GDF15 Analog 1 at days -2, 6, and 13; (Group 3) EAE mice administered 1 mg/kg of GDF15 Analog 1 at days -2,

6, and 13; (Group 4) EAE mice administered 3 mg/kg of GDF15 Analog 1 at days -2,

6, and 13; (Group 5) EAE mice administered 3 mg/kg of anti-GFRAL antibody at days -3, 5, and 12 and 3 mg/kg of GDF15 Analog 1 at days -2, 6, and 13. (Group 6) non-EAE mice.

[0027] FIG. 2 is a pair of graphs showing body weight (left) and body weight changes (right) of mice at the indicated time (days post immunization) as described in FIG. 1.

[0028] FIG. 3 is a graph showing the EAE score at the indicated time (days post immunization) induced by MOG35-55 immunization in C57BL/6 mice (n=10). Wild type mice or GFRAL knockout mice were administered with prophylactic dosing of GDF15 Analog 1 or control (anti-KLH antibody).

[0029] FIG. 4 is a graph showing the EAE score at the indicated time (days post immunization) induced by MOG35-55 immunization in C57BL/6 mice administered on day -2, 5, and 12 with 0.3 mg/kg anti-KLH antibody, 0.03 mg/kg of GDF15 Analog 1, 0.1 mg/kg of GDF15 Analog 1, or 0.3 mg/kg of GDF15 Analog 1.

[0030] FIG. 5 is a graph showing the EAE score at the indicated time (days post immunization) induced by MOG35-55 immunization in C57BL/6 mice. Mice were administered prophylactically (on day -2, 6, and 13) with 3 mg/kg anti-KLH antibody or 1 mg/kg of GDF15 Analog 1, or administered therapeutically (on day 12) with 3 mg/kg of GDF15 Analog 1.

[0031] FIG. 6A depicts representative FACS plots of Thl7 (T-helper 17) cells within the spinal tissue from the mice of Group 1 (left), Group 4 (middle), and Group 5 (right) as described in FIG. 1. Y-axis: cluster of differentiation 4 (CD4); X-axis: retinoic acid receptor-related orphan receptor gamma t (RORgt).

[0032] FIG. 6B is a pair of graphs showing the relative proportion (left) and absolute numbers (right) of Thl7 cells (RORgt CD4+ T cells) within the spinal tissue for the mice of Group 1, Group 4, and Group 5 as described in FIG. 1.

[0033] FIG. 7A depicts representative FACS plots of CD45 expressing cells within the spinal tissue from the mice of Groups 1 and 4-6 as described in FIG. 1. Y- axis: L-D (Live-Dead); X-axis: cluster of differentiation 45 (CD45) expression. CD45hi marks all leukocytes. CD45mid marks microglia.

[0034] FIG. 7B is a pair of graphs showing the relative proportion (left) and absolute numbers (right) of leukocytes (CD45hi cells) within the spinal tissue for the mice of Group 1, Group 4, and Group 5 as described in FIG. 1.

[0035] FIG. 8A depicts representative FACS plots of CD4+T cells within the spinal tissue from the mice of Groups 1 and 4-6 as described in FIG. 1. Y-axis: CD45 ; X-axis: CD4.

[0036] FIG. 8B is a pair of graphs showing the relative proportion (left) and absolute numbers (right) of CD4+ T cells within the spinal tissue for the mice of Group 1, Group 4, and Group 5 as described in FIG. 1.

[0037] FIG. 9A depicts representative FACS plots of MOG-specific CD4+ T cells within the spinal tissue from the mice of Groups 1 and 4-6 as described in FIG. 1. Y-axis: CD4 ; X-axis: MOG35-55 Tetramer. [0038] FIG. 9B is a pair of graphs showing the relative proportion (left) and absolute numbers (right) of MOG-specific T cells within the spinal tissue for the mice of Group 1, Group 4, and Group 5 as described in FIG. 1.

[0039] FIG. 10 is a graph showing the EAE score of mice at the indicated days post receiving immune cells from donor mice. C57BL/6 donor mice (n=20) were administered with 1 mg/kg anti-KLH or GDF15 Analog 1 (on day -15) followed by MOG35 -55 immunization (on day -14). Lymphocytes (from lymph nodes) and spleen cells from the mice were isolated (on day -3) and cultured ex vivo with anti-IL4 and IL12 for three days before being injected intraperitoneally into the recipient C57BL/6 mice.

[0040] FIG. 11 is a graph showing the EAE score of mice at the indicated days post receiving immune cells from donor mice. The donor C57BL/6 mice were immunized with MOG35-55 (on day -14). The lymphocytes (from lymph nodes) and spleen cells were isolated (on day -3) from the mice, and cultured ex vivo with anti- IL4 and IL12 for three days before being injected intraperitoneally into the recipient C57BL/6 mice. The recipient mice were administered with 1 mg/kg anti-KLH or GDF15 Analog 1 at day 10, 17, and 24 post immune cell transfer.

[0041] FIG. 12 is a graph showing the EAE score in mice immunized with human MOG protein (1-120) at the indicated time. Mice were administered with control, 1 mg/kg GDF15 Analog 1, or anti-CD20 (lOOpg) weekly starting on day 7 post immunization with the human MOG protein.

[0042] FIG. 13 is a graph showing the EAE score in SJL mice (n=10) immunized with PLP139-151 at the indicated time. Mice were administered prophylactically with 1 mg/kg anti-KLH control or GDF15 Analog 1 (once weekly for 8 weeks starting on day -2), or therapeutically with 1 mg/kg GDF1 Analog 1 (once weekly for 5 weeks starting on day 19).

[0043] FIG. 14 is a pair of graphs depicting the spleen weight (left) and spleen/body weight ratio (right) in a mouse model of inflammatory bowel disease (IBD), wherein CD4+ naive T cells from Balb/c donor mice were injected intraperitoneally into SCID Balb/c recipient mice. GDF15 Analog 1 or controls were administered weekly starting from the day of T cell transfer. Tissues were harvest and weighted at day 44 after cell transfer. Key: (Group 1) Naive mice administered with vehicle HBSS; (Group 2) IBD mice administered with 1 mg/kg anti-KLH; (Group 3) IBD mice administered with 0.3 mg/kg GDF15 Analog 1; (Group 4) IBD mice administered with 1 mg/kg GDF15 Analog 1.

[0044] FIG. 15 shows the colon length (left), the colon weight (middle), and the colon weight/colon length ratio (right) of the mice as described in FIG.14

[0045] FIG. 16A is representative images of hematoxylin and eosin staining of longitudinal colon sections from the mice as described in FIG14. The arrows illustrate the length defined as the thickness of the colonic epithelium.

[0046] FIG. 16B is a graph illustrating the sum of the averaged mucosal thickness in proximal, mid, and distal colon (pm) of the mice as described in FIG. 14

[0047] FIG. 17 is a pair of graphs showing the body weight (BW, gram) and body weight changes of SCID Balb/c mice at the indicated days post receiving CD4+ naive T cells from donor Balb/c mice as described in FIG.14.

[0048] FIG. 18 is a pair of graphs depicting the blood glucose level (mg/dL) (top) and the percentage of diabetes-free mice (bottom) at the indicated age (weeks) in the spontaneous type 1 diabetes NOD mouse model. 6-week old female NOD mice were administered once weekly with 1 mg/kg anti-KLH, 0.1 mg/kg GDF15 Analog 1, or 1 mg/kg GDF15 Analog 1. Diabetes was called when a mouse with blood glucose exceeding 250 mg/dL for two consecutive weeks.

[0049] FIG. 19 is a pair of graphs showing the body weight (top) and body weight changes (bottom) of mice at the indicated age as described in FIG. 18.

[0050] FIG. 20A is representative images of Luxol fast blue (LFB) staining (left) and IBA1 staining (right) of brain areas containing the corpus callosum from mice fed with 0.2% cuprizone (CPZ) diet for 3 or 5 weeks and being concomitantly administered with 0.3 mg/kg anti-KLH or GDF15 Analog 1 weekly.

[0051] FIG. 20B is a pair of graphs showing the quantification (mean +/- SEM) of LFB staining (left) and IBA1 staining (right) in corpus callosum from mice as described in FIG. 19A. Higher LFB grade shows more severe loss myelin sheath. * indicates p<0.05. [0052] FIG. 21 is a graph depicting the quantification (mean +/- SEM) of IBA1 staining in hippocampus from mice as described in FIG. 20A. * indicates p<0.05, *** indicates p<0.001.

[0053] FIG. 22 is a pair of graphs showing the quantification (mean +/- SEM) of GFAP staining in corpus callosum (left) or cortex (right) from mice as described in FIG. 19A. * indicates p<0.05, **** indicates pO.OOOl.

[0054] FIG. 23 is a pair of graphs showing the quantification (mean +/- SEM) of LFB grade (left) and IBA1 staining (right) in the corpus callosum of mice that were fed with 0.2% CPZ-containing diet for 5 weeks followed by 2 weeks of normal chow to allow for remyelination. CPZ + anti-KLH group and CPZ + GDF15 Analog 1 (Prophylactic, “Pro”) group received weekly dosing of anti-KLH or GDF15 Analog 1, respectively concomitant with the initiation of CPZ diet. CPZ + GDF15 Analog 1 (Therapeutic, “Ther”) group received two weekly dosing of GDF15 Analog 1 at the onset of normal chow diet. * indicates p<0.05, *** indicates p<0.001.

[0055] FIG. 24 is a graph depicting the quantification (mean +/- SEM) of IB Al staining in hippocampus from mice as described in FIG. 22. ** indicates p<0.01.

[0056] FIG. 25 is a pair of graphs showing the quantification (mean +/- SEM) of GFAP staining in corpus callosum (left) or cortex (right) from mice as described in FIG. 22. **** indicates pO.OOOl.

[0057] FIG. 26 is a graph showing EAE score at the indicated time induced by MOG35 -55 immunization in C57BL/6 mice. Each of Img/kg of wild-type GDF15 and Img/kg of GDF15 Analog 2 was administered daily starting one day before the immunization. 0.3 mg/kg anti-KLH antibody was used as a control.

DETAILED DESCRIPTION

[0058] The present disclosure is based on surprising and unexpected the finding in the working examples herein that GDF15 and a GDF15 analog is capable of immune suppression in autoimmune diseases. Both prophylactic and therapeutic administration of a GDF15 analog attenuated autoimmune disease of the CNS in an animal model of MS. Administration of the GDF15 analog blocked pathogenic immune cell infiltration in the central nervous system in an animal model of MS and modulated the immune cell profile. The GDF15/GFRAL pathway mediated the role of GDF15 in attenuating autoimmune disease. These data support targeting GDF15, e.g., via administration of a GDF15 polypeptide, in the treatment and prevention of autoimmune diseases (e.g., relapsing-remitting MS, primary progressive MS, secondary progressive MS). Thus, provided herein are methods of treating and preventing an autoimmune disease (e.g., relapsing-remitting MS, primary progressive MS, secondary progressive MS) in a subject (e.g., a human) in need thereof, the method comprising administering to the subject a therapeutically effective amount of a GDF15 polypeptide (or a pharmaceutical composition comprising the same). In some instances, the subject is at risk of developing the autoimmune disease.

Autoimmune Diseases

[0059] Autoimmune diseases are diseases in which a subject’s immune system mistakenly attacks the subject’s body. Normally, the immune system guards against foreign invaders such as bacteria and viruses. However, in autoimmune diseases the immune system mistakenly identifies the subject’s own cells as foreign invaders and mounts an immune response against the subject itself. Autoreactive T cells play an important effector role in the cell and tissue damage that encompasses autoimmune diseases, while under normal circumstances T cells maintain tolerance to self. Signs and symptoms of autoimmune diseases are known in the art, as are methods of diagnosing autoimmune diseases. GDF15 polypeptides described herein can block infiltration of pathogenic immune cells into tissues. This is at least one mechanism via which GDF15 polypeptides can prevent or delay onset of autoimmune diseases, slow down progression of autoimmune diseases or treat autoimmune diseases.

[0060] Non-limiting examples of autoimmune diseases that can be treated according to the methods described herein include: MS, relapsing-remitting MS, primary progressive MS, secondary progressive MS, rheumatoid arthritis, psoriasis, psoriatic arthritis, systemic lupus erythematosus (SLE), cutaneous lupus erythematosus (CLE), discoid lupus erythematosus (DLE), inflammatory bowel disease (e.g., Crohn’s disease, ulcerative colitis), polymyositis, dermatopolymyositis, Addison’s disease, Graves’ disease, Sjogren’s syndrome, Hashimoto’s thyroiditis, myasthenia gravis, autoimmune vasculitis, pernicious anemia, scleroderma, cytokine release syndrome, and celiac disease. In some instances, the autoimmune disease is type 1 diabetes. In some instances, the autoimmune disease is not Type 1 diabetes. In some instances, the autoimmune disease is not a glucose metabolism disorder.

[0061] In some instances, the autoimmune disease is MS. MS is an autoimmune disease of the CNS that affects over two million people. MS is an immune-mediated inflammatory disease affecting brain, nerve, and spinal cord tissue which causes demyelination of neurons, axonal damage, and neurodegeneration. CD4 (Thl, Thl7), CD8, and B cells are believed to play a role in MS. The disease primarily affects white matter, but demyelination in gray matter also plays an important role. In most cases (>80%) the disease is relapsing/remitting. The main symptoms of MS include CNS symptoms (e.g., fatigue, cognitive impairment, depression, anxiety, unstable mood), visual symptoms (e.g., nystagmus, optic neuritis, diplopia), speech symptoms (e.g., dysarthria), throat symptoms (e.g., dysphagia), musculoskeletal symptoms (e.g., weakness, spasms, ataxia), sensation symptoms (e.g., pain, hypoesthesias, paraesthesias), bowel symptoms (e.g., incontinence, diarrhea or constipation), and urinary symptoms (e.g., incontinence, frequency or retention). Other symptoms include: numbness or weakness in one or more limbs that typically occurs on one side of the body at a time, or your legs and trunk; electric-shock sensations that occur with certain neck movements, especially bending the neck forward; tremor, lack of coordination or unsteady gait; partial or complete loss of vision, usually in one eye at a time, often with pain during eye movement; prolonged double vision; blurry vision; slurred speech; fatigue; dizziness; tingling or pain in parts of your body; and problems with sexual, bowel and bladder function.

[0062] In some instances, the autoimmune disease is relapsing-remitting MS. Relapsing-remitting MS is a type of MS where one has relapses followed by recovery and there is no new disability between attacks.

[0063] In some instances, the autoimmune disease is primary progressive MS.

[0064] In some instances, the autoimmune disease is secondary progressive MS.

[0065] Subjects may be genetically predisposed to develop an autoimmune disease. For instance, SLE and MS tend to run in families. In some instances, a subject is at risk of developing an autoimmune disease (e.g., MS) if one or more relatives (e.g., parent, sibling, grandparent, cousin, aunt, uncle) have been diagnosed with the autoimmune disease. In some instances, a subject is at risk of developing an autoimmune disease (e.g., MS) if the subject has one or more (e.g., 1, 2, 3, 4, 5) markers of the autoimmune disease (e.g., autoantibodies or genetic markers).

GDF15

[0066] Provided herein are GDF15 polypeptides for use with the methods of treatment described herein. GDF15, also known as MIC-1 (macrophage inhibitory cytokine- 1), PDF (prostate differentiation factor), PLAB (placental bone morphogenetic protein), NAG-1 (non-steroidal anti-inflammatory drugs (NSAIDs) activated gene), TGF-PL, and PTGFB, is a member of the transforming growth factor P (TGF- ) super-family. Human GDF15 is synthesized as a 62 kDa intracellular precursor protein that is subsequently cleaved by a furin-like protease and is secreted as a 25 kDa disulfide-linked protein (see, e.g., Fairlie et al., J. Leukoc. Biol 65:2-5 (1999)). GDF15 mRNA is seen in several tissues, including liver, kidney, pancreas, colon and placenta, and GDF15 expression in liver can be significantly up-regulated during injury of organs such as the liver, kidneys, heart and lungs.

[0067] The human GDF15 precursor is a 308 amino acid polypeptide (NCBI Ref. Seq.NP_004855.2; SEQ ID NO:2) containing a 29 amino acid signal peptide, a 167 amino acid pro-domain, and a mature domain of 112 amino acids which is excised from the pro-domain by furin-like proteases. The human GDF15 precursor amino acid sequence — referred to as “full-length” human GDF15 — is set forth in SEQ ID NO:2:

MPGQELRTVNGSQMLLVLLVLSWLPHGGALSLAEASRASFPGPSE LHSEDSRFRELRKRYEDLLTRLRANQSWEDSNTDLVPAPAVRILTP EVRLGSGGHLHLRISRAALPEGLPEASRLHRALFRLSPTASRSWDV TRPLRRQLSLARPQAPALHLRLSPPPSQSDQLLAESSSARPQLELHL RPQAARGRRRARARNGDHCPLGPGRCCRLHTVRASLEDLGWAD WVLSPREVQVTMCIGACPSQFRAANMHAQIKTSLHRLKPDTVPAP CCVPASYNPMVLIQKTDTGVSLQTYDDLLAKDCHCI (SEQ ID NO: 2) The 112-amino acid human wild type GDF15 (amino acids 197-308 of SEQ ID NO:2) — referred to as “mature” human GDF15 — is set forth in SEQ ID NO: 1:

ARNGDHCPLGPGRCCRLHTVRASLEDLGWADWVLSPREVQVTMC IGACPSQFRAANMHAQIKTSLHRLKPDTVPAPCCVPASYNPMVLIQ KTDTGVSLQTYDDLLAKDCHCI (SEQ ID NO:1)

[0068] Numerical references to particular GDF15 residues refer to the 112 amino acid mature sequence (i.e., residue 1 is Ala (A), and residue 112 is He (I); see SEQ ID NO: 1). Of note, while the GDF15 precursor amino acid sequence predicts three excision sites, resulting in three putative forms of "mature" human GDF15 (i.e., 110, 112 and 115 amino acids), the 112 amino acid mature sequence is accepted as being correct.

[0069] The scope of the present disclosure includes GDF15 orthologs, and modified forms thereof, from other mammalian species, and their use, including mouse (NP_035949), chimpanzee (XP_524157), orangutan (XP_002828972), Rhesus monkey (EHH29815), giant panda (XP_002912774), gibbon (XP_003275874), guinea pig (XP_003465238), ferret (AER98997), cow (NP_001193227), pig (NP_001167527), dog (XP_541938) and platypus (Omithorhynchus anatinus;

AFV61279). The mature form of human GDF15 has approximately 67% amino acid identity to the mouse ortholog. The sequences of the orthologs can be used to identify and modify non-conserved residues in human GDF15. Thus, variants of human GF 15 are encompassed by this disclosure. These variants may differ from SEQ ID NO:1 at 1 to 30, 1 to 25, 1 to 20, 1 to 15, 1 to 10, 10, 9, 8, 7, 6, 5, 4, 3, 2 or 1 amino acid(s). These variants are still capable of being GDF15 agonists. In some instances, the variants able to bind GFRAL.

GDF15 Polypeptides

[0070] GDF15 polypeptides that are used in the methods of treatment and prevention described herein include all existing and known GDF15 proteins, homologues, muteins, fragments, modified forms thereof, fusion proteins, and analogues in the art. See, e.g., International Patent Application Publication Nos. WO 2016/069921, WO 2016/018931, WO 2014/120619, WO 2013/148117, WO 2015/017710, WO 2013/113008, WO 2012/138919, WO 2017/109706, WO 2015/200078, WO 2019/195091, WO 2020/185533, WO 2018/215525, WO 2020/084496, WO 2020/104948, WO 2020/218827 and WO 2018/071493; and US Patent Application Publication Nos. US 2016/0120999, US 2016/0031960, US 2014/0213511, US 2016/0200787, US 2015/0023960, US 2017/0291929, US 2021/0147500, US 2017/0327560, and US 2019/0309033, each of which is incorporated by reference herein in its entirety.

[0071] In some instance, the GDF15 polypeptide comprises AMG 171. In some instance, the GDF15 polypeptide comprises LY3463251. In some instances, the GDF15 polypeptide comprises NN9215. In some instances, the GDF15 polypeptide comprises CIN-109.

[0072] In some instances, the GDF15 polypeptides used herein retain one or more functions of wild-type GDF15. For example, in some instances, the GDF15 polypeptides used herein bind GFRAL. In some instances, the GDF15 polypeptides activate GFRAL. Methods of determining binding of a GDF15 polypeptide described herein are known in the art, such as, e.g., co-immunoprecipitation assays, ELISA assays, BIACORE, and co-immunofluorescence assays. Methods of determining whether a GDF15 polypeptide described herein activates GFRAL are known in the art, such as, e.g., signaling reporter assays and detection of downstream signaling components.

[0073] In some instances, the GDF15 polypeptides used herein exhibit at least 50% (e.g., at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95%, or at least 100%) of the activity of wild type GDF15. For example, in some instances, the GDF15 polypeptide used herein activates GFRAL to a level at least 50% of the level of GFRAL activation by wild type GDF15. In some instances, the GDF15 polypeptides used herein exhibit the same activity as wild type GDF15. In some instances, the GDF15 polypeptides used herein have an improved activity as compared to wild type GDF15, such as, e.g., increased solubility, increased stability, increased half-life, and/or increased expression yield compared to wild type GDF15. In some instances, the GDF15 polypeptide used herein have improved therapeutic efficacy compared to wild type GDF15 (e.g., at least 5%, at least 10%, at least 15%, 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 95%, or at least 100% increased efficacy compared to wild type GDF15).

[0074] In some instances, a GDF15 polypeptide used herein induces weight loss in an animal model. In some instances, when administered into rodents (e.g., mice or rats) and/or non-human primates (e.g., cynomolgus monkeys), the GDF15 polypeptides are capable of inducing weight loss (e.g., a reduction in weight by at least 2%, at least 3%, at least 4%, at least 5%, at least 6%, at least 7%, at least 8%, at least 9%, at least 10%, at least 15%, at least 20%, at least 25%, or at least 30% as compared to the weight prior to the administration).

[0075] In some instances, the GDF15 polypeptide comprises wild type human GDF15 (SEQ ID NO: 1).

[0076] In some instances, the GDF15 polypeptide comprises a GDF15 mutein. GDF15 muteins are polypeptides comprising an artificially introduced change in the amino acid sequence of a wild type GDF15 (e.g., SEQ ID NO: 1), e.g., a change in amino acid sequence generated in the laboratory or other facility by human intervention (“hand of man”). GDF15 muteins carry single or multiple (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10) amino acid substitutions and/or deletions relative to the amino acid sequence of SEQ ID NO: 1. A GDF15 mutein can be derived from a cloned gene (e.g., a wild type GDF15 gene) that has been subjected to site-directed or random mutagenesis, or from a completely synthetic gene. In some instances, a GDF15 mutein comprises one or more (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10) amino acid substitutions and/or amino acid deletions relative to a reference polypeptide, e.g., relative to the amino acid sequence of SEQ ID NO: 1. In some instances, the GDF15 mutein has one or more substitutions at non-conserved residues in human GDF15 when it is aligned with one or more of its orthologs. In some instances, a GDF15 mutein comprises a contiguous amino acid sequence that is at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical to the amino acid sequence of SEQ ID NO: 1. In some instances, the GDF15 mutein includes one or more (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10) amino acid substitutions and/or deletions relative to the amino acid sequence of SEQ ID NO: 1. In some instances, the GDF15 mutein includes one or more (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10) amino acid substitutions relative to the amino acid sequence of SEQ ID NO: 1. In some instances, the GDF15 mutein includes one or more (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10) amino acid deletions relative to the amino acid sequence of SEQ ID NO: 1. In some instances, the GDF15 mutein includes one or more (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10) amino acid substitutions relative to the amino acid sequence of SEQ ID NO: 1 and one or more (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10) amino acid deletions relative to the amino acid sequence of SEQ ID NO: 1. Modifications (e.g., substitutions, additions, and deletions) to GDF15 are well known in the art (see, e.g., International Patent Application Publication Nos. WO 2016/069921, WO 2016/018931, WO 2014/120619, WO 2013/148117, WO 2015/017710, WO 2013/113008, WO 2012/138919, and WO 2017/109706, and US Patent Application Publication Nos. US 2016/0120999, US 2016/0031960, US 2014/0213511, US 2016/0200787, and US 2015/0023960, each of which is incorporated by reference herein in its entirety). The GDF15 muteins used in the methods described herein bind and activate GFRAL (e.g., human GFRAL). In some instances, the GDF15 mutein exhibits at least 50% (e.g., at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95%, or at least 100%) of the activity of wild type GDF15.

[0077] For convenience and clarity, the amino acid sequence of SEQ ID NO: 1 is used as a reference sequence for the GDF15 muteins presented herein. Therefore, the amino acid residue positions are numbered herein with reference to SEQ ID NO: 1. [0078] In some instances, the GDF15 mutein includes one, two, three or more (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10) amino acid substitutions, additions, and/or deletions that introduce one or more (e.g., 1, 2, 3, 4, 5) N-linked glycosylation consensus site(s) at a location where such a site is not present in SEQ ID NO: 1. The N-linked glycosylation consensus site includes the sequence NXS/T, where N is Asn; X is an amino acid other than proline; followed by either Ser (S) or Thr (T). See, e.g., International Patent Application Publication Nos. WO 2016/069921 and WO 2016/018931, each of which is incorporated by reference herein in its entirety, for exemplary substitutions that introduce one or more N-linked glycosylation consensus site(s). In some instances, the GDF15 mutein comprises a contiguous amino acid sequence having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, or at least 98% sequence identity to the amino acid sequence of SEQ ID NO: 1, where the contiguous amino acid sequence has the substitution D5T/S or R21N. In some instances, the GDF15 mutein comprises a contiguous amino acid sequence having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, or at least 98% sequence identity to the amino acid sequence of SEQ ID NO: 1, where the contiguous amino acid sequence has at least one of the following pairs of substitutions relative to the corresponding amino acids in SEQ ID NO: 1: (i) R16N and H18T or R16N and H18S; (ii) S23N and E25T or S23N and E25S; (iii) L24N and D26T or L24N and D26S; (iv) S50N and F52T or S50N and F52S; (v) F52N and A54T or F52N and A54S; (vi) Q51N and R53T or Q51N and R53S; (vii) R53N and A55T or R53N and A55S; (viii) S64N and H66T or S64N and H66S; (ix) L65N and R67T or L65N and R67S; (x) S82N and N84T or S82N and N84S; (xi) K91N and D93T or K91N and D93S; (xii) D93N and G95T or D93N and G95S; (xiii) T94N and V96T or T94N and V96S; (xiv) V96N and L98T or V96N and L98S; (xv) S97N and Q99T or S97N and Q99S; or (xvi) A106N and D108T or A106N and D108S, wherein the contiguous amino acid sequence has at least one of the pairs of substitutions of (i)-(xvi), respectively. For example, the substitutions in (i) above, denotes that the polypeptide has a threonine (T) or serine (S) at an amino acid position that corresponds to amino acid position 18 in SEQ ID NO:1, wherein in SEQ ID NO: 1 a histidine (H) is present at the amino acid position 18. The position of the corresponding amino acid in a polypeptide relative to SEQ ID NO: 1 may be determined by aligning the amino acid sequences. In some instances, the GDF15 mutein comprises the amino acid sequence of SEQ ID NO: 1, except for one or more (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10) substitutions and/or deletions, optionally wherein one or more of the substitutions is selected from the following pairs of substitutions relative to the corresponding amino acids in SEQ ID NO:1 (i) R16N and H18T or R16N and H18S; (ii) S23N and E25T or S23N and E25S; (iii) L24N and D26T or L24N and D26S; (iv) S50N and F52T or S50N and F52S; (v) F52N and A54T or F52N and A54S; (vi) Q51N and R53T or Q51N and R53S; (vii) R53N and A55T or R53N and A55S; (viii) S64N and H66T or S64N and H66S; (ix) L65N and R67T or L65N and R67S; (x) S82N and N84T or S82N and N84S; (xi) K91N and D93T or K91N and D93S; (xii) D93N and G95T or D93N and G95S; (xiii) T94N and V96T or T94N and V96S; (xiv) V96N and L98T or V96N and L98S; (xv) S97N and Q99T or S97N and Q99S; or (xvi) A106N and D108T or A106N and D108S.

[0079] In some instances, the GDF15 mutein lacks the first amino acid of SEQ ID NO: 1. In some instances, the GDF15 mutein lacks the first two amino acids of SEQ ID NO: 1. In some instances, the GDF15 mutein lacks the first three amino acids of SEQ ID NO: 1. In some instances, the GDF15 mutein lacks the first six amino acids of SEQ ID NO: 1. In some instances, the GDF15 mutein lacks the first 14 amino acids of SEQ ID NO: 1. In some instances, the GDF15 mutein has at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, or at least 98% sequence identity to the amino acid sequence of SEQ ID NO: 3, except wherein the GDF15 mutein comprises a D5T/S substitution (relative to SEQ ID NO: 1) and lacks the first two amino acids of SEQ ID NO: 1. In some instances, the GDF15 mutein has at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, or at least 98% sequence identity to the amino acid sequence of SEQ ID NO: 5, except wherein the GDF15 mutein comprises a K91N substitution (relative to SEQ ID NO: 1) and a D93T/S substitution (relative to SEQ ID NO: 1). In some instances, the GDF15 mutein has at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, or at least 98% sequence identity to the amino acid sequence of SEQ ID NO: 5, except wherein the GDF15 mutein comprises a K91N substitution (relative to SEQ ID NO: 1), a D93T/S substitution (relative to SEQ ID NO: 1), and lacks the first three amino acids of SEQ ID NO: 1. In some instances, the GDF15 mutein has at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, or at least 98% sequence identity to the amino acid sequence of SEQ ID NO: 5, except wherein the GDF15 mutein comprises a D93N substitution (relative to SEQ ID NO: 1) and a G95T/S substitution (relative to SEQ ID NO: 1). In some instances, the GDF15 mutein has at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, or at least 98% sequence identity to the amino acid sequence of SEQ ID NO: 5, except wherein the GDF15 mutein comprises a D93N substitution (relative to SEQ ID NO: 1), a G95T/S substitution (relative to SEQ ID NO: 1), and lacks the first three amino acids of SEQ ID NO: 1. In some instances, the GDF15 mutein binds to GFRAL.

[0080] The present disclosure also includes GDF15 polypeptides that comprise active fragments (e.g., subsequences) of the GDF15 muteins described above. The length of active fragments or subsequences may be 40 amino acids to 111 amino acids, e.g., 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 98, 106, 109, or up to 111 amino acids. Exemplary active fragments of the GDF15 muteins disclosed herein include GDF15 muteins that have deletions of amino acids (e.g., of 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 amino acids) relative to SEQ ID NO: 1. For example, the polypeptides may have N-terminal truncations and/or C-terminal truncations relative to SEQ ID NO: 1. The truncations may be of 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more amino acids relative to a reference polypeptide, e.g., SEQ ID NO: 1. Truncated, active, forms of GDF15 are known in the art, see, e.g., International Patent Application Publication Nos. WO 2016/069921, WO 2016/018931, and WO 2014/120619, and US Patent Application Publication Nos. US 2016/0120999, US 2016/0031960, US 2014/0213511, and US 2016/0200787, each of which is incorporated by reference herein in its entirety. In some instances, the active fragment of the GDF15 mutein is lacking amino acid 1 of the amino acid sequence of SEQ ID NO: 1. In some instances, the active fragment of the GDF15 mutein is lacking amino acids 1 and 2 of the amino acid sequence of SEQ ID NO:1. In some instances, the active fragment of the GDF15 mutein is lacking amino acids 1 -3 of the amino acid sequence of SEQ ID NO: 1. In some instances, the active fragment of the GDF15 mutein is lacking amino acids 1-6 of the amino acid sequence of SEQ ID NO: 1. In some instances, the active fragment of the GDF15 mutein binds GFRAL.

[0081] In some instances, the GDF15 muteins have a defined sequence identity compared to a reference sequence over a defined length of contiguous amino acids (e.g., a "comparison window"). Methods of alignment of sequences for comparison are well-known in the art. Optimal alignment of sequences for comparison can be conducted, e.g., by the local homology algorithm of Smith & Waterman, Adv. Appl. Math. 2:482 (1981), by the homology alignment algorithm of Needleman & Wunsch, J. Mol. Biol. 48:443 (1970), by the search for similarity method of Pearson & Lipman, Proc. Nat'l. Acad. Sci. USA 85:2444 (1988), by computerized implementations of these algorithms (GAP, BESTFIT, FASTA, and TFASTA in the Wisconsin Genetics Software Package, Madison, Wis.), or by manual alignment and visual inspection (see, e.g., Current Protocols in Molecular Biology (Ausubel et al., eds. 1995 supplement). As an example, a GDF15 mutein can comprise an amino acid sequence having at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 98%, or at least about 99%, amino acid sequence identity to a contiguous stretch of 40, 45, 50, 55, 60, 65, 70, 75, 80, 85 or up to 112 amino acids in SEQ ID NO: 1. In some instances, the GDF15 mutein comprises an amino acid sequence having at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99%, or at least 100% amino acid sequence identity to the entire length of the amino acid sequence of SEQ ID NO: 1. [0082] In some instances, the GDF15 mutein is 112, 111, 110, 109, 108, 107, or 106 amino acids in length. In some instances, the GDF15 mutein is 112 amino acids in length. In some instances, the GDF15 mutein is 109 amino acids in length (e.g., lacks the first three amino acids of SEQ ID NO: 1). In some instances, the GDF15 mutein is 106 amino acids in length (e.g., lacks the first 6 amino acids of SEQ ID NO:1).

[0083] GDF15 in its active form is a dimer. Thus, in some instances, a GDF15 polypeptide comprises a dimer, wherein the dimer comprises a first polypeptide and a second polypeptide, wherein each of the first polypeptide and the second polypeptide comprises a GDF15 mutein or wild type GDF15. In some instances, the first polypeptide is identical to the second polypeptide. In some instances, the first polypeptide is not identical to the second polypeptide. In some instances, the GDF15 polypeptide comprises a dimer, wherein the dimer comprises a first polypeptide and a second polypeptide, wherein each of the first polypeptide and the second polypeptide comprises a GDF15 mutein, e.g., a GDF15 mutein described herein. In some instances, the GDF15 polypeptide comprises a dimer, wherein the dimer comprises a first polypeptide and a second polypeptide, wherein each of the first polypeptide and the second polypeptide comprises an amino acid sequence having at least 90%, at least 95%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to the amino acid sequence the amino acid sequence of any one of SEQ ID NOs:3-12. In some instances, the GDF15 polypeptide comprises a dimer, wherein the dimer comprises a first polypeptide and a second polypeptide, wherein each of the first polypeptide and the second polypeptide comprises an amino acid sequence having one or more (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10) substitutions or deletions relative to the amino acid sequence of any one of SEQ ID NOs:3-10). In some instances, each of the first polypeptide and the second polypeptide lacks the first two amino acids of SEQ ID NO: 1. In some instances, each of the first polypeptide and the second polypeptide lacks the first three amino acids of SEQ ID NO: 1. In some instances, each of the first polypeptide and the second polypeptide lacks the first six amino acids of SEQ ID NO:1. In some instances, the GDF15 polypeptide comprises a dimer, wherein the dimer comprises a first polypeptide and a second polypeptide, wherein each of the first polypeptide and the second polypeptide comprises the amino acid sequence of SEQ ID NO:7. In some instances, the GDF15 polypeptide comprises a dimer, wherein the dimer comprises a first polypeptide and a second polypeptide, wherein each of the first polypeptide and the second polypeptide comprises the amino acid sequence of SEQ ID NO:7, wherein each of the first polypeptide and the second polypeptide lacks the first three amino acids of SEQ ID NO: 1. In some instances, the GDF15 polypeptide comprises a dimer, wherein the dimer comprises a first polypeptide and a second polypeptide, wherein each of the first polypeptide and the second polypeptide consists of the amino acid sequence of SEQ ID NO:7.

[0084] In some instances, the GDF15 polypeptide comprises a wild type GDF15 or a GDF15 mutein attached to one or more molecules or moi eties. In some instances, the one or more molecules or moi eties extends the half-life of the GDF15 polypeptide. In some instances, the GDF15 polypeptide comprises a wild type GDF15 or a GDF15 mutein modified through, for example, pegylation (covalent attachment of one or more molecules of polyethylene glycol (PEG), or derivatives thereof); glycosylation (e.g., N-glycosylation); polysialylation; albumin fusion molecules comprising serum albumin (e.g., human serum albumin (HSA), cyno serum albumin, or bovine serum albumin (BSA)); albumin binding through, for example, a conjugated fatty acid chain (acylation); Fc-fusion; or fusion with a PEG mimetic. In some instances, the GDF15 polypeptide comprises a wild type GDF15 or a GDF15 mutein linked to HSA as exemplified in US 2017/0327560, which is incorporated by reference herein in its entirety. In some instances, the GDF15 polypeptide is CIN-109. In some instances, the GDF15 polypeptide comprises a wild type GDF15 or a GDF15 mutein linked to a fatty acid chain, as exemplified in the Al, A2, or A3 formulae in WO 2015/200078, which is incorporated by reference herein in its entirety. In some instances, the GDF15 polypeptide comprises a wild type GDF15 or a GDF15 mutein, a N-terminal amino acid extensiON and a protractor, as exemplified in wo 2018/215525, which is incorporated by reference herein in its entirety. In some instances, the modifications are introduced in a site-specific manner. In other instances, the modifications include a linker. The linker may conjugate the modifying moiety to the polypeptide.

[0085] In some instances, the GDF15 polypeptide comprises an Fc domain fused (either directly or indirectly, e.g., indirectly via a linker) to a GDF15 mutein or wild type GDF15, i.e., an Fc-GDF15 fusion polypeptide. In some instances, the Fc domain is from an IgG molecule (e.g., a human IgG moiety). In some instances, the human IgG Fc domain is from human IgGl, human IgG2, human IgG3, or human IgG4. In some instances, the Fc domain has reduced effector function or no effector function. In some instances, the Fc domain is a hinge-deleted Fc fragment, a charge-paired mutant of Fc fragment, a hinge deleted charge charge-paired Fc fragment, or a singlechain Fc fragment, as exemplified in Xiong et al., Sci. Transl. Med. 9, eaan8732 (2017), which is incorporated by reference herein in its entirety. In some instances, the GDF15 polypeptide comprises a GDF15 Fc fusion protein as exemplified in WO/2013/113008, which is incorporated by reference herein in its entirety. In some instances, the GDF15 polypeptide is AMG 171. In some instances, the GDF15 polypeptide is LY3463251 (see, e.g., Benichou et al., 2023, Cell Metabolism 35, 1- 13, which is incorporated by reference herein in its entirety).

[0086] In some instances, the GDF15 polypeptide comprises a complex comprising a first heterodimer and a second heterodimer, wherein each of the first heterodimer and the second heterodimer comprises a first polypeptide and a second polypeptide; wherein the first polypeptide comprises a first IgG Fc domain (e.g., an IgGl Fc domain, an IgG2 Fc domain, an IgG3 Fc domain, an IgG4 Fc domain, e.g., a human IgGl Fc domain, a human IgG2 Fc domain, a human IgG3 Fc domain, a human IgG4 Fc domain) optionally a linker, and a GDF15 mutein or a wild type GDF15; wherein the second polypeptide comprises a second IgG Fc domain (e.g., an IgGl Fc domain, an IgG2 Fc domain, an IgG3 Fc domain, an IgG4 Fc domain, e.g., a human IgGl Fc domain, a human IgG2 Fc domain, a human IgG3 Fc domain, a human IgG4 Fc domain); wherein the GDF15 polypeptide binds to GFRAL. In some instances, the first IgG Fc domain comprises a human IgG Fc domain (e.g., a human IgGl Fc domain, a human IgG2 Fc domain, a human IgG3 Fc domain, a human IgG4 Fc domain). In some instances, the first and/or second IgG Fc domain has reduced or no effector function. The first polypeptide in the first heterodimer dimerizes with the second polypeptide in the first heterodimer, the first polypeptide in the second heterodimer dimerizes with the second polypeptide in the second heterodimer, and the GDF15 in the first heterodimer dimerizes with the GDF15 in the second heterodimer. In some instances, the first IgG Fc domain comprises a human IgGl Fc domain. In some instances, the second IgG Fc domain comprises a human IgG Fc domain (e.g., a human IgGl Fc domain, a human IgG2 Fc domain, a human IgG3 Fc domain, a human IgG4 Fc domain). In some instances, the second IgG Fc domain comprises a human IgGl Fc domain. In some instances, the first IgG Fc domain comprises a first hinge region, a first CH2 domain, and a first CH3 domain. In some instances, the second IgG Fc domain comprises a second hinge region, a second CH2 domain, and a second CH3 domain. In some instances, the first IgG Fc domain and the second IgG Fc domain have the same amino acid sequence as each other. In some instances, the first IgG Fc domain and the second IgG Fc domain have different amino acid sequences compared to each other. In some instances, the first IgG Fc domain is fused (e.g., directly or indirectly, e.g., indirectly via a linker) to the N-terminus of the GDF15 mutein or wild type GDF15. In some instances, the first IgG Fc domain is fused (e.g., directly or indirectly, e.g., indirectly via a linker) to the C-terminus of the GDF15 mutein or wild type GDF15. In some instances, the first IgG Fc domain is fused directly to the GDF15 mutein or wild type GDF15. In some instances, the first IgG Fc domain is fused to the GDF15 mutein or wild type GDF15 via a linker.

Linkers suitable for fusing an IgG Fc domain to a GDF15 mutein or wild type GDF15 are known in the art, see, e.g., the linkers disclosed in International Patent Application Publication No. WO 2016/069921 and US Patent Application Publication No. US 2016/0120999, each of which is incorporated by reference herein in its entirety. In some instances, the linker comprises the amino acid sequence (G4S)n, wherein n=2-5 (SEQ ID NO: 17). In some instances, the linker comprises the amino acid sequence (G4S)2 (SEQ ID NO: 18). In some instances, the linker comprises the amino acid sequence (G4S)s (SEQ ID NO: 19). In some instances, the linker comprises the amino acid sequence (G4S)s (SEQ ID NO:20). In some instances, the linker consists of the amino acid sequence of any one of SEQ ID NOs: 18-20.

[0087] Any IgG Fc domain sequence described herein or known in the art can be a component of a GDF15 polypeptide described herein. The IgG Fc domain used in a GDF15 polypeptide described herein maintains the ability to bind to an Fc domain receptor when dimerized with another IgG Fc domain. In some instances, an IgG Fc domain is a human IgG Fc domain (e.g., a human IgGl, a human IgG2, a human IgG3, or a human IgG4) or a variant thereof (e.g., a human IgG Fc domain having one or more (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10) substitutions and/or deletions while still maintaining the ability to bind to an Fc receptor when dimerized). In some instances, an IgG Fc domain is a human IgGl Fc domain. The amino acid sequence of an exemplary human IgGl Fc domain is provided as SEQ ID NO: 16. The numbering of the position of an amino acid in the Fc domain sequence is according to the EU numbering (Edelman, G.M. et al., Proc. Natl. Acad. USA, 63, 78-85 (1969)). Thus, the glutamic acid residue "E" at position 1 in SEQ ID NO: 16 is numbered as 216; CH2 domain starts with alanine (A) which is numbered 231; CH3 domain starts at glycine (G) which is numbered 341, according to EU numbering.

[0088] In some instances, an IgG Fc domain of a GDF15 polypeptide has at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to the amino acid sequence of SEQ ID NO: 16, wherein the IgG Fc domain binds to an Fc receptor. In some instances, an IgG Fc domain of a GDF15 polypeptide comprises the amino acid sequence of SEQ ID NO: 16 except having one or more (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10) amino acid substitutions relative to the amino acid sequence of SEQ ID NO: 16, wherein the IgG Fc domain binds to an Fc receptor. In some instances, an IgG Fc domain of a GDF15 polypeptide is a fragment of an IgG Fc domain, wherein the fragment comprises a CH3 domain, wherein the fragment binds to an Fc receptor. In some instances, an IgG Fc domain of a GDF15 polypeptide is a fragment of an IgG Fc domain, wherein the fragment comprises a CH2 domain and a CH3 domain, wherein the fragment binds to an Fc receptor. In some instances, an IgG Fc domain of a GDF15 polypeptide is a fragment of an IgG Fc domain, wherein the fragment comprises a partial hinge region, CH2 domain and a CH3 domain, wherein the fragment binds to an Fc receptor. In some instances, an IgG Fc domain of a GDF15 polypeptide is a fragment of an IgG Fc domain, wherein the fragment comprises a hinge region, CH2 domain and a CH3 domain, wherein the fragment binds to an Fc receptor. In some instances, the CH3 domain has at least 90%, at least 95%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to the CH3 domain of the amino acid sequence of SEQ ID NO: 16. In some instances, the CH2 domain has at least 90%, at least 95%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to the CH2 domain of the amino acid sequence of SEQ ID NO: 16. In some instances, the hinge region has at least 90%, at least 95%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to the hinge region of the amino acid sequence of SEQ ID NO: 16. In some instances, the partial hinge region has at least 90%, at least 95%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to the partial hinge region of the amino acid sequence of SEQ ID NO: 16.

[0089] In some instances, a first IgG Fc domain and a second IgG Fc domain of a GDF15 polypeptide are modified (relative to a corresponding wild type IgG Fc domain) to be partners of a charged Fc domain pair (see, e.g., International Patent Application Publication No. WO 2013/113008, which is incorporated by reference herein in its entirety). All existing Fc heterodimerization technologies in the art can be used, including but not limited to steric complementary design, charge-to-charge swap design, charge-to-steric complementarity swap, long-range electrostatic interaction design, and isotype strand swap design (See, for example, Ha et al., Front. Immunol. 7:394, which is incorporated by reference herein in its entirety). [0090] In some instances, a first IgG Fc domain and a second IgG Fc domain of a GDF15 polypeptide are modified (relative to a corresponding wild type IgG Fc domain) to be partners of a protuberance/ cavity pair (also known in the art as a “knob/hole” pair) (see, e.g., the Fc domains and Fc domain substitutions disclosed in International Patent Application Publication No. WO 2016/069921 and US Patent Application Publication No. US 2016/0120999, each of which is incorporated by reference herein in its entirety). In some instances, the first IgG Fc domain comprises an engineered protuberance (also known in the art as a “knob”) and the second IgG Fc domain comprises an engineered cavity (also known in the art as a “hole”). In some instances, the first IgG Fc domain comprises an engineered cavity and the second IgG Fc domain comprises an engineered protuberance. Exemplary protuberances and cavities are disclosed in US 8,216,805, which is incorporated by reference herein in its entirety, and include substitutions at the following amino acid positions: 347, 366, 368, 394, 405, and 407 according to the EU numbering. See, e.g., the Fc-GDF15 fusion polypeptides disclosed in International Patent Application Publication No. WO 2016/069921 and US Patent Application Publication No. US 2016/0120999, each of which is incorporated by reference herein in its entirety, for exemplary Fc domains comprising engineered protuberances/cavities. In some instances, the IgG Fc domain comprising an engineered protuberance comprises at least one (e.g., 1, 2, 3) substitution selected from the group consisting of Q347W/Y, T366W/Y, and T394W/Y according to the EU numbering. In some instances, the IgG Fc domain comprising an engineered cavity comprises at least one (e.g. 1, 2, 3, 4, 5) substitution selected from the group consisting of T366S, L368A, T394S, F405I7V/A, and Y407T/V/A according to the EU numbering. In some instances, the IgG Fc domain comprising an engineered protuberance comprises the substitution T366W/Y according to the EU numbering and the IgG Fc domain comprising an engineered cavity comprises the substitutions T366S, L368A, and Y407I7V/A according to the EU numbering. In some instances, the IgG Fc domain comprising an engineered protuberance comprising the substitution T366W/Y according to the EU numbering and the IgG Fc domain comprising an engineered cavity comprises the substitution Y407T/V/A according to the EU numbering. In some instances, the IgG Fc domain comprising an engineered protuberance comprises the substitution T366Y according to the EU numbering and the IgG Fc domain comprising an engineered cavity comprises the substitution Y407T according to the EU numbering. In some instances, the IgG Fc domain comprising an engineered protuberance comprises the substitution T366W according to the EU numbering and the IgG Fc domain comprising an engineered cavity comprises the substitution Y407A according to the EU numbering. In some instances, the IgG Fc domain comprising an engineered protuberance comprises the substitution T394Y according to the EU numbering and the IgG Fc domain comprising an engineered cavity comprises the substitution Y407T according to the EU numbering. In some instances, the IgG Fc domain comprising an engineered protuberance comprises the substitution T366W according to the EU numbering and the IgG Fc domain comprising an engineered cavity comprises the substitutions T366S, L368A, and Y407V according to the EU numbering.

[0091] In some instances, an IgG Fc domain of a GDF15 polypeptide comprises one or more (e.g., 1, 2, 3, 4, 5) mutation(s) that improve a property of the fusion polypeptide, e.g., reduction or abrogation of an IgG effector function such as complement-dependent cytotoxicity (CDC), antibody-dependent cellular cytotoxicity (ADCC) or antibody-dependent cell phagocytosis (ADCP). Non-limiting examples of modifications to an Fc domain that abrogate an IgG effector function are described in Armour KL. et al., 1999. Eur J Immunol. 29(8):2613-24; Shields RL et al., 2001, J Biol Chem. 276(9):6591-604; Idusogie EE. et al., 2000. J Immunol. 164(8):4178-84; Steurer W. et al., 1995. J Immunol. 155(3): 1165- 74; each of which is incorporated by reference herein in its entirety.

[0092] In some instances, the GDF15 polypeptide comprises a complex comprising a first heterodimer and a second heterodimer; wherein each of the first heterodimer and the second heterodimer comprises a first polypeptide and a second polypeptide; wherein the first polypeptide comprises from the N-terminus to the C- terminus a first human IgGl Fc domain, a linker, and a GDF15 mutein, wherein the first human IgGl Fc domain comprises a first hinge region, a first CH2 domain, and a first CH3 domain; wherein the second polypeptide comprises from the N-terminus to the C-terminus a second human IgGl Fc domain, wherein the second human IgGl Fc domain comprises a second hinge region, a second CH2 domain, and a second CH3 domain; and wherein the GDF15 polypeptide binds to GFRAL. In some instances, the first human IgGl Fc domain comprises an engineered protuberance and the second human IgGl Fc domain comprises an engineered cavity. In some instances, the engineered protuberance comprises the substitution T366W according to the EU numbering and the engineered cavity comprises the substitutions T366S, L368A, and Y407V according to the EU numbering. In some instances, the GDF15 mutein comprises an amino acid sequence having at least 90%, at least 95%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to the amino acid sequence of any one of SEQ ID NOs:3-12 (except wherein the GDF15 mutein retains the one or more differences in any one of SEQ ID NOs:3-12, respectively, relative to SEQ ID NO: 1). In some instances, the GDF15 mutein comprises the amino acid sequence of any one of SEQ ID NOs:3-12 except for one or more (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10) amino acid substitutions (except wherein the GDF15 mutein retains the one or more differences in any one of SEQ ID NOs:3-12, respectively, relative to SEQ ID NO: 1). In some instances, the GDF15 mutein comprises an amino acid sequence having at least 90%, at least 95%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to the amino acid sequence of SEQ ID NO:3, except wherein the GDF15 mutein comprises a D5T/S substitution relative to SEQ ID NO:1 and lacks the first two amino acids of SEQ ID NO:1. In some instances, the GDF15 mutein comprises the amino acid sequence of SEQ ID NO:3 except for one or more (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10) amino acid substitutions, except wherein the GDF15 mutein comprises a D5T/S substitution relative to SEQ ID NO:1 and lacks the first two amino acids of SEQ ID NO:1. In some instances, the GDF15 mutein comprises the amino acid sequence of SEQ ID NO:3 and lacks the first two amino acids of SEQ ID NO: 1. In some instances, the GDF15 mutein consists of the amino acid sequence of SEQ ID NO:3. In some instances, the first polypeptide comprises an amino acid sequence having at least 90%, at least 95%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to the amino acid sequence of SEQ ID NO: 13 or 14. In some instances, the first polypeptide comprises the amino acid sequence of SEQ ID NO: 13 or 14 except for one or more (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10) amino acid substitutions. In some instances, the first polypeptide comprises an amino acid sequence having at least 90%, at least 95%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to the amino acid sequence of SEQ ID NO: 13. In some instances, the first polypeptide comprises an amino acid sequence having one or more (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10) amino acid substitutions relative to the amino acid sequence of SED ID NO: 13, except wherein the GDF15 mutein in SEQ ID NO: 13 comprises a D5T/S substitution relative to SEQ ID NO: 1 and lacks the first two amino acids of SEQ ID NO:1. In some instances, the first polypeptide comprises the amino acid sequence of SEQ ID NO: 13. In some instances, the first polypeptide consists of the amino acid sequence of SEQ ID NO: 13. In some instances, the second polypeptide comprises an amino acid sequence having at least 90%, at least 95%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to the amino acid sequence of SEQ ID NO: 15. In some instances, the first polypeptide comprises the amino acid sequence of SEQ ID NO: 13, and the second polypeptide comprises the amino acid sequence of SEQ ID NO: 15. In some instances, the first polypeptide consists of the amino acid sequence of SEQ ID NO: 13, and the second polypeptide consists of the amino acid sequence of SEQ ID NO: 15. In some instances, the complex binds to GFRAL.

[0093] In some instances, the GDF15 polypeptide described herein comprises a GDF15 mutein or wild type GDF15 and hemiFc polypeptides (see, e.g., International Patent Application Publication NO. WO 2013/113008, which is incorporated by reference herein in its entirety).

[0094] In some instances, the GDF15 polypeptide described herein retains one or more functions of wild type GDF15 (e.g., wild type human GDF15). For example, in some instances, a GDF15 polypeptide described herein binds to GFRAL (e.g., human GFRAL) and/or mediates GDF15 signaling. Methods for determining the ability of a GDF15 polypeptide described herein to bind to GFRAL (e.g., human GFRAL) are known in the art, such as, e.g., radiolabeled antigen binding assay, Biacore, biolayer interferometry, enzyme-linked immunosorbent assay, immunohistochemistry, colocalization, co-immunoprecipitation, and signaling reporter assays (see, e.g., International Patent Application Publication No. WO 2017/172260, which is incorporated by reference herein in its entirety). Methods for determining GDF15 signaling are known in the art, such as, e.g., ELK1 -luciferase reporter assays (see, e.g., International Patent Application Publication No. WO 2017/172260, which is incorporated by reference herein in its entirety).

[0095] In some instances, the GDF15 polypeptide comprises a GDF15 polypeptide or a complex described in International Patent Application Publication No. WO 2016/069921 or in US Patent Application Publication No. US 2016/0120999, each of which is incorporated by reference herein in its entirety. In some instances, the GDF15 polypeptide comprises a GDF15 mutein, a modified GDF15 mutein, or a modified GDF15 described in International Patent Application Publication No. WO/2016/01893 lor in US Patent Application Publication No. US 2016/0031960, each of which is incorporated by reference herein in its entirety. In some instances, the GDF15 polypeptide comprises a GDF15 mutein, a modified GDF15 mutein, or a modified GDF15 described in International Patent Application Publication No. WO 2014/120619, in US Patent Application No. US 2014/0213511-Al, or in US Patent Application No. US 2016/0200787, each of which is incorporated by reference herein in its entirety. In some instances, the GDF15 polypeptide comprises a GDF15 mutein, a modified GDF15 mutein, or a modified GDF15 described in International Patent Application Publication No. WO 2013/148117 or in US Patent Application Publication No. US 2015/0023960, each of which is incorporated by reference herein in its entirety. In some instances, the GDF15 polypeptide comprises a fusion protein described in WO 2015/017710, which is incorporated by reference herein in its entirety. In some instances, the GDF15 polypeptide comprises a GDF15 polypeptide or construct comprising a GDF15 polypeptide described in WO 2013/113008, which is incorporated by reference herein in its entirety. In some instances, the GDF15 polypeptide comprises a GDF15 polypeptide described in WO 2012/138919, which is incorporated by reference herein in its entirety. In some instances, the GDF15 polypeptide comprises a GDF15 variant, a GDF15 fusion protein, or a GDF15 conjugate described in WO 2017/109706, which is incorporated by reference herein in its entirety. In some instances, the GDF15 polypeptide comprises a compound or homodimer described in US Patent Application Publication No. US 2019/030903 or International Publication No. W02019195091A1, each of which is incorporated by reference herein in its entirety. In some instances, the GDF15 polypeptide comprises a GDF15 molecule described in International Publication No. WO 2020/185533, which is incorporated by reference herein in its entirety. In some instances, the GDF15 polypeptide comprises a GDF15 agonist identified using a screening assay described in International Publication No. WO/2018/071493, which is incorporated by reference herein in its entirety.

[0096] The GDF15 polypeptides described herein can be produced by any suitable method known in the art. Such methods range from direct protein synthesis methods to constructing a DNA sequence encoding polypeptide sequences and expressing those sequences in a suitable host. See, e.g., International Patent Application Publication No. WO 2016/069921, which is incorporated by reference herein in its entirety, for a description of various methods for producing GDF15 polypeptides.

[0097] In some instances, the GDF15 polypeptide described herein is administered to a subject (e.g., human) as a pharmaceutical composition (e.g., a sterile pharmaceutical composition) in a method of treatment described herein. Thus, also provided herein are pharmaceutical compositions comprising a GDF15 polypeptide described herein. In some instances, the pharmaceutical composition comprises: a GDF15 polypeptide, and a pharmaceutically acceptable carrier. Pharmaceutically acceptable carriers are known in the art. In some instances, the pharmaceutical composition is a sterile pharmaceutical composition.

Methods of Treating Autoimmune Diseases

[0098] A GDF15 polypeptide described herein or a pharmaceutical composition comprising the same can be used to treat an autoimmune disease (e.g., relapsingremitting MS, primary progressive MS, secondary progressive MS) in a subject (e.g., a human) in need thereof. Thus, provided herein is a method for treating an autoimmune disease (e.g., relapsing-remitting MS, primary progressive MS, secondary progressive MS) in a subject (e.g., a human) in need thereof, the method comprising administering to the subject (e.g., human) a therapeutically effective amount of a GDF15 polypeptide (or composition comprising the same) to the subject. In some instances, the autoimmune disease is relapsing-remitting MS. In some instances, the autoimmune disease is primary progressive MS. In some instances, the autoimmune disease is secondary progressive MS. In some cases, the MS is benign MS. In some instances, the autoimmune diseases is psoriasis, psoriatic arthritis, or an inflammatory bowel disease (e.g., Crohn’s disease, ulcerative colitis). In some instances, the subject is a human.

[0099] A GDF15 polypeptide described herein or a pharmaceutical composition comprising the same can be used to slow down progression of an autoimmune disease (e.g., MS) in a subject (e.g., a human) in need thereof. Thus, also provided herein is a method for slowing down progression of an autoimmune disease (e.g., MS) in a subject (e.g., a human) in need thereof, the method comprising administering to the subject (e.g., human) a therapeutically effective amount of a GDF15 polypeptide (or composition comprising the same) to the subject. In some instances, the autoimmune disease is relapsing-remitting MS. In some instances, the autoimmune disease is primary progressive MS. In some instances, the autoimmune disease is secondary progressive MS. In some cases, the MS is benign MS. In some instances, the autoimmune diseases is psoriasis, psoriatic arthritis, or an inflammatory bowel disease (e.g., Crohn’s disease, ulcerative colitis). In some instances, the subject is a human. [00100] In some instances, the autoimmune disease is MS. In some cases, the MS is relapsing-remitting MS. In some cases, the MS is primary progressive MS. In some cases, the MS is secondary progressive MS. In some cases, the MS is benign MS.

[00101] In some instances, the autoimmune diseases is selected from the group consisting of MS (e.g., relapsing-remitting MS, primary progressive MS, secondary progressive MS), rheumatoid arthritis, psoriasis, psoriatic arthritis, systemic lupus erythematosus (SLE), cutaneous lupus erythematosus (CLE), discoid lupus erythematosus (DLE), inflammatory bowel disease (e.g., Crohn’s disease, ulcerative colitis), polymyositis, dermatopolymyositis, Addison’s disease, Graves’ disease, Sjogren’s syndrome, Hashimoto’s thyroiditis, myasthenia gravis, autoimmune vasculitis, pernicious anemia, scleroderma, cytokine release syndrome, and celiac disease. In some instances, the autoimmune disease is MS. In some cases, the MS is relapsing-remiting MS. In some cases, the MS is primary progressive MS. In some cases, the MS is secondary progressive MS. In some cases, the MS is benign MS. [00102] In some instances, the autoimmune disease is type 1 diabetes. In some instances, the autoimmune disease is not Type 1 diabetes. In some instances, the autoimmune disease is not a glucose metabolism disorder.

[00103] In some instances, the method comprises administering to the subject a therapeutically effective amount of a GDF15 polypeptide (e.g., a GDF15 polypeptide described herein).

[00104] In some instances, the GDF15 polypeptide comprises a complex comprising a first heterodimer and a second heterodimer; wherein each of the first heterodimer and the second heterodimer comprises a first polypeptide and a second polypeptide; wherein the first polypeptide comprises from the N-terminus to the C- terminus a first human IgGl Fc domain, a linker, and a GDF15 mutein, wherein the first human IgGl Fc domain comprises a first hinge region, a first CH2 domain, and a first CH3 domain; wherein the second polypeptide comprises from the N-terminus to the C-terminus a second human IgGl Fc domain, wherein the second human IgGl Fc domain comprises a second hinge region, a second CH2 domain, and a second CH3 domain; and wherein the GDF15 polypeptide binds to GFRAL. In some instances, the first human IgGl Fc domain comprises one or more (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10) substitutions that reduces or abrogates an Fc effector function (e.g., as described herein above). In some instances, the first human IgGl Fc domain comprises an engineered protuberance and the second human IgGl Fc domain comprises an engineered cavity. In some instances, the engineered protuberance comprises the substitution T366W according to the EU numbering and the engineered cavity comprises the substitutions T366S, L368A, and Y407V according to the EU numbering. In some instances, the GDF15 mutein comprises an amino acid sequence having at least 90%, at least 95%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to the amino acid sequence of any one of SEQ ID NOs:3-12 (except wherein the GDF15 mutein retains the one or more differences in any one of SEQ ID NOs:3-12, respectively, relative to SEQ ID NO: 1). In some instances, the GDF15 mutein comprises the amino acid sequence of any one of SEQ ID NOs:3-12 except for one or more (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10) amino acid substitutions (except wherein the GDF15 mutein retains the one or more differences in any one of SEQ ID NOs:3- 12, respectively, relative to SEQ ID NO: 1). In some instances, the GDF15 mutein comprises an amino acid sequence having at least 90%, at least 95%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to the amino acid sequence of SEQ ID NO:3, except wherein the GDF15 mutein comprises a D5T/S substitution relative to SEQ ID NO: 1 and lacks the first two amino acids of SEQ ID NO: 1. In some instances, the GDF15 mutein comprises the amino acid sequence of SEQ ID NO:3 except for one or more (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10) amino acid substitutions, except wherein the GDF15 mutein comprises a D5T/S substitution relative to SEQ ID NO: 1 and lacks the first two amino acids of SEQ ID NO: 1. In some instances, the GDF15 mutein comprises the amino acid sequence of SEQ ID NO:3 and lacks the first two amino acids of SEQ ID NO: 1. In some instances, the GDF15 mutein consists of the amino acid sequence of SEQ ID NO:3. In some instances, the first polypeptide comprises an amino acid sequence having at least 90%, at least 95%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to the amino acid sequence of SEQ ID NO: 13 or 14. In some instances, the first polypeptide comprises the amino acid sequence of SEQ ID NO: 13 or 14 except for one or more (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10) amino acid substitutions. In some instances, the first polypeptide comprises an amino acid sequence having at least 90%, at least 95%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to the amino acid sequence of SEQ ID NO: 13. In some instances, the first polypeptide comprises an amino acid sequence having one or more (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10) amino acid substitutions relative to the amino acid sequence of SED ID NO: 13, except wherein the GDF15 mutein in SEQ ID NO: 13 comprises a D5T/S substitution relative to SEQ ID NO: 1 and lacks the first two amino acids of SEQ ID NO: 1. In some instances, the first polypeptide comprises the amino acid sequence of SEQ ID NO: 13. In some instances, the first polypeptide consists of the amino acid sequence of SEQ ID NO: 13. In some instances, the second polypeptide comprises an amino acid sequence having at least 90%, at least 95%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to the amino acid sequence of SEQ ID NO: 15. In some instances, the first polypeptide comprises the amino acid sequence of SEQ ID NO: 13, and the second polypeptide comprises the amino acid sequence of SEQ ID NO: 15. In some instances, the first polypeptide consists of the amino acid sequence of SEQ ID NO: 13, and the second polypeptide consists of the amino acid sequence of SEQ ID NO: 15. In some instances, the complex binds to GFRAL.

[00105] In some instances, the GDF15 polypeptide comprises a complex comprising a first heterodimer and a second heterodimer, wherein each of the first heterodimer and the second heterodimer comprises a first polypeptide and a second polypeptide, wherein the first polypeptide comprises the amino acid sequence of SEQ ID NO: 13 and the second polypeptide comprises the amino acid sequence of SEQ ID NO: 15; wherein the first heterodimer dimerizes with the second heterodimer. In some instances, the first polypeptide consists of the amino acid sequence of SEQ ID NO: 13. In some instances, the second polypeptide consists of the amino acid sequence of SEQ ID NO: 15. In some instances, the first polypeptide consists of the amino acid sequence of SEQ ID NO: 13 and the second polypeptide consists of the amino acid sequence of SEQ ID NO: 15. In some instances, the subject is a human. In some instances, the autoimmune disease is MS. In some cases, the MS is relapsingremitting MS. In some cases, the MS is primary progressive MS. In some cases, the MS is secondary progressive MS. In some cases, the MS is benign MS. In some instances, the autoimmune diseases is psoriasis, psoriatic arthritis, or an inflammatory bowel disease (e.g., Crohn’s disease, ulcerative colitis).

[00106] In some instances, the GDF15 polypeptide comprises a dimer, wherein the dimer comprises a first polypeptide and a second polypeptide, wherein each of the first polypeptide and the second polypeptide comprises a GDF15 mutein or wild type GDF15. In some instances, the first polypeptide is identical to the second polypeptide. In some instances, the first polypeptide is not identical to the second polypeptide. In some instances, the GDF15 polypeptide comprises a dimer, wherein the dimer comprises a first polypeptide and a second polypeptide, wherein each of the first polypeptide and the second polypeptide comprises a GDF15 mutein, e.g., a GDF15 mutein described herein. In some instances, the GDF15 polypeptide comprises a dimer, wherein the dimer comprises a first polypeptide and a second polypeptide, wherein each of the first polypeptide and the second polypeptide comprises an amino acid sequence having at least 90%, at least 95%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to the amino acid sequence the amino acid sequence of any one of SEQ ID NOs:3-12. In some instances, the GDF15 polypeptide comprises a dimer, wherein the dimer comprises a first polypeptide and a second polypeptide, wherein each of the first polypeptide and the second polypeptide comprises an amino acid sequence having one or more (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10) substitutions or deletions relative to the amino acid sequence of any one of SEQ ID NOs:3-10). In some instances, each of the first polypeptide and the second polypeptide lacks the first two amino acids of SEQ ID NO: 1. In some instances, each of the first polypeptide and the second polypeptide lacks the first three amino acids of SEQ ID NO: 1. In some instances, each of the first polypeptide and the second polypeptide lacks the first six amino acids of SEQ ID NO: 1. In some instances, the GDF15 polypeptide comprises a dimer, wherein the dimer comprises a first polypeptide and a second polypeptide, wherein each of the first polypeptide and the second polypeptide comprises the amino acid sequence of SEQ ID NO:7. In some instances, the GDF15 polypeptide comprises a dimer, wherein the dimer comprises a first polypeptide and a second polypeptide, wherein each of the first polypeptide and the second polypeptide comprises the amino acid sequence of SEQ ID NO:7, wherein each of the first polypeptide and the second polypeptide lacks the first three amino acids of SEQ ID NO:1. In some instances, the GDF15 polypeptide comprises a dimer, wherein the dimer comprises a first polypeptide and a second polypeptide, wherein each of the first polypeptide and the second polypeptide consists of the amino acid sequence of SEQ ID NO:7.

[00107] In some instances, the GDF15 polypeptide comprises a dimer, wherein the dimer comprises a first polypeptide and a second polypeptide, wherein each of the first polypeptide and the second polypeptide comprises the amino acid sequence of SEQ ID NO:7, wherein each of the first polypeptide and second polypeptide lacks the first three amino acids of SEQ ID NO: 1. In some instances, each of the first polypeptide and the second polypeptide consists of the amino acid sequence of SEQ ID NO:7. In some instances, the subject is a human. In some instances, the autoimmune disease is MS. In some cases, the MS is relapsing-remitting MS. In some cases, the MS is primary progressive MS. In some cases, the MS is secondary progressive MS. In some cases, the MS is benign MS. In some instances, the autoimmune diseases is psoriasis, psoriatic arthritis, or an inflammatory bowel disease (e.g., Crohn’s disease, ulcerative colitis).

[00108] The GDF15 polypeptides (or pharmaceutical compositions comprising the same) can be administered in any number of ways for either local or systemic delivery. Further, the GDF15 polypeptides (or pharmaceutical compositions comprising the same) used in the methods of treatment described herein can be administered to the subject in a therapeutically effective amount. In this context, the term “therapeutically effective amount” as used herein refers to the amount of an agent (e.g., GDF15 polypeptide) that is sufficient to reduce and/or ameliorate the severity and/or duration of (i) a disease, disorder or condition in a subject, and/or (ii) a symptom in a subject.

[00109] In some instances, the method of treating obtains a desired pharmacological and/or physiological effect in the subject being administered the GDF15 polypeptide. Thus, in some instances, the method of treating inhibits the autoimmune disease, e.g., arrests its development. In some instances, the method of treating relieves the autoimmune disease, e.g., causes a regression of the autoimmune disease. In some instances, the method of treating slows down progression of the autoimmune disease. In some instances, the method of treating prolongs survival of the subject administered the GDF15 polypeptide as compared to expected survival if not receiving the treatment.

[00110] In some instances, the method of treating reduces one or more (e.g., 1, 2, 3, 4, 5) symptoms of the autoimmune disease.

[00111] In some instances in which the method is for treating MS (e.g., (e.g., benign MS, primary progressive MS, secondary progressive MS, relapsing-remitting MS), the method of treating reduces one or more (e.g., 1, 2, 3, 4, 5) symptoms of MS. In some instances, the MS is relapsing-remitting MS. In some instances, the MS is primary progressive MS. In some instances, the MS is secondary progressive MS. Symptoms of MS and clinical endpoints for treatment of MS are known in the art (see, e.g., van Munster and Uitdehaag, CNS Drugs, 2017; 31(3):217-236, which is incorporated by reference herein in its entirety).

[00112] In some instances, the method for treatment reduces (e.g., by at least 5%, at least 10%, at least 15%, at least 20%, at least 30%, at least 40%, or at least 50%, 10-50%, 10-25%, 10-15%, 25-50%, or 5-25%) infiltration of pathogenic immune cells into the tissues or organs (e.g., CNS) in the subject. In some instances, the reduction is compared to a level of pathogenic immune cells in the tissues or organs (e.g., CNS) prior to administration of the GDF15 polypeptide. Methods for determining infiltration of pathogenic immune cells into the CNS are known in the art. For example, CNS biopsy lumbar puncture can be performed to obtain spinal tissue or spinal fluid, which is then tested for pathogenic immune cells and autoantibodies. [00113] In some instances, the GDF15 polypeptide is administered to the subject (e.g., human) as a pharmaceutical composition (e.g., a sterile pharmaceutical composition). In some instances, the pharmaceutical composition comprises: a GDF15 polypeptide, and a pharmaceutically acceptable carrier. Pharmaceutically acceptable carriers are known in the art. In some instances, the pharmaceutical composition is a sterile pharmaceutical composition.

Methods of Preventing Autoimmune Diseases

[00114] A GDF15 polypeptide described herein or a pharmaceutical composition comprising the same can be used to prevent an autoimmune disease (e.g., MS) in a subject (e.g., a human) in need thereof. Thus, provided herein is a method for preventing an autoimmune disease in a subject (e.g., a human) in need thereof, the method comprising administering to the subject (e.g., human) a therapeutically effective amount of a GDF15 polypeptide (or composition comprising the same) to the subject. In some instances, the autoimmune disease is relapsing-remitting MS. In some instances, the autoimmune disease is primary progressive MS. In some instances, the autoimmune disease is secondary progressive MS. In some cases, the MS is benign MS. In some instances, the autoimmune diseases is psoriasis, psoriatic arthritis, or an inflammatory bowel disease (e.g., Crohn’s disease, ulcerative colitis). In some instances, the subject is a human. [00115] In some instances, a subject is at risk of developing an autoimmune disease if one or more relatives (e.g., parent, sibling, grandparent, cousin, aunt, uncle) of the subject have been diagnosed with the autoimmune disease. In some instances, a subject is at risk of developing an autoimmune disease if the subject has one or more markers of the autoimmune disease (e.g., autoantibodies or genetic marker(s).

[00116] In some instances, the therapeutically effective amount of the GDF15 polypeptide is administered to the subject prior to onset of one or more symptoms of the autoimmune disease. In some instances, the therapeutically effective amount of the GDF15 polypeptide is administered to the subject prior to a diagnosis of the autoimmune disease in the subject. In some instances, the autoimmune disease is relapsing-remitting MS. In some instances, the autoimmune disease is primary progressive MS. In some instances, the autoimmune disease is secondary progressive MS. In some cases, the MS is benign MS. In some instances, the autoimmune diseases is psoriasis, psoriatic arthritis, or inflammatory bowel disease (e.g., Crohn’s disease, ulcerative colitis).

[00117] In some instances, the autoimmune disease is MS. In some cases, the MS is relapsing-remitting MS. In some cases, the MS is primary progressive MS. In some cases, the MS is secondary progressive MS. In some cases, the MS is benign MS.

[00118] In some instances, the autoimmune diseases is selected from the group consisting of MS (e.g., relapsing-remitting MS, primary progressive MS, secondary progressive MS), rheumatoid arthritis, psoriasis, psoriatic arthritis, systemic lupus erythematosus (SLE), cutaneous lupus erythematosus (CLE), discoid lupus erythematosus (DLE), inflammatory bowel disease (e.g., Crohn’s disease, ulcerative colitis), polymyositis, dermatopolymyositis, Addison’s disease, Graves’ disease, Sjogren’s syndrome, Hashimoto’s thyroiditis, myasthenia gravis, autoimmune vasculitis, pernicious anemia, scleroderma, cytokine release syndrome, and celiac disease. In some instances, the autoimmune disease is MS. In some cases, the MS is relapsing-remitting MS. In some cases, the MS is primary progressive MS. In some cases, the MS is secondary progressive MS. In some cases, the MS is benign MS. [00119] In some instances, the autoimmune disease is type 1 diabetes. In some instances, the autoimmune disease is not Type 1 diabetes. In some instances, the autoimmune disease is not a glucose metabolism disorder.

[00120] In some instances, the GDF15 polypeptide comprises a complex comprising a first heterodimer and a second heterodimer; wherein each of the first heterodimer and the second heterodimer comprises a first polypeptide and a second polypeptide; wherein the first polypeptide comprises from the N-terminus to the C- terminus a first human IgGl Fc domain, a linker, and a GDF15 mutein, wherein the first human IgGl Fc domain comprises a first hinge region, a first CH2 domain, and a first CH3 domain; wherein the second polypeptide comprises from the N-terminus to the C-terminus a second human IgGl Fc domain, wherein the second human IgGl Fc domain comprises a second hinge region, a second CH2 domain, and a second CH3 domain; and wherein the GDF15 polypeptide binds to GFRAL. In some instances, the first human IgGl Fc domain comprises an engineered protuberance and the second human IgGl Fc domain comprises an engineered cavity. In some instances, the engineered protuberance comprises the substitution T366W according to the EU numbering and the engineered cavity comprises the substitutions T366S, L368A, and Y407V according to the EU numbering. In some instances, the GDF15 mutein comprises an amino acid sequence having at least 90%, at least 95%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to the amino acid sequence of any one of SEQ ID NOs:3-12 (except wherein the GDF15 mutein retains the one or more differences in any one of SEQ ID NOs:3-12, respectively, relative to SEQ ID NO: 1). In some instances, the GDF15 mutein comprises the amino acid sequence of any one of SEQ ID NOs:3-12 except for one or more (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10) amino acid substitutions (except wherein the GDF15 mutein retains the one or more differences in any one of SEQ ID NOs:3-12, respectively, relative to SEQ ID NO: 1). In some instances, the GDF15 mutein comprises an amino acid sequence having at least 90%, at least 95%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to the amino acid sequence of SEQ ID NO:3, except wherein the GDF15 mutein comprises a D5T/S substitution relative to SEQ ID NO:1 and lacks the first two amino acids of SEQ ID NO:1. In some instances, the GDF15 mutein comprises the amino acid sequence of SEQ ID NO:3 except for one or more (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10) amino acid substitutions, except wherein the GDF15 mutein comprises a D5T/S substitution relative to SEQ ID NO: 1 and lacks the first two amino acids of SEQ ID NO: 1. In some instances, the GDF15 mutein comprises the amino acid sequence of SEQ ID NO:3 and lacks the first two amino acids of SEQ ID NO: 1. In some instances, the GDF15 mutein consists of the amino acid sequence of SEQ ID NO:3. In some instances, the first polypeptide comprises an amino acid sequence having at least 90%, at least 95%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to the amino acid sequence of SEQ ID NO: 13 or 14. In some instances, the first polypeptide comprises the amino acid sequence of SEQ ID NO: 13 or 14 except for one or more (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10) amino acid substitutions. In some instances, the first polypeptide comprises an amino acid sequence having at least 90%, at least 95%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to the amino acid sequence of SEQ ID NO: 13. In some instances, the first polypeptide comprises an amino acid sequence having one or more (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10) amino acid substitutions relative to the amino acid sequence of SED ID NO: 13, except wherein the GDF15 mutein in SEQ ID NO: 13 comprises a D5T/S substitution relative to SEQ ID NO: 1 and lacks the first two amino acids of SEQ ID NO: 1. In some instances, the first polypeptide comprises the amino acid sequence of SEQ ID NO: 13. In some instances, the first polypeptide consists of the amino acid sequence of SEQ ID NO: 13. In some instances, the second polypeptide comprises an amino acid sequence having at least 90%, at least 95%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to the amino acid sequence of SEQ ID NO: 15. In some instances, the first polypeptide comprises the amino acid sequence of SEQ ID NO: 13, and the second polypeptide comprises the amino acid sequence of SEQ ID NO: 15. In some instances, the first polypeptide consists of the amino acid sequence of SEQ ID NO: 13, and the second polypeptide consists of the amino acid sequence of SEQ ID NO: 15. In some instances, the complex binds to GFRAL.

[00121] In some instances, the GDF15 polypeptide comprises a complex comprising a first heterodimer and a second heterodimer, wherein each of the first heterodimer and the second heterodimer comprises a first polypeptide and a second polypeptide, wherein the first polypeptide comprises the amino acid sequence of SEQ ID NO: 13 and the second polypeptide comprises the amino acid sequence of SEQ ID NO: 15; wherein the first heterodimer dimerizes with the second heterodimer. In some instances, the first polypeptide consists of the amino acid sequence of SEQ ID NO: 13. In some instances, the second polypeptide consists of the amino acid sequence of SEQ ID NO: 15. In some instances, the first polypeptide consists of the amino acid sequence of SEQ ID NO: 13 and the second polypeptide consists of the amino acid sequence of SEQ ID NO: 15. In some instances, the subject is a human. In some instances, the autoimmune disease is MS. In some cases, the MS is relapsingremitting MS. In some cases, the MS is primary progressive MS. In some cases, the MS is secondary progressive MS. In some cases, the MS is benign MS. In some instances, the autoimmune diseases is psoriasis, psoriatic arthritis, or an inflammatory bowel disease (e.g., Crohn’s disease, ulcerative colitis).

[00122] In some instances, the GDF15 polypeptide comprises a dimer, wherein the dimer comprises a first polypeptide and a second polypeptide, wherein each of the first polypeptide and the second polypeptide comprises a GDF15 mutein or wild type GDF15. In some instances, the first polypeptide is identical to the second polypeptide. In some instances, the first polypeptide is not identical to the second polypeptide. In some instances, the GDF15 polypeptide comprises a dimer, wherein the dimer comprises a first polypeptide and a second polypeptide, wherein each of the first polypeptide and the second polypeptide comprises a GDF15 mutein, e.g., a GDF15 mutein described herein. In some instances, the GDF15 polypeptide comprises a dimer, wherein the dimer comprises a first polypeptide and a second polypeptide, wherein each of the first polypeptide and the second polypeptide comprises an amino acid sequence having at least 90%, at least 95%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to the amino acid sequence the amino acid sequence of any one of SEQ ID NOs:3-12. In some instances, the GDF15 polypeptide comprises a dimer, wherein the dimer comprises a first polypeptide and a second polypeptide, wherein each of the first polypeptide and the second polypeptide comprises an amino acid sequence having one or more (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10) substitutions or deletions relative to the amino acid sequence of any one of SEQ ID NOs:3-10). In some instances, each of the first polypeptide and the second polypeptide lacks the first two amino acids of SEQ ID NO: 1. In some instances, each of the first polypeptide and the second polypeptide lacks the first three amino acids of SEQ ID NO: 1. In some instances, each of the first polypeptide and the second polypeptide lacks the first six amino acids of SEQ ID NO: 1. In some instances, the GDF15 polypeptide comprises a dimer, wherein the dimer comprises a first polypeptide and a second polypeptide, wherein each of the first polypeptide and the second polypeptide comprises the amino acid sequence of SEQ ID NO:7. In some instances, the GDF15 polypeptide comprises a dimer, wherein the dimer comprises a first polypeptide and a second polypeptide, wherein each of the first polypeptide and the second polypeptide comprises the amino acid sequence of SEQ ID NO:7, wherein each of the first polypeptide and the second polypeptide lacks the first three amino acids of SEQ ID NO:1. In some instances, the GDF15 polypeptide comprises a dimer, wherein the dimer comprises a first polypeptide and a second polypeptide, wherein each of the first polypeptide and the second polypeptide consists of the amino acid sequence of SEQ ID NO:7.

[00123] In some instances, the GDF15 polypeptide comprises a dimer, wherein the dimer comprises a first polypeptide and a second polypeptide, wherein each of the first polypeptide and the second polypeptide comprises the amino acid sequence of SEQ ID NO:7, wherein each of the first polypeptide and second polypeptide lacks the first three amino acids of SEQ ID NO: 1. In some instances, each of the first polypeptide and the second polypeptide consists of the amino acid sequence of SEQ ID NO:7. In some instances, the subject is a human. In some instances, the autoimmune disease is MS. In some cases, the MS is relapsing-remitting MS. In some cases, the MS is primary progressive MS. In some cases, the MS is secondary progressive MS. In some cases, the MS is benign MS. In some instances, the autoimmune diseases is psoriasis, psoriatic arthritis, or an inflammatory bowel disease (e.g., Crohn’s disease, ulcerative colitis). The GDF15 polypeptides (or pharmaceutical compositions comprising the same) can be administered in any number of ways for either local or systemic delivery. Further, the GDF15 polypeptides (or pharmaceutical compositions comprising the same) used in the methods of prevention described herein can be administered to the subject in a therapeutically effective amount. In this context, the term “therapeutically effective amount” as used herein refers to the amount of an agent (e.g., GDF15 polypeptide) that is sufficient to reduce and/or ameliorate the severity and/or duration of (i) a disease, disorder or condition in a subject, and/or (ii) a symptom in a subject.

[00124] In some instances, the method of prevention obtains a desired pharmacological and/or physiological effect in the subject being administered the GDF15 polypeptide. Thus, in some instances, the method of prevention inhibits the autoimmune disease, e.g., arrests its development or prolongs or delays its onset as compared to expected development or onset if not receiving the treatment. In some instances, the method of prevention prolongs survival of the subject administered the GDF15 polypeptide as compared to expected survival if not receiving the treatment. [00125] The GDF15 polypeptides (or pharmaceutical compositions comprising the same) can be administered in any number of ways for either local or systemic prevention. Further, the GDF15 polypeptides (or pharmaceutical compositions comprising the same) used in the methods of prevention described herein can be administered to the subject in a therapeutically effective amount. In this context, the term “therapeutically effective amount” encompasses an amount of an agent (e.g., GDF15 polypeptide) necessary for the (i) reduction or amelioration of the development, or onset of a given disease, disorder, or condition, and/or (ii) the improvement or enhancement of the prophylactic or therapeutic effect(s) of another agent or therapy (e.g., an agent other than the binding agents provided herein).

[00126] In some instances, the method of prevention prevents or delays onset of one or more (e.g., 1, 2, 3, 4, 5) symptoms of the autoimmune disease.

[00127] In some instances in which the method is for preventing MS (e.g., benign MS, primary progressive MS, secondary progressive MS, relapsing-remitting MS), the method of prevention prevents or delays onset of one or more (e.g., 1, 2, 3, 4, 5) symptoms of MS. In some instances, the MS is relapsing-remitting MS. In some instances, the MS is primary progressive MS. In some instances, the MS is secondary progressive MS. Symptoms of MS and clinical endpoints for treatment of MS are known in the art (see, e.g., van Munster and Uitdehaag, CNS Drugs, 2017; 31(3):217- 236, which is incorporated by reference herein in its entirety).

[00128] In some instances, the method for prevention reduces (e.g., by at least 5%, at least 10%, at least 15%, at least 20%, at least 30%, at least 40%, or at least 50%, 10-50%, 10-25%, 10-15%, 25-50%, or 5-25%) infiltration of pathogenic immune cells into the tissues or organs (e.g., CNS) in the subject. In some instances, the reduction is compared to a level of pathogenic immune cells in the tissues or organs (e.g., CNS) prior to administration of the GDF15 polypeptide. Methods for determining infiltration of pathogenic immune cells into the tissues or organs are known in the art. For example, for CNS biopsy lumbar puncture can be performed to obtain spinal tissue or spinal fluid, which is then tested for pathogenic immune cells and autoantibodies.

[00129] In some instances, the GDF15 polypeptide is administered to the subject (e.g., human) as a pharmaceutical composition (e.g., a sterile pharmaceutical composition). In some instances, the pharmaceutical composition comprises: a GDF15 polypeptide, and a pharmaceutically acceptable carrier. Pharmaceutically acceptable carriers are known in the art. In some instances, the pharmaceutical composition is a sterile pharmaceutical composition.

Methods of Reducing or Blocking Pathogenic Immune Cell Infiltration into Tissues or Organs

[00130] A GDF15 polypeptide described herein or a pharmaceutical composition comprising the same can be used to reduce or block pathogenic immune cell infiltration into tissue(s) or organ(s) (e.g., CNS) of a subject suffering from an autoimmune disease. In some instances, a GDF15 polypeptide described herein or a pharmaceutical composition comprising the same is used to reduce or block pathogenic immune cell infiltration into tissue(s) or organ(s) (e.g., CNS) of a subject suffering from relapsing-remitting MS. In some instances, a GDF15 polypeptide described herein or a pharmaceutical composition comprising the same is used to reduce or block pathogenic immune cell infiltration into tissue(s) or organ(s) (e.g., CNS) of a subject suffering from primary progressive MS. In some instances, a GDF15 polypeptide described herein or a pharmaceutical composition comprising the same is used to reduce or block pathogenic immune cell infiltration into tissue(s) or organ(s) (e.g., CNS) of a subject suffering from secondary progressive MS. In some instances, a GDF15 polypeptide described herein or a pharmaceutical composition comprising the same is used to reduce or block pathogenic immune cell infiltration into one or more joints of a subject suffering from psoriatic arthritis. In some instances, a GDF15 polypeptide described herein or a pharmaceutical composition comprising the same is used to reduce or block pathogenic immune cell infiltration into the skin, nails or joints of a subject suffering from psoriasis. In some instances, a GDF15 polypeptide described herein or a pharmaceutical composition comprising the same is used to reduce or block pathogenic immune cell infiltration into the digestive tract of a subject suffering from inflammatory bowel disease.

[00131] Thus, provided herein is a method for reducing or blocking pathogenic immune cell infiltration into tissue(s) or organ(s) of a subject (e.g., a subject, e.g., a human, suffering from an autoimmune disease), the method comprising administering to the subject (e.g., human) a therapeutically effective amount of a GDF15 polypeptide (or composition comprising the same) to the subject. In some instances, the autoimmune disease is relapsing-remitting MS. In some instances, the autoimmune disease is primary progressive MS. In some instances, the autoimmune disease is secondary progressive MS. In some instances, the autoimmune disease is benign MS. In some instances, the autoimmune disease is psoriasis, psoriatic arthritis, or inflammatory bowel disease (e.g., Crohn’s disease, ulcerative colitis). In some instances, the autoimmune disease is type 1 diabetes. In some instances, the autoimmune disease is not Type 1 diabetes. In some instances, the autoimmune disease is not a glucose metabolism disorder. In some instances, the subject is a human.

[00132] In some instances, the pathogenic immune cell is a pathogenic T cell. In some instances, the T cell is a CD4+ T cell. In some instances, the T cell is a Th-1 cell. In some instances, the T cell is a Th-17 cell. In some instances, the T cell is a myelin oligodendrocyte glycoprotein (MOG)-specific CD4+ T cell. Methods of identifying pathogenic immune cells are known in the art, see, e.g., the working examples herein. For instance, the amount of white blood cells may be quantitated in spinal fluid.

[00133] Methods for determining infiltration of pathogenic immune cells into tissues and organs are known in the art. For example, to determine infiltration into the CNS, a biopsy can be taken via lumbar puncture to obtain spinal tissue or spinal fluid, which is then tested for pathogenic immune cells and autoantibodies.

[00134] In some instances, the subject has an autoimmune disease. In some instances, the autoimmune disease is MS. In some cases, the MS is relapsingremitting MS. In some cases, the MS is primary progressive MS. In some cases, the MS is secondary progressive MS. In some cases, the MS is benign MS.

[00135] In some instances, the autoimmune disease is selected from the group consisting of MS, rheumatoid arthritis, psoriasis, psoriatic arthritis, systemic lupus erythematosus (SLE), cutaneous lupus erythematosus (CLE), discoid lupus erythematosus (DLE), inflammatory bowel disease (e.g., Crohn’s disease, ulcerative colitis), polymyositis, dermatopolymyositis, Addison’s disease, Graves’ disease, Sjogren’s syndrome, Hashimoto’s thyroiditis, myasthenia gravis, autoimmune vasculitis, pernicious anemia, scleroderma, cytokine release syndrome, and celiac disease. In some instances, the autoimmune disease is MS. In some cases, the MS is relapsing-remitting MS. In some cases, the MS is primary progressive MS. In some cases, the MS is secondary progressive MS. In some cases, the MS is benign MS. [00136] In some instances, the autoimmune disease is type 1 diabetes. In some instances, the autoimmune disease is not Type 1 diabetes. In some instances, the autoimmune disease is not a glucose metabolism disorder.

[00137] In some instances, the GDF15 polypeptide comprises a complex comprising a first heterodimer and a second heterodimer; wherein each of the first heterodimer and the second heterodimer comprises a first polypeptide and a second polypeptide; wherein the first polypeptide comprises from the N-terminus to the C- terminus a first human IgGl Fc domain, a linker, and a GDF15 mutein, wherein the first human IgGl Fc domain comprises a first hinge region, a first CH2 domain, and a first CH3 domain; wherein the second polypeptide comprises from the N-terminus to the C-terminus a second human IgGl Fc domain, wherein the second human IgGl Fc domain comprises a second hinge region, a second CH2 domain, and a second CH3 domain; and wherein the GDF15 polypeptide binds to GFRAL. In some instances, the first human IgGl Fc domain comprises one or more (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10) substitutions that reduces or abrogates an Fc effector function (e.g., as described herein above). In some instances, the first human IgGl Fc domain comprises an engineered protuberance and the second human IgGl Fc domain comprises an engineered cavity. In some instances, the engineered protuberance comprises the substitution T366W according to the EU numbering and the engineered cavity comprises the substitutions T366S, L368A, and Y407V according to the EU numbering. In some instances, the GDF15 mutein comprises an amino acid sequence having at least 90%, at least 95%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to the amino acid sequence of any one of SEQ ID NOs:3-12 (except wherein the GDF15 mutein retains the one or more differences in any one of SEQ ID NOs:3-12, respectively, relative to SEQ ID NO:1). In some instances, the GDF15 mutein comprises the amino acid sequence of any one of SEQ ID NOs:3-12 except for one or more (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10) amino acid substitutions (except wherein the GDF15 mutein retains the one or more differences in any one of SEQ ID NOs:3- 12, respectively, relative to SEQ ID NO:1). In some instances, the GDF15 mutein comprises an amino acid sequence having at least 90%, at least 95%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to the amino acid sequence of SEQ ID NO:3, except wherein the GDF15 mutein comprises a D5T/S substitution relative to SEQ ID NO: 1 and lacks the first two amino acids of SEQ ID NO: 1. In some instances, the GDF15 mutein comprises the amino acid sequence of SEQ ID NO:3 except for one or more (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10) amino acid substitutions, except wherein the GDF15 mutein comprises a D5T/S substitution relative to SEQ ID NO: 1 and lacks the first two amino acids of SEQ ID NO: 1. In some instances, the GDF15 mutein comprises the amino acid sequence of SEQ ID NO:3 and lacks the first two amino acids of SEQ ID NO: 1. In some instances, the GDF15 mutein consists of the amino acid sequence of SEQ ID NO:3. In some instances, the first polypeptide comprises an amino acid sequence having at least 90%, at least 95%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to the amino acid sequence of SEQ ID NO: 13 or 14. In some instances, the first polypeptide comprises the amino acid sequence of SEQ ID NO: 13 or 14 except for one or more (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10) amino acid substitutions. In some instances, the first polypeptide comprises an amino acid sequence having at least 90%, at least 95%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to the amino acid sequence of SEQ ID NO: 13. In some instances, the first polypeptide comprises an amino acid sequence having one or more (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10) amino acid substitutions relative to the amino acid sequence of SED ID NO: 13, except wherein the GDF15 mutein in SEQ ID NO: 13 comprises a D5T/S substitution relative to SEQ ID NO: 1 and lacks the first two amino acids of SEQ ID NO: 1. In some instances, the first polypeptide comprises the amino acid sequence of SEQ ID NO: 13. In some instances, the first polypeptide consists of the amino acid sequence of SEQ ID NO: 13. In some instances, the second polypeptide comprises an amino acid sequence having at least 90%, at least 95%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to the amino acid sequence of SEQ ID NO: 15. In some instances, the first polypeptide comprises the amino acid sequence of SEQ ID NO: 13, and the second polypeptide comprises the amino acid sequence of SEQ ID NO: 15. In some instances, the first polypeptide consists of the amino acid sequence of SEQ ID NO: 13, and the second polypeptide consists of the amino acid sequence of SEQ ID NO: 15. In some instances, the complex binds to GFRAL.

[00138] In some instances, the GDF15 polypeptide comprises a complex comprising a first heterodimer and a second heterodimer, wherein each of the first heterodimer and the second heterodimer comprises a first polypeptide and a second polypeptide, wherein the first polypeptide comprises the amino acid sequence of SEQ ID NO: 13 and the second polypeptide comprises the amino acid sequence of SEQ ID NO: 15; wherein the first heterodimer dimerizes with the second heterodimer. In some instances, the first polypeptide consists of the amino acid sequence of SEQ ID NO: 13. In some instances, the second polypeptide consists of the amino acid sequence of SEQ ID NO: 15. In some instances, the first polypeptide consists of the amino acid sequence of SEQ ID NO: 13 and the second polypeptide consists of the amino acid sequence of SEQ ID NO: 15. In some instances, the subject is a human. In some instances, the autoimmune disease is MS. In some instances, the MS is relapsingremitting MS. In some instances, the MS is primary progressive MS. In some instances, the MS is secondary progressive MS. In some cases, the MS is benign MS. In some instances, the autoimmune disease is psoriasis, psoriatic arthritis, or inflammatory bowel disease (e.g., Crohn’s disease, ulcerative colitis). In some instances, the autoimmune disease is type 1 diabetes. In some instances, the autoimmune disease is not Type 1 diabetes. In some instances, the autoimmune disease is not a glucose metabolism disorder.

[00139] In some instances, the GDF15 polypeptide comprises a dimer, wherein the dimer comprises a first polypeptide and a second polypeptide, wherein each of the first polypeptide and the second polypeptide comprises a GDF15 mutein or wild type GDF15. In some instances, the first polypeptide is identical to the second polypeptide. In some instances, the first polypeptide is not identical to the second polypeptide. In some instances, the GDF15 polypeptide comprises a dimer, wherein the dimer comprises a first polypeptide and a second polypeptide, wherein each of the first polypeptide and the second polypeptide comprises a GDF15 mutein, e.g., a GDF15 mutein described herein. In some instances, the GDF15 polypeptide comprises a dimer, wherein the dimer comprises a first polypeptide and a second polypeptide, wherein each of the first polypeptide and the second polypeptide comprises an amino acid sequence having at least 90%, at least 95%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to the amino acid sequence the amino acid sequence of any one of SEQ ID NOs:3-12. In some instances, the GDF15 polypeptide comprises a dimer, wherein the dimer comprises a first polypeptide and a second polypeptide, wherein each of the first polypeptide and the second polypeptide comprises an amino acid sequence having one or more (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10) substitutions or deletions relative to the amino acid sequence of any one of SEQ ID NOs:3-10). In some instances, each of the first polypeptide and the second polypeptide lacks the first two amino acids of SEQ ID NO: 1. In some instances, each of the first polypeptide and the second polypeptide lacks the first three amino acids of SEQ ID NO: 1. In some instances, each of the first polypeptide and the second polypeptide lacks the first six amino acids of SEQ ID NO: 1. In some instances, the GDF15 polypeptide comprises a dimer, wherein the dimer comprises a first polypeptide and a second polypeptide, wherein each of the first polypeptide and the second polypeptide comprises the amino acid sequence of SEQ ID NO:7. In some instances, the GDF15 polypeptide comprises a dimer, wherein the dimer comprises a first polypeptide and a second polypeptide, wherein each of the first polypeptide and the second polypeptide comprises the amino acid sequence of SEQ ID NO:7, wherein each of the first polypeptide and the second polypeptide lacks the first three amino acids of SEQ ID NO:1. In some instances, the GDF15 polypeptide comprises a dimer, wherein the dimer comprises a first polypeptide and a second polypeptide, wherein each of the first polypeptide and the second polypeptide consists of the amino acid sequence of SEQ ID NO:7.

[00140] In some instances, the GDF15 polypeptide comprises a dimer, wherein the dimer comprises a first polypeptide and a second polypeptide, wherein each of the first polypeptide and the second polypeptide comprises the amino acid sequence of SEQ ID NO:7, wherein each of the first polypeptide and second polypeptide lacks the first three amino acids of SEQ ID NO: 1. In some instances, each of the first polypeptide and the second polypeptide consists of the amino acid sequence of SEQ ID NO:7. In some instances, the subject is a human. In some instances, the autoimmune disease is MS. In some instances, the MS is relapsing-remitting MS. In some instances, the MS is primary progressive MS. In some instances, the MS is secondary progressive MS. In some instances, the autoimmune disease is benign MS. In some instances, the autoimmune disease is psoriasis, psoriatic arthritis, or inflammatory bowel disease (e.g., Crohn’s disease, ulcerative colitis). In some instances, the autoimmune disease is type 1 diabetes. In some instances, the autoimmune disease is not Type 1 diabetes. In some instances, the autoimmune disease is not a glucose metabolism disorder.

[00141] The GDF15 polypeptides (or pharmaceutical compositions comprising the same) can be administered in any number of ways for either local or systemic delivery. Further, the GDF15 polypeptides (or pharmaceutical compositions comprising the same) used in the methods reducing or blocking pathogenic immune cell infiltration into the tissue(s) or organ(s) (e.g., CNS) of a subject described herein can be administered to the subject in a therapeutically effective amount. In this context, the term “therapeutically effective amount” as used herein refers to the amount of an agent (e.g., GDF15 polypeptide) that is sufficient to reduce and/or block pathogenic immune cell infiltration into the tissue(s) or organ(s) (e.g., CNS) of the subject.

[00142] In some instances, the method of reducing or blocking pathogenic immune cell infiltration into the tissue(s) or organ(s) (e.g., CNS) of a subject reduces or blocks the pathogenic immune cell infiltration into the tissue(s) or organ(s) (e.g., CNS) of the subject by 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%, or 100 % as compared to pathogenic immune cell infiltration into the tissue(s) or organ(s) (e.g., CNS) of a subject suffering from an autoimmune disease and not receiving the GDF15 polypeptide. In some instances, the method of reducing or blocking pathogenic immune cell infiltration into the tissue(s) or organ(s) (e.g., CNS) of a subject reduces or blocks the pathogenic immune cell infiltration into the tissue(s) or organ(s) (e.g., CNS) of the subject by 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%, or 100 % as compared to pathogenic immune cell infiltration into the tissue(s) or organ(s) (e.g., CNS) of the subject prior to (e.g., within 1 day, within 1 week, within 1 month) being administered the GDF15 polypeptide.

[00143] In some instances, the GDF15 polypeptide is administered to the subject (e.g., human) as a pharmaceutical composition (e.g., a sterile pharmaceutical composition). In some instances, the pharmaceutical composition comprises: a GDF15 polypeptide, and a pharmaceutically acceptable carrier. Pharmaceutically acceptable carriers are known in the art. In some instances, the pharmaceutical composition is a sterile pharmaceutical composition.

EXAMPLES

Materials and Methods

Animal Studies Experimental autoimmune encephalomyelitis (EAE)induced by MOG35-55 [00144] Female C57BL/6 mice around 22 grams and approximately 16 weeks old (The Jackson Laboratory, ME, USA) were used. The animals were kept under a 12- hour-light/dark cycle under controlled temperature (22 ± 2°C) and humidity (50% ± 20%) with access to food and water ad libitum.

[00145] Experimental autoimmune encephalomyelitis (EAE) in the mice was induced by immunization with myelin oligodendrocyte glycoprotein/CFA Emulsion PTX kit from Hooke Labs (EK-2110, Hooke Laboratories, Lawrence, MA, USA). Sequence MEVGWYRSPFSRVVHLYRNGK (SEQ ID NO:23).

[00146] Briefly, under 2% isoflurane anesthesia, MOG/CFA emulsion in prefilled syringes were injected subcutaneously on two sites dorsally, 0.1 mL/site (0.2 mL/mouse total) with a total of 0.2 mg of MOGss-sspeptide. Subsequently, the mice were injected intraperitoneally with 85-110 ng of pertussis toxin (PTX) at 4 hours and then 1 day post MOG injection. Mice were weighed and assessed daily to record behavioral and neurological signs starting on the eighth day after the first immunization. The signs of EAE were scored as described previously (Selvaraj et al., J. Biol. Chem. 2009, 284:26070-84) and summarized in Table 1. For prophylactic GDF15 ANALOGS treatment in C57BL/6 mice, the protein was injected subcutaneously 1-2 days prior to MOG immunization followed by weekly subcutaneous injection. In some studies, the mice were sacrificed at day 16 after MOG immunization and their inguinal lymph nodes, spleen and spinal cords harvested for FACS analyses. For therapeutic treatment, GDF15 ANALOGS were injected 7-10 days post immunization when at least 20% of mice reached EAE score of 0.5-l.

EAE induced by MOG 1-125

[00147] Female C57BL/6 mice around 22 grams and approximately 16 weeks old (The Jackson Laboratory, ME, USA) were used. The animals were kept under a 12- hour-light/dark cycle under controlled temperature (22 ± 2°C) and humidity (50% ± 20%) with access to food and water ad libitum. [00148] EAE in these mice were induced by using the MOG1-125/CFA Emulsion PTX kit (EK-2160, Hooke Laboratories, Lawrence, MA, USA). Sequence MGQFRVIGPRHPIRALVGDEVELPCRISPGKNATGMEVGWYRPPFSRVVHLY RNGKDQDGDQAPEYRGRTELLKDAIGEGKVTLRIRNVRFSDEGGFTCFFRDH SYQEEAAMELKVEDPFYWVSPGHHHHHH (SEQ ID NO:24). Animal handling was identical to that of MOG35-55 induced EAE except that a total of 0.1 mg of MOG1-125 protein was injected.

[00149] Therapeutic GDF15 ANALOG 1 (1 mg/kg) or anti-CD20 (100 ug) (MB20-11, BioXCell, Catalog #BP0356) weekly dosing (subcutaneous) was initiated 7-10 days post immunization when at least 20% of mice reached EAE score at 0.5-1.

EAE induced by proteolipid protein (PEP)

[00150] Female SJL mice around 20 grams and approximately 8 weeks old (The Jackson Laboratory, ME, USA) were used. The animals were kept under a 12-hour- light/dark cycle under controlled temperature (22 ± 2°C) and humidity (50% ± 20%) with access to food and water ad libitum.

[00151] EAE in these mice were induced by using the [Serl40]-PLP 139-151/CFA Emulsion kit (EK0120, Hooke Laboratories, Lawrence, MA, USA). Sequence HSLGKWLGHPDKF (SEQ ID NO: 25). PLP139-151 emulsion prefilled syringes were injected subcutaneously on four sites dorsally, 0.05 mL/site (0.2 mL/mouse total) with a total of 0.1 mg of PLP139-151 peptide.

[00152] For prophylactic GDF15 ANALOG 1 treatment, the protein was injected subcutaneously 1-2 days prior to PLP immunization followed by weekly administration. For therapeutic treatment in SJL mice, GDF15 ANALOG 1 injection was initiated 19 days post immunization when EAE remitted from the peak score above 2 to the nadir score at 1 with all the mice showing signs of EAE. [00153] Table 1

Single Cell preparation for FACS

[00154] Single cell suspensions were prepared from the lumbar spinal cord, both inguinal lymph nodes, and spleen (cut into small pieces) for FACS. Isolated tissues were weighed and kept in ice cold phosphate buffered saline (PBS). Tissues were then transferred to 1 mL of digestion media (human tumor dissociation kit, Miltenyi; 130-110-201) in a 15 mL conical tube and incubated in a 37 °C water bath for 45 minutes, during which the tissues were gently mixed every 15 minutes by repetitive pipetting. The digested single cell suspension was filtered and transferred to a tube with 5 mL RPMI + 2% fetal bovine serum (FS) and centrifuged for 5 minutes at 1300 revolutions per minute (rpm) at 4 °C. Supernatant was discarded and the cell pellet was resuspended in ~ 1 mL RBC lysis buffer (ebiosciences). Cells were centrifuged for 5 minutes at 1300 rpm at 4 °C. Supernatant was discarded and the cell pellet was resuspended in 500 pL (for spinal cord and lymph nodes) or 2 mL (for spleen) of RPMI + 2% FBS. Cells were counted on Vi-cell cell viability analyzer (Beckman Coulter). 150 pl of cell suspension was placed in 96 well U bottom plates for FACS staining.

Antibody staining and FACS analysis

[00155] Plated cells were centrifuged for 4 minutes at 1300 rpm at 4 °C. Human Fc block (1:200) and Live/Dead fixable dead stain (Thermo Fisher scientific) was added in 100 pL volume of PBS, and cells were incubated for 15 minutes at 4 °C.

Cells were centrifuged and resuspended in 50 pL of BD cell staining buffer (FBS; BD biosciences). 10 pL of MOG tetramer (MOG 35-55, MBL international corporation) was added to each well and the cells were incubated for 40 minutes at 4°C. 50 pL of cell staining buffer containing surface antibodies was added and the cells were further incubated for 20 minutes at 4 °C. Cells were centrifuged and washed twice in 150 pL volume of BD cell staining buffer. For surface staining panel the cells were incubated in 100 pL of BD cytofix (BD biosciences) for 10 minutes at 4 °C. Cells were centrifuged and washed twice in 150 pL volume of BD cell staining buffer. Cells were resuspended in 250 pL of BD cell staining buffer for FACS analysis.

[00156] Intracellular Staining: Following surface staining as described above the cells were centrifuged and washed twice in 150 pL volume of BD cell staining buffer. Following which intracellular staining was carried out using FoxP3/Transcription factor staining buffer set according to manufacturer’s protocol (Thermofisher). Cells were stained with intracellular antibodies for 30 minutes at 4 °C. Cells were centrifuged and washed twice in 150 pL volume of BD cell staining buffer. Cells were resuspended in 250 pL of BD cell staining buffer for FACS analysis.

[00157] FACS was performed on BD LSRII and analysis performed on FlowJo software (BD biosciences).

Antibodies

[00158] Antibodies other than those to RORgt and 3P10 were procured from Biolegend. Antibody to RORgt (B2D) was procured from BD Biosciences.

[00159] 3P 10 is an inhibitory anti-GFRAL antibody that blocks GFRAL binding to

RET to antagonize GDF15 signaling without affecting GDF15 binding to GFRAL and comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:21 and a light chain variable region comprising the amino acid sequence of SEQ ID NO:22.

GDF15 Analog [00160] GDF15 Analog 1 is a complex comprising a first heterodimer and a second heterodimer, wherein each of the first heterodimer and the second heterodimer consists of a first polypeptide and a second polypeptide, wherein the first polypeptide consists of the amino acid sequence of SEQ ID NO: 13 and the second polypeptide consists of the amino acid sequence of SEQ ID NO: 15, and wherein the first heterodimer dimerizes with the second heterodimer. GDF15 Analog 2 is a homodimer comprising a first polypeptide and a second polypeptide, wherein each of the first polypeptide and the second polypeptide consists of the amino acid sequence of SEQ ID NO: 7.

Adoptive transfer of EAE

[00161] Donor mice (female B6.PL-Thyla/CyJ) and recipient mice (female C57BL/6), purchased from The Jackson Laboratory, ME, USA, were around 20 grams and approximately 9-week-old at the beginning of the study. The animals were kept under a 12-hour-light/dark cycle under controlled temperature (22 ± 2°C) and humidity (50% ± 20%) with access to food and water ad libitum. Donor mice were immunized with MOG/CFA (from Hooke lab) to induce experimental autoimmune encephalomyelitis (EAE) using a standard protocol. The signs of EAE were scored as described previously (Selvaraj et al., J. Biol. Chem. 2009, 284:26070-84, which is incorporated by reference herein in its entirety) and summarized in Table 1. On day 12, spleens and inguinal lymph nodes from MOG-immunized donor mice were harvested and collected in a single-cell suspension and red blood cells were lysed. These splenocytes and lymphocytes were cultured in T cell medium (DMEM media enriched with 10% FBS, 2 mM L-glutamine, 50 pM 2-mercaptoethanol, 1 mM nonessential amino acids, sodium pyruvate, and penicillin/streptomycin) at the density of 150 mm TC dish with 50 mL of 7E6/mL in the presence of 30 pg/ml MOG (Tocris Bioscience) plus recombinant murine IL-12 (25 ng/ml), murine IL-23 (25 ng/ml), and anti-IL-4 ( I Opg/ml). Three days after initiation of the cultures, cells were harvested, washed in PBS, and 4E6 cell/mouse in 100 pL PBS were injected intraperitoneally into recipients. PTX administration as described above for EAE induction and mice were weighed and assessed daily to record behavioral and neurological signs on the seventh day after cell transfer. For the study described in FIG. 9, the donor mice were dosed with 1 mg/kg GDF15 Analog 1 or anti-KLH one day prior to MOG/CFA immunization in donor mice. For the study described in FIG. 10, the recipient mice were dosed with 1 mg/kg GDF15 Analog 1 or anti-KLH at day 10, 17, and 24 post receiving donor immune cells.

T cell adoptive transfer-induced inflammatory bowel disease (IBD) model

[00162] On Study Day 0, spleens were harvested from BALB/c mice and processed by the GentleMACS Octodissociator (Miltenyi, 130-093-235) to generate single cell suspension. CD4+CD62L+ T-cells were then sorted using the Stemcell negative selection Kit (Cat# 19765) per the manufacturer’s instructions. 0.5 x 10⁶ of CD4+CD62L+ T-cells were transferred per BALB/c SCID female animal via intraperitoneal injection. Body weight were recorded once a week from day 0 to 21 post T-cell transfer, and then three times a week from Day 21 until Day 42. Mice were euthanized via cervical dislocation and tissues were weighted and processed for immunohistochemistry and pathology analyses.

Spontaneous type I diabetes model

[00163] Female NOD, NOD/ShiLtJ mice (Stock No: 001976) around 19 grams approximately 6 weeks old (The Jackson Laboratory, ME, USA) were used. The animals were kept in the clean room under a 12-hour-light/dark cycle under controlled temperature (22 ± 2°C) and humidity (50% ± 20%) with access to food and water ad libitum.

Female NOD mice spontaneously develop autoimmune diabetes at about 12-14 weeks of age. Starting from the age of 6 weeks, mice were subcutaneously injected with GDF15 Analog 1 or mouse anti-KLH proteins weekly for 18 weeks. Body weight and blood glucose were monitored weekly. Mice with blood glucose level at 250 mg/dL or above for two consecutive weeks were called diabetic and euthanized.

Cuprizone model [00164] 6-week-old male C57BL/6 mice from Jackson Lab were used for the studies. The diet containing 0.2% cuprizone (CPZ) was purchased from Envigo (Envigo, TD.140803). Mice were fed with CPZ-containing diet for 5 weeks, during which fresh food pellets were replaced every 3 to 5 days. The mice were euthanized on week 3 or week 5 and the whole brain were harvested for immunohistochemistry and pathology analyses. In some studies, the mice were fed with CPZ-containing diet for 5 weeks, followed by feeding with normal chow for 2 weeks to allow for remyelination. The mice were then euthanized on week 7 and the whole brain were harvested for immunohistochemistry and pathology analyses.

Immunohistochemistry and pathology readouts

[00165] For the IBD study, the whole colon was fixed as longitudinal segments in 10% neutral buffered formalin, processed to paraffin, and embedded creating strips to visualize the full height of the colonic glands. The slides were stained for hematoxylin and eosin on the Leica Autostainer XL (Leica Biosystems, Buffalo Grove, IL).

[00166] The colon was evaluated for the thickness of the colonic mucosa from the surface epithelium to the deep lamina propria at the junction with the muscularis mucosa using the Visiopharm software (Visiopharm A/S, Horsholm, Denmark). This assessment was performed in representative areas of the proximal, mid and distal colon.

[00167] For the cuprizone study, the whole brain was fixed in 10% neutral buffered formalin, processed to paraffin, and embedded. Cross sections of the brain centered on the corpus callosum and including the hippocampus (Bregma -2.12 mm to Bregma -5.30 mm) were stained for Luxol Fast Blue (LFB, StatLab, McKinney, TX), Ionized Calcium-Binding Adaptor Molecule 1 (IB Al, Clone E404W, Cell Signaling Technology, Danvers, MA), or Glial Fibrillary Acidic Protein (GFAP, rabbit polyclonal antibody catalog g9269, Millipore Sigma, Burlington, MA). The LFB staining was performed by hand as per the manufacturer’s instructions. The staining for IBA1 and GFAP was performed on the Leica Bond Rx autostainer (Leica Biosystems, Buffalo Grove, IL). [00168] The slides were evaluated visually for differences between the control group and the CPZ model. IBA1 and GFAP expression was assessed by image analysis in hand-drawn specific areas of the brain using the Visiopharm software (Visiopharm A/S, Horsholm, Denmark). The slides stained with LFB were graded twice blindly for demyelination which was graded from 0 (none) to 4 (complete demyelination).

Results

Example 1: Effect of GDF15 Polypeptide in MS

[00169] Prophylactic dosing of EAE mice with 0.3 mg/kg, 1.0 mg/kg, or 3.0 mg/kg of GDF15 Analog 1 blocked MOG35-55 peptide-induced EAE at day 16 (FIG. 1). Anti-KLH treated mice showed a sharp decline of body weight around day 10 post the immunization (FIG. 2) coinciding with the onset of EAE development (FIG. 1). GDF15 Analog 1 treated animals maintained stable body weight from 10 days onwards post immunization despite the initial does-dependent weight loss, which was driven by the well-documented metabolic effect of GDF15 (FIG. 2). Prophylactic dosing of EAE mice with anti-GFRAL antibody 3P10 blocked the effect of GDF15 Analog 1, indicating that it is a GFRAL-dependent mechanism (FIG. 1 and FIG. 2). Prophylactic dosing of GDF15 Analog 1 did not show EAE blocking effect when dosed in GFRAL knockout mice, confirming the GFRAL-dependent mechanism (FIG. 3). Prophylactic dosing of GDF15 Analog 1 as low as 0.1 mg/kg potently prevented disease from developing in EAE mice (FIG 4).

[00170] Therapeutic dosing with GDF15 Analog 1 on day 12 post immunization and at disease onset attenuated and prevented severe EAE symptom development as well as stabilizing EAE progression (FIG. 5). This attenuation of EAE score was mediated by the GDF15/GFRAL pathway (data not shown). In summary, administration of GDF15 Analog 1 prophylactically and therapeutically robustly improved the outcome of autoimmune encephalomyelitis in a well-validated animal model of multiple sclerosis.

[00171] Prophylactic administration of GDF15 Analog 1 blocked infiltration of Th-

17 cells (FIG. 6A and FIG. 6B) and immune cells (FIG. 7A and FIG. 7B) into the CNS; this effect was reversed by treatment with anti-GFRAL antibody. Additionally, prophylactic administration of GDF15 Analog 1 blocked CD4+ T cell infiltration into the CNS; this effect was reversed by treatment with anti-GFRAL antibody (FIG. 8A and FIG. 8B). Finally, prophylactic administration of GDF15 Analog 1 blocked MOG-specific T cell infiltration in the CNS; this effect was reversed by treatment with anti-GFRAL antibody (FIG. 9A and FIG. 9B).

[00172] In an adoptive transfer EAE model in which mice developed disease upon receiving immune cells from donor mice that were immunized with MOG35-55, treating the donor mice with GDF15 Analog 1 prior to immunization delayed the disease onset of the recipient mice (FIG. 10). In the same model, treating the recipient mice with GDF15 Analog 1 at the disease onset ameliorated disease progression (FIG. 11). These data demonstrated mechanistically that GDF15 Analog 1 can attenuate the generation of EAE-causing immune cells as well as inhibit the pathological activity of these pathogenic immune cells in the CNS.

[00173] Early therapeutic dosing of 1 mg/kg GDF15 Analog 1 in a human M0G1- 120 protein-induced EAE model attenuated disease progression to a similar degree as that of B cell depletion by dosing the mice with anti-CD20 antibody (FIG. 12).

[00174] In a relapsing-remitting EAE model in which SJL mice were immunized with PLP139-151 peptide, prophylactic dosing of GDF15 Analog 1 blocked the onset of disease. In the same model, therapeutic dosing of GDF15 Analog 1 ameliorated disease progression and reduced EAE severity despite pathogenic immune cell infiltration into the CNS (FIG. 13).

[00175] The effect of wildtype GDF15 and GDF15 Analog 2 were tested in the EAE model wherein C57BL/6 mice were immunized with MOG35-55 peptide. Prophylactic dosing of the mice with 1.0 mg/kg GDF15 Analog 2 or 1.0 mg/kg wildtype GDF15 blocked the EAE development compared to the mice dosed with the control anti-KLH antibody (FIG. 26), consistent with the effect seen from GDF15 Analog 1.

[00176] Taken together, therapeutic administration of a GDF15 polypeptide in multiple animal models of MS showed attenuation or blockage of autoimmune disease progression in the CNS. Further, prophylactic administration of a GDF15 polypeptide completely blocked pathogenic immune cell infiltration in the CNS and prevented onset of autoimmune disease in EAE mice. Anti-GFRAL antibody completely reversed the GDF 15 -mediated effects, suggesting that this effect is mediated by GDF15-GFRAL pathway. These data support use of a GDF 15 polypeptide in the treatment and prevention of autoimmune diseases such as MS.

Example 2: Effects of GDF 15 Polypeptide in Inflammatory Bowel Disease [00177] The effect of a GDF 15 polypeptide on autoimmune disease was tested in an inflammatory bowel disease (IBD) model, wherein the IBD was induced by adoptive transfer of naive CD4+ T cells from wild type Balb/c donor mice to SCID Balb/c recipient mice. Administration of 1 mg/kg of GDF15 Analog 1 significantly reduced the spleen weight and spleen weight/body weight ratio (FIG. 14), suggesting a systemic reduction of inflammation by GDF 15 Analog 1. GDF 15 Analog 1 dosing at 1 mg/kg also reduced the colon weight and colon weight/body weight ratio without affecting colon length (FIG. 15). GDF 15 Analog 1 also reduced the colon mucosal thickening, a hallmark of epithelium proliferation in response to inflammation (FIG. 16A and 16B). As expected, GDF 15 Analog 1 caused a dose-dependent weight loss at the onset of the study demonstrating that the metabolic effect of GDF 15 is preserved in this model (FIG. 17). All these data show that a GDF 15 polypeptide including a GDF 15 analog in the treatment and prevention of autoimmune diseases such as inflammatory bowel disease.

Example 3: Effects of GDF15 Polypeptide in Type I Diabetes

[00178] The effect of a GDF 15 polypeptide on autoimmune disease was tested in a spontaneous diabetes model NOD mice. Administration of GDF 15 Analog 1 lowered blood glucose levels, lowered body weight, and reduced the incidence of diabetes development in a dose-dependent manner (FIG. 18 and FIG. 19). These data show that a GDF 15 polypeptide including a GDF 15 analog in the treatment and prevention of autoimmune diseases such as type I diabetes.

Example 4: Effects of GDF15 Polypeptide in CNS Demyelination [00179] The effect of GDF15 polypeptides on CNS demyelination was studied. CNS demyelination happens in progressive MS, including primary progressive MS and secondary progressive MS.

In a mouse model of demyelinated diseases, mice was fed with 0.2% cuprizone (CPZ)-containing diet. Cuprizone resulted in a loss of myeline sheath, which was detected by loss of Luxol fast blue (LFB) staining, in the corpus collosum (CC) of the brain. Cuprizone also resulted in activation of microglia, detected by IBA1 staining. Prophylactic treatment of mice with GDF15 Analog 1 protected the animals from losing myeline sheath and suppressed the microglia activation (FIG. 20A and 20B). [00180] In addition to corpus collosum, activation of microglia (IBA1) in hippocampus observed in CPZ -treated mice was also suppressed by prophylactic treatment of GDF15 Analog 1 (FIG. 21).

[00181] Activation of astrocytes, measured by GFAP staining, was observed in corpus callosum and cortex in cuprizone-treated mice and this effect was suppressed by prophylactic treatment of GDF15 Analog 1 (FIG. 22).

[00182] All these results show the beneficial effects of GDF15 Analog 1 including prevention of toxin-induced demyelination and restoration of glia homeostasis in the CNS.

[00183] To examine the effect of GDF15 Analog 1 during remyelination, the mice were fed with 5 weeks of CPZ diet and then switched to normal chow for an additional 2 weeks. Some mice were treated with GDF15 Analog 1 throughout the study (Prophylactic treatment) or only after the withdrawal of CPZ-diet (Therapeutic treatment). Two weeks after CPZ withdrawal, the animals in all treatment groups recovered more LFB staining than in the control (anti-KLH) group. In fact, most of the animals in all treatment groups recovered LFB staining (LFB grade 0) in CC (FIG. 23). This indicates GDF15 Analog 1 plays a beneficial role in remyelination. Furthermore, the activated microglia marker IBA1 staining in CC (FIG. 23) and hippocampus (FIG. 24) remained high in the control group (anti-KLH) but was reduced in all groups prophylactically and therapeutically treated with GDF15 Analog 1. In addition, the number of activated astrocytes in the brain, detected by GFAP staining, was also reduced in all treatment groups than in the control (anti-KLH) group (FIG. 25). These data support better recovery from demyelination in mice undergone prophylactic and therapeutic treatment of GDF15 Analog 1.

[00184] In summary, all these data in the cuprizone treated mice model demonstrate both prophylactic and therapeutic effect of GDF15 Analog 1 for reducing demyelination, recovering from demyelination and improve remyelination after demyelination

[00185] Although the foregoing present disclosure has been described in some detail by way of illustration and example for purposes of clarity of understanding, the descriptions and examples should not be construed as limiting the scope of the present disclosure. The embodiments of the present disclosure described herein are intended to be merely exemplary, and those skilled in the art will recognize numerous equivalents to the specific procedures described herein. All such equivalents are considered to be within the scope of the present disclosure and are covered by the embodiments.

[00186] All publications, patents, patent applications, internet sites, and accession numbers/ database sequences including both polynucleotide and polypeptide sequences cited herein are hereby incorporated by reference in their entirety for all purposes to the same extent as if each individual publication, patent, patent application, internet site, or accession number/database sequence were specifically and individually indicated to be so incorporated by reference.

[00187] Following are sequences disclosed in the application.

Mature wild type human GDF15

ARNGDHCPLGPGRCCRLHTVRASLEDLGWADWVLSPREVQVTMCIGACPSQ FRAANMHAQIKTSLHRLKPDTVPAPCCVPASYNPMVLIQKTDTGVSLQTYDD LLAKDCHCI (SEQ ID NO:1)

Full-length wild type human GDF15

MPGQELRTVNGSQMLLVLLVLSWLPHGGALSLAEASRASFPGPSELHSEDSR FRELRKRYEDLLTRLRANQSWEDSNTDLVPAPAVRILTPEVRLGSGGHLHLRI SRAALPEGLPEASRLHRALFRLSPTASRSWDVTRPLRRQLSLARPQAPALHLR LSPPPSQSDQLLAESSSARPQLELHLRPQAARGRRRARARNGDHCPLGPGRCC RLHTVRASLEDLGWADWVLSPREVQVTMCIGACPSQFRAANMHAQIKTSLH RLKPDTVPAPCCVPASYNPMVLIQKTDTGVSLQTYDDLLAKDCHCI (SEQ ID

NO: 2)

GDF15 Muteins

AN2-GDF15 (N3-I112) D5T

NGTHCPLGPGRCCRLHTVRASLEDLGWADWVLSPREVQVTMCIGACPSQFR AANMHAQIKTSLHRLKPDTVPAPCCVPASYNPMVLIQKTDTGVSLQTYDDLL AKDCHCI (SEQ ID NO: 3)

AN2-GDF15 (N3-I112) D5S

NGSHCPLGPGRCCRLHTVRASLEDLGWADWVLSPREVQVTMCIGACPSQFR AANMHAQIKTSLHRLKPDTVPAPCCVPASYNPMVLIQKTDTGVSLQTYDDLL AKDCHCI (SEQ ID NO: 4)

GDF15-K91N-D93T

ARNGDHCPLGPGRCCRLHTVRASLEDLGWADWVLSPREVQVTMCIGACPSQ FRAANMHAQIKTSLHRLKPDTVPAPCCVPASYNPMVLIQNTTTGVSLQTYDD LLAKDCHCI (SEQ ID NO:5)

GDF15-D93N-G95T

ARNGDHCPLGPGRCCRLHTVRASLEDLGWADWVLSPREVQVTMCIGACPSQ FRAANMHAQIKTSLHRLKPDTVPAPCCVPASYNPMVLIQKTNTTVSLQTYDD LLAKDCHCI (SEQ ID NO:6)

GDF15-AN3-K91N-D93T

GDHCPLGPGRCCRLHTVRASLEDLGWADWVLSPREVQVTMCIGACPSQFRA ANMHAQIKTSLHRLKPDTVPAPCCVPASYNPMVLIQNTTTGVSLQTYDDLLA KDCHCI (SEQ ID NO:7)

GDF15-AN3-D93N-G95T

GDHCPLGPGRCCRLHTVRASLEDLGWADWVLSPREVQVTMCIGACPSQFRA ANMHAQIKTSLHRLKPDTVPAPCCVPASYNPMVLIQKTNTTVSLQTYDDLLA KDCHCI (SEQ ID NO: 8) GDF15-K91N-D93S

ARNGDHCPLGPGRCCRLHTVRASLEDLGWADWVLSPREVQVTMCIGACPSQ

FRAANMHAQIKTSLHRLKPDTVPAPCCVPASYNPMVLIQNTSTGVSLQTYDD LLAKDCHCI (SEQ ID N0:9)

GDF15-D93N-G95S

ARNGDHCPLGPGRCCRLHTVRASLEDLGWADWVLSPREVQVTMCIGACPSQ

FRAANMHAQIKTSLHRLKPDTVPAPCCVPASYNPMVLIQKTNTSVSLQTYDD LLAKDCHCI (SEQ ID NO: 10)

GDF 15- AN3 -K91N-D93 S

GDHCPLGPGRCCRLHTVRASLEDLGWADWVLSPREVQVTMCIGACPSQFRA

ANMHAQIKTSLHRLKPDTVPAPCCVPASYNPMVLIQNTSTGVSLQTYDDLLA KDCHCI (SEQ ID NO: 11)

GDF 15- AN3 -D93N-G95 S

GDHCPLGPGRCCRLHTVRASLEDLGWADWVLSPREVQVTMCIGACPSQFRA

ANMHAQIKTSLHRLKPDTVPAPCCVPASYNPMVLIQKTNTSVSLQTYDDLLA

KDCHCI (SEQ ID NO: 12)

Fc-GDF15 Fusion Polypeptides - First Polypeptide

First Polypeptide: h!gGl-Fc(AA)(T366W)-(G₄S)5-AN2-GDF15 (N3-H 12) (D5T)

DKTHTCPPCPAPALAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEV

KFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKV

SNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLWCLVKGFYPSD

IAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVM

HEALHNHYTQKSLSLSPGKGGGGSGGGGSGGGGSGGGGSGGGGSNGTHCPL

GPGRCCRLHTVRASLEDLGWADWVLSPREVQVTMCIGACPSQFRAANMHA

QIKTSLHRLKPDTVPAPCCVPASYNPMVLIQKTDTGVSLQTYDDLLAKDCHCI (SEQ ID NO: 13)

First Polypeptide: h!gGl-Fc(AA)(T366W)-(G₄S)2-AN2-GDF15 (N3-H 12) (D5T)

DKTHTCPPCPAPALAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEV

KFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKV

SNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLWCLVKGFYPSD

IAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVM

HEALHNHYTQKSLSLSPGKGGGGSGGGGSNGTHCPLGPGRCCRLHTVRASLE

DLGWADWVLSPREVQVTMCIGACPSQFRAANMHAQIKTSLHRLKPDTVPAP

CCVPASYNPMVLIQKTDTGVSLQTYDDLLAKDCHCI (SEQ ID NO: 14)

Fc-GDF15 Fusion Polypeptides - Second Polypeptide h!gGl-Fc(AA)(T366S)(L368A)(Y407V)

DKTHTCPPCPAPALAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEV

KFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKV

SNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLSCAVKGFYPSDI

10

Claims

WHAT IS CLAIMED:

1. A method for treating an autoimmune disease in a subject in need thereof, the method comprising: administering to the subject a therapeutically effective amount of a growth differentiation factor 15 (GDF15) polypeptide, wherein the autoimmune disorder is not type 1 diabetes.

2. The method of claim 1, wherein the GDF15 polypeptide binds GFRAL and activates GFRAL.

3. The method of claim 1 or 2, wherein the autoimmune disease is MS.

4. The method of claim 3, wherein the MS is benign MS, primary progressive MS, secondary progressive MS, or relapsing-remitting MS.

5. The method of claim 1 or 2, wherein the autoimmune disease is relapsingremitting multiple sclerosis (MS).

6. The method of claim 1 or 2, wherein the autoimmune disease is psoriasis, psoriatic arthritis, or an inflammatory bowel disease (e.g., Crohn’s disease, ulcerative colitis).

7. The method of claim 1 or 2, wherein the autoimmune disease is selected from the group consisting of MS, rheumatoid arthritis, psoriasis, psoriatic arthritis, systemic lupus erythematosus (SLE), cutaneous lupus erythematosus (CLE), discoid lupus erythematosus (DLE), inflammatory bowel disease (e.g., Crohn’s disease, ulcerative colitis), polymyositis, dermatopolymyositis, Addison’s disease, Graves’ disease, Sjogren’s syndrome, Hashimoto’s thyroiditis, myasthenia gravis, autoimmune vasculitis, pernicious anemia, scleroderma, cytokine release syndrome, and celiac disease.

8. The method of claim 1 or 2, wherein the autoimmune disease is not a glucose metabolism disorder.

9. A method for preventing an autoimmune disease in a subject in need thereof, the method comprising: administering to the subject a therapeutically effective amount of a GDF15 polypeptide, wherein the subject is at risk of developing the autoimmune disease, and wherein the autoimmune disease is not type 1 diabetes.

10. A method for slowing down the onset of an autoimmune disease in a subject in need thereof, the method comprising: administering to the subject a therapeutically effective amount of a GDF15 polypeptide, wherein the subject is at risk of developing the autoimmune disease, and wherein the autoimmune disease is not type 1 diabetes.

11. The method of claim 9 or 10, wherein the autoimmune disease is MS.

12. The method of claim 11, wherein the MS is benign MS, primary progressive MS, secondary progressive MS, or relapsing-remitting MS.

13. The method of claim 9 or 10, wherein the autoimmune disease is relapsingremitting MS.

14. The method of claim 9 or 10, wherein the autoimmune disease is psoriasis, psoriatic arthritis, or an inflammatory bowel disease (e.g., Crohn’s disease, ulcerative colitis).

15. The method of claim 9 or 10, wherein the autoimmune disease is selected from the group consisting of MS, rheumatoid arthritis, psoriasis, psoriatic arthritis, systemic lupus erythematosus (SLE), cutaneous lupus erythematosus (CLE), discoid lupus erythematosus (DLE), inflammatory bowel disease (e.g., Crohn’s disease, ulcerative colitis), polymyositis, dermatopolymyositis, Addison’s disease, Graves’ disease, Sjogren’s syndrome, Hashimoto’s thyroiditis, myasthenia gravis, autoimmune vasculitis, pernicious anemia, scleroderma, cytokine release syndrome, and celiac disease.

16. The method of claim 9 or 10, wherein the autoimmune disease is not a glucose metabolism disorder.

17. A method of reducing or blocking pathogenic immune cell infiltration into a tissue or an organ of a subject, the method comprising administering to the subject a therapeutically effective amount of a GDF15 polypeptide.

18. The method of claim 17, wherein the tissue or organ is the central nervous system.

19. The method of claim 17 or 18, wherein the pathogenic immune cell is a pathogenic T cell.

20. The method of claim 19, wherein the T cell is a CD4+ T cell.

21. The method of claim 19, wherein the T cell is a Th-17 cell.

22. The method of claim 19, wherein the T cell is a Th-1 cell.

23. The method of claim 19, wherein the T cell is a myelin oligodendrocyte glycoprotein (MOG)-specific CD4+ T cell.

24. The method of any one of claims 1 to 23, wherein the GDF15 polypeptide comprises a complex comprising a first heterodimer and a second heterodimer; wherein each of the first heterodimer and the second heterodimer comprises a first polypeptide and a second polypeptide; wherein the first polypeptide comprises from the N-terminus to the C-terminus a first human IgGl Fc domain, a linker, and a GDF15 mutein, wherein the first human IgGl Fc domain comprises a first hinge region, a first CH2 domain, and a first CH3 domain; wherein the second polypeptide comprises from the N-terminus to the C-terminus a second human IgGl Fc domain, wherein the second human IgGl Fc domain comprises a second hinge region, a second CH2 domain, and a second CH3 domain; and wherein the GDF15 polypeptide binds to GFRAL.

25. The method of claim 24, wherein the first human IgGl Fc domain comprises an engineered protuberance and the second human IgGl Fc domain comprises an engineered cavity.

26. The method of claim 25, wherein the engineered protuberance comprises at least one substitution of the corresponding amino acid in a human IgGl Fc sequence selected from the group consisting of amino acid residues 347, 366 and 394, according to EU numbering; and wherein the engineered cavity comprises at least one substitution of the corresponding amino acid in a human IgGl Fc sequence selected from the group consisting of consisting of amino acid residues 366, 368, 394, 405, and 407, according to EU numbering.

27. The method of claim 25, wherein the engineered protuberance comprises at least one substitution of the corresponding amino acid in a human IgGl Fc sequence selected from the group consisting of Q347W/Y, T366W/Y. and T394W/Y. according to EU numbering; and wherein the engineered cavity comprises at least one substitution of the corresponding amino acid in a human IgGl Fc sequence selected from the group consisting of T366S, L368A, T394S, F405T/V/A, and Y407T/V/A, according to EU numbering.

28. The method of claim 27, wherein the engineered protuberance comprises the substitution T366W, according to EU numbering, and wherein the engineered cavity comprises the substitutions T366S, L368A, and Y407V, according to EU numbering.

29. The method of claim 27, wherein the engineered protuberance comprises the substitution T366Y, according to EU numbering, and wherein the engineered cavity comprises the substitution Y407T, according to EU numbering.

30. The method of any one of claims 24 to 29, wherein the linker comprises the sequence Glycine-Glycine-Glycine-Glycine-Ser (G4S)n, wherein n=2-5 (SEQ ID NO: 17).

31. The method of any one of claims 24 to 30, wherein the GDF15 mutein comprises an amino acid sequence having at least 90%, at least 95%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to the amino acid sequence of any one of SEQ ID NOs:3-12.

32. The method of claim 24 or 25, wherein the GDF15 mutein comprises an amino acid sequence having at least 90%, at least 95%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to the amino acid sequence of SEQ ID NO:3.

33. The method of claim 24 or 25, wherein the GDF15 mutein comprises the amino acid sequence of SEQ ID NO:3.

34. The method of claim 24 or 25, wherein the GDF15 mutein consists of the amino acid sequence of SEQ ID NO:3.

35. The method of claim 24 or 25, wherein the first polypeptide comprises an amino acid sequence having at least 90%, at least 95%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to the amino acid sequence of SEQ ID NO: 13 or 14.

36. The method of claim 24 or 25, wherein the first polypeptide comprises an amino acid sequence having at least 90%, at least 95%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to the amino acid sequence of SEQ ID NO: 13.

37. The method of claim 24 or 25, wherein the first polypeptide comprises the amino acid sequence of SEQ ID NO: 13.

38. The method of claim 24 or 25, wherein the first polypeptide consists of the amino acid sequence of SEQ ID NO: 13.

39. The method of any one of claims 24 to 38, wherein the second polypeptide comprises an amino acid sequence having at least 90%, at least 95%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to the amino acid sequence of SEQ ID NO: 15.

40. The method of claim 24, wherein the first polypeptide comprises the amino acid sequence of SEQ ID NO: 13, and wherein the second polypeptide comprises the amino acid sequence of SEQ ID NO: 15.

41. The method of claim 24, wherein the first polypeptide consists of the amino acid sequence of SEQ ID NO: 13, and wherein the second polypeptide consists of the amino acid sequence of SEQ ID NO: 15.

42. The method of any one of claims 1 to 23, wherein the GDF15 polypeptide comprises a GDF15 mutein, wherein the GDF15 mutein comprises an amino acid sequence having at least 90%, at least 95%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to the amino acid sequence the amino acid sequence of any one of SEQ ID NOs:3-12.

43. The method of claim 42, wherein the GDF15 mutein comprises the amino acid sequence of SEQ ID NO:7.

44. The method of claim 42, wherein the GDF15 mutein consists of the amino acid sequence of SEQ ID NO:7.

45. The method of any one of claims 1 to 23, wherein the GDF15 mutein comprises a contiguous amino acid sequence that is at least 90% identical to the amino acid sequence of wild type GDF15 (SEQ ID NO: 1).

46. The method of claim 45, wherein the GDF15 mutein comprises at least one substitution of the corresponding amino acid in SEQ ID NO: 1 that creates the N- linked glycosylation consensus site.

47. The method of claim 46, wherein the GDF15 mutein comprises at least one pair of substitutions of the corresponding amino acids in SEQ ID NO: 1 selected from the group consisting of:(i) K91N and D93T, (ii) K91N and D93S, (iii) D93N and G95T, or (iv) D93N and G95S.

48. The method of any of claims 45 to 47, wherein the GDF15 mutein comprises an N-terminal deletion of one or more amino acids relative to the amino acid sequence of SEQ ID NO:1.

49. The method of any one of claims 1 to 23, wherein the GDF15 polypeptide comprises a dimer, wherein the dimer comprises a first polypeptide and a second polypeptide, wherein each of the first polypeptide and the second polypeptide comprises a GDF15 mutein.

50. The method of claim 49, wherein the GDF15 mutein in the first polypeptide is fused to a first heterologous polypeptide, and wherein the GDF15 mutein in the second polypeptide is fused to a second heterologous polypeptide.

51. The method of claim 50, wherein the first heterologous polypeptide is serum albumin, maltose binding protein, or immunoglobulin Fc polypeptide, and wherein the second heterologous polypeptide is serum albumin, maltose binding protein, or immunoglobulin Fc polypeptide.

52. The method of any of claims 49 to 51, wherein the GDF15 mutein comprises a contiguous amino acid sequence that is at least 90% identical to the amino acid sequence of wild type GDF15 (SEQ ID NO: 1).

53. The method of claim 52, wherein the GDF15 mutein comprises at least one substitution of the corresponding amino acid in SEQ ID NO: 1 that creates the N- linked glycosylation consensus site.

54. The method of claim 53, wherein the GDF15 mutein comprises at least one of the following pairs of substitutions of the corresponding amino acids in SEQ ID NO: 1 :(1) K91N and D93T, (2) K91N and D93S, (3) D93N and G95T or (4) D93N and G95S.

55. The method of any of claims 49 to 54, wherein the GDF15 mutein comprises an N-terminal deletion of one or more amino acids that corresponds to the amino acids of SEQ ID NO: 1.

56. The method of any one of claims 49 to 54, wherein the GDF15 mutein comprises an N-terminal deletion of one, two, three, six or fourteen amino acids corresponds to the amino acids of SEQ ID NO: 1.

57. The method of any one of claims 1 to 23, wherein the GDF15 polypeptide comprises a dimer, wherein the dimer comprises a first polypeptide and a second polypeptide, wherein each of the first polypeptide and the second polypeptide comprises an amino acid sequence having at least 90%, at least 95%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to the amino acid sequence the amino acid sequence of any one of SEQ ID NOs:3-12, wherein the first polypeptide is identical to the second polypeptide.

58. The method of any one of claims 1 to 23, wherein the GDF15 polypeptide comprises a dimer, wherein the dimer comprises a first polypeptide and a second polypeptide, wherein each of the first polypeptide and the second polypeptide comprises the amino acid sequence of SEQ ID NO:7.

59. The method of any one of claims 1 to 23, wherein the GDF15 polypeptide comprises a dimer, wherein the dimer comprises a first polypeptide and a second polypeptide, wherein each of the first polypeptide and the second polypeptide consists of the amino acid sequence of SEQ ID NO:7.

60. The method of any one of claims 1 to 23, wherein the GDF15 polypeptide comprises AMG 171.

61. The method of any one of claims 1 to 23, wherein the GDF15 polypeptide comprises LY3463251.

62. The method of any one of claims 1 to 23, wherein the GDF15 polypeptide comprises NN9215.

63. The method of any one of claims 1 to 23, wherein the GDF15 polypeptide comprises CIN-109.

64. The method of any one of claims 1 to 63, wherein the subject is a human.