Anti-TMPRSS6 antibodies and uses thereof

Abstract

Aspects of the present disclosure provide anti-TMPRSS6 antibodies and methods of using the same to promote hepcidin expression and treat iron overload related disorders such as hemopigmentation, sickle cell disease, thalassemia, hemolysis, Diamond-Blackfan anemia, myelodysplastic syndrome (MDS), and blood transfusion.

Description

[0001] Cross-reference to related applications

[0002] This application claims priority to U.S. Provisional Application No. 63 / 457,081, filed April 4, 2023, which is incorporated herein by reference in its entirety.

[0003] Reference to electronic sequence listing

[0004] The contents of the electronic serial number (D084270011WO00-SEQ-LJG.xml; size: 95,921 bytes; creation date: April 1, 2024) are incorporated herein by reference in their entirety. Background Technology

[0005] Transmembrane serine protease 6 (TMPRSS6; also known as matriptase-2 or MTP-2) is a type II transmembrane serine protease. TMPRSS6 is highly expressed in the liver and plays a role in iron homeostasis by negatively regulating hepcidin expression. TMPRSS6 cleaves BMP co-receptors (e.g., hemojuvelin (HJV)), which leads to decreased hepcidin expression. Decreased hepcidin expression levels result in iron overload under various conditions. Improved compositions and methods are needed to treat iron overload syndromes. Summary of the Invention

[0006] Certain aspects of this disclosure relate to the understanding that iron overload occurs in a variety of conditions (e.g., hemochromatosis, sickle cell disease (SCD), thalassemia, hemolysis, African iron overload, Diamond-Blackfan anemia, myelodysplastic syndrome (MDS), transfusion, etc.). In some embodiments, the methods and related compositions provided herein can be used to treat a variety of diseases and conditions (e.g., hemochromatosis, sickle cell disease (SCD), thalassemia, hemolysis, African iron overload, Diamond-Blackfan anemia, myelodysplastic syndrome (MDS), etc.). In some embodiments, the methods and related compositions provided herein can be used to inhibit TMPRSS6 to reduce iron overload in the diseases and conditions described herein. In some embodiments, reducing iron overload improves disease severity. Some aspects of this disclosure provide anti-TMPRSS6 antibodies that have high binding affinity and specificity to TMPRSS6 and inhibit TMPRSS6 activity. Therefore, in some embodiments, this disclosure provides methods and related anti-TMPRSS6 antibody compositions for treating iron overload-related diseases and conditions, such as hemochromatosis, sickle cell disease (SCD), thalassemia, hemolysis, Diamond-Blackfan anemia, African iron overload, myelodysplastic syndrome (MDS), blood transfusions, etc. In some embodiments, the methods provided herein reduce iron overload in the subject. In some embodiments, the methods provided herein reduce systemic iron levels. In some embodiments, the methods provided herein reduce mean corpuscular hemoglobin concentration (MCHC) in the subject.

[0007] In some embodiments, this disclosure provides a method for treating sickle cell disease (SCD) by administering an effective amount of anti-TMPRSS6 antibody to a subject. In some aspects, this disclosure provides a method and composition for treating iron overload in a subject suffering from SCD by administering an effective amount of anti-TMPRSS6 antibody to said subject. In some embodiments, the methods provided herein result in a reduction in the frequency of painful crises (i.e., vascular occlusive crises, VOCs), the severity of VOCs (e.g., hospitalization and duration), systemic iron levels, mean corpuscular hemoglobin concentration (MCHC), hemoglobin S (HbS) polymerization, transfusions, and / or hemolysis in subjects suffering from SCD (e.g., hemoglobin SC disease). In some embodiments, the methods provided herein reduce the frequency of painful crises (VOCs) in hemoglobin SC disease.

[0008] In some aspects, this disclosure provides a method for treating sickle cell disease (SCD), the method comprising administering to a subject an effective amount of an anti-transmembrane serine protease 6 (TMPRSS6) antibody, said antibody comprising:

[0009] (a) HC CDR1, HC CDR2 and HCCDR3 having the amino acid sequence of SEQ ID NO: 49, and LC CDR1, LC CDR2 and LCCDR3 having the amino acid sequence of SEQ ID NO: 50;

[0010] (b) HC CDR1, HC CDR2 and HCCDR3 having the heavy chain variable domain of the amino acid sequence of SEQ ID NO: 7, and LC CDR1, LC CDR2 and LCCDR3 having the light chain variable domain of the amino acid sequence of SEQ ID NO: 8;

[0011] (c) HC CDR1, HC CDR2 and HC CDR3 having the amino acid sequence of SEQ ID NO: 19 or 78, and LC CDR1, LC CDR2 and LC CDR3 having the amino acid sequence of SEQ ID NO: 20;

[0012] (d) HC CDR1, HC CDR2 and HCCDR3 having the amino acid sequence of SEQ ID NO: 30, and LC CDR1, LC CDR2 and LCCDR3 having the amino acid sequence of SEQ ID NO: 31;

[0013] (e) HC CDR1, HC CDR2, and HCCDR3 having the heavy chain variable domain of the amino acid sequence of SEQ ID NO: 37, and LC CDR1, LC CDR2, and LCCDR3 having the light chain variable domain of the amino acid sequence of SEQ ID NO: 38; or

[0014] (f) HC CDR1, HC CDR2 and HCCDR3 having the amino acid sequence of SEQ ID NO: 44 and the light chain variable domain having the amino acid sequence of SEQ ID NO: 45.

[0015] In some embodiments, the anti-TMPRSS6 antibody comprises HC CDR1, HC CDR2, and HC CDR3, which have the amino acid sequence of SEQ ID NO: 49, and LC CDR1, LC CDR2, and LC CDR3, which have the amino acid sequence of SEQ ID NO: 50.

[0016] In some implementations, the anti-TMPRSS6 antibody comprises:

[0017] (a) HC CDR1 having the amino acid sequence of SEQ ID NO: 24, HC CDR2 having the amino acid sequence of SEQ ID NO: 48, HC CDR3 having the amino acid sequence of SEQ ID NO: 26, LC CDR1 having the amino acid sequence of SEQ ID NO: 27, LC CDR2 having the amino acid sequence of WAT, and LCCDR3 having the amino acid sequence of SEQ ID NO: 29;

[0018] (b) HC CDR1 having the amino acid sequence of SEQ ID NO: 1, HC CDR2 having the amino acid sequence of SEQ ID NO: 2, HC CDR3 having the amino acid sequence of SEQ ID NO: 3, LC CDR1 having the amino acid sequence of SEQ ID NO: 4, LC CDR2 having the amino acid sequence of RAN, and LCCDR3 having the amino acid sequence of SEQ ID NO: 6;

[0019] (c) HC CDR1 having the amino acid sequence of SEQ ID NO: 13, HC CDR2 having the amino acid sequence of SEQ ID NO: 14, HC CDR3 having the amino acid sequence of SEQ ID NO: 15, LC CDR1 having the amino acid sequence of SEQ ID NO: 16, LC CDR2 having the amino acid sequence of WAF, and LCCDR3 having the amino acid sequence of SEQ ID NO: 18;

[0020] (d) HC CDR1 having the amino acid sequence of SEQ ID NO: 24, HC CDR2 having the amino acid sequence of SEQ ID NO: 25, HC CDR3 having the amino acid sequence of SEQ ID NO: 26, LC CDR1 having the amino acid sequence of SEQ ID NO: 27, LC CDR2 having the amino acid sequence of WAT, and LCCDR3 having the amino acid sequence of SEQ ID NO: 29;

[0021] (e) HC CDR1 having the amino acid sequence of SEQ ID NO: 1, HC CDR2 having the amino acid sequence of SEQ ID NO: 2, HC CDR3 having the amino acid sequence of SEQ ID NO: 35, LC CDR1 having the amino acid sequence of SEQ ID NO: 36, LC CDR2 having the amino acid sequence of RAN, and LCCDR3 having the amino acid sequence of SEQ ID NO: 6; or

[0022] (f) HC CDR1 having the amino acid sequence of SEQ ID NO: 13, HC CDR2 having the amino acid sequence of SEQ ID NO: 43, HC CDR3 having the amino acid sequence of SEQ ID NO: 15, LC CDR1 having the amino acid sequence of SEQ ID NO: 16, LC CDR2 having the amino acid sequence of WAF, and LCCDR3 having the amino acid sequence of SEQ ID NO: 18.

[0023] In some embodiments, the anti-TMPRSS6 antibody comprises HCCDR1 having the amino acid sequence of SEQ ID NO: 24, HCCDR2 having the amino acid sequence of SEQ ID NO: 48, HCCDR3 having the amino acid sequence of SEQ ID NO: 26, LC CDR1 having the amino acid sequence of SEQ ID NO: 27, LC CDR2 having the amino acid sequence of WAT, and LC CDR3 having the amino acid sequence of SEQ ID NO: 29.

[0024] In some implementations, the anti-TMPRSS6 antibody comprises:

[0025] (a) A heavy chain variable domain having the amino acid sequence of SEQ ID NO: 49 and a light chain variable domain having the amino acid sequence of SEQ ID NO: 50;

[0026] (b) A heavy chain variable domain having the amino acid sequence of SEQ ID NO: 7 and a light chain variable domain having the amino acid sequence of SEQ ID NO: 8;

[0027] (c) A heavy chain variable domain having an amino acid sequence of SEQ ID NO: 19 or 78 and a light chain variable domain having an amino acid sequence of SEQ ID NO: 20;

[0028] (d) A heavy chain variable domain having the amino acid sequence of SEQ ID NO: 30 and a light chain variable domain having the amino acid sequence of SEQ ID NO: 31;

[0029] (e) A heavy chain variable domain having the amino acid sequence of SEQ ID NO: 37 and a light chain variable domain having the amino acid sequence of SEQ ID NO: 38; or

[0030] (f) A heavy chain variable domain having the amino acid sequence of SEQ ID NO: 44 and a light chain variable domain having the amino acid sequence of SEQ ID NO: 45.

[0031] In some embodiments, the anti-TMPRSS6 antibody comprises a heavy chain variable domain having the amino acid sequence of SEQ ID NO: 49 and a light chain variable domain having the amino acid sequence of SEQ ID NO: 50.

[0032] In some implementations, the anti-TMPRSS6 antibody comprises:

[0033] (a) The heavy chain containing the amino acid sequence of SEQ ID NO: 52 and the light chain containing the amino acid sequence of SEQ ID NO: 53;

[0034] (b) The heavy chain containing the amino acid sequence of SEQ ID NO: 11 and the light chain containing the amino acid sequence of SEQ ID NO: 12;

[0035] (c) A heavy chain containing the amino acid sequence of SEQ ID NO: 17 or 22 and a light chain containing the amino acid sequence of SEQ ID NO: 23;

[0036] (d) The heavy chain containing the amino acid sequence of SEQ ID NO: 33 and the light chain containing the amino acid sequence of SEQ ID NO: 34;

[0037] (e) A heavy chain containing the amino acid sequence of SEQ ID NO: 41 and a light chain containing the amino acid sequence of SEQ ID NO: 42; or

[0038] (f) The heavy chain containing the amino acid sequence of SEQ ID NO: 46 and the light chain containing the amino acid sequence of SEQ ID NO: 47.

[0039] In some embodiments, the anti-TMPRSS6 antibody comprises a heavy chain containing the amino acid sequence of SEQ ID NO: 52 and a light chain containing the amino acid sequence of SEQ ID NO: 53.

[0040] In some embodiments, antibody administration reduces iron overload in the subject. In some embodiments, antibody administration reduces hemolysis in the subject relative to the state prior to administration. In some embodiments, the subject has recurrent moderate to severe vasoclusive crisis (VOC). In some embodiments, antibody administration reduces the frequency of VOCs in the subject relative to the state prior to administration. In some embodiments, the administration reduces the severity of VOCs relative to the state prior to administration. In some embodiments, the severity of VOCs is measured by hospitalization frequency and / or hospitalization duration.

[0041] In some embodiments, antibody administration reduces systemic iron in the subject prior to administration. In some embodiments, antibody administration reduces mean corpuscular hemoglobin concentration (MCHC) relative to the subject prior to administration. In some embodiments, the administration reduces hemoglobin S (HbS) polymerization relative to the subject prior to administration. In some embodiments, the administration reduces the frequency of blood transfusions required by the subject relative to the subject prior to administration. In some embodiments, the administration reduces inflammation in the subject relative to the subject prior to administration. In some embodiments, sickle cell disease is hemoglobin SS disease. In some embodiments, sickle cell disease is hemoglobin SC disease.

[0042] In some implementations, the anti-TMPRSS6 antibody is administered as a co-treatment in combination with the following: hemoglobin S-polymerization inhibitors (e.g., voxelotor), therapeutic agents for reducing VOCs (e.g., hydroxyurea, L-glutamine oral powder, Crizanlizumab, selective pyruvate kinase-R (PKR) activators), analgesics (e.g., anesthetics, opioids, gabapentin, cannabis), blood transfusions, stem cell transplantation, Exagmglogene autotemcel (exa-cel), LentiGlobin, GMI-1070, VIT-2763 (vamifeport), ticagrelor, vitamin D, simvastatin, AG-348 (mitapivat sulfate). Sulfate), propranolol, adrenaline, regadenoson, atorvastatin, prasugrel, L-arginine, OTQ923, oxygen therapy, or gene therapy.

[0043] In some aspects, this disclosure provides a method for treating iron overload-related conditions in subjects who do not suffer from intermediate β-thalassemia or type I hemochromatosis, the method comprising administering to the subject an effective amount of an anti-transmembrane serine protease 6 (TMPRSS6) antibody, the antibody comprising:

[0044] (a) HC CDR1, HC CDR2 and HCCDR3 having the amino acid sequence of SEQ ID NO: 49, and LC CDR1, LC CDR2 and LCCDR3 having the amino acid sequence of SEQ ID NO: 50;

[0045] (b) HC CDR1, HC CDR2 and HCCDR3 having the heavy chain variable domain of the amino acid sequence of SEQ ID NO: 7, and LC CDR1, LC CDR2 and LCCDR3 having the light chain variable domain of the amino acid sequence of SEQ ID NO: 8;

[0046] (c) HC CDR1, HC CDR2 and HC CDR3 having the amino acid sequence of SEQ ID NO: 19 or 78, and LC CDR1, LC CDR2 and LC CDR3 having the amino acid sequence of SEQ ID NO: 20;

[0047] (d) HC CDR1, HC CDR2 and HCCDR3 having the amino acid sequence of SEQ ID NO: 30, and LC CDR1, LC CDR2 and LCCDR3 having the amino acid sequence of SEQ ID NO: 31;

[0048] (e) HC CDR1, HC CDR2, and HCCDR3 having the heavy chain variable domain of the amino acid sequence of SEQ ID NO: 37, and LC CDR1, LC CDR2, and LCCDR3 having the light chain variable domain of the amino acid sequence of SEQ ID NO: 38; or

[0049] (f) HC CDR1, HC CDR2 and HCCDR3 having the amino acid sequence of SEQ ID NO: 44 and the light chain variable domain having the amino acid sequence of SEQ ID NO: 45.

[0050] In some embodiments, the anti-TMPRSS6 antibody comprises HC CDR1, HC CDR2, and HC CDR3, which have the amino acid sequence of SEQ ID NO: 49, and LC CDR1, LC CDR2, and LC CDR3, which have the amino acid sequence of SEQ ID NO: 50.

[0051] In some embodiments, the anti-TMPRSS6 antibody comprises:

[0052] (a) HC CDR1 having the amino acid sequence of SEQ ID NO: 24, HC CDR2 having the amino acid sequence of SEQ ID NO: 48, HC CDR3 having the amino acid sequence of SEQ ID NO: 26, LC CDR1 having the amino acid sequence of SEQ ID NO: 27, LC CDR2 having the amino acid sequence of WAT, and LCCDR3 having the amino acid sequence of SEQ ID NO: 29;

[0053] (b) HC CDR1 having the amino acid sequence of SEQ ID NO: 1, HC CDR2 having the amino acid sequence of SEQ ID NO: 2, HC CDR3 having the amino acid sequence of SEQ ID NO: 3, LC CDR1 having the amino acid sequence of SEQ ID NO: 4, LC CDR2 having the amino acid sequence of RAN, and LCCDR3 having the amino acid sequence of SEQ ID NO: 6;

[0054] (c) HC CDR1 having the amino acid sequence of SEQ ID NO: 13, HC CDR2 having the amino acid sequence of SEQ ID NO: 14, HC CDR3 having the amino acid sequence of SEQ ID NO: 15, LC CDR1 having the amino acid sequence of SEQ ID NO: 16, LC CDR2 having the amino acid sequence of WAF, and LCCDR3 having the amino acid sequence of SEQ ID NO: 18;

[0055] (d) HC CDR1 having the amino acid sequence of SEQ ID NO: 24, HC CDR2 having the amino acid sequence of SEQ ID NO: 25, HC CDR3 having the amino acid sequence of SEQ ID NO: 26, LC CDR1 having the amino acid sequence of SEQ ID NO: 27, LC CDR2 having the amino acid sequence of WAT, and LCCDR3 having the amino acid sequence of SEQ ID NO: 29;

[0056] (e) HC CDR1 having the amino acid sequence of SEQ ID NO: 1, HC CDR2 having the amino acid sequence of SEQ ID NO: 2, HC CDR3 having the amino acid sequence of SEQ ID NO: 35, LC CDR1 having the amino acid sequence of SEQ ID NO: 36, LC CDR2 having the amino acid sequence of RAN, and LCCDR3 having the amino acid sequence of SEQ ID NO: 6; or

[0057] (f) HC CDR1 having the amino acid sequence of SEQ ID NO: 13, HC CDR2 having the amino acid sequence of SEQ ID NO: 43, HC CDR3 having the amino acid sequence of SEQ ID NO: 15, LC CDR1 having the amino acid sequence of SEQ ID NO: 16, LC CDR2 having the amino acid sequence of WAF, and LCCDR3 having the amino acid sequence of SEQ ID NO: 18.

[0058] In some embodiments, the anti-TMPRSS6 antibody comprises HCCDR1 having the amino acid sequence of SEQ ID NO: 24, HCCDR2 having the amino acid sequence of SEQ ID NO: 48, HCCDR3 having the amino acid sequence of SEQ ID NO: 26, LC CDR1 having the amino acid sequence of SEQ ID NO: 27, LC CDR2 having the amino acid sequence of WAT, and LC CDR3 having the amino acid sequence of SEQ ID NO: 29.

[0059] In some implementations, the anti-TMPRSS6 antibody comprises:

[0060] (a) A heavy chain variable domain having the amino acid sequence of SEQ ID NO: 49 and a light chain variable domain having the amino acid sequence of SEQ ID NO: 50;

[0061] (b) A heavy chain variable domain having the amino acid sequence of SEQ ID NO: 7 and a light chain variable domain having the amino acid sequence of SEQ ID NO: 8;

[0062] (c) A heavy chain variable domain having an amino acid sequence of SEQ ID NO: 19 or 78 and a light chain variable domain having an amino acid sequence of SEQ ID NO: 20;

[0063] (d) A heavy chain variable domain having the amino acid sequence of SEQ ID NO: 30 and a light chain variable domain having the amino acid sequence of SEQ ID NO: 31;

[0064] (e) A heavy chain variable domain having the amino acid sequence of SEQ ID NO: 37 and a light chain variable domain having the amino acid sequence of SEQ ID NO: 38; or

[0065] (f) A heavy chain variable domain having the amino acid sequence of SEQ ID NO: 44 and a light chain variable domain having the amino acid sequence of SEQ ID NO: 45.

[0066] In some embodiments, the anti-TMPRSS6 antibody comprises a heavy chain variable domain having the amino acid sequence of SEQ ID NO: 49 and a light chain variable domain having the amino acid sequence of SEQ ID NO: 50.

[0067] In some implementations, the anti-TMPRSS6 antibody comprises:

[0068] (a) The heavy chain containing the amino acid sequence of SEQ ID NO: 52 and the light chain containing the amino acid sequence of SEQ ID NO: 53;

[0069] (b) The heavy chain containing the amino acid sequence of SEQ ID NO: 11 and the light chain containing the amino acid sequence of SEQ ID NO: 12;

[0070] (c) A heavy chain containing the amino acid sequence of SEQ ID NO: 17 or 22 and a light chain containing the amino acid sequence of SEQ ID NO: 23;

[0071] (d) The heavy chain containing the amino acid sequence of SEQ ID NO: 33 and the light chain containing the amino acid sequence of SEQ ID NO: 34;

[0072] (e) A heavy chain containing the amino acid sequence of SEQ ID NO: 41 and a light chain containing the amino acid sequence of SEQ ID NO: 42; or

[0073] (f) The heavy chain containing the amino acid sequence of SEQ ID NO: 46 and the light chain containing the amino acid sequence of SEQ ID NO: 47.

[0074] In some embodiments, the anti-TMPRSS6 antibody comprises a heavy chain containing the amino acid sequence of SEQ ID NO: 52 and a light chain containing the amino acid sequence of SEQ ID NO: 53.

[0075] In some embodiments, antibody administration reduces systemic iron in the subject relative to prior administration. In some embodiments, antibody administration reduces mean corpuscular hemoglobin concentration (MCHC) relative to prior administration. In some embodiments, antibody administration reduces iron overload in the subject relative to prior administration.

[0076] In some implementations, the subject suffers from a hereditary hemochromatosis. In some implementations, the hereditary hemochromatosis is type 3 hereditary hemochromatosis.

[0077] In some embodiments, the subject has or is suspected of having thalassemia. In some embodiments, the thalassemia is alpha-thalassemia. In some embodiments, the alpha-thalassemia is severe alpha-thalassemia, intermediate alpha-thalassemia, or mild alpha-thalassemia. In some embodiments, the thalassemia is beta-thalassemia. In some embodiments, the beta-thalassemia is severe beta-thalassemia or mild beta-thalassemia.

[0078] In some implementations, the subject has or is suspected of having transfusion-related iron overload. In some implementations, the subject receives a transfusion due to blood loss. In some implementations, the subject receives repeated transfusions due to anemia.

[0079] In some implementations, the subject has hemolytic anemia. In some implementations, the subject has transfusion-dependent hemolytic anemia, pyruvate kinase deficiency hemolytic anemia, thalassemia-associated hemolytic anemia, or sickle cell disease-associated hemolytic anemia.

[0080] In some implementation schemes, the subjects suffer from African iron overload.

[0081] In some implementations, the subject suffers from Diamond-Blackfan anemia.

[0082] In some implementations, the subject has myelodysplastic syndrome (MDS). In some implementations, MDS is refractory anemia with ring-sideroblasts (RARS). In some implementations, the subject has an SF3B1 mutation.

[0083] The foregoing and other aspects, implementation methods, functions, features, and schemes of this teaching can be more fully understood by taking into account the following description in conjunction with the accompanying drawings. Attached Figure Description

[0084] Figures 1A to 1E The dose-dependent effects of treatment of mice with inhibitory anti-TMPRSS6 antibody were demonstrated. Mice were injected intraperitoneally with 0 (loador), 2, 5, or 10 mg / kg of anti-TMPRSS6 antibody, and monitored for 10 days. Serum antibody levels were measured over the 10-day period following injection. Figure 1A ),iron( Figure 1B ) and hepcidin ( Figure 1C )concentration. Figure 1D The expression of Hamp in the liver was shown, and Figure 1E The TSAT of mice during the study is shown.

[0085] Figures 2A to 2D The body weight, spleen weight, and liver weight of sickle cell (Townes model) mice administered with anti-TMPRSS6 antibody are shown. Figure 2A The progression of total body weight in mice administered 0, 3, or 10 mg / kg of anti-TMPRSS6 antibody, divided into male and female groups (labeled M and F, respectively), is shown over an 8-week period. Figure 2B The spleen weight / body weight measurements of male and female mice 56 days after administration of 0, 3, or 10 mg / kg anti-TMPRSS6 antibody are shown. Figure 2C The liver weight / body weight measurements of male and female mice 56 days after administration of 0, 3, or 10 mg / kg anti-TMPRSS6 antibody are shown. Figure 2D The liver weight / body weight of all mice (male and female combination) are shown 56 days after administration of 0 or 10 mg / kg anti-TMPRSS6 antibody.

[0086] Figures 3A to 3C This study showed a decrease in hemolytic markers in sickle cell mice administered anti-TMPRSS6 antibody. Lactate dehydrogenase (LDH) was measured in sickle cell mice 56 days after administration of 0, 3, or 10 mg / kg anti-TMPRSS6 antibody. Figure 3A ), direct bilirubin (DBili) Figure 3B ) and total bilirubin (TBili) Figure 3C ) level.

[0087] Figures 4A to 4C This study demonstrated a reduction in systemic inflammatory markers in sickle cell mice administered anti-TMPRSS6 antibody. White blood cells (WBCs) in the blood of sickle cell mice were measured by peroxidase assay 56 days after administration of 0, 3, or 10 mg / kg anti-TMPRSS6 antibody. Figure 4A ), neutrophils ( Figure 4B ) and lymphocytes ( Figure 4C ) level.

[0088] Figures 5A to 5C This study demonstrated a decrease in HbS polymerization in sickle cell mice treated with anti-TMPRSS6 antibody. HbS (CHCM) levels in the blood of sickle cell mice were measured 56 days after administration of 0, 3, or 10 mg / kg anti-TMPRSS6 antibody. Figure 5A ), hypochromic erythrocytes ( Figure 5B ) and microcytic erythrocytes ( Figure 5C ) level. Detailed Implementation

[0089] This disclosure relates at least in part to the understanding that iron overload occurs in a variety of conditions (e.g., hemochromatosis, sickle cell disease, thalassemia, hemolysis, Diamond-Blackfan anemia, African iron overload, myelodysplastic syndrome (MDS), blood transfusion, etc.). In some embodiments, the methods and related compositions provided herein can be used to treat a variety of diseases and conditions (e.g., hemochromatosis, sickle cell disease (SCD), thalassemia, hemolysis, African iron overload, Diamond-Blackfan anemia, myelodysplastic syndrome (MDS), etc.). In some embodiments, methods and related compositions for inhibiting TMPRSS6 to reduce iron overload are provided, which improve the severity of the disease. Some aspects of this disclosure provide anti-TMPRSS6 antibodies that have high binding affinity and specificity to TMPRSS6 and inhibit TMPRSS6 activity. Therefore, in some embodiments, this disclosure provides methods and associated anti-TMPRSS6 antibody compositions for treating iron overload-related diseases and conditions such as hemochromatosis, sickle cell disease (SCD), thalassemia, hemolysis, Diamond-Blackfan anemia, African iron overload, myelodysplastic syndrome (MDS), and blood transfusions. In some embodiments, the methods provided herein reduce iron overload.

[0090] In some embodiments, this disclosure provides a method for treating sickle cell disease (SCD) by administering an effective amount of anti-TMPRSS6 antibody to a subject. In some aspects, this disclosure provides methods and compositions for treating iron overload in a subject suffering from SCD by administering an effective amount of anti-TMPRSS6 antibody to said subject. In some embodiments, the methods provided herein reduce the frequency of painful crises (i.e., vascular occlusive crises, VOCs), the severity of VOCs (e.g., hospitalization and duration), hemoglobin S (HbS) polymerization, transfusions, and / or hemolysis in subjects suffering from SCD (e.g., hemoglobin-SC disease). In some embodiments, the methods provided herein reduce the frequency of painful crises (VOCs) in hemoglobin-sickle cell disease (SC).

[0091] The foregoing and other aspects, implementation methods, functions, features, and schemes of this teaching can be more fully understood by taking into account the following description in conjunction with the accompanying drawings.

[0092] I. Definition

[0093] And / or: The term “and / or” as used herein should be considered as an explicit disclosure of each of two or more specified features or components, with or without the other features or components. Thus, the term “and / or” as used in phrases such as “A and / or B” herein is intended to include “A and B”, “A or B”, “A” (alone), and “B” (alone). Similarly, the term “and / or” as used in phrases such as “A, B, and / or C” is intended to cover each of the following: A, B, and C; A, B, or C; A or C; A or B; B or C; A and C; A and B; B and C; A (alone); B (alone); and C (alone).

[0094] Application: As used herein, the term “application” and its variations mean to provide an antibody or a combination thereof to a subject in a manner useful for physiological and / or pharmacological purposes (e.g., to treat a condition in the subject).

[0095] Affinity-matured antibodies: "Affinity-matured antibody" is used herein to refer to an antibody with one or more alterations in one or more CDRs, resulting in improved affinity (i.e., KD, kd, or ka) for the target antigen compared to a parent antibody without alterations. Exemplary affinity-matured antibodies will have nanomolar or even picomolar affinity for the target antigen. Various methods for generating affinity-matured antibodies are known in the art, including screening combinatorial antibody libraries prepared using biodisplay. For example, Marks et al., BioTechnology, 10: 779-783 (1992) describes affinity maturation through the co-encoding of VH and VL domains. Random mutagenesis of CDR and / or framework residues is described in the following publications: Barbas et al., Proc. Nat. Acad. Sci. USA, 91: 3809-3813 (1994); Schier et al., Gene, 169: 147-155 (1995); Yelton et al., J. Immunol., 155: 1994-2004 (1995); Jackson et al., J. Immunol., 154(7): 3310-3319 (1995); and Hawkins et al., J. Mol. Biol., 226: 889-896 (1992). Selective mutagenesis at selective mutagenic sites and at contact or highly mutagenic sites using activity-enhancing amino acid residues is described in U.S. Patent No. 6,914,128 B1.

[0096] Antibody: As used herein, the term "antibody" refers to a polypeptide comprising at least one immunoglobulin variable domain or at least one site (e.g., a complementary site) that specifically binds to an antigen. In some embodiments, the antibody comprises a complementary site. In some embodiments, the complementary site comprises one or more complementarity determining regions (CDRs). In some embodiments, the antibody is a full-length antibody. In some embodiments, the antibody is a chimeric antibody. In some embodiments, the antibody is a humanized antibody. However, in some embodiments, the antibody is a Fab fragment, an F(ab')2 fragment, an Fv fragment, or a scFv fragment. In some embodiments, the antibody is a nanobody derived from a camel antibody or a nanobody derived from a shark antibody. In some embodiments, the antibody is a biantibody. In some embodiments, the antibody comprises a framework having a human germline sequence. In another embodiment, the antibody comprises a heavy chain constant domain selected from IgG, IgG1, IgG2, IgG2A, IgG2B, IgG2C, IgG3, IgG4, IgA1, IgA2, IgD, IgM, and IgE constant domains. In some embodiments, the antibody comprises a heavy (H) chain variable region (hereinafter abbreviated as VH), and / or a light (L) chain variable region (hereinafter abbreviated as VL). In some embodiments, the antibody comprises a constant domain, such as an Fc region. An immunoglobulin constant domain refers to a heavy chain or light chain constant domain. The amino acid sequences and functional variations of the human IgG heavy and light chain constant domains are known. Regarding the heavy chain, in some embodiments, the heavy chain of the antibody described herein may be an alpha (α), delta (Δ), epsilon (ε), gamma (γ), or mu (µ) heavy chain. In some embodiments, the heavy chain of the antibody described herein may comprise a human alpha (α), delta (Δ), epsilon (ε), gamma (γ), or mu (µ) heavy chain. In a specific embodiment, the antibody described herein comprises human γ1 CH1, CH2, and / or CH3 domains. In some embodiments, the amino acid sequence of the VH domain comprises the amino acid sequence of the human gamma (γ) heavy chain constant region, such as any sequence known in the art. Several non-limiting examples of human constant region sequences have been described in the art, see, for example, U.S. Patent No. 5,693,780 and Kabat E A et al., (1991), ibid. In some embodiments, the VH domain comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 95%, 98%, or at least 99% identity with any variable chain constant region provided herein. In some embodiments, the antibody is modified, for example, by glycosylation, phosphorylation, threonization, and / or methylation.In some embodiments, the antibody is a glycosylated antibody conjugated to one or more sugar or carbohydrate molecules. In some embodiments, one or more sugar or carbohydrate molecules are conjugated to the antibody via N-glycosylation, O-glycosylation, C-glycosylation, glycosylphosphatidylinositylation (GPI anchor attachment), and / or phosphoglycosylation. In some embodiments, one or more sugar or carbohydrate molecules are monosaccharides, disaccharides, oligosaccharides, or glycans. In some embodiments, one or more sugar or carbohydrate molecules are branched oligosaccharides or branched glycans. In some embodiments, one or more sugar or carbohydrate molecules include mannose units, glucose units, N-acetylglucosamine units, or phospholipid units. In some embodiments, the antibody is a construct comprising a polypeptide containing one or more antigen-binding fragments of this disclosure linked to a linker polypeptide or an immunoglobulin constant domain. The linker polypeptide comprises two or more amino acid residues linked by peptide bonds and is used to link one or more antigen-binding moieties. Several examples of linker peptides have been reported (see, for example, Holliger, P., et al. (1993) Proc. Natl. Acad. Sci. USA 90:6444-6448; Poljak, RJ, et al. (1994) Structure 2:1121-1123). Furthermore, antibodies can be part of larger immunoadhesion molecules formed by the covalent or non-covalent association of the antibody or antibody portion with one or more other proteins or peptides. Some examples of such immunoadhesion molecules include the use of streptavidin core regions to prepare tetrameric scFv molecules (Kipriyanov, SM, et al. (1995) Human Antibodies and Hybridomas 6:93-101) and the use of cysteine ​​residues, labeled peptides, and C-terminal multihistidine tags to prepare divalent and biotinylated scFv molecules (Kipriyanov, SM, et al. (1994) Mol. Immunol. 31:1047-1058).

[0097] Approximately / About: As used herein, the term “approximately” or “about” when applied to one or more target values ​​refers to a value similar to the stated reference value. In some embodiments, the term “approximately” or “about” refers to a range of values ​​falling within (greater than or less than) 15%, 14%, 13%, 12%, 11%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1%, or less in any direction of the stated reference value, unless otherwise stated or obvious from the context (unless such a figure exceeds 100% of the possible value).

[0098] CDR: As used herein, the term "CDR" refers to the complementarity-determining region within the variable sequence of an antibody. A typical antibody molecule contains a heavy chain variable region (VH) and a light chain variable region (VL), which are typically involved in antigen binding. The VH and VL regions can be further subdivided into hypervariable regions, also known as "complementarity-determining regions" ("CDRs"), interspersed with more conserved regions called "framework regions" ("FRs"). Each VH and VL typically consists of three CDRs and four FRs, arranged in the following order from the amino terminus to the carboxyl terminus: FR1, CDR1, FR2, CDR2, FR3, CDR3, FR4. The extent of the frame region and CDRs can be precisely determined using methods known in the art, such as the Kabat definition, IMGT definition, Chothia definition, AbM definition, and / or contact definition, all of which are well-known in the art.See, e.g., Kabat, EA, et al. (1991) Sequences of Proteins of Immunological Interest, Fifth Edition, US Department of Health and Human Services, NIH Publication No. 91-3242; IMGT®, the international ImMunoGeneTics information system® http: / / www.imgt.org, Lefranc, M.-P. et al., Nucleic Acids Res., 27:209-212 (1999); Ruiz, M. et al., Nucleic Acids Res., 28:219-221 (2000); Lefranc, M.-P., Nucleic Acids Res., 29:207-209 (2001); Lefranc, M.-P., Nucleic Acids Res., 31:307-310 (2003); Lefranc, M.-P. et al., In Silico Biol., 5, 0006 (2004) (Epub), 5:45-60 (2005); Lefranc, M.-P. et al., Nucleic Acids Res., 33:D593-597 (2005); Res., 37:D1006-1012 (2009); Lefranc, M.-P. et al., Nucleic Acids Res., 43:D413-422 (2015); Chothia et al., (1989) Nature 342:877; Chothia, C. etal. (1987) J. Mol. Biol. 196:901-917, Al-lazikani et al (1997) J. Molec. Biol. 273:927-948; and Almagro, J. Mol. Recognit. 17:132-143 (2004). See also hgmp.mrc.ac.uk and bioinf.org.uk / abs. The CDR used herein may refer to a CDR defined by any method known in the art.Two antibodies with the same CDR mean that the amino acid sequence of that CDR is the same for both antibodies, as determined by the same method (e.g., IMGT definition).

[0099] In some implementations, there are three CDRs in each variable region of the heavy and light chains, which are named CDR1, CDR2, and CDR3 for each variable region. As used herein, the term "CDR group" refers to a group of three CDRs capable of binding the antigen that appear within a single variable region. The exact boundaries of these CDRs are defined differently depending on the system. The system described by Kabat (Kabat et al., Sequences of Proteins of Immunological Interest (National Institutes of Health, Bethesda, Md. (1987) and (1991)) not only provides a definitive residue numbering system applicable to any variable region of an antibody, but also provides precise residue boundaries defining three CDRs. These CDRs can be referred to as Kabat CDRs. Subregions of a CDR can be referred to as L1, L2, and L3 or H1, H2, and H3, where “L” and “H” represent the light chain and heavy chain regions, respectively. These regions can be referred to as Chothia CDRs, which have boundaries that overlap with Kabat CDRs. Padlan (FASEB J. 9:133-139 (1995)) and MacCallum (J Mol Biol 262(5):732-45 (1996)) describe other boundaries defining CDRs that overlap with Kabat CDRs. Other CDR boundary definitions may not strictly follow one of the systems described above, but will still be consistent with Kabat. CDR overlap, although they can be shortened or lengthened based on predictions or experimental findings that a particular residue or group of residues, or even the entire CDR, does not significantly affect antigen binding. While the preferred embodiment uses CDRs defined by Kabat or Chothia, the methods used herein may utilize CDRs defined according to either of these systems.

[0100] In some embodiments, the CDR of an antibody can have different amino acid sequences when different definition systems (e.g., IMGT definition, Kabat definition, or Chothia definition) are used. The definition systems annotate each amino acid in a given antibody sequence (e.g., a VH or VL sequence) with numbers, and Table 2 provides the numbers corresponding to the heavy chain CDR and light chain CDR. The CDRs listed in Table 1 are defined according to the Kabat definition. Those skilled in the art can derive the CDR sequences of the anti-TMPRSS6 antibodies provided in Table 1 using different numbering systems.

[0101] Table 2. CDR Definition

[0102]

[0103] CDR-grafted antibody: The term "CDR-grafted antibody" refers to an antibody that contains heavy and light chain variable region sequences from one species, but in which one or more CDR regions of VH and / or VL are replaced by CDR sequences from another species, such as an antibody with one or more mouse CDRs (e.g., CDR3) in the mouse heavy and light chain variable regions that have been replaced by human CDR sequences.

[0104] Chimeric antibody: The term "chimeric antibody" refers to an antibody that contains heavy and light chain variable region sequences from one species and constant region sequences from another species, such as a mouse heavy and light chain variable region linked to a human constant region.

[0105] Complementarity: As used herein, the term “complementarity” refers to the ability to precisely pair between two nucleotides or groups of nucleotides. Specifically, complementarity is a term characterizing the degree of hydrogen bond pairing that causes the binding between two nucleotides or groups of nucleotides. For example, if a base at a position of an oligonucleotide is able to hydrogen bond with a base at a corresponding position of a target nucleic acid (e.g., mRNA), the bases at that position are considered complementary to each other. Base pairing can include both typical Watson-Crick base pairing and non-Watson-Crick base pairing (e.g., Wobble base pairing and Hoogsteen base pairing). For example, in some embodiments, for complementary base pairing, an adenosine base (A) is complementary to a thymidine base (T) or a uracil base (U), a cytosine base (C) is complementary to a guanosine base (G), and universal bases such as 3-nitropyrrole or 5-nitroindole can hybridize with any A, C, U, or T and are considered complementary to any A, C, U, or T. Inosine (I) is also considered a universal base in the art and is complementary to any A, C, U or T.

[0106] Conservative amino acid substitutions: As used herein, “conservative amino acid substitution” refers to an amino acid substitution that does not alter the relative charge or size characteristics of the protein to which the substitution is made. Variants can be prepared according to methods known to those skilled in the art for altering polypeptide sequences, methods which can be found, for example, in compilations of such methods, such as *Molecular Cloning: A Laboratory Manual*, J. Sambrook, et al., eds., Fourth Edition, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York, 2012, or *Current Protocols in Molecular Biology*, FM Ausubel, et al., eds., John Wiley & Sons, Inc., New York. Conservative substitutions of amino acids include substitutions between amino acids within the following groups: (a) M, I, L, V; (b) F, Y, W; (c) K, R, H; (d) A, G; (e) S, T; (f) Q, N; and (g) E, D.

[0107] Cross-reactivity: As used herein and in the context of a target substance (e.g., an antibody), "cross-reactivity" refers to the property of a substance to specifically bind with more than one antigen of similar type or class (e.g., multiple homologs, paralogs, or orthologs) with similar affinity or coherence. For example, in some embodiments, antibodies that are cross-reactive against similar types or classes of human and non-human primate antigens (e.g., human TMPRSS6 and non-human TMPRSS6) are capable of binding with human and non-human primate antigens with similar affinity or coherence. In some embodiments, antibodies are cross-reactive against similar types or classes of human and rodent antigens. In some embodiments, antibodies are cross-reactive against similar types or classes of rodent and non-human primate antigens. In some embodiments, antibodies are cross-reactive against similar types or classes of human, non-human primate, and rodent antigens.

[0108] Effective Amount: As used herein, “effective amount” refers to the amount of each active substance (e.g., anti-TMPRSS6 antibody) required to impart a therapeutic effect to a subject, alone or in combination with one or more other active substances. In some embodiments, the therapeutic effect is a reduction in TMPRSS6 levels or activity, an increase in hepcidin levels or activity, and / or a reduction in disease conditions (e.g., iron overload in sickle cell disease, thalassemia, hemochromatosis, etc.).

[0109] Frame: As used herein, the term “frame” or “frame sequence” refers to the sequence remaining after subtracting the CDR from the variable region. Because the exact definition of a CDR sequence can be determined by different systems, the meaning of a frame sequence can be interpreted accordingly. The six CDRs (CDR-L1, CDR-L2, and CDR-L3 for the light chain and CDR-H1, CDR-H2, and CDR-H3 for the heavy chain) also distinguish frames on both the light and heavy chains into four sub-regions (FR1, FR2, FR3, and FR4) on each chain, where CDR1 is located between FR1 and FR2, CDR2 between FR2 and FR3, and CDR3 between FR3 and FR4. Where no specific sub-region is designated as FR1, FR2, FR3, or FR4, as otherwise mentioned, a frame region represents a combination of FRs within the variable region of a single naturally occurring immunoglobulin chain. As used herein, FR represents one of the four sub-regions, and FR represents two or more of the four sub-regions constituting a frame region. Human heavy and light chain receptor sequences are known in the art. In one embodiment, an acceptor sequence known in the art can be used in the antibody disclosed herein. In some embodiments, the sequence of the disclosed framework region may differ from the disclosed sequence by up to 20% (e.g., the sequence of the disclosed framework region has about 80% (e.g., 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%) identity with the disclosed sequence).

[0110] Human Antibodies: As used herein, the term "human antibody" is intended to include antibodies having variable and constant regions derived from human germline immunoglobulin sequences. Human antibodies of this disclosure may contain amino acid residues not encoded by human germline immunoglobulin sequences (e.g., mutations introduced through random or site-specific mutagenesis in vitro or through somatic mutations in vivo), such as in CDRs, and particularly in CDR3. However, the term "human antibody" as used herein is not intended to include antibodies in which a CDR sequence derived from another mammalian species (e.g., mouse) has been grafted onto a human frame sequence.

[0111] Humanized Antibody: The term "humanized antibody" refers to an antibody containing heavy and light chain variable region sequences from a non-human species (e.g., mouse), but in which at least a portion of the VH and / or VL sequences has been altered to be more "human-like" (i.e., more similar to human germline variable sequences). One type of humanized antibody is a CDR-grafted antibody, in which a human CDR sequence is introduced into non-human VH and VL sequences to replace the corresponding non-human CDR sequences. In one embodiment, a humanized anti-TMPRSS6 antibody and an antigen-binding moiety are provided. Such antibodies can be produced by obtaining a mouse anti-TMPRSS6 monoclonal antibody using conventional hybridoma techniques followed by humanization using in vitro genetic modification, such as those disclosed in Kasaian et al. PCT Publication No. WO 2005 / 123126 A2.

[0112] Humanized antibodies are human immunoglobulins (acceptor antibodies) in which residues of the complementarity-determining region (CDR) from the acceptor are replaced by residues of the CDR from a non-human species (donor antibody) having the desired specificity, affinity, and capability, such as mouse, rat, or rabbit. In some embodiments, Fv frame region (FR) residues of the human immunoglobulin are replaced by corresponding non-human residues. Furthermore, humanized antibodies may include residues not present in the acceptor antibody or the introduced CDR or frame sequence but included to further refine and optimize antibody performance. Generally, humanized antibodies will contain at least one, and typically substantially all, of two variable domains, wherein all or substantially all of the CDR regions correspond to those of non-human immunoglobulins, and all or substantially all of the FR regions are those of human immunoglobulin common sequences. Humanized antibodies will preferably also contain at least a portion of the immunoglobulin constant region or domain (Fc), typically those of human immunoglobulins. The antibody may have an Fc region modified as described in WO 99 / 58572. Other forms of humanized antibodies have one or more CDRs (one, two, three, four, five, or six) that are altered relative to the original antibody; these are also referred to as one or more CDRs derived from one or more CDRs of the original antibody. Humanized antibodies may also involve affinity maturation.

[0113] In some embodiments, humanization is achieved by grafting a CDR (e.g., as shown in Table 1) into a human variable domain (e.g., the IGKV1-NL1*01 and IGHV1-3*01 human variable domains). In some embodiments, the anti-TMPRSS6 antibody of this disclosure is a humanized variant that contains one or more amino acid substitutions (e.g., in the VH frame region) compared to any of the VHs listed in Table 1; and / or contains one or more amino acid substitutions (e.g., in the VL frame region) compared to any of the VLs listed in Table 1.

[0114] Isolated Antibodies: As used herein, “isolated antibodies” are intended to refer to antibodies that are substantially free of other antibodies with different antigen specificities (e.g., isolated antibodies that specifically bind to TMPRSS6 are substantially free of antibodies that specifically bind to antigens other than TMPRSS6). However, isolated antibodies that specifically bind to TMPRSS6 may have cross-reactivity with other antigens. Furthermore, isolated antibodies may be substantially free of other cellular material and / or chemicals.

[0115] Kabat Numbering: The terms “Kabat numbering,” “Kabat definition,” and “Kabat labeling” are used interchangeably herein. These terms, as accepted in the art, refer to a system for numbering amino acid residues in the variable regions of the heavy and light chains of antibodies or their antigen-binding moieties that are more variable (i.e., hypervariable) than other amino acid residues (Kabat et al. (1971) Ann. NY Acad, Sci. 190:382-391 and, Kabat, EA, et al. (1991) Sequences of Proteins of Immunological Interest, Fifth Edition, US Department of Health and Human Services, NIH Publication No. 91-3242). For the heavy chain variable regions, the hypervariable regions are amino acids 31 to 35 of CDR1, amino acids 50 to 65 of CDR2, and amino acids 95 to 102 of CDR3. For the light chain variable regions, the hypervariable regions are amino acids 24 to 34 of CDR1, amino acids 50 to 56 of CDR2, and amino acids 89 to 97 of CDR3.

[0116] Percentage (%) Sequence Identity: The terms “percentage (%) sequence identity,” “percentage (%) identity,” and “percentage (%) identical” relative to a reference polypeptide (or nucleotide) sequence are defined as the percentage of amino acid residues (or nucleic acids) in a candidate sequence that are identical to amino acid residues (or nucleic acids) in a reference polypeptide (or nucleotide) sequence after sequence alignment and the introduction of vacancies (if necessary) to achieve maximum percentage sequence identity, without considering any conservative substitutions as part of sequence identity. Alignment for determining the percentage of sequence identity can be performed in various ways within the art, for example, using publicly available computer software such as BLAST, BLAST-2, ALIGN, or Megalign (DNASTAR) software. Those skilled in the art can determine appropriate parameters for sequence alignment, including any algorithm required to achieve maximum alignment across the full length of the compared sequences. However, for the purposes of this document, the sequence comparison computer program ALIGN-2 is used to generate the % amino acid (or nucleic acid) sequence identity value. The ALIGN-2 sequence comparison computer program was written by Genentech, Inc., and its source code is filed with the US Copyright Office, Washington DC, 20559, under US Copyright Registration No. TXU510087. The ALIGN-2 program is publicly available from Genentech, Inc., South San Francisco, California, or can be compiled from the source code. The ALIGN-2 program should be compiled for use with UNIX operating systems, including digital UNIX V4.0D. All sequence comparison parameters are set by the ALIGN-2 program and remain unchanged.

[0117] Recombinant Antibody: As used herein, the term “recombinant antibody” is intended to include all antibodies prepared, expressed, generated, or isolated by recombinant means, such as antibodies expressed using recombinant expression vectors transfected into host cells (described in more detail in this disclosure), including, for example, antibodies isolated from recombinant human antibody libraries (Hoogenboom HR, (1997) TIB Tech. 15:62-70; Azzazy H., and Highsmith WE, (2002) Clin. Biochem. 35:425-445; Gavilondo JV, and Larrick JW (2002) BioTechniques29:128-145; Hoogenboom H., and Chames P. (2000) Immunology Today 21:371-378), and antibodies isolated from animals transgenic with human immunoglobulin genes (e.g., mice) (see, for example, Taylor, LD, et al. (1992) Nucl. Acids Res.). 20:6287-6295; Kellermann SA., and Green LL (2002) Current Opinion in Biotechnology 13:593-597; Little M. et al (2000) Immunology Today 21:364-370) or antibodies prepared, expressed, generated, or isolated by any other means involving splicing human immunoglobulin gene sequences into other DNA sequences. In some embodiments, recombinant human antibodies are provided herein. In some embodiments, such recombinant human antibodies have variable and constant regions derived from human germline immunoglobulin sequences. However, in some embodiments, such recombinant human antibodies are mutagenized in vitro (or, when using animals transgenic to target human Ig sequences, in vivo somatic cell mutagenization), and thus the amino acid sequences of the VH and VL regions of the recombinant antibody are sequences that, while derived from and associated with human germline VH and VL sequences, may not be naturally present in a human antibody germline library in vivo. One embodiment of this disclosure provides a fully human antibody capable of binding to human TMPRSS6, which can be generated using techniques known in the art, such as, but not limited to, using human Ig phage libraries, such as those disclosed in Jermutuset al., PCT Publication No. WO 2005 / 007699 A2.

[0118] Selectivity: As used herein, the term "selectivity" or "selectively" refers to the ability of a molecule to influence its target molecule (e.g., inhibit, antagonize, agonize, etc.) compared to a reference molecule. For example, a molecule that selectively inhibits its target molecule means that the molecule is able to inhibit its target molecule to a degree that is distinguishable from a reference molecule in an inhibition assay or other inhibitory setting. For example, with respect to inhibitors, the term "selective inhibition" refers to the ability of an inhibitor to inhibit its target molecule to a degree that is distinguishable from a reference molecule that is substantially uninhibited in an inhibition assay, for example, to the extent that selective inhibition of the target molecule is permitted, as described herein. Once the reaction is terminated, the signal generated by inhibiting the target molecule can be measured. The half-maximum inhibitor concentrations of the target molecule and the reference molecule can be calculated. In some embodiments, the molecules described herein selectively bind to the target molecule. In some embodiments, the molecules described herein selectively inhibit the target molecule. In some embodiments, the molecules described herein selectively antagonize the target molecule.

[0119] Specific binding: As used herein, the term "specific binding" refers to the ability of a molecule to bind to a binding partner with a degree of affinity or cohesion, enabling the molecule to be used to distinguish the binding partner from a suitable control in a binding assay or other binding setting. Regarding antibodies, the term "specific binding" refers to the ability of an antibody to bind to a specific antigen with a degree of affinity or cohesion compared to one or more suitable reference antigens, enabling the antibody to be used to distinguish the specific antigen from other antigens, as described herein. In some embodiments, if the antibody binds to the K of the target... D For at least about 10 -4 M, 10 -5 M, 10 -6 M, 10 -7 M, 10 -8 M, 10 -9 M, 10 -10 M, 10 -11 M, 10 -12 M, 10 -13 If M is low or lower, the antibody binds specifically to the target. In some implementations, the antibody specifically binds to TMPRSS6.

[0120] Object: As used herein, the term "object" refers to a mammal. In some embodiments, the object is a non-human primate or rodent. In some embodiments, the object is a human being. In some embodiments, the object is a patient, such as a person who has or is suspected of having a disease. In some embodiments, the object is a person who has or is suspected of having iron overload and / or one or more conditions caused by iron overload.

[0121] Treatment: As used herein, the term "treatment" and its variations refer to the application or administration of a composition comprising one or more active substances (e.g., an anti-TMPRSS6 antibody) to a subject suffering from a target disease or condition, having symptoms of that disease / condition, or being predisposed to that disease / condition, with the aim of curing, treating, alleviating, relieving, altering, remedying, improving, or influencing the condition, symptoms of the disease, or predisposition to the disease or condition. Alleviating a target disease / condition includes delaying or preventing the onset, development, or progression of the disease, or reducing the severity of the disease. In some embodiments, the target disease or condition of this disclosure includes, but is not limited to, hemochromatosis, sickle cell disease (SCD), thalassemia, hemolysis, African iron overload, Diamond-Blackfan anemia, myelodysplastic syndromes (MDS), and blood transfusions.

[0122] II. Anti-TMPRSS6 antibody

[0123] In some embodiments, the anti-TMPRSS6 antibody is an antibody specific to TMPRSS6. In some aspects, this document provides antibodies that bind to human TMPRSS6 with high specificity and affinity. In some embodiments, the anti-TMPRSS6 antibody described herein specifically binds to any extracellular epitope of TMPRSS6 or an epitope exposed to the antibody. In some embodiments, the anti-TMPRSS6 antibody provided herein specifically binds to TMPRSS6 derived from humans, non-human primates, mice, rats, etc. In some embodiments, the anti-TMPRSS6 antibody provided herein binds to human TMPRSS6. In some embodiments, the anti-TMPRSS6 antibody described herein binds to an amino acid segment of human or non-human primate TMPRSS6. In some embodiments, the anti-TMPRSS6 antibody described herein specifically binds to an epitope on human TMPRSS6.

[0124] TMPRSS6, also known as protein lyase 2, is a serine protease belonging to the type II transmembrane serine protease (TTSP) family. Enzymes in this family share common structural features, including a serine protease domain, a variable-length stem region containing a mosaic of structural domains, and a short cytoplasmic tail (Hooper et al., Type II transmembrane serine proteases: Insights into an emerging class of cell surface proteolytic enzymes. J BiolChem. 2001; 276:857-60). The TTSP family of proteases comprises several subfamilies, including the protein lyase subfamily containing TMPRSS 2 through 5. TMPRSS6, which binds specifically to antibodies in some embodiments of this disclosure, is highly conserved across mammalian species. In some embodiments, the antibody of this disclosure specifically binds to the TMPRSS6 protein encoded by the human TMPRSS6 gene (e.g., GenBank: AAH39082.1, e.g., NCBI Gene ID: 164656). In some embodiments, the antibody of this disclosure specifically binds to the TMPRSS6 protein encoded by the mouse TMPRSS6 gene (e.g., GenBank: AAH57674.1, e.g., NCBI Gene ID: 71753). In other embodiments, the antibody of this disclosure specifically binds to the TMPRSS6 protein encoded by a non-human primate TMPRSS6 gene (e.g., cynomolgus monkey (i.e., Macacafascicularis), e.g., NCBI Gene ID: 102139590). The TMPRSS6 gene spans 18 exons and 17 intercalated introns, and the boundaries of the proteolytic enzyme-2 protein domain correspond to intron / exon connections in the encoding genes across all species. Structurally, protein lyase-2 contains typical TTSP features, such as a short cytoplasmic amino-terminal tail, a transmembrane region, a stem region containing two complement protein subcomponents (C1r / C1s, urchin embryonic growth factor and bone morphogenetic protein 1 (CUB) domains), three low-density lipoprotein receptor class A (LDLR) domains, and a carboxyl-terminal trypsin-like serine protease domain.All of these structural features are strictly conserved in humans, macaques, dogs, cattle, mice, and rats, with human proteins sharing 95.6%, 91.1%, 85.6%, 80.1%, and 80.4% identity with proteolytic enzyme-2 from these species, respectively (Ramsay et al., The type II transmembrane serine protease matriptase-2—identification, structural features, enzymology, expression pattern and potential roles. Front Biosci. 2008; 13:259-79).

[0125] The proteolytic domain of Matriptase-2 contains a serine protease triplet of H, D, and S residues required for catalytic activity; and an SWG motif expected to be located at the top of the substrate S1 binding pocket, which orients the substrate's cleavable bonds correctly. Proteolytic activation of Matriptase-2 is expected to occur within the motif (RIVGG (SEQ ID NO: 80)) at the junction of the predomain and catalytic domain, a characteristic feature of serine proteases and conserved across species (Velasco et al., Matriptase-2, a membrane-bound mosaic serine proteinase predominantly expressed in human liver and showing degrading activity against extracellular matrix proteins. J Biol Chem. 2002; 277:37637-46). In adult and mouse tissues, the predominant site of Matriptase-2 mRNA expression is the liver. Specifically, it has been shown that Matriptase-2 mRNA expression is limited to hepatocytes of the liver.

[0126] Functionally, proteolytic enzyme-2 cleaves hemojuvelin (HJV), a protein essential for iron regulation through its positive regulation of hepcidin expression. Hepcidin is known to be a negative regulator of iron uptake, release, and recycling. In the absence of proteolytic enzyme-2, HJV levels increase, leading to increased hepcidin levels and thus inhibiting iron transport. The establishment of proteolytic enzyme-2 as an essential component of iron homeostasis is based on the phenotype of iron-refractory iron deficiency anemia in human patients with loss-of-function TMPRSS6 mutations (Cui et al., Iron-refractory iron deficiency anemia: new molecular mechanisms. Kidney Int. 2009; 76(11):1137-1141). Therefore, in some embodiments, the antibody of this disclosure inhibits proteolytic enzyme-2 activity to regulate iron homeostasis. In some embodiments, the antibody of this disclosure inhibits proteolytic enzyme-2 activity to reduce iron transport by increasing hepcidin activity.

[0127] In some embodiments, the anti-TMPRSS6 antibody described herein can bind to a fragment of human TMPRSS6. The TMPRSS6 fragment length can be about 5 to about 425 amino acids, about 10 to about 400 amino acids, about 50 to about 350 amino acids, about 100 to about 300 amino acids, about 150 to about 250 amino acids, about 200 to about 300 amino acids, or about 75 to about 150 amino acids. In some embodiments, the fragment may contain a sequential number of amino acids from TMPRSS6. In other embodiments, the fragment may contain a non-sequential number of amino acids from TMPRSS6.

[0128] In some embodiments, the anti-TMPRSS6 antibody disclosed herein specifically binds to mouse TMPRSS6. In some embodiments, the anti-TMPRSS6 antibody disclosed herein specifically binds to rat TMPRSS6. In some embodiments, the anti-TMPRSS6 antibody disclosed herein specifically binds to non-human primate TMPRSS6 (e.g., TMPRSS6 from cynomolgus monkeys, rhesus monkeys, African green monkeys, vervet monkeys, squirrel monkeys, owl monkeys, pig-tailed monkeys, or baboons). In some embodiments, the anti-TMPRSS6 antibody disclosed herein specifically binds to human TMPRSS6.

[0129] In some embodiments, the anti-TMPRSS6 antibody described herein is an affinity-matured clone. In some embodiments, the anti-TMPRSS6 antibody has an affinity of at least about 10... -4 M, 10 -5M, 10 -6 M, 10 -7 M, 8 -7 M, 6 -7 M, 4 -7 M, 2 -7 M, 10 -8 M, 8 -8 M, 6 -8 M, 4 -8 M, 2 -8 M, 10 -9 M, 10 -10 M, 10 -11 M, 10 -12 M, 10 -13 M or less binding affinity (e.g., as by K) D The antibodies described herein specifically bind to TMPRSS6 (e.g., human or non-human primate TMPRSS6). For example, the anti-TMPRSS6 antibodies of this disclosure can bind to the TMPRSS6 protein (e.g., human TMPRSS6) with an affinity of 5 pM to 750 nM, such as 1 nM to 500 nM, such as 10 nM to 450 nM, such as 50 pM to 100 nM, such as 500 pM to 50 nM. This disclosure also includes antibodies that compete with any antibodies described herein for binding to the TMPRSS6 protein (e.g., human TMPRSS6) and have an affinity of 500 nM or less (e.g., 400 nM or less, 100 nM or less, 80 nM or less, 50 nM or less, 20 nM or less, 10 nM or less, 500 pM or less, 50 pM or less, or 5 pM or less). In some implementations, the anti-TMPRSS6 antibody described herein is in the sub-nanomolar range of K D Combined with TMPRSS6.

[0130] The affinity and binding kinetics of anti-TMPRSS6 antibodies can be tested using any suitable method. For example, binding affinity (or binding specificity) can be determined by a variety of methods, including but not limited to biosensor technologies (e.g., OCTET or BIACORE), equilibrium dialysis, equilibrium binding, gel filtration, ELISA, surface plasmon resonance (SPR), fluorescence activated cell sorting (FACS), or spectroscopy (e.g., using fluorescence assays). Exemplary conditions for evaluating binding affinity are in HBS-P buffer (10 mM HEPES pH 7.4, 150 mM NaCl, 0.005% (v / v) surfactant P20) and PBS buffer (10 mM PO4-3, 137 mM NaCl, and 2.7 mM KCl). These techniques can be used to measure the concentration of the bound protein as a function of the target protein concentration. The concentration of the bound protein ([[bound]]) is generally correlated with the concentration of the free target protein ([[free]]) by the following formula:

[0131] [[Bound]] = [[Free]] / (Kd + [[Free]])

[0132] However, it is not always necessary to target K. A Precise determination is necessary because sometimes obtaining a quantitative measurement of affinity is sufficient, for example, by using methods such as ELISA or FACS analysis to determine the affinity with K. A Proportional and therefore can be used for comparison, such as determining whether a higher affinity is, for example, twice as high, to obtain a qualitative measurement of affinity or to obtain an inference of affinity, such as by functional assays (e.g., in vitro or in vivo assays) of activity.

[0133] Table 1 provides some non-restrictive examples of heavy chain (HC) and light chain (LC) sequences, heavy chain variable domain (VH) and light chain variable domain (VL) sequences, CDR sequences, and heavy chain and light chain constant region sequences for anti-TMPRSS6 antibodies.

[0134] Table 1. Some examples of anti-TMPRSS6 antibodies (according to Kabat's definition of CDR)

[0135]

[0136]

[0137]

[0138]

[0139]

[0140]

[0141]

[0142]

[0143]

[0144]

[0145]

[0146]

[0147] In some embodiments, the anti-TMPRSS6 antibody of this disclosure comprises one or more HC CDRs (e.g., HC CDR1, HC CDR2, or HC CDR3) amino acid sequences from any anti-TMPRSS6 antibody selected from Table 1. In some embodiments, the anti-TMPRSS6 antibody of this disclosure comprises HCCDR1, HC CDR2, and HC CDR3 as provided for any antibody selected from Table 1. In some embodiments, the anti-TMPRSS6 antibody of this disclosure comprises one or more LC CDRs (e.g., LC CDR1, LC CDR2, or LC CDR3) amino acid sequences from any anti-TMPRSS6 antibody selected from Table 1. In some embodiments, the anti-TMPRSS6 antibody of this disclosure comprises LC CDR1, LC CDR2, and LC CDR3 as provided for any anti-TMPRSS6 antibody selected from Table 1.

[0148] In some embodiments, the anti-TMPRSS6 antibody of this disclosure comprises HC CDR1, HC CDR2, HC CDR3, LC CDR1, LC CDR2, and LC CDR3, as provided by any of the anti-TMPRSS6 antibodies selected from Table 1. In some embodiments, the antibody heavy chain CDR3 domain and the light chain CDR3 domain may play a particularly important role in the antibody's binding specificity / affinity to the antigen. Therefore, the anti-TMPRSS6 antibody of this disclosure may at least comprise the heavy chain CDR3 and / or light chain CDR3 of any of the anti-TMPRSS6 antibodies selected from Table 1.

[0149] In some embodiments, the anti-TMPRSS6 antibody includes a heavy chain variable region comprising heavy chain CDR1 (HC CDR1), heavy chain CDR2 (HC CDR2), and heavy chain CDR3 (HC CDR3). In some embodiments, the anti-TMPRSS6 antibody includes a light chain variable region comprising light chain CDR1 (LC CDR1), light chain CDR2 (LC CDR2), and light chain CDR3 (LC CDR3).

[0150] Functional variants of any exemplary anti-TMPRSS6 antibodies disclosed herein are also within the scope of this disclosure. Relative to the reference antibody, the functional variant may contain one or more amino acid residue variations in VH and / or VL, or in one or more HC CDRs and / or one or more LC CDRs, while retaining substantially similar binding and biological activities (e.g., substantially similar binding affinity, binding specificity, inhibitory activity, or combinations thereof) to the reference antibody.

[0151] In some embodiments, any anti-TMPRSS6 antibody of this disclosure has one or more CDR (e.g., HC CDR or LC CDR) sequences substantially similar to any HC CDR1, HC CDR2, HC CDR3, LC CDR1, LC CDR2 and / or LC CDR3 sequence from an anti-TMPRSS6 antibody selected from Table 1. In some embodiments, the positions of one or more CDRs (e.g., HC CDR1, HC CDR2, or HC CDR3) along the VH region and / or one or more CDRs (e.g., LC CDR1, LC CDR2, or LC CDR3) along the VL region of the antibody described herein may vary by one, two, three, four, five, or six amino acid positions, as long as immune-specific binding to TMPRSS6 (e.g., human TMPRSS6) is maintained (e.g., substantially maintained, for example, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95% of the binding to the original antibody from which it is derived). For example, in some embodiments, the location of the CDR of any antibody described herein can be varied by shifting the N-terminal and / or C-terminal boundary of the CDR by one, two, three, four, five, or six amino acids relative to the CDR location of any antibody described herein, as long as immune-specific binding to TMPRSS6 (e.g., human TMPRSS6) is maintained (e.g., substantially maintained, for example, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95% of the binding to the original antibody from which it is derived). In another embodiment, the length of one or more CDRs (e.g., HC CDR1, HC CDR2, or HC CDR3) along the VH region and / or one or more CDRs (e.g., LC CDR1, LCCDR2, or LC CDR3) along the VL region of the antibody described herein may vary (shorter or longer) by one, two, three, four, five, or more amino acids, as long as immune-specific binding to TMPRSS6 (e.g., human TMPRSS6) is maintained (e.g., substantially maintained, for example, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95% of the binding to the original antibody from which it is derived).

[0152] Therefore, in some embodiments, the HC CDR1, HC CDR2, HC CDR3, LC CDR1, LCCDR2 and / or LC CDR3 described herein may be one, two, three, four, five or more amino acids shorter than one or more of the CDRs described herein (e.g., CDRs from any anti-TMPRSS6 antibody selected from Table 1), as long as the immune-specific binding to TMPRSS6 (e.g., human TMPRSS6) is maintained (e.g., substantially maintained, for example, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95% of the binding relative to the original antibody from which it is derived). In some embodiments, the HC CDR1, HC CDR2, HC CDR3, LC CDR1, LC CDR2 and / or LC CDR3 described herein may be one or more amino acids longer than the CDRs described herein (e.g., CDRs from any anti-TMPRSS6 antibody selected from Table 1), provided that an immune-specific binding to TMPRSS6 (e.g., human TMPRSS6) is maintained (e.g., substantially maintained, for example, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95% of the binding relative to the original antibody from which it is derived). In some embodiments, the amino moiety of HC CDR1, HC CDR2, HC CDR3, LCCDR1, LC CDR2 and / or LC CDR3 described herein may be extended by one, two, three, four, five or more amino acids than one or more of the CDRs described herein (e.g., CDRs from any anti-TMPRSS6 antibody selected from Table 1), as long as the immune-specific binding to TMPRSS6 (e.g., human TMPRSS6) is maintained (e.g., substantially maintained, for example, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95% of the binding relative to the original antibody from which it is derived). In some embodiments, the carboxyl moiety of HC CDR1, HC CDR2, HC CDR3, LC CDR1, LC CDR2 and / or LCCDR3 described herein may be extended by one, two, three, four, five or more amino acids than one of the CDRs described herein (e.g., CDRs from any anti-TMPRSS6 antibody selected from Table 1), as long as the immune-specific binding to TMPRSS6 (e.g., human TMPRSS6) is maintained (e.g., substantially maintained, for example, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95% of the binding relative to the original antibody from which it is derived).In some embodiments, the amino moiety of HC CDR1, HC CDR2, HC CDR3, LC CDR1, LC CDR2 and / or LC CDR3 described herein may be shortened by one, two, three, four, five or more amino acids compared to one or more of the CDRs described herein (e.g., CDRs from any anti-TMPRSS6 antibody selected from Table 1), as long as immune-specific binding to TMPRSS6 (e.g., human TMPRSS6) is maintained (e.g., substantially maintained, for example, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95% of the binding relative to the original antibody from which it is derived). In some embodiments, the carboxyl moiety of HC CDR1, HCCDR2, HC CDR3, LC CDR1, LC CDR2, and / or LC CDR3 described herein may be shortened by one, two, three, four, five, or more amino acids compared to one or more of the CDRs described herein (e.g., CDRs from any anti-TMPRSS6 antibody selected from Table 1), as long as immune-specific binding to TMPRSS6 (e.g., human TMPRSS6) is maintained (e.g., substantially maintained, for example, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95% of the binding relative to the original antibody from which it is derived). Any method may be used to determine whether immune-specific binding to TMPRSS6 (e.g., human TMPRSS6) is maintained, for example using binding assays and conditions described in the art.

[0153] In some instances, any anti-TMPRSS6 antibody of this disclosure has one or more CDR (e.g., HC CDR or LC CDR) sequences substantially similar to any of the anti-TMPRSS6 antibodies selected from Table 1. For example, an antibody may comprise one or more CDR sequences from any of the anti-TMPRSS6 antibodies selected from Table 1, containing up to 5, 4, 3, 2, or 1 amino acid residue variations compared to the corresponding CDR region in any of the CDRs provided herein (e.g., CDRs from any of the anti-TMPRSS6 antibodies selected from Table 1), provided that immune-specific binding to TMPRSS6 (e.g., human TMPRSS6) is maintained (e.g., substantially maintained, for example, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95% of the binding relative to the original antibody from which it is derived). In some embodiments, any amino acid variation in any CDR provided herein may be a conserved variation. Conserved variations can be introduced into the CDR at locations where residues are unlikely to interact with the TMPRSS6 protein (e.g., human TMPRSS6 protein) (e.g., as determined based on crystal structure). Some aspects of this disclosure provide anti-TMPRSS6 antibodies comprising one or more heavy chain variable (VH) domains and / or light chain variable (VL) domains provided herein. In some embodiments, any VH domain provided herein comprises one or more HC CDR sequences (e.g., HC CDR1, HC CDR2, and HC CDR3) provided herein, such as any CDR-H sequence provided in any of the anti-TMPRSS6 antibodies listed in Table 1. In some embodiments, any VL domain provided herein comprises one or more CDR-L sequences (e.g., LC CDR1, LC CDR2, and LC CDR3) provided herein, such as any LC CDR sequence provided in any of the anti-TMPRSS6 antibodies listed in Table 1.

[0154] In some embodiments, the anti-TMPRSS6 antibody of this disclosure includes any antibody comprising a heavy chain variable domain and / or a light chain variable domain selected from any of the anti-TMPRSS6 antibodies in Table 1, and variants thereof. In some embodiments, the anti-TMPRSS6 antibody of this disclosure includes any antibody comprising a heavy chain variable and light chain variable pair selected from any of the anti-TMPRSS6 antibodies in Table 1.

[0155] Some aspects of this disclosure provide anti-TMPRSS6 antibodies having amino acid sequences homologous to any of those described herein, including heavy chain variable (VH) domains and / or light chain variable (VL) domains. In some embodiments, the anti-TMPRSS6 antibody comprises a heavy chain variable sequence or light chain variable sequence having at least 75% (e.g., at least 80%, at least 85%, 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%, or at least 99%) identity to a heavy chain variable sequence and / or any light chain variable sequence selected from any of the anti-TMPRSS6 antibodies in Table 1. In some embodiments, the heavy chain variable and / or light chain variable amino acid sequences do not vary within any CDR sequences provided herein. For example, in some embodiments, the degree of sequence variation (e.g., at least 75%, at least 80%, at least 85%, 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%, or at least 99%) may occur within the heavy chain variable region and / or light chain variable region sequences, except for any CDR sequences provided herein. In some embodiments, any anti-TMPRSS6 antibody provided herein comprises a heavy chain variable sequence and a light chain variable sequence comprising a frame sequence having at least 75%, at least 80%, at least 85%, 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%, or at least 99% identity with a frame sequence selected from any anti-TMPRSS6 antibody in Table 1.

[0156] In some embodiments, the anti-TMPRSS6 antibody of this disclosure is an antibody containing a VH, which contains no more than 20 amino acid variations (e.g., no more than 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1 amino acid variation) compared to the VH of any anti-TMPRSS6 antibody listed in Table 1. Alternatively or additionally, the anti-TMPRSS6 antibody of this disclosure is a humanized antibody containing a VL, which contains no more than 20 amino acid variations (e.g., no more than 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 9, 8, 7, 6, 5, 4, 3, 2, or 1 amino acid variation) compared to the VL of any anti-TMPRSS6 antibody listed in Table 1. In some embodiments, the heavy chain variable and / or light chain variable amino acid sequences do not vary within any CDR sequences provided herein. For example, in some embodiments, the number of amino acid variations (e.g., no more than 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 10, 9, 8, 7, 6, 5, 4, 3, 2 or 1 amino acid variation) may appear in the heavy chain variable and / or light chain variable sequences in places other than any CDR sequence provided herein. In some embodiments, any anti-TMPRSS6 antibody provided herein comprises a heavy chain variable sequence and / or a light chain variable sequence, wherein the heavy chain variable sequence comprises a frame sequence having a variation of no more than 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 10, 9, 8, 7, 6, 5, 4, 3, 2 or 1 amino acid compared to the frame sequence of the VH of any anti-TMPRSS6 antibody selected from Table 1, and the light chain variable sequence comprises a frame sequence having a variation of no more than 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 10, 9, 8, 7, 6, 5, 4, 3, 2 or 1 amino acid compared to the frame sequence of the VL of any anti-TMPRSS6 antibody selected from Table 1.

[0157] In some embodiments, the anti-TMPRSS6 antibody of this disclosure is a humanized antibody (e.g., containing humanized variants of one or more CDRs listed in Table 1). In some embodiments, the anti-TMPRSS6 antibody of this disclosure contains the same HC CDR1, HC CDR2, HC CDR3, LC CDR1, LC CDR2, and LC CDR3 as shown in Table 1, and contains humanized heavy chain variable regions and / or humanized light chain variable regions. In some embodiments, the humanized heavy chain variable and / or humanized light chain variable amino acid sequences do not vary within any CDR sequences provided herein. For example, in some embodiments, the degree of sequence variation (e.g., at least 75%, at least 80%, at least 85%, 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%, or at least 99%) may occur within the heavy chain variable and / or light chain variable sequences, except for any CDR sequences provided herein. In some embodiments, the humanized anti-TMPRSS6 antibody provided herein comprises a heavy chain variable sequence and a light chain variable sequence comprising a frame sequence having at least 75%, at least 80%, at least 85%, 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%, or at least 99% identity with a frame sequence selected from any anti-TMPRSS6 antibody in Table 1. In some embodiments, the humanized heavy chain variable and / or humanized light chain variable amino acid sequences do not vary within any CDR sequences provided herein. For example, in some embodiments, the number of amino acid variations (e.g., no more than 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 10, 9, 8, 7, 6, 5, 4, 3, 2 or 1 amino acid variation) may appear in the heavy chain variable and / or light chain variable sequences in places other than any CDR sequences provided herein. In some embodiments, the humanized anti-TMPRSS6 antibody provided herein comprises a heavy chain variable sequence and / or a light chain variable sequence, wherein the heavy chain variable sequence comprises a frame sequence having a variation of no more than 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 10, 9, 8, 7, 6, 5, 4, 3, 2 or 1 amino acid variation compared to the frame sequence of the VH of any anti-TMPRSS6 antibody selected from Table 1, and the light chain variable sequence comprises a frame sequence having a variation of no more than 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 10, 9, 8, 7, 6, 5, 4, 3, 2 or 1 amino acid variation compared to the frame sequence of the VL of any anti-TMPRSS6 antibody selected from Table 1.

[0158] In some embodiments, the anti-TMPRSS6 antibody of this disclosure comprises HC CDR1, HC CDR2, and HC CDR3, which are heavy chain variable domains having the amino acid sequence of SEQ ID NO: 7. Alternatively or supplementally, the anti-TMPRSS6 antibody of this disclosure comprises LC CDR1, LCCDR2, and LC CDR3, which are light chain variable domains having the amino acid sequence of SEQ ID NO: 8.

[0159] In some embodiments, according to the Kabat definition system, the anti-TMPRSS6 antibody of this disclosure comprises HC CDR1 having the amino acid sequence of SEQ ID NO: 1, HC CDR2 having the amino acid sequence of SEQ ID NO: 2, HC CDR3 having the amino acid sequence of SEQ ID NO: 3, LC CDR1 having the amino acid sequence of SEQ ID NO: 4, LC CDR2 having the amino acid sequence of RAN, and LC CDR3 having the amino acid sequence of SEQ ID NO: 6.

[0160] In some embodiments, the anti-TMPRSS6 antibody of this disclosure comprises HC CDR1, HC CDR2, and HCCDR3, which, compared to HC CDR1 having the amino acid sequence of SEQ ID NO: 1, HCCDR2 having the amino acid sequence of SEQ ID NO: 2, and HC CDR3 having the amino acid sequence of SEQ ID NO: 3, collectively contain no more than 5 amino acid variations (e.g., no more than 5, 4, 3, 2, or 1 amino acid variation). As used anywhere in this disclosure, "commonly" means that the total number of amino acid variations in all three heavy chain CDRs is within a defined range. Alternatively or supplementally, the anti-TMPRSS6 antibody of this disclosure comprises LC CDR1, LC CDR2, and LC CDR3, which, compared to LC CDR1 having the amino acid sequence of SEQ ID NO: 4, LC CDR2 having the amino acid sequence of RAN, and LC CDR3 having the amino acid sequence of SEQ ID NO: 6, collectively contain no more than 5 amino acid variations (e.g., no more than 5, 4, 3, 2, or 1 amino acid variation).

[0161] In some embodiments, the anti-TMPRSS6 antibody of this disclosure comprises HC CDR1, HC CDR2, and HCCDR3, which together have at least 80% (e.g., at least 80%, at least 85%, 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%, or at least 99%) identity with HC CDR1 having the amino acid sequence of SEQ ID NO: 1, HC CDR2 having the amino acid sequence of SEQ ID NO: 2, and HC CDR3 having the amino acid sequence of SEQ ID NO: 3. As an alternative or supplement, the anti-TMPRSS6 antibody of this disclosure comprises LC CDR1, LC CDR2 and LC CDR3, which together have at least 80% (e.g. at least 80%, at least 85%, 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% or at least 99%) identity with LC CDR1 having the amino acid sequence of SEQ ID NO: 4, LC CDR2 having the amino acid sequence of RAN and LC CDR3 having the amino acid sequence of SEQ ID NO: 6.

[0162] In some embodiments, the anti-TMPRSS6 antibody of this disclosure comprises: HC CDR1 having no more than 3 amino acid variations (e.g., no more than 3, 2, or 1 amino acid variation) compared to HC CDR1 having the amino acid sequence of SEQ ID NO: 1; HC CDR2 having no more than 3 amino acid variations (e.g., no more than 3, 2, or 1 amino acid variation) compared to HC CDR2 having the amino acid sequence of SEQ ID NO: 2; and / or HCCDR3 having no more than 3 amino acid variations (e.g., no more than 3, 2, or 1 amino acid variation) compared to HC CDR3 having the amino acid sequence of SEQ ID NO: 3. As an alternative or supplement, the anti-TMPRSS6 antibody of this disclosure comprises: LCCDR1 having no more than 3 amino acid variations (e.g., no more than 3, 2, or 1 amino acid variation) compared to LCCDR1 having the amino acid sequence of SEQ ID NO: 4; LCCDR2 having no more than 3 amino acid variations (e.g., no more than 3, 2, or 1 amino acid variation) compared to LCCDR2 having the amino acid sequence of RAN; and / or LCCDR3 having no more than 3 amino acid variations (e.g., no more than 3, 2, or 1 amino acid variation) compared to LCCDR3 having the amino acid sequence of SEQ ID NO: 6.

[0163] In some embodiments, the anti-TMPRSS6 antibody of this disclosure comprises: an HC CDR1 having at least 80% (e.g., at least 80%, at least 85%, 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%, or at least 99%) identity with an HC CDR1 having the amino acid sequence SEQ ID NO: 1; an HC CDR2 having at least 80% (e.g., at least 80%, at least 85%, 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%, or at least 99%) identity with an HC CDR2 having the amino acid sequence SEQ ID NO: 2; and / or an HC CDR2 having the amino acid sequence SEQ ID NO: 3. CDR3 having at least 80% (e.g., at least 80%, at least 85%, 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%, or at least 99%) identity with HC CDR3. As an alternative or supplement, the anti-TMPRSS6 antibody of this disclosure comprises: an LC CDR1 having at least 80% (e.g., at least 80%, at least 85%, 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%, or at least 99%) identity with an LC CDR1 having the amino acid sequence of SEQ ID NO: 4; an LC CDR2 having at least 80% (e.g., at least 80%, at least 85%, 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%, or at least 99%) identity with an LC CDR2 having the amino acid sequence of SEQ ID NO: 6; and / or an LC CDR2 having the amino acid sequence of SEQ ID NO: 6. CDR3 having at least 80% (e.g., at least 80%, at least 85%, 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%, or at least 99%) identity with LC CDR3.

[0164] In some embodiments, the anti-TMPRSS6 antibody of this disclosure comprises a VH containing the amino acid sequence of SEQ ID NO: 7. Alternatively or supplementally, the anti-TMPRSS6 antibody of this disclosure comprises a VL containing the amino acid sequence of SEQ ID NO: 8.

[0165] In some embodiments, the anti-TMPRSS6 antibody of this disclosure comprises a VH containing no more than 20 amino acid variations (e.g., no more than 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1 amino acid variation) compared to the VH shown in SEQ ID NO: 7. Alternatively or supplementally, the anti-TMPRSS6 antibody of this disclosure comprises a VL containing no more than 20 amino acid variations (e.g., no more than 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 9, 8, 7, 6, 5, 4, 3, 2, or 1 amino acid variation) compared to the VL shown in SEQ ID NO: 8. In some embodiments, the number of amino acid variations (e.g., not exceeding 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 10, 9, 8, 7, 6, 5, 4, 3, 2 or 1 amino acid variation) may appear in VH of SEQ ID NO: 7 and / or VL of SEQ ID NO: 8, except for any CDR sequences therein. In some embodiments, the anti-TMPRSS6 antibody provided herein comprises a heavy chain variable sequence and / or a light chain variable sequence, wherein the heavy chain variable sequence comprises a frame sequence having a variation of no more than 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 10, 9, 8, 7, 6, 5, 4, 3, 2 or 1 amino acid variation compared to the frame sequence of VH in SEQ ID NO: 7, and the light chain variable sequence comprises a frame sequence having a variation of no more than 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 10, 9, 8, 7, 6, 5, 4, 3, 2 or 1 amino acid variation compared to the frame sequence of VL in SEQ ID NO: 8.

[0166] In some embodiments, the anti-TMPRSS6 antibody of this disclosure comprises VH, which contains an amino acid sequence having at least 80% (e.g., at least 80%, at least 85%, 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%, or at least 99%) identity with the VH shown in SEQ ID NO: 7. Alternatively or supplementally, the anti-TMPRSS6 antibody of this disclosure comprises VL, which contains an amino acid sequence having at least 80% (e.g., at least 80%, at least 85%, 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%, or at least 99%) identity with the VL shown in SEQ ID NO: 8. In some implementations, the degree of sequence variation (e.g., at least 75%, at least 80%, at least 85%, 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%, or at least 99%) may appear in VH of SEQ ID NO: 7 and / or VL of SEQ ID NO: 8, except for any CDR sequences therein. In some embodiments, the anti-TMPRSS6 antibody provided herein comprises a heavy chain variable sequence and / or a light chain variable sequence, wherein the heavy chain variable sequence has at least 75%, at least 80%, at least 85%, 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%, or at least 99% identity with the frame sequence of VH in SEQ ID NO: 7, and the light chain variable sequence comprises a frame sequence having at least 75%, at least 80%, at least 85%, 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%, or at least 99% identity with the frame sequence of VL in SEQ ID NO: 8.

[0167] In some embodiments, the anti-TMPRSS6 antibody of this disclosure comprises HC CDR1, HC CDR2, and HC CDR3, which are heavy chain variable domains having the amino acid sequence of SEQ ID NO: 19 or 78. Alternatively or supplementally, the anti-TMPRSS6 antibody of this disclosure comprises LC CDR1, LC CDR2, and LC CDR3, which are light chain variable domains having the amino acid sequence of SEQ ID NO: 20.

[0168] In some embodiments, according to the Kabat definition system, the anti-TMPRSS6 antibody of this disclosure comprises HC CDR1 having the amino acid sequence of SEQ ID NO: 13, HC CDR2 having the amino acid sequence of SEQ ID NO: 14, HC CDR3 having the amino acid sequence of SEQ ID NO: 15, LC CDR1 having the amino acid sequence of SEQ ID NO: 16, LC CDR2 having the amino acid sequence of WAF, and LC CDR3 having the amino acid sequence of SEQ ID NO: 18.

[0169] In some embodiments, the anti-TMPRSS6 antibody of this disclosure comprises HC CDR1, HC CDR2, and HCCDR3, which, compared to HC CDR1 having the amino acid sequence of SEQ ID NO: 13, HC CDR2 having the amino acid sequence of SEQ ID NO: 14, and HC CDR3 having the amino acid sequence of SEQ ID NO: 15, collectively contain no more than 5 amino acid variations (e.g., no more than 5, 4, 3, 2, or 1 amino acid variation). Alternatively or supplementarily, the anti-TMPRSS6 antibody of this disclosure comprises LC CDR1, LC CDR2, and LC CDR3, which, compared to LC CDR1 having the amino acid sequence of SEQ ID NO: 16, LC CDR2 having the WAF amino acid sequence, and LC CDR3 having the amino acid sequence of SEQ ID NO: 18, collectively contain no more than 5 amino acid variations (e.g., no more than 5, 4, 3, 2, or 1 amino acid variation).

[0170] In some embodiments, the anti-TMPRSS6 antibody of this disclosure comprises HC CDR1, HC CDR2, and HCCDR3, which together have at least 80% (e.g., at least 80%, at least 85%, 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%, or at least 99%) identity with HC CDR1 having the amino acid sequence of SEQ ID NO: 13, HC CDR2 having the amino acid sequence of SEQ ID NO: 14, and HC CDR3 having the amino acid sequence of SEQ ID NO: 15. As an alternative or supplement, the anti-TMPRSS6 antibody of this disclosure comprises LC CDR1, LC CDR2 and LC CDR3, which together have at least 80% (e.g. at least 80%, at least 85%, 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% or at least 99%) identity with LC CDR1 having the amino acid sequence of SEQ ID NO: 16, LC CDR2 having the amino acid sequence of WAF and LC CDR3 having the amino acid sequence of SEQ ID NO: 18.

[0171] In some embodiments, the anti-TMPRSS6 antibody of this disclosure comprises: HC CDR1 having no more than 3 amino acid variations (e.g., no more than 3, 2, or 1 amino acid variation) compared to HC CDR1 having the amino acid sequence of SEQ ID NO: 13; HC CDR2 having no more than 3 amino acid variations (e.g., no more than 3, 2, or 1 amino acid variation) compared to HC CDR2 having the amino acid sequence of SEQ ID NO: 14; and / or HCCDR3 having no more than 3 amino acid variations (e.g., no more than 3, 2, or 1 amino acid variation) compared to HC CDR3 having the amino acid sequence of SEQ ID NO: 15. As an alternative or supplement, the anti-TMPRSS6 antibody of this disclosure comprises: LCCDR1 having no more than 3 amino acid variations (e.g., no more than 3, 2, or 1 amino acid variation) compared to LCCDR1 having the amino acid sequence of SEQ ID NO: 16; LCCDR2 having no more than 3 amino acid variations (e.g., no more than 3, 2, or 1 amino acid variation) compared to LCCDR2 having the amino acid sequence of WAF; and / or LCCDR3 having no more than 3 amino acid variations (e.g., no more than 3, 2, or 1 amino acid variation) compared to LCCDR3 having the amino acid sequence of SEQ ID NO: 18.

[0172] In some embodiments, the anti-TMPRSS6 antibody of this disclosure comprises: an HC CDR1 having at least 80% (e.g., at least 80%, at least 85%, 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%, or at least 99%) identity with an HC CDR1 having the amino acid sequence of SEQ ID NO: 13; an HC CDR2 having at least 80% (e.g., at least 80%, at least 85%, 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%, or at least 99%) identity with an HC CDR2 having the amino acid sequence of SEQ ID NO: 14; and / or an HC CDR2 having the amino acid sequence of SEQ ID NO: 15. CDR3 having at least 80% (e.g., at least 80%, at least 85%, 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%, or at least 99%) identity with HC CDR3. As an alternative or supplement, the anti-TMPRSS6 antibody of this disclosure comprises: an LC CDR1 having at least 80% (e.g., at least 80%, at least 85%, 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%, or at least 99%) identity with the amino acid sequence having SEQ ID NO: 16; an LC CDR2 having at least 80% (e.g., at least 80%, at least 85%, 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%, or at least 99%) identity with the amino acid sequence having SEQ ID NO: 16; and / or ... The LCCDR3 of the 18 amino acid sequence has at least 80% (e.g., at least 80%, at least 85%, 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%, or at least 99%) identity.

[0173] In some embodiments, the anti-TMPRSS6 antibody of this disclosure comprises a VH containing the amino acid sequence of SEQ ID NO: 19 or 78. Alternatively or supplementally, the anti-TMPRSS6 antibody of this disclosure comprises a VL containing the amino acid sequence of SEQ ID NO: 20.

[0174] In some embodiments, the anti-TMPRSS6 antibody of this disclosure comprises a VH containing no more than 20 amino acid variations (e.g., no more than 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1 amino acid variation) compared to the VH shown in SEQ ID NO: 19 or 78. Alternatively or supplementally, the anti-TMPRSS6 antibody of this disclosure comprises a VL containing no more than 20 amino acid variations (e.g., no more than 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 9, 8, 7, 6, 5, 4, 3, 2, or 1 amino acid variation) compared to the VL shown in SEQ ID NO: 20. In some embodiments, the number of amino acid variations (e.g., no more than 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 10, 9, 8, 7, 6, 5, 4, 3, 2 or 1 amino acid variation) may appear in VH of SEQ ID NO: 19 or 78 and / or VL of SEQ ID NO: 20, except for any CDR sequences therein. In some embodiments, the anti-TMPRSS6 antibody provided herein comprises a heavy chain variable sequence and / or a light chain variable sequence, wherein the heavy chain variable sequence comprises a frame sequence having a variation of no more than 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 10, 9, 8, 7, 6, 5, 4, 3, 2 or 1 amino acid variation compared to the frame sequence of VH in SEQ ID NO: 19 or 78, and the light chain variable sequence comprises a frame sequence having a variation of no more than 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 10, 9, 8, 7, 6, 5, 4, 3, 2 or 1 amino acid variation compared to the frame sequence of VL in SEQ ID NO: 20.

[0175] In some embodiments, the anti-TMPRSS6 antibody of this disclosure comprises a VH containing an amino acid sequence having at least 80% (e.g., at least 80%, at least 85%, 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%, or at least 99%) identity with the VH shown in SEQ ID NO: 19 or 78. Alternatively or supplementally, the anti-TMPRSS6 antibody of this disclosure comprises a VL containing an amino acid sequence having at least 80% (e.g., at least 80%, at least 85%, 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%, or at least 99%) identity with the VL shown in SEQ ID NO: 20. In some embodiments, the degree of sequence variation (e.g., at least 75%, at least 80%, at least 85%, 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%, or at least 99%) may appear in VH of SEQ ID NO: 19 or 78 and / or VL of SEQ ID NO: 20, except for any CDR sequences therein. In some embodiments, the anti-TMPRSS6 antibody provided herein comprises a heavy chain variable sequence and / or a light chain variable sequence, wherein the heavy chain variable sequence has at least 75%, at least 80%, at least 85%, 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%, or at least 99% identity with the frame sequence of VH of SEQ ID NO: 19 or 78, and the light chain variable sequence comprises a frame sequence having at least 75%, at least 80%, at least 85%, 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%, or at least 99% identity with the frame sequence of VL of SEQ ID NO: 20.

[0176] In some embodiments, the anti-TMPRSS6 antibody of this disclosure comprises HC CDR1, HC CDR2, and HC CDR3, which are heavy chain variable domains having the amino acid sequence of SEQ ID NO: 30. Alternatively or supplementally, the anti-TMPRSS6 antibody of this disclosure comprises LC CDR1, LCCDR2, and LC CDR3, which are light chain variable domains having the amino acid sequence of SEQ ID NO: 31.

[0177] In some embodiments, according to the Kabat definition system, the anti-TMPRSS6 antibody of this disclosure comprises HC CDR1 having the amino acid sequence of SEQ ID NO: 24, HC CDR2 having the amino acid sequence of SEQ ID NO: 25, HC CDR3 having the amino acid sequence of SEQ ID NO: 26, LC CDR1 having the amino acid sequence of SEQ ID NO: 27, LC CDR2 having the amino acid sequence of WAT, and LC CDR3 having the amino acid sequence of SEQ ID NO: 29.

[0178] In some embodiments, the anti-TMPRSS6 antibody of this disclosure comprises HC CDR1, HC CDR2, and HCCDR3, which, compared to HC CDR1 having the amino acid sequence of SEQ ID NO: 24, HC CDR2 having the amino acid sequence of SEQ ID NO: 25, and HC CDR3 having the amino acid sequence of SEQ ID NO: 26, collectively contain no more than 5 amino acid variations (e.g., no more than 5, 4, 3, 2, or 1 amino acid variation). Alternatively or supplementarily, the anti-TMPRSS6 antibody of this disclosure comprises LC CDR1, LC CDR2, and LC CDR3, which, compared to LC CDR1 having the amino acid sequence of SEQ ID NO: 27, LC CDR2 having the amino acid sequence of WAT, and LC CDR3 having the amino acid sequence of SEQ ID NO: 29, collectively contain no more than 5 amino acid variations (e.g., no more than 5, 4, 3, 2, or 1 amino acid variation).

[0179] In some embodiments, the anti-TMPRSS6 antibody of this disclosure comprises HC CDR1, HC CDR2, and HCCDR3, which together have at least 80% (e.g., at least 80%, at least 85%, 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%, or at least 99%) identity with HC CDR1 having the amino acid sequence of SEQ ID NO: 24, HC CDR2 having the amino acid sequence of SEQ ID NO: 25, and HC CDR3 having the amino acid sequence of SEQ ID NO: 26. As an alternative or supplement, the anti-TMPRSS6 antibody of this disclosure comprises LC CDR1, LC CDR2 and LC CDR3, which together have at least 80% (e.g. at least 80%, at least 85%, 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% or at least 99%) identity with LC CDR1 having the amino acid sequence of SEQ ID NO: 27, LC CDR2 having the amino acid sequence of WAT and LC CDR3 having the amino acid sequence of SEQ ID NO: 29.

[0180] In some embodiments, the anti-TMPRSS6 antibody of this disclosure comprises: HC CDR1 having no more than 3 amino acid variations (e.g., no more than 3, 2, or 1 amino acid variation) compared to HC CDR1 having the amino acid sequence of SEQ ID NO: 24; HC CDR2 having no more than 3 amino acid variations (e.g., no more than 3, 2, or 1 amino acid variation) compared to HC CDR2 having the amino acid sequence of SEQ ID NO: 25; and / or HCCDR3 having no more than 3 amino acid variations (e.g., no more than 3, 2, or 1 amino acid variation) compared to HC CDR3 having the amino acid sequence of SEQ ID NO: 26. As an alternative or supplement, the anti-TMPRSS6 antibody of this disclosure comprises: LCCDR1 having no more than 3 amino acid variations (e.g., no more than 3, 2, or 1 amino acid variation) compared to LCCDR1 having the amino acid sequence of SEQ ID NO: 27; LCCDR2 having no more than 3 amino acid variations (e.g., no more than 3, 2, or 1 amino acid variation) compared to LCCDR2 having the amino acid sequence of WAT; and / or LCCDR3 having no more than 3 amino acid variations (e.g., no more than 3, 2, or 1 amino acid variation) compared to LCCDR3 having the amino acid sequence of SEQ ID NO: 29.

[0181] In some embodiments, the anti-TMPRSS6 antibody of this disclosure comprises: HC CDR1 having at least 80% (e.g., at least 80%, at least 85%, 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%, or at least 99%) identity with HC CDR1 having the amino acid sequence of SEQ ID NO: 24; HC CDR2 having at least 80% (e.g., at least 80%, at least 85%, 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%, or at least 99%) identity with HC CDR2 having the amino acid sequence of SEQ ID NO: 25; and / or HC CDR2 having the amino acid sequence of SEQ ID NO: 26. CDR3 having at least 80% (e.g., at least 80%, at least 85%, 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%, or at least 99%) identity with HC CDR3. As an alternative or supplement, the anti-TMPRSS6 antibody of this disclosure comprises: an LC CDR1 having at least 80% (e.g., at least 80%, at least 85%, 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%, or at least 99%) identity with an LC CDR1 having the amino acid sequence of SEQ ID NO: 27; an LC CDR2 having at least 80% (e.g., at least 80%, at least 85%, 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%, or at least 99%) identity with an LC CDR2 having the amino acid sequence of SEQ ID NO: 27; and / or an LC CDR2 having the amino acid sequence of SEQ ID NO: 27. The LCCDR3 of the amino acid sequence 29 has at least 80% (e.g., at least 80%, at least 85%, 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%, or at least 99%) identity.

[0182] In some embodiments, the anti-TMPRSS6 antibody of this disclosure comprises VH containing the amino acid sequence of SEQ ID NO: 30. Alternatively or supplementally, the anti-TMPRSS6 antibody of this disclosure comprises VL containing the amino acid sequence of SEQ ID NO: 31.

[0183] In some embodiments, the anti-TMPRSS6 antibody of this disclosure comprises a VH containing no more than 20 amino acid variations (e.g., no more than 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1 amino acid variation) compared to the VH shown in SEQ ID NO: 30. Alternatively or supplementally, the anti-TMPRSS6 antibody of this disclosure comprises a VL containing no more than 20 amino acid variations (e.g., no more than 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 9, 8, 7, 6, 5, 4, 3, 2, or 1 amino acid variation) compared to the VL shown in SEQ ID NO: 31. In some embodiments, the number of amino acid variations (e.g., not exceeding 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 10, 9, 8, 7, 6, 5, 4, 3, 2 or 1 amino acid variation) may appear in VH of SEQ ID NO: 30 and / or VL of SEQ ID NO: 31, except for any CDR sequences therein. In some embodiments, the anti-TMPRSS6 antibody provided herein comprises a heavy chain variable sequence and / or a light chain variable sequence, wherein the heavy chain variable sequence comprises a frame sequence having a variation of no more than 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 10, 9, 8, 7, 6, 5, 4, 3, 2 or 1 amino acid variation compared to the frame sequence of VH in SEQ ID NO: 30, and the light chain variable sequence comprises a frame sequence having a variation of no more than 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 10, 9, 8, 7, 6, 5, 4, 3, 2 or 1 amino acid variation compared to the frame sequence of VL in SEQ ID NO: 31.

[0184] In some embodiments, the anti-TMPRSS6 antibody of this disclosure comprises a VH containing an amino acid sequence having at least 80% (e.g., at least 80%, at least 85%, 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%, or at least 99%) identity with the VH shown in SEQ ID NO: 30. Alternatively or supplementally, the anti-TMPRSS6 antibody of this disclosure comprises a VL containing an amino acid sequence having at least 80% (e.g., at least 80%, at least 85%, 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%, or at least 99%) identity with the VL shown in SEQ ID NO: 31. In some implementations, the degree of sequence variation (e.g., at least 75%, at least 80%, at least 85%, 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%, or at least 99%) may appear in VH of SEQ ID NO: 30 and / or VL of SEQ ID NO: 31, except for any CDR sequences therein. In some embodiments, the anti-TMPRSS6 antibody provided herein comprises a heavy chain variable sequence and / or a light chain variable sequence, wherein the heavy chain variable sequence has at least 75%, at least 80%, at least 85%, 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%, or at least 99% identity with the frame sequence of VH in SEQ ID NO: 30, and the light chain variable sequence comprises a frame sequence having at least 75%, at least 80%, at least 85%, 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%, or at least 99% identity with the frame sequence of VL in SEQ ID NO: 31.

[0185] In some embodiments, the anti-TMPRSS6 antibody of this disclosure comprises HC CDR1, HC CDR2, and HC CDR3, which are heavy chain variable domains having the amino acid sequence of SEQ ID NO: 37. Alternatively or supplementally, the anti-TMPRSS6 antibody of this disclosure comprises LC CDR1, LCCDR2, and LC CDR3, which are light chain variable domains having the amino acid sequence of SEQ ID NO: 38.

[0186] In some embodiments, according to the Kabat definition system, the anti-TMPRSS6 antibody of this disclosure comprises HC CDR1 having the amino acid sequence of SEQ ID NO: 1, HC CDR2 having the amino acid sequence of SEQ ID NO: 2, HC CDR3 having the amino acid sequence of SEQ ID NO: 35, LC CDR1 having the amino acid sequence of SEQ ID NO: 36, LC CDR2 having the amino acid sequence of RAN, and LC CDR3 having the amino acid sequence of SEQ ID NO: 6.

[0187] In some embodiments, the anti-TMPRSS6 antibody of this disclosure comprises HC CDR1, HC CDR2, and HCCDR3, which, compared to HC CDR1 having the amino acid sequence of SEQ ID NO: 1, HCCDR2 having the amino acid sequence of SEQ ID NO: 2, and HC CDR3 having the amino acid sequence of SEQ ID NO: 35, collectively contain no more than 5 amino acid variations (e.g., no more than 5, 4, 3, 2, or 1 amino acid variation). Alternatively or supplementarily, the anti-TMPRSS6 antibody of this disclosure comprises LC CDR1, LC CDR2, and LC CDR3, which, compared to LC CDR1 having the amino acid sequence of SEQ ID NO: 36, LC CDR2 having the amino acid sequence of RAN, and LC CDR3 having the amino acid sequence of SEQ ID NO: 6, collectively contain no more than 5 amino acid variations (e.g., no more than 5, 4, 3, 2, or 1 amino acid variation).

[0188] In some embodiments, the anti-TMPRSS6 antibody of this disclosure comprises HC CDR1, HC CDR2, and HCCDR3, which together have at least 80% (e.g., at least 80%, at least 85%, 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%, or at least 99%) identity with HC CDR1 having the amino acid sequence of SEQ ID NO: 1, HC CDR2 having the amino acid sequence of SEQ ID NO: 2, and HC CDR3 having the amino acid sequence of SEQ ID NO: 35. As an alternative or supplement, the anti-TMPRSS6 antibody of this disclosure comprises LC CDR1, LCCDR2, and LC CDR3, which together have at least 80% (e.g., at least 80%, at least 85%, 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%, or at least 99%) identity with LC CDR1 having the amino acid sequence of SEQ ID NO: 36, LC CDR2 having the amino acid sequence of RAN, and LC CDR3 having the amino acid sequence of SEQ ID NO: 6.

[0189] In some embodiments, the anti-TMPRSS6 antibody of this disclosure comprises: HC CDR1 having no more than 3 amino acid variations (e.g., no more than 3, 2, or 1 amino acid variation) compared to HC CDR1 having the amino acid sequence of SEQ ID NO: 1; HC CDR2 having no more than 3 amino acid variations (e.g., no more than 3, 2, or 1 amino acid variation) compared to HC CDR2 having the amino acid sequence of SEQ ID NO: 2; and / or HCCDR3 having no more than 3 amino acid variations (e.g., no more than 3, 2, or 1 amino acid variation) compared to HC CDR3 having the amino acid sequence of SEQ ID NO: 35. As an alternative or supplement, the anti-TMPRSS6 antibody of this disclosure comprises: LCCDR1 having no more than 3 amino acid variations (e.g., no more than 3, 2, or 1 amino acid variation) compared to LCCDR1 having the amino acid sequence of SEQ ID NO: 36; LCCDR2 having no more than 3 amino acid variations (e.g., no more than 3, 2, or 1 amino acid variation) compared to LCCDR2 having the amino acid sequence of RAN; and / or LCCDR3 having no more than 3 amino acid variations (e.g., no more than 3, 2, or 1 amino acid variation) compared to LCCDR3 having the amino acid sequence of SEQ ID NO: 6.

[0190] In some embodiments, the anti-TMPRSS6 antibody of this disclosure comprises: an HC CDR1 having at least 80% (e.g., at least 80%, at least 85%, 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%, or at least 99%) identity with an HC CDR1 having the amino acid sequence of SEQ ID NO: 1; an HC CDR2 having at least 80% (e.g., at least 80%, at least 85%, 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%, or at least 99%) identity with an HC CDR2 having the amino acid sequence of SEQ ID NO: 2; and / or an HC CDR2 having the amino acid sequence of SEQ ID NO: 35. CDR3 having at least 80% (e.g., at least 80%, at least 85%, 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%, or at least 99%) identity with HC CDR3. As an alternative or supplement, the anti-TMPRSS6 antibody of this disclosure comprises: an LC CDR1 having at least 80% (e.g., at least 80%, at least 85%, 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%, or at least 99%) identity with an LC CDR1 having the amino acid sequence of SEQ ID NO: 36; an LC CDR2 having at least 80% (e.g., at least 80%, at least 85%, 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%, or at least 99%) identity with an LC CDR2 having the amino acid sequence of SEQ ID NO: 6; and / or an LC CDR2 having the amino acid sequence of SEQ ID NO: 6. CDR3 having at least 80% (e.g., at least 80%, at least 85%, 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%, or at least 99%) identity with LC CDR3.

[0191] In some embodiments, the anti-TMPRSS6 antibody of this disclosure comprises a VH containing the amino acid sequence of SEQ ID NO: 37. Alternatively or supplementally, the anti-TMPRSS6 antibody of this disclosure comprises a VL containing the amino acid sequence of SEQ ID NO: 38.

[0192] In some embodiments, the anti-TMPRSS6 antibody of this disclosure comprises a VH containing no more than 20 amino acid variations (e.g., no more than 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1 amino acid variation) compared to the VH shown in SEQ ID NO: 37. Alternatively or supplementally, the anti-TMPRSS6 antibody of this disclosure comprises a VL containing no more than 20 amino acid variations (e.g., no more than 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 9, 8, 7, 6, 5, 4, 3, 2, or 1 amino acid variation) compared to the VL shown in SEQ ID NO: 38. In some embodiments, the number of amino acid variations (e.g., not exceeding 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 10, 9, 8, 7, 6, 5, 4, 3, 2 or 1 amino acid variation) may appear in VH of SEQ ID NO: 37 and / or VL of SEQ ID NO: 38, except for any CDR sequences therein. In some embodiments, the anti-TMPRSS6 antibody provided herein comprises a heavy chain variable sequence and / or a light chain variable sequence, wherein the heavy chain variable sequence comprises a frame sequence having a variation of no more than 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 10, 9, 8, 7, 6, 5, 4, 3, 2 or 1 amino acid variation compared to the frame sequence of VH in SEQ ID NO: 37, and the light chain variable sequence comprises a frame sequence having a variation of no more than 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 10, 9, 8, 7, 6, 5, 4, 3, 2 or 1 amino acid variation compared to the frame sequence of VL in SEQ ID NO: 38.

[0193] In some embodiments, the anti-TMPRSS6 antibody of this disclosure comprises a VH containing an amino acid sequence having at least 80% (e.g., at least 80%, at least 85%, 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%, or at least 99%) identity with the VH shown in SEQ ID NO: 37. Alternatively or supplementally, the anti-TMPRSS6 antibody of this disclosure comprises a VL containing an amino acid sequence having at least 80% (e.g., at least 80%, at least 85%, 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%, or at least 99%) identity with the VL shown in SEQ ID NO: 38. In some implementations, the degree of sequence variation (e.g., at least 75%, at least 80%, at least 85%, 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%, or at least 99%) may appear in VH of SEQ ID NO: 37 and / or VL of SEQ ID NO: 38, except for any CDR sequences therein. In some embodiments, the anti-TMPRSS6 antibody provided herein comprises a heavy chain variable sequence and / or a light chain variable sequence, wherein the heavy chain variable sequence has at least 75%, at least 80%, at least 85%, 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%, or at least 99% identity with the frame sequence of VH in SEQ ID NO: 37, and the light chain variable sequence comprises a frame sequence having at least 75%, at least 80%, at least 85%, 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%, or at least 99% identity with the frame sequence of VL in SEQ ID NO: 38.

[0194] In some embodiments, the anti-TMPRSS6 antibody of this disclosure comprises HC CDR1, HC CDR2, and HC CDR3, which are heavy chain variable domains having the amino acid sequence of SEQ ID NO: 44. Alternatively or supplementally, the anti-TMPRSS6 antibody of this disclosure comprises LC CDR1, LCCDR2, and LC CDR3, which are light chain variable domains having the amino acid sequence of SEQ ID NO: 45.

[0195] In some embodiments, according to the Kabat definition system, the anti-TMPRSS6 antibody of this disclosure comprises HC CDR1 having the amino acid sequence of SEQ ID NO: 13, HC CDR2 having the amino acid sequence of SEQ ID NO: 43, HC CDR3 having the amino acid sequence of SEQ ID NO: 15, LC CDR1 having the amino acid sequence of SEQ ID NO: 16, LC CDR2 having the amino acid sequence of WAF, and LC CDR3 having the amino acid sequence of SEQ ID NO: 18.

[0196] In some embodiments, the anti-TMPRSS6 antibody of this disclosure comprises HC CDR1, HC CDR2, and HCCDR3, which, compared to HC CDR1 having the amino acid sequence of SEQ ID NO: 13, HC CDR2 having the amino acid sequence of SEQ ID NO: 43, and HC CDR3 having the amino acid sequence of SEQ ID NO: 15, collectively contain no more than 5 amino acid variations (e.g., no more than 5, 4, 3, 2, or 1 amino acid variation). Alternatively or supplementarily, the anti-TMPRSS6 antibody of this disclosure comprises LC CDR1, LC CDR2, and LC CDR3, which, compared to LC CDR1 having the amino acid sequence of SEQ ID NO: 16, LC CDR2 having the WAF amino acid sequence, and LC CDR3 having the amino acid sequence of SEQ ID NO: 18, collectively contain no more than 5 amino acid variations (e.g., no more than 5, 4, 3, 2, or 1 amino acid variation).

[0197] In some embodiments, the anti-TMPRSS6 antibody of this disclosure comprises HC CDR1, HC CDR2, and HCCDR3, which together have at least 80% (e.g., at least 80%, at least 85%, 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%, or at least 99%) identity with HC CDR1 having the amino acid sequence of SEQ ID NO: 13, HC CDR2 having the amino acid sequence of SEQ ID NO: 43, and HC CDR3 having the amino acid sequence of SEQ ID NO: 15. As an alternative or supplement, the anti-TMPRSS6 antibody of this disclosure comprises LC CDR1, LC CDR2 and LC CDR3, which together have at least 80% (e.g. at least 80%, at least 85%, 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% or at least 99%) identity with LC CDR1 having the amino acid sequence of SEQ ID NO: 16, LC CDR2 having the amino acid sequence of WAF and LC CDR3 having the amino acid sequence of SEQ ID NO: 18.

[0198] In some embodiments, the anti-TMPRSS6 antibody of this disclosure comprises: HC CDR1 having no more than 3 amino acid variations (e.g., no more than 3, 2, or 1 amino acid variation) compared to HC CDR1 having the amino acid sequence of SEQ ID NO: 13; HC CDR2 having no more than 3 amino acid variations (e.g., no more than 3, 2, or 1 amino acid variation) compared to HC CDR2 having the amino acid sequence of SEQ ID NO: 43; and / or HCCDR3 having no more than 3 amino acid variations (e.g., no more than 3, 2, or 1 amino acid variation) compared to HC CDR3 having the amino acid sequence of SEQ ID NO: 15. As an alternative or supplement, the anti-TMPRSS6 antibody of this disclosure comprises: LCCDR1 having no more than 3 amino acid variations (e.g., no more than 3, 2, or 1 amino acid variation) compared to LCCDR1 having the amino acid sequence of SEQ ID NO: 16; LCCDR2 having no more than 3 amino acid variations (e.g., no more than 3, 2, or 1 amino acid variation) compared to LCCDR2 having the amino acid sequence of WAF; and / or LCCDR3 having no more than 3 amino acid variations (e.g., no more than 3, 2, or 1 amino acid variation) compared to LCCDR3 having the amino acid sequence of SEQ ID NO: 18.

[0199] In some embodiments, the anti-TMPRSS6 antibody of this disclosure comprises: HC CDR1 having at least 80% (e.g., at least 80%, at least 85%, 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%, or at least 99%) identity with HC CDR1 having the amino acid sequence of SEQ ID NO: 13; HC CDR2 having at least 80% (e.g., at least 80%, at least 85%, 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%, or at least 99%) identity with HC CDR2 having the amino acid sequence of SEQ ID NO: 43; and / or HC CDR2 having the amino acid sequence of SEQ ID NO: 15. CDR3 having at least 80% (e.g., at least 80%, at least 85%, 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%, or at least 99%) identity with HC CDR3. As an alternative or supplement, the anti-TMPRSS6 antibody of this disclosure comprises: an LC CDR1 having at least 80% (e.g., at least 80%, at least 85%, 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%, or at least 99%) identity with the amino acid sequence having SEQ ID NO: 16; an LC CDR2 having at least 80% (e.g., at least 80%, at least 85%, 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%, or at least 99%) identity with the amino acid sequence having SEQ ID NO: 16; and / or ... The LCCDR3 of the 18 amino acid sequence has at least 80% (e.g., at least 80%, at least 85%, 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%, or at least 99%) identity.

[0200] In some embodiments, the anti-TMPRSS6 antibody of this disclosure comprises a VH containing the amino acid sequence of SEQ ID NO: 44. Alternatively or supplementally, the anti-TMPRSS6 antibody of this disclosure comprises a VL containing the amino acid sequence of SEQ ID NO: 45.

[0201] In some embodiments, the anti-TMPRSS6 antibody of this disclosure comprises a VH containing no more than 20 amino acid variations (e.g., no more than 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1 amino acid variation) compared to the VH shown in SEQ ID NO: 44. Alternatively or supplementally, the anti-TMPRSS6 antibody of this disclosure comprises a VL containing no more than 20 amino acid variations (e.g., no more than 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 9, 8, 7, 6, 5, 4, 3, 2, or 1 amino acid variation) compared to the VL shown in SEQ ID NO: 45. In some embodiments, the number of amino acid variations (e.g., not exceeding 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 10, 9, 8, 7, 6, 5, 4, 3, 2 or 1 amino acid variation) may appear in VH of SEQ ID NO: 44 and / or VL of SEQ ID NO: 45, except for any CDR sequences therein. In some embodiments, the anti-TMPRSS6 antibody provided herein comprises a heavy chain variable sequence and / or a light chain variable sequence, wherein the heavy chain variable sequence comprises a frame sequence having a variation of no more than 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 10, 9, 8, 7, 6, 5, 4, 3, 2 or 1 amino acid variation compared to the frame sequence of VH in SEQ ID NO: 44, and the light chain variable sequence comprises a frame sequence having a variation of no more than 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 10, 9, 8, 7, 6, 5, 4, 3, 2 or 1 amino acid variation compared to the frame sequence of VL in SEQ ID NO: 45.

[0202] In some embodiments, the anti-TMPRSS6 antibody of this disclosure comprises a VH containing an amino acid sequence having at least 80% (e.g., at least 80%, at least 85%, 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%, or at least 99%) identity with the VH shown in SEQ ID NO: 44. Alternatively or supplementally, the anti-TMPRSS6 antibody of this disclosure comprises a VL containing an amino acid sequence having at least 80% (e.g., at least 80%, at least 85%, 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%, or at least 99%) identity with the VL shown in SEQ ID NO: 45. In some implementations, the degree of sequence variation (e.g., at least 75%, at least 80%, at least 85%, 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%, or at least 99%) may appear in VH of SEQ ID NO: 44 and / or VL of SEQ ID NO: 45, except for any CDR sequences therein. In some embodiments, the anti-TMPRSS6 antibody provided herein comprises a heavy chain variable sequence and / or a light chain variable sequence, wherein the heavy chain variable sequence has at least 75%, at least 80%, at least 85%, 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%, or at least 99% identity with the frame sequence of VH in SEQ ID NO: 44, and the light chain variable sequence comprises a frame sequence having at least 75%, at least 80%, at least 85%, 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%, or at least 99% identity with the frame sequence of VL in SEQ ID NO: 45.

[0203] In some embodiments, the anti-TMPRSS6 antibody of this disclosure comprises HC CDR1, HC CDR2, and HC CDR3, which are heavy chain variable domains having the amino acid sequence of SEQ ID NO: 49. Alternatively or supplementally, the anti-TMPRSS6 antibody of this disclosure comprises LC CDR1, LCCDR2, and LC CDR3, which are light chain variable domains having the amino acid sequence of SEQ ID NO: 50.

[0204] In some embodiments, according to the Kabat definition system, the anti-TMPRSS6 antibody of this disclosure comprises HC CDR1 having the amino acid sequence of SEQ ID NO: 24, HC CDR2 having the amino acid sequence of SEQ ID NO: 48, HC CDR3 having the amino acid sequence of SEQ ID NO: 26, LC CDR1 having the amino acid sequence of SEQ ID NO: 27, LC CDR2 having the amino acid sequence of WAT, and LC CDR3 having the amino acid sequence of SEQ ID NO: 29.

[0205] In some embodiments, the anti-TMPRSS6 antibody of this disclosure comprises HC CDR1, HC CDR2, and HCCDR3, which, compared to HC CDR1 having the amino acid sequence of SEQ ID NO: 24, HC CDR2 having the amino acid sequence of SEQ ID NO: 48, and HC CDR3 having the amino acid sequence of SEQ ID NO: 26, collectively contain no more than 5 amino acid variations (e.g., no more than 5, 4, 3, 2, or 1 amino acid variation). Alternatively or supplementarily, the anti-TMPRSS6 antibody of this disclosure comprises LC CDR1, LC CDR2, and LC CDR3, which, compared to LC CDR1 having the amino acid sequence of SEQ ID NO: 27, LC CDR2 having the amino acid sequence of WAT, and LC CDR3 having the amino acid sequence of SEQ ID NO: 29, collectively contain no more than 5 amino acid variations (e.g., no more than 5, 4, 3, 2, or 1 amino acid variation).

[0206] In some embodiments, the anti-TMPRSS6 antibody of this disclosure comprises HC CDR1, HC CDR2, and HCCDR3, which together have at least 80% (e.g., at least 80%, at least 85%, 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%, or at least 99%) identity with HC CDR1 having the amino acid sequence of SEQ ID NO: 24, HC CDR2 having the amino acid sequence of SEQ ID NO: 48, and HC CDR3 having the amino acid sequence of SEQ ID NO: 26. As an alternative or supplement, the anti-TMPRSS6 antibody of this disclosure comprises LC CDR1, LC CDR2 and LC CDR3, which together have at least 80% (e.g. at least 80%, at least 85%, 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% or at least 99%) identity with LC CDR1 having the amino acid sequence of SEQ ID NO: 27, LC CDR2 having the amino acid sequence of WAT and LC CDR3 having the amino acid sequence of SEQ ID NO: 29.

[0207] In some embodiments, the anti-TMPRSS6 antibody of this disclosure comprises: HC CDR1 having no more than 3 amino acid variations (e.g., no more than 3, 2, or 1 amino acid variation) compared to HC CDR1 having the amino acid sequence of SEQ ID NO: 24; HC CDR2 having no more than 3 amino acid variations (e.g., no more than 3, 2, or 1 amino acid variation) compared to HC CDR2 having the amino acid sequence of SEQ ID NO: 48; and / or HCCDR3 having no more than 3 amino acid variations (e.g., no more than 3, 2, or 1 amino acid variation) compared to HC CDR3 having the amino acid sequence of SEQ ID NO: 26. As an alternative or supplement, the anti-TMPRSS6 antibody of this disclosure comprises: LCCDR1 having no more than 3 amino acid variations (e.g., no more than 3, 2, or 1 amino acid variation) compared to LCCDR1 having the amino acid sequence of SEQ ID NO: 27; LCCDR2 having no more than 3 amino acid variations (e.g., no more than 3, 2, or 1 amino acid variation) compared to LCCDR2 having the amino acid sequence of WAT; and / or LCCDR3 having no more than 3 amino acid variations (e.g., no more than 3, 2, or 1 amino acid variation) compared to LCCDR3 having the amino acid sequence of SEQ ID NO: 29.

[0208] In some embodiments, the anti-TMPRSS6 antibody of this disclosure comprises: HC CDR1 having at least 80% (e.g., at least 80%, at least 85%, 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%, or at least 99%) identity with HC CDR1 having the amino acid sequence of SEQ ID NO: 24; HC CDR2 having at least 80% (e.g., at least 80%, at least 85%, 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%, or at least 99%) identity with HC CDR2 having the amino acid sequence of SEQ ID NO: 48; and / or HC CDR2 having the amino acid sequence of SEQ ID NO: 26. CDR3 having at least 80% (e.g., at least 80%, at least 85%, 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%, or at least 99%) identity with HC CDR3. As an alternative or supplement, the anti-TMPRSS6 antibody of this disclosure comprises: an LC CDR1 having at least 80% (e.g., at least 80%, at least 85%, 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%, or at least 99%) identity with an LC CDR1 having the amino acid sequence of SEQ ID NO: 27; an LC CDR2 having at least 80% (e.g., at least 80%, at least 85%, 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%, or at least 99%) identity with an LC CDR2 having the amino acid sequence of SEQ ID NO: 27; and / or an LC CDR2 having the amino acid sequence of SEQ ID NO: 27. The LCCDR3 of the amino acid sequence 29 has at least 80% (e.g., at least 80%, at least 85%, 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%, or at least 99%) identity.

[0209] In some embodiments, the anti-TMPRSS6 antibody of this disclosure comprises a VH containing the amino acid sequence of SEQ ID NO: 49. Alternatively or supplementally, the anti-TMPRSS6 antibody of this disclosure comprises a VL containing the amino acid sequence of SEQ ID NO: 50.

[0210] In some embodiments, the anti-TMPRSS6 antibody of this disclosure comprises a VH containing no more than 20 amino acid variations (e.g., no more than 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1 amino acid variation) compared to the VH described in SEQ ID NO: 49. Alternatively or supplementally, the anti-TMPRSS6 antibody of this disclosure comprises a VL containing no more than 20 amino acid variations (e.g., no more than 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 9, 8, 7, 6, 5, 4, 3, 2, or 1 amino acid variation) compared to the VL shown in SEQ ID NO: 50. In some embodiments, the number of amino acid variations (e.g., no more than 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 10, 9, 8, 7, 6, 5, 4, 3, 2 or 1 amino acid variation) may appear in VH of SEQ ID NO: 49 and / or VL of SEQ ID NO: 50, except for any CDR sequences therein. In some embodiments, the anti-TMPRSS6 antibody provided herein comprises a heavy chain variable sequence and / or a light chain variable sequence, wherein the heavy chain variable sequence comprises a frame sequence having a variation of no more than 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 10, 9, 8, 7, 6, 5, 4, 3, 2 or 1 amino acid variation compared to the frame sequence of VH in SEQ ID NO: 49, and the light chain variable sequence comprises a frame sequence having a variation of no more than 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 10, 9, 8, 7, 6, 5, 4, 3, 2 or 1 amino acid variation compared to the frame sequence of VL in SEQ ID NO: 50.

[0211] In some embodiments, the anti-TMPRSS6 antibody of this disclosure comprises a VH having at least 80% (e.g., at least 80%, at least 85%, 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%, or at least 99%) amino acid sequence identity with the VH shown in SEQ ID NO: 49. Alternatively or supplementally, the anti-TMPRSS6 antibody of this disclosure comprises a VL having at least 80% (e.g., at least 80%, at least 85%, 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%, or at least 99%) amino acid sequence identity with the VL shown in SEQ ID NO: 50. In some implementations, the degree of sequence variation (e.g., at least 75%, at least 80%, at least 85%, 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%, or at least 99%) may occur within VH of SEQ ID NO: 49 and / or VL of SEQ ID NO: 50, except for any CDR sequences therein. In some embodiments, the anti-TMPRSS6 antibody provided herein comprises a heavy chain variable sequence having at least 75%, at least 80%, at least 85%, 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%, or at least 99% identity with the frame sequence of VH of SEQ ID NO: 49, and / or comprises a light chain variable sequence containing a frame sequence having at least 75%, at least 80%, at least 85%, 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%, or at least 99% identity with the frame sequence of VL of SEQ ID NO: 50.

[0212] In some embodiments, the anti-TMPRSS6 antibody described herein is a chimeric antibody, which may comprise a heavy constant region and a light constant region derived from a human antibody. A chimeric antibody is an antibody having a variable region or a portion thereof derived from a first species and a constant region derived from a second species. Typically, in these chimeric antibodies, the variable regions of both the light and heavy chains mimic the variable regions derived from an antibody of one mammalian species (e.g., a non-human mammal, such as a mouse, rabbit, or rat), while the constant regions are sequence homologous to those of an antibody derived from another mammalian species (e.g., a human). In some embodiments, amino acid modifications may be made in the variable and / or constant regions.

[0213] In some embodiments, the anti-TMPRSS6 antibody of this disclosure comprises a VH domain and / or a VL domain selected from any of the anti-TMPRSS6 antibodies in Table 1, and includes a constant region comprising the amino acid sequence of a constant region of an IgG, IgE, IgM, IgD, IgA, or IgY immunoglobulin molecule, any class of immunoglobulin molecules (e.g., IgG1, IgG2, IgG3, IgG4, IgA1, and IgA2), or any subclass (e.g., IgG2a and IgG2b). Some non-limiting examples of human constant regions are described in the art, for example, see above, Kabat EA et al., (1991).

[0214] In some embodiments, the light chain of any anti-TMPRSS6 antibody described herein may also include a light chain constant region (CL), which may be any CL known in the art. In some instances, the CL is a κ light chain. In other instances, the CL is a λ light chain. In some embodiments, the CL is a κ light chain.

[0215] Other antibody heavy and light chain constant regions are well known in the art, for example, those available in the IMGT database (www.imgt.org) or in www.vbase2.org / vbstat.php, both of which are incorporated herein by reference.

[0216] In some embodiments, the anti-TMPRSS6 antibody described herein comprises a heavy chain comprising any VH or any variant thereof listed in Table 1 and a heavy chain constant region comprising at least 80%, at least 85%, 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%, or at least 99% of SEQ ID NO: 9. In some embodiments, the anti-TMPRSS6 antibody described herein comprises a heavy chain comprising any VH or any variant thereof listed in Table 1 and a heavy chain constant region containing no more than 20 amino acid variations (e.g., no more than 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1 amino acid variation) compared to SEQ ID NO: 9. In some embodiments, the anti-TMPRSS6 antibody described herein comprises a heavy chain comprising any VH or any variant thereof listed in Table 1 and the heavy chain constant region shown in SEQ ID NO: 9.

[0217] In some embodiments, the anti-TMPRSS6 antibody described herein comprises a heavy chain comprising any VH or any variant thereof listed in Table 1 and a heavy chain constant region having at least 80%, at least 85%, 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%, or at least 99% identity with SEQ ID NO: 21. In some embodiments, the anti-TMPRSS6 antibody described herein comprises a heavy chain comprising any VH or any variant thereof listed in Table 1 and a heavy chain constant region containing no more than 20 amino acid variations (e.g., no more than 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1 amino acid variation) compared to SEQ ID NO: 21. In some embodiments, the anti-TMPRSS6 antibody described herein comprises a heavy chain comprising any VH or any variant thereof listed in Table 1 and the heavy chain constant region shown in SEQ ID NO: 21.

[0218] In some embodiments, the anti-TMPRSS6 antibody described herein comprises a heavy chain comprising any VH or any variant thereof listed in Table 1 and a heavy chain constant region having at least 80%, at least 85%, 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%, or at least 99% identity with SEQ ID NO: 39. In some embodiments, the anti-TMPRSS6 antibody described herein comprises a heavy chain comprising any VH or any variant thereof listed in Table 1 and a heavy chain constant region containing no more than 20 amino acid variations (e.g., no more than 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1 amino acid variation) compared to SEQ ID NO: 39. In some embodiments, the anti-TMPRSS6 antibody described herein comprises a heavy chain comprising any VH or any variant thereof listed in Table 1 and the heavy chain constant region shown in SEQ ID NO: 39.

[0219] In some embodiments, the anti-TMPRSS6 antibody described herein comprises a heavy chain comprising any VH or any variant thereof listed in Table 1 and a heavy chain constant region having at least 80%, at least 85%, 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%, or at least 99% identity with SEQ ID NO: 51. In some embodiments, the anti-TMPRSS6 antibody described herein comprises a heavy chain comprising any VH or any variant thereof listed in Table 1 and a heavy chain constant region containing no more than 20 amino acid variations (e.g., no more than 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1 amino acid variation) compared to SEQ ID NO: 51. In some embodiments, the anti-TMPRSS6 antibody described herein comprises a heavy chain comprising any VH or any variant thereof listed in Table 1 and the heavy chain constant region shown in SEQ ID NO: 51.

[0220] In some embodiments, the anti-TMPRSS6 antibody described herein comprises a light chain comprising any VL or any variant thereof listed in Table 1 and a light chain constant region comprising at least 80%, at least 85%, 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%, or at least 99% of SEQ ID NO: 10. In some embodiments, the anti-TMPRSS6 antibody described herein comprises a light chain comprising any VL or any variant thereof listed in Table 1 and a light chain constant region containing no more than 20 amino acid variations (e.g., no more than 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1 amino acid variation) compared to SEQ ID NO: 10. In some embodiments, the anti-TMPRSS6 antibody described herein comprises a light chain comprising any of the VLs listed in Table 1 or any variant thereof, and the light chain constant region shown in SEQ ID NO: 10.

[0221] In some embodiments, the anti-TMPRSS6 antibody described herein comprises a light chain comprising any VL or any variant thereof listed in Table 1 and a light chain constant region comprising at least 80%, at least 85%, 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%, or at least 99% of SEQ ID NO: 40. In some embodiments, the anti-TMPRSS6 antibody described herein comprises a light chain comprising any VL or any variant thereof listed in Table 1 and a light chain constant region containing no more than 20 amino acid variations (e.g., no more than 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1 amino acid variation) compared to SEQ ID NO: 40. In some embodiments, the anti-TMPRSS6 antibody described herein comprises a light chain containing any of the VLs or any variants thereof listed in Table 1, as well as the light chain constant region shown in SEQ ID NO: 40. Table 1 above provides some examples of the amino acid sequences of the IgG heavy and light chains of the anti-TMPRSS6 antibody.

[0222] In some embodiments, the anti-TMPRSS6 antibody of this disclosure comprises a heavy chain containing no more than 20 amino acid variations (e.g., no more than 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1 amino acid variation) compared to the heavy chain shown in SEQ ID NO: 11. Alternatively or supplementally, the anti-TMPRSS6 antibody of this disclosure comprises a light chain containing no more than 20 amino acid variations (e.g., no more than 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 9, 8, 7, 6, 5, 4, 3, 2, or 1 amino acid variation) compared to the light chain shown in SEQ ID NO: 12. In some embodiments, the anti-TMPRSS6 antibody described herein comprises a heavy chain containing an amino acid sequence that is at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO: 11. Alternatively or supplementally, the anti-TMPRSS6 antibody described herein comprises a light chain containing an amino acid sequence that is at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO: 12. In some embodiments, the anti-TMPRSS6 antibody described herein comprises a heavy chain containing the amino acid sequence of SEQ ID NO: 11. Alternatively or supplementally, the anti-TMPRSS6 antibody described herein comprises a light chain containing the amino acid sequence of SEQ ID NO: 12.

[0223] In some embodiments, the anti-TMPRSS6 antibody of this disclosure comprises a heavy chain containing no more than 20 amino acid variations (e.g., no more than 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1 amino acid variation) compared to the heavy chain shown in SEQ ID NO: 23. Alternatively or supplementally, the anti-TMPRSS6 antibody of this disclosure comprises a light chain containing no more than 20 amino acid variations (e.g., no more than 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 9, 8, 7, 6, 5, 4, 3, 2, or 1 amino acid variation) compared to the light chain shown in SEQ ID NO: 23. In some embodiments, the anti-TMPRSS6 antibody described herein comprises a heavy chain containing an amino acid sequence having at least 80% (e.g., 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%) identity with SEQ ID NO: 17 or 22. Alternatively or supplementally, the anti-TMPRSS6 antibody described herein comprises a light chain containing an amino acid sequence having at least 80% (e.g., 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%) identity with SEQ ID NO: 23. In some embodiments, the anti-TMPRSS6 antibody described herein comprises a heavy chain containing the amino acid sequence of SEQ ID NO: 17 or 22. Alternatively or supplementally, the anti-TMPRSS6 antibody described herein comprises a light chain containing the amino acid sequence of SEQ ID NO: 23.

[0224] In some embodiments, the anti-TMPRSS6 antibody of this disclosure comprises a heavy chain containing no more than 20 amino acid variations (e.g., no more than 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1 amino acid variation) compared to the heavy chain shown in SEQ ID NO: 33. Alternatively or supplementally, the anti-TMPRSS6 antibody of this disclosure comprises a light chain containing no more than 20 amino acid variations (e.g., no more than 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 9, 8, 7, 6, 5, 4, 3, 2, or 1 amino acid variation) compared to the light chain shown in SEQ ID NO: 34. In some embodiments, the anti-TMPRSS6 antibody described herein comprises a heavy chain containing an amino acid sequence having at least 80% (e.g., 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%) identity with SEQ ID NO: 33. Alternatively or supplementally, the anti-TMPRSS6 antibody described herein comprises a light chain containing an amino acid sequence having at least 80% (e.g., 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%) identity with SEQ ID NO: 34. In some embodiments, the anti-TMPRSS6 antibody described herein comprises a heavy chain containing the amino acid sequence of SEQ ID NO: 33. Alternatively or supplementally, the anti-TMPRSS6 antibody described herein comprises a light chain containing the amino acid sequence of SEQ ID NO: 34.

[0225] In some embodiments, the anti-TMPRSS6 antibody of this disclosure comprises a heavy chain containing no more than 20 amino acid variations (e.g., no more than 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1 amino acid variation) compared to the heavy chain shown in SEQ ID NO: 42. Alternatively or supplementally, the anti-TMPRSS6 antibody of this disclosure comprises a light chain containing no more than 20 amino acid variations (e.g., no more than 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 9, 8, 7, 6, 5, 4, 3, 2, or 1 amino acid variation) compared to the light chain shown in SEQ ID NO: 42. In some embodiments, the anti-TMPRSS6 antibody described herein comprises a heavy chain containing an amino acid sequence having at least 80% (e.g., 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%) identity with SEQ ID NO: 41. Alternatively or supplementally, the anti-TMPRSS6 antibody described herein comprises a light chain containing an amino acid sequence having at least 80% (e.g., 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%) identity with SEQ ID NO: 42. In some embodiments, the anti-TMPRSS6 antibody described herein comprises a heavy chain containing the amino acid sequence of SEQ ID NO: 41. Alternatively or supplementally, the anti-TMPRSS6 antibody described herein comprises a light chain containing the amino acid sequence of SEQ ID NO: 42.

[0226] In some embodiments, the anti-TMPRSS6 antibody of this disclosure comprises a heavy chain containing no more than 20 amino acid variations (e.g., no more than 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1 amino acid variation) compared to the heavy chain shown in SEQ ID NO: 47. Alternatively or supplementally, the anti-TMPRSS6 antibody of this disclosure comprises a light chain containing no more than 20 amino acid variations (e.g., no more than 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 9, 8, 7, 6, 5, 4, 3, 2, or 1 amino acid variation) compared to the light chain shown in SEQ ID NO: 47. In some embodiments, the anti-TMPRSS6 antibody described herein comprises a heavy chain containing an amino acid sequence having at least 80% (e.g., 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%) identity with SEQ ID NO: 46. Alternatively or supplementally, the anti-TMPRSS6 antibody described herein comprises a light chain containing an amino acid sequence having at least 80% (e.g., 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%) identity with SEQ ID NO: 47. In some embodiments, the anti-TMPRSS6 antibody described herein comprises a heavy chain containing the amino acid sequence of SEQ ID NO: 46. Alternatively or supplementally, the anti-TMPRSS6 antibody described herein comprises a light chain containing the amino acid sequence of SEQ ID NO: 47.

[0227] In some embodiments, the anti-TMPRSS6 antibody of this disclosure comprises a heavy chain containing no more than 20 amino acid variations (e.g., no more than 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1 amino acid variation) compared to the heavy chain shown in SEQ ID NO: 53. Alternatively or supplementally, the anti-TMPRSS6 antibody of this disclosure comprises a light chain containing no more than 20 amino acid variations (e.g., no more than 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 9, 8, 7, 6, 5, 4, 3, 2, or 1 amino acid variation) compared to the light chain shown in SEQ ID NO: 53. In some embodiments, the anti-TMPRSS6 antibody described herein comprises a heavy chain containing an amino acid sequence having at least 80% (e.g., 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%) identity with SEQ ID NO: 52. Alternatively or supplementally, the anti-TMPRSS6 antibody described herein comprises a light chain containing an amino acid sequence having at least 80% (e.g., 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%) identity with SEQ ID NO: 53. In some embodiments, the anti-TMPRSS6 antibody described herein comprises a heavy chain containing the amino acid sequence of SEQ ID NO: 52. Alternatively or supplementally, the anti-TMPRSS6 antibody described herein comprises a light chain containing the amino acid sequence of SEQ ID NO: 53.

[0228] The anti-TMPRSS6 antibody described herein may be in any antibody form, including but not limited to full-length (i.e., complete) antibodies, their antigen-binding fragments (e.g., Fab, F(ab'), F(ab')2, Fv), single-chain antibodies, bispecific antibodies, or nanobodies. In some embodiments, the anti-TMPRSS6 antibody described herein is an scFv. In some embodiments, the anti-TMPRSS6 antibody described herein is an scFv-Fab (e.g., an scFv fused to a portion of a constant region).

[0229] In some embodiments, conserved mutations may be introduced into the antibody sequence (e.g., CDR or framework sequence) at locations where residues are unlikely to interact with the target antigen (e.g., TMPRSS6) (e.g., as determined based on crystal structure). In some embodiments, one, two, or more mutations (e.g., amino acid substitutions) may be introduced into the Fc region of the anti-TMPRSS6 antibody described herein (e.g., in the CH2 domain (residues 231 to 340 of human IgG1) and / or the CH3 domain (residues 341 to 447 of human IgG1) and / or the hinge region, numbered according to the Kabat numbering system (e.g., the EU index in Kabat) to alter one or more functional properties of the antibody, such as serum half-life, complement fixation, Fc receptor binding, and / or antigen-dependent cytotoxicity.

[0230] In some embodiments, one, two, or more mutations (e.g., amino acid substitutions) are introduced into the hinge region of the Fc region (CH1 domain) to alter (e.g., increase or decrease) the number of cysteine ​​residues in the hinge region, as described, for example, in U.S. Patent No. 5,677,425. Altering the number of cysteine ​​residues in the hinge region of the CH1 domain can, for example, promote the assembly of light and heavy chains, or alter (e.g., increase or decrease) antibody stability or promote linker conjugation.

[0231] In some embodiments, one, two, or more mutations (e.g., amino acid substitutions) are introduced into the Fc region of the antibody described herein (e.g., in the CH2 domain (residues 231 to 340 of human IgG1) and / or the CH3 domain (residues 341 to 447 of human IgG1) and / or the hinge region, numbered according to the Kabat numbering system (e.g., the EU index in Kabat)) to increase or decrease the antibody's affinity for Fc receptors (e.g., activated Fc receptors) on the surface of effector cells. Mutations in the Fc region of antibodies that decrease or increase the antibody's affinity for Fc receptors, and techniques for introducing such mutations into the Fc receptor or fragments thereof, are known to those skilled in the art. Some examples of mutations in the Fc receptor that can be used to alter the affinity of an antibody for the Fc receptor are described below: for example, Smith P et al., (2012) PNAS 109: 6181-6186, U.S. Patent No. 6,737,056, and International Publication No. WO 97 / 34631, which are incorporated herein by reference.

[0232] In some embodiments, one, two, or more amino acid mutations (i.e., substitutions, insertions, or deletions) are introduced into the constant domain of IgG or its FcRn binding fragment (preferably, an Fc or hinge-Fc domain fragment) to alter (e.g., decrease or increase) the half-life of the antibody in vivo. See, for example, International Publication No. WO 97 / 34631; and U.S. Patent Nos. 5,869,046, 6,121,022, 6,277,375, and 6,165,745, for example, mutations that alter (e.g., decrease or increase) the half-life of the antibody in vivo.

[0233] In some embodiments, one, two, or more amino acid mutations (i.e., substitutions, insertions, or deletions) are introduced into the constant domain of IgG or its FcRn binding fragment (preferably, an Fc or hinge-Fc domain fragment) to reduce the half-life of the anti-TMPRSS6 antibody in vivo. In some embodiments, one, two, or more amino acid mutations (i.e., substitutions, insertions, or deletions) are introduced into the constant domain of IgG or its FcRn binding fragment (preferably, an Fc or hinge-Fc domain fragment) to increase the half-life of the antibody in vivo. In some embodiments, the antibody may have one or more amino acid mutations (e.g., substitutions) in the second constant (CH2) domain (residues 231 to 340 of human IgG1) and / or the third constant (CH3) domain (residues 341 to 447 of human IgG1) (numbered according to the EU index in Kabat (Kabat EA et al., (1991) ibid.)). In some embodiments, the constant region of the IgG1 of the antibody described herein includes a methionine (M) to tyrosine (Y) substitution at position 252, a serine (S) to threonine (T) substitution at position 254, and a threonine (T) to glutamic acid (E) substitution at position 256, the positions of which are indexed according to the EU index as in Kabat. See U.S. Patent No. 7,658,921, which is incorporated herein by reference. This type of mutant IgG (referred to as the “YTE mutant”) has been shown to exhibit a half-life increase of up to 4-fold compared to the wild-type form of the same antibody (see Dall'Acqua WF et al., (2006) J Biol Chem 281: 23514-24).

[0234] In some embodiments, one, two, or more amino acid substitutions are introduced into the Fc region of the constant domain of IgG to alter the effector function of the anti-TMPRSS6 antibody. The effector ligand with altered affinity can be, for example, an Fc receptor or a C1 component of complement. This approach is described in more detail in U.S. Patent Nos. 5,624,821 and 5,648,260. In some embodiments, one or more amino acid substitutions may be introduced into the Fc region of the antibody described herein to remove potential glycosylation sites on the Fc region, which can reduce Fc receptor binding (see, for example, Shields RL et al., (2001) J Biol Chem 276: 6591-604).

[0235] In some embodiments, one or more amino groups in the constant region of the anti-TMPRSS6 antibody described herein may be replaced with different amino acid residues, resulting in altered C1q binding and / or reduced or eliminated complement-dependent cytotoxicity (CDC). This method is described in more detail in U.S. Patent No. 6,194,551 (Idusogie et al). In some embodiments, one or more amino acid residues in the N-terminal region of the CH2 domain of the antibody described herein are modified to alter the antibody's complement-fixing ability. This method is further described in International Publication No. WO 94 / 29351. In some embodiments, the Fc region of the antibody described herein is modified to enhance the antibody's ability to mediate antibody-dependent cellular cytotoxicity (ADCC) and / or increase the antibody's affinity for the Fcγ receptor. This method is further described in International Publication No. WO 00 / 42072.

[0236] In some embodiments, the heavy chain and / or light chain variable domain sequences of the antibodies provided herein can be used to generate, for example, CDR-grafted, chimeric, humanized, or complexed human antibodies or antigen-binding fragments, as described in other sections herein. As will be understood by those skilled in the art, any variant, CDR-grafted, chimeric, humanized, or complexed antibody derived from any antibody provided herein can be used in the compositions and methods described herein and will maintain the ability to specifically bind to TMPRSS6, such that the variant, CDR-grafted, chimeric, humanized, or complexed antibody has at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95%, or higher binding to TMPRSS6 relative to the original antibody from which it is derived.

[0237] In some embodiments, the antibodies provided herein contain mutations that confer the desired properties to the antibody. For example, to avoid potential complications due to Fab-arm exchanges known to occur with native IgG4 mAb, the antibodies provided herein may contain a stabilizing 'Adair' mutation (Angal S., et al., A single amino acid substitution abolishes the heterogeneity of chimeric mouse / human (IgG4) antibody, MolImmunol 30, 105-108; 1993), in which serine at position 228 (EU number, residue 241 according to Kabat number) is converted to proline, thereby producing an IgG1-like hinge sequence. Therefore, any antibody may contain a stabilizing 'Adair' mutation.

[0238] In some embodiments, the antibody is modified, for example, by glycosylation, phosphorylation, SUMOylation, and / or methylation. In some embodiments, the antibody is a glycosylated antibody conjugated to one or more sugar or carbohydrate molecules. In some embodiments, one or more sugar or carbohydrate molecules are conjugated to the antibody via N-glycosylation, O-glycosylation, C-glycosylation, glycosylphosphatidylinositol (GPI-anchored attachment), and / or phosphoglycosylation. In some embodiments, one or more sugar or carbohydrate molecules are monosaccharides, disaccharides, oligosaccharides, or glycans. In some embodiments, one or more sugar or carbohydrate molecules are branched oligosaccharides or branched glycans. In some embodiments, one or more sugar or carbohydrate molecules comprise mannose units, glucose units, N-acetylglucosamine units, N-acetylgalactosamine units, galactose units, fucose units, or phospholipid units. In some embodiments, about 1 to 10, about 1 to 5, about 5 to 10, about 1 to 4, about 1 to 3, or about 2 sugar molecules are present. In some embodiments, the glycosylated antibody is fully or partially glycosylated. In some embodiments, the antibody is glycosylated by a chemical reaction or by an enzymatic means. In some embodiments, the antibody is glycosylated in vitro or intracellularly, optionally lacking enzymes in the N- or O-glycosylation pathway, such as glycosyltransferases. In some embodiments, the antibody is functionalized with sugar or carbohydrate molecules, as described in International Patent Application Publication WO2014065661, published May 1, 2014, entitled "Modified antibody, antibody-conjugate and process for the preparation thereof".

[0239] In some embodiments, any of the anti-TMPRSS6 antibodies described herein may include a signal peptide (e.g., an N-terminal signal peptide) in the heavy chain sequence and / or light chain sequence. In some embodiments, the anti-TMPRSS6 antibody described herein comprises any of the VH and VL sequences described herein, any of the IgG heavy chain and light chain sequences, or any of the F(ab') heavy chain and light chain sequences, and further comprises a signal peptide (e.g., an N-terminal signal peptide).

[0240] III. Preparation of anti-TMPRSS6 antibody

[0241] The antibodies described herein that bind to TMPRSS6 can be prepared by any method known in the art. See, for example, Harlow and Lane, (1998) Antibodies: A Laboratory Manual, Cold SpringHarbor Laboratory, New York.

[0242] In some embodiments, antibodies specific to a target antigen (e.g., TMPRSS6) can be prepared using conventional hybridoma techniques. Full-length target antigens or fragments thereof, optionally conjugated to a carrier protein such as KLH, can be used to immunize host animals to generate antibodies that bind to that antigen. As further described herein, the routes and protocols for immunizing host animals are generally consistent with established and conventional techniques for antibody stimulation and generation. General techniques for generating mouse antibodies, humanized antibodies, and human antibodies are known in the art and described herein. Any mammalian subject (including humans) or antibody-producing cells derived from such subjects can be manipulated to serve as the basis for generating mammalian (including human) hybridoma cell lines. Typically, an immunogen (including those described herein) is inoculated into the host animal intraperitoneally, intramuscularly, orally, subcutaneously, intraplantarly, and / or intradermally.

[0243] If desired, the target antibody (e.g., generated via hybridoma), whether monoclonal or polyclonal, can be sequenced, and the polynucleotide sequence can subsequently be cloned into a vector for expression or proliferation. The sequence encoding the target antibody can be maintained in the vector within host cells, which can then be amplified and frozen for future use. Alternatively, the polynucleotide sequence can be used for genetic manipulation to “humanize” the antibody or improve its affinity (affinity maturation) or other characteristics. For example, if the antibody is to be used in human clinical trials and treatments, the constant region can be modified to more closely resemble the human constant region to avoid an immune response. Genetic manipulation of the antibody sequence can be expected to obtain greater affinity for the target antigen and greater potency. It will be apparent to those skilled in the art that one or more polynucleotide alterations can be made to the antibody while still maintaining its binding specificity to the target antigen.

[0244] In other embodiments, fully human antibodies can be obtained using commercially available mice that have been engineered to express specific human immunoglobulins. Transgenic animals designed to produce a more desired (e.g., fully human antibody) or more robust immune response can also be used to generate humanized antibodies or human antibodies. Some examples of such techniques are Xenomouse RTM from Amgen, Inc. (Fremont, CA) and HuMAb-Mouse RTM and TC Mouse™ from Medarex, Inc. (Princeton, NJ), or H2L2 mice from Harbour Antibodies BV (Holland). In another alternative, antibodies can be recombinantly prepared using phage display or yeast technology. See, for example, U.S. Patent Nos. 5,565,332, 5,580,717, 5,733,743, and 6,265,150, and Winter et al., (1994) Annu.Rev. Immunol. 12:433-455. Alternatively, phage display technology (McCafferty et al., (1990) Nature 348:552-553) can be used to generate human antibodies and antibody fragments in vitro from a gene library of immunoglobulin variable (V) domains from unimmunized donors.

[0245] The antigen-binding fragment of a complete antibody (full-length antibody) can be prepared using conventional methods. For example, the F(ab')2 fragment can be produced by pepsin digestion of the antibody molecule, and the Fab fragment can be produced by reducing the disulfide bridge of the F(ab')2 fragment. Genetically modified antibodies, such as humanized antibodies, chimeric antibodies, single-chain antibodies, and bispecific antibodies, can be produced, for example, using conventional recombinant techniques. In one instance, DNA encoding a monoclonal antibody specific to a target antigen can be readily isolated and sequenced using conventional procedures (e.g., by using oligonucleotide probes capable of specifically binding to genes encoding the heavy and light chains of the monoclonal antibody). Hybridoma cells serve as a preferred source of such DNA. Once isolated, the DNA can be placed into one or more expression vectors, which are then transfected into host cells (e.g., E. coli cells, simian COS cells, Chinese hamster ovary (CHO) cells, human HEK293 cells, or myeloma cells that do not otherwise produce immunoglobulins) to obtain monoclonal antibody synthesis in recombinant host cells. See, for example, PCT Publication No. WO 87 / 04462. The DNA can then be modified, for example, by replacing the coding sequences for the human heavy chain constant domain and light chain constant domain with homologous mouse sequences, Morrison et al., (1984) Proc. Nat. Acad. Sci. 81:6851, or by covalently linking all or part of the coding sequence of a non-immunoglobulin polypeptide to an immunoglobulin coding sequence. In this way, genetically modified antibodies with binding specificity to target antigens, such as “chimeric” or “hybrid” antibodies, can be prepared.

[0246] Single-chain antibodies can be prepared using recombinant technology by linking nucleotide sequences encoding the heavy chain variable region and the light chain variable region. Preferably, a flexible linker is incorporated between the two variable regions.

[0247] Alternatively, the described techniques for generating single-chain antibodies (US Patent Nos. 4,946,778 and 4,704,692) can be adapted to generate phage or yeast scFv libraries, and scFv clones specific for TMPRSS6 can be identified from the libraries according to standard procedures. Positive clones can be further screened to identify those with high TMPRSS6 binding affinity.

[0248] Antibodies obtained according to methods known in the art and described herein can be characterized using methods well-known in the art. For example, one approach is to identify epitopes that bind to antigens, or "epitope mapping." Numerous methods are known in the art for mapping and characterizing the location of epitopes on proteins, including resolving the crystal structure of antibody-antigen complexes, competition assays, gene fragment expression assays, and synthetic peptide-based assays, as described, for example, in Chapter 11 of Harlow and Lane, Using Antibodies, a Laboratory Manual, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY, 1999. In one instance, epitope mapping can be performed using H / D-Ex (hydrogen-deuterium exchange) coupled with proteolysis and mass spectrometry. In another instance, epitope mapping can be used to determine the sequence that binds to the antibody. Epitopes can be linear epitopes, i.e., contained in a single amino acid segment, or they can be conformational epitopes formed through three-dimensional interactions of amino acids, which may not necessarily be contained in a single segment (a linear sequence of primary structure). Peptides of varying lengths (e.g., at least 4 to 6 amino acids long) can be isolated or synthesized (e.g., recombinantly isolated or synthesized) and used for antibody binding assays. In another example, antibody-bound epitopes can be identified in systematic screening by using overlapping peptides derived from target antigen sequences and determining antibody binding. Based on gene fragment expression assays, open reading frames encoding target antigens are randomly fragmented or fragmented via specific genetic constructs, and the reactivity of the expressed antigen fragments with the test antibody is determined. For example, gene fragments can be generated by PCR, subsequently transcribed, and translated into proteins in vitro in the presence of radioactive amino acids. Antibody binding to radiolabeled antigen fragments is then determined by immunoprecipitation and gel electrophoresis. Some epitopes can also be identified using large libraries of random peptide sequences displayed on the surface of phage particles (phage libraries). Alternatively, the binding of an identified library of overlapping peptide fragments to the test antibody can be tested in a simple binding assay. In another example, mutagenesis of antigen-binding domains, domain exchange experiments, and alanine scanning mutagenesis can be performed to identify residues required, sufficient, and / or essential for epitope binding. Alternatively, a competitive assay can be performed using other antibodies known to bind to the same antigen to determine whether the antibody binds to the same epitope as other antibodies. Competitive assays are well known to those skilled in the art.

[0249] In some instances, anti-TMPRSS6 antibodies are prepared using recombinant techniques exemplified below. Nucleic acids encoding the heavy and light chains of the anti-TMPRSS6 antibody as described herein can be cloned into an expression vector, each nucleotide sequence operatively linked to a suitable promoter. In one instance, each nucleotide sequence encoding the heavy and light chains is operatively linked to a different promoter. Alternatively, the nucleotide sequences encoding the heavy and light chains can be operatively linked to a single promoter, such that both the heavy and light chains are expressed by the same promoter. If necessary, an internal ribosomal entry site (IRES) can be inserted between the heavy and light chain coding sequences.

[0250] In some instances, the nucleotide sequences encoding the two chains of an antibody are cloned into two vectors, which can be introduced into the same or different cells. When the two chains are expressed in different cells, they can be isolated from the host cells in which they are expressed, and the isolated heavy and light chains can be mixed and incubated under suitable conditions that allow for antibody formation.

[0251] Typically, a nucleic acid sequence encoding one or all strands of an antibody can be cloned into a suitable expression vector operatively linked to a suitable promoter using methods known in the art. For example, the nucleotide sequence and the vector can be contacted with a restriction enzyme under suitable conditions to generate complementary ends on each molecule that can pair with each other and be linked together with a ligase. Alternatively, synthetic nucleic acid adapters can be ligated to the ends of the gene. These synthetic adapters contain nucleic acid sequences corresponding to specific restriction sites in the vector. The choice of expression vector / promoter will depend on the type of host cell used to produce the antibody.

[0252] A variety of promoters can be used to express the antibodies described herein, including but not limited to the intermediate early promoter of cytomegalovirus (CMV), viral LTRs (e.g., Rous sarcoma virus LTR, HIV-LTR, HTLV-1 LTR), the early promoter of simian virus 40 (SV40), the Escherichia coli lac UV promoter, and the herpes simplex virus TK promoter.

[0253] Adjustable promoters can also be used. Such adjustable promoters include those that use lac repressors from E. coli as transcription regulators to regulate transcription from mammalian cell promoters with lac operons (Brown, M. et al., Cell, 49:603-612 (1987)), and those that use tetracycline repressors (tetR) (Gossen, M., and Bujard, H., Proc. Natl. Acad. Sci. USA 89:5547-555115 (1992); Yao, F. et al., Human Gene Therapy, 9:1939-1950 (1998); Shockelt, P., et al., Proc. Natl. Acad. Sci. USA, 92:6522-6526 (1995)). Other systems include: FK506 dimer, VP16 or p65 using astradiol, RU486, diphenolmurislerone, or rapamycin. Inducible systems are available from Invitrogen, Clontech, and Ariad, among others.

[0254] Adjustable promoters, including repressors with operons, can be used. In one embodiment, a lac repressor from *E. coli* can act as a transcription regulator to regulate transcription from mammalian cell promoters with lac operons (Brown et al., *Cell*, 49:603-612 (1987)); Gossen and Bujard (1992); (M. Gossen et al., *Natl. Acad. Sci. USA*, 89:5547-5551 (1992)) combine a tetracycline repressor (tetR) with a transcription activator (VP 16) to produce a tetR-mammalian cell transcription activator fusion protein tTa (tetR-VP 16), wherein tetO carries a minimal promoter derived from a human cytomegalovirus (hCMV) promoter, to generate the tetR-tet operon system to control gene expression in mammalian cells. In one embodiment, a tetracycline-inducible switch is used. When the tetracycline operon is properly positioned downstream of the TATA element of the CMVIE promoter, the tetracycline repressor alone (tetR), rather than a tetR-mammalian cell transcription factor fusion derivative, can act as a potent trans-regulator to regulate gene expression in mammalian cells (Yao et al., Human Gene Therapy). A particular advantage of this tetracycline-inducible switch is that it does not require the use of tetracycline repressor-mammalian cell transactivators or repressor fusion proteins, which may be cellularly toxic in some cases (Gossen 5 et al., Natl. Acad. Sci. USA, 89:5547-5551 (1992); Shockett et al., Proc. Natl. Acad. Sci. USA, 92:6522-6526 (1995)), to achieve its modulatory effect.

[0255] Additionally, the vector may contain some or all of the following: selectable marker genes, such as the neomycin gene for selecting stable or transient transfectants in mammalian cells; enhancer / promoter sequences from early genes derived from human CMV for high-level transcription; transcription termination and RNA processing signals from SV40 for mRNA stability; SV40 polynodular origin of replication and ColE1 for appropriate appendage replication; internal ribosome binding sites (IRES), multifunctional multiple cloning sites; and T7 and SP6 RNA promoters for in vitro transcription of sense and antisense RNA. Suitable vectors and methods for generating vectors containing transgenes are well known and available in the art. Some examples of polyadenylation signals that can be used to practice the methods described herein include, but are not limited to, human collagen I polyadenylation signals, human collagen II polyadenylation signals, and SV40 polyadenylation signals.

[0256] One or more vectors (e.g., expression vectors) containing a nucleic acid encoding any antibody (e.g., nucleic acid coding sequences listed in Table 3) can be introduced into suitable host cells to produce antibodies. Some non-limiting examples of host cells include Chinese hamster ovary (CHO) cells, dhfr-CHO cells, human embryonic kidney (HEK)-293 cells, verda reno (VERO) cells, non-secretory null (NS0) cells, human embryonic retina (PER.C6) cells, Sp2 / 0 cells, baby hamster kidney (BHK) cells, Madin-Darby canine kidney (MDCK) cells, Madin-Darby bovine kidney (MDBK) cells, and the monkey kidney CV1 line transformed from SV40 (COS) cells. In some embodiments, the host cell expressing the anti-TMPRSS6 antibody is a CHO cell. Host cells can be cultured under conditions suitable for expressing the antibody or any of its polypeptide chains. Such antibodies or their polypeptide chains can be recovered from cultured cells (e.g., from said cells or culture supernatant) using conventional methods (e.g., affinity purification). If necessary, the antibody polypeptide chains can be incubated under suitable conditions for a suitable period of time to allow antibody production. In some embodiments, the host cell contains nucleic acid encoding the heavy chain of the anti-TMPRSS6 antibody. In some embodiments, the host cell contains nucleic acid encoding the light chain of the anti-TMPRSS6 antibody. In some embodiments, the host cell contains nucleic acid encoding both the heavy chain and the light chain.

[0257] In some embodiments, the methods for preparing the antibodies described herein involve a recombinant expression vector encoding both the heavy and light chains of an anti-TMPRSS6 antibody, as also described herein. The recombinant expression vector can be introduced into suitable host cells (e.g., dhfr-CHO cells) by conventional methods, such as calcium phosphate-mediated transfection. Positive transformant host cells can be selected and cultured under suitable conditions that allow expression of the two polypeptide chains that form the antibody, which can be recovered from the cells or from the culture medium. If necessary, the two chains recovered from the host cells can be incubated under suitable conditions that allow antibody formation.

[0258] In one example, two recombinant expression vectors are provided: one encoding the heavy chain of an anti-TMPRSS6 antibody, and the other encoding the light chain of an anti-TMPRSS6 antibody. Both recombinant expression vectors can be introduced into suitable host cells (e.g., dhfr-CHO cells) via conventional methods such as calcium phosphate-mediated transfection.

[0259] Alternatively, each expression vector can be introduced into a suitable host cell. Positive transformants can be selected and cultured under suitable conditions that allow for antibody expression of the polypeptide chain. When both expression vectors are introduced into the same host cell, the antibodies produced therein can be recovered from the host cell or from the culture medium. If necessary, the polypeptide chain can be recovered from the host cell or from the culture medium and subsequently incubated under suitable conditions that allow for antibody formation. When the two expression vectors are introduced into different host cells, they can each be recovered from the respective host cell or from the respective culture medium. The two polypeptide chains can then be incubated under suitable conditions to form antibodies.

[0260] Standard molecular biology techniques are used to prepare recombinant expression vectors, transfect host cells, select transformants, culture host cells, and recover antibodies from culture media. For example, some antibodies can be separated by affinity chromatography using a protein A or protein G conjugated matrix.

[0261] Any nucleic acid (e.g., as provided in Table 3) encoding the heavy chain, light chain, or both of the anti-TMPRSS6 antibody described herein, the vector containing it (e.g., an expression vector), and the host cell containing said vector are all within the scope of this disclosure.

[0262] Table 3: Nucleic acid sequences encoding the VH / VL of the anti-TMPRSS6 antibody listed in Table 1

[0263]

[0264]

[0265]

[0266]

[0267]

[0268]

[0269]

[0270]

[0271]

[0272]

[0273]

[0274]

[0275]

[0276]

[0277]

[0278] In some embodiments, this disclosure provides isolated nucleic acids comprising a sequence having at least 60% (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) identity with SEQ ID NO: 54, and / or comprising isolated nucleic acids comprising a sequence having at least 60% (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) identity with SEQ ID NO: 55. In some embodiments, this disclosure provides an expression vector comprising: an isolated nucleic acid containing a sequence having at least 60% (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) identity with SEQ ID NO: 54, and / or an isolated nucleic acid containing a sequence having at least 60% (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) identity with SEQ ID NO: 55.

[0279] In some embodiments, this disclosure provides isolated nucleic acids comprising a sequence having at least 60% (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) identity with SEQ ID NO: 56, and / or comprising isolated nucleic acids comprising a sequence having at least 60% (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) identity with SEQ ID NO: 57. In some embodiments, this disclosure provides an expression vector comprising: an isolated nucleic acid containing a sequence having at least 60% (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) identity with SEQ ID NO: 56, and / or an isolated nucleic acid containing a sequence having at least 60% (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) identity with SEQ ID NO: 57.

[0280] In some embodiments, this disclosure provides isolated nucleic acids comprising a sequence having at least 60% (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) identity with SEQ ID NO: 58, and / or comprising isolated nucleic acids comprising a sequence having at least 60% (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) identity with SEQ ID NO: 59. In some embodiments, this disclosure provides an expression vector comprising: isolated nucleic acid containing a sequence having at least 60% (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) identity with SEQ ID NO: 58, and / or containing isolated nucleic acid containing a sequence having at least 60% (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) identity with SEQ ID NO: 59.

[0281] In some embodiments, this disclosure provides isolated nucleic acids comprising a sequence having at least 60% (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) identity with SEQ ID NO: 60, and / or comprising isolated nucleic acids comprising a sequence having at least 60% (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) identity with SEQ ID NO: 61. In some embodiments, this disclosure provides an expression vector comprising: an isolated nucleic acid containing a sequence having at least 60% (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) identity with SEQ ID NO: 60, and / or an isolated nucleic acid containing a sequence having at least 60% (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) identity with SEQ ID NO: 61.

[0282] In some embodiments, this disclosure provides isolated nucleic acids comprising a sequence having at least 60% (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) identity with SEQ ID NO: 62, and / or comprising isolated nucleic acids comprising a sequence having at least 60% (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) identity with SEQ ID NO: 63. In some embodiments, this disclosure provides an expression vector comprising: an isolated nucleic acid containing a sequence having at least 60% (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) identity with SEQ ID NO: 62, and / or an isolated nucleic acid containing a sequence having at least 60% (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) identity with SEQ ID NO: 63.

[0283] In some embodiments, this disclosure provides isolated nucleic acids comprising a sequence having at least 60% (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) identity with SEQ ID NO: 5, and / or comprising isolated nucleic acids comprising a sequence having at least 60% (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) identity with SEQ ID NO: 32. In some embodiments, this disclosure provides an expression vector comprising: an isolated nucleic acid containing a sequence having at least 60% (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) identity with SEQ ID NO: 5, and / or an isolated nucleic acid containing a sequence having at least 60% (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) identity with SEQ ID NO: 32.

[0284] In some embodiments, this disclosure provides isolated nucleic acids comprising a sequence having at least 60% (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) identity with SEQ ID NO: 79, and / or comprising isolated nucleic acids comprising a sequence having at least 60% (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) identity with SEQ ID NO: 28. In some embodiments, this disclosure provides an expression vector comprising: isolated nucleic acid containing a sequence having at least 60% (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) identity with SEQ ID NO: 79, and / or containing isolated nucleic acid containing a sequence having at least 60% (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) identity with SEQ ID NO: 28.

[0285] In some embodiments, this disclosure provides isolated nucleic acids comprising a sequence having at least 60% (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) identity with SEQ ID NO: 64, and / or comprising isolated nucleic acids comprising a sequence having at least 60% (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) identity with SEQ ID NO: 65. In some embodiments, this disclosure provides an expression vector comprising: an isolated nucleic acid containing a sequence having at least 60% (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) identity with SEQ ID NO: 64, and / or an isolated nucleic acid containing a sequence having at least 60% (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) identity with SEQ ID NO: 65.

[0286] In some embodiments, this disclosure provides isolated nucleic acids comprising a sequence having at least 60% (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) identity with SEQ ID NO: 66, and / or comprising isolated nucleic acids comprising a sequence having at least 60% (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) identity with SEQ ID NO: 67. In some embodiments, this disclosure provides an expression vector comprising: an isolated nucleic acid containing a sequence having at least 60% (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) identity with SEQ ID NO: 66, and / or an isolated nucleic acid containing a sequence having at least 60% (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) identity with SEQ ID NO: 67.

[0287] In some embodiments, this disclosure provides isolated nucleic acids comprising a sequence having at least 60% (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) identity with SEQ ID NO: 68, and / or comprising isolated nucleic acids comprising a sequence having at least 60% (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) identity with SEQ ID NO: 69. In some embodiments, this disclosure provides an expression vector comprising: an isolated nucleic acid containing a sequence having at least 60% (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) identity with SEQ ID NO: 68, and / or an isolated nucleic acid containing a sequence having at least 60% (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) identity with SEQ ID NO: 69.

[0288] In some embodiments, this disclosure provides isolated nucleic acids comprising a sequence having at least 60% (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) identity with SEQ ID NO: 70, and / or comprising isolated nucleic acids comprising a sequence having at least 60% (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) identity with SEQ ID NO: 71. In some embodiments, this disclosure provides an expression vector comprising: an isolated nucleic acid containing a sequence having at least 60% (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) identity with SEQ ID NO: 70, and / or an isolated nucleic acid containing a sequence having at least 60% (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) identity with SEQ ID NO: 71.

[0289] In some embodiments, this disclosure provides isolated nucleic acids comprising a sequence having at least 60% (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) identity with SEQ ID NO: 72, and / or comprising isolated nucleic acids comprising a sequence having at least 60% (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) identity with SEQ ID NO: 73. In some embodiments, this disclosure provides an expression vector comprising: an isolated nucleic acid containing a sequence having at least 60% (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) identity with SEQ ID NO: 72, and / or an isolated nucleic acid containing a sequence having at least 60% (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) identity with SEQ ID NO: 73.

[0290] In some embodiments, this disclosure provides isolated nucleic acids comprising a sequence having at least 60% (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) identity with SEQ ID NO: 74, and / or comprising isolated nucleic acids comprising a sequence having at least 60% (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) identity with SEQ ID NO: 75. In some embodiments, this disclosure provides an expression vector comprising: isolated nucleic acid containing a sequence having at least 60% (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) identity with SEQ ID NO: 74, and / or isolated nucleic acid containing a sequence having at least 60% (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) identity with SEQ ID NO: 75.

[0291] In some embodiments, this disclosure provides isolated nucleic acids comprising a sequence having at least 60% (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) identity with SEQ ID NO: 76, and / or comprising isolated nucleic acids comprising a sequence having at least 60% (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) identity with SEQ ID NO: 77. In some embodiments, this disclosure provides an expression vector comprising: isolated nucleic acid containing a sequence having at least 60% (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) identity with SEQ ID NO: 76, and / or isolated nucleic acid containing a sequence having at least 60% (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) identity with SEQ ID NO: 77.

[0292] In some embodiments, the anti-TMPRSS6 described herein is generated by expressing: (i) a nucleic acid having at least 60% (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) identity with SEQ ID NO: 54, and / or (ii) a nucleic acid having at least 60% (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) identity with SEQ ID NO: 55.

[0293] In some embodiments, the anti-TMPRSS6 described herein is generated by expressing: (i) a nucleic acid having at least 60% (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) identity with SEQ ID NO: 57, and / or (ii) a nucleic acid having at least 60% (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) identity with SEQ ID NO: 57.

[0294] In some embodiments, the anti-TMPRSS6 described herein is generated by expressing: (i) a nucleic acid having at least 60% (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) identity with SEQ ID NO: 58, and / or (ii) a nucleic acid having at least 60% (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) identity with SEQ ID NO: 59.

[0295] In some embodiments, the anti-TMPRSS6 described herein is generated by expressing: (i) a nucleic acid having at least 60% (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) identity with SEQ ID NO: 60, and / or (ii) a nucleic acid having at least 60% (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) identity with SEQ ID NO: 61.

[0296] In some embodiments, the anti-TMPRSS6 described herein is generated by expressing: (i) a nucleic acid having at least 60% (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) identity with SEQ ID NO: 62, and / or (ii) a nucleic acid having at least 60% (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) identity with SEQ ID NO: 63.

[0297] In some embodiments, the anti-TMPRSS6 described herein is generated by expressing: (i) a nucleic acid having at least 60% (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) identity with SEQ ID NO: 5, and / or (ii) a nucleic acid having at least 60% (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) identity with SEQ ID NO: 32.

[0298] In some embodiments, the anti-TMPRSS6 described herein is generated by expressing: (i) a nucleic acid having at least 60% (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) identity with SEQ ID NO: 79, and / or (ii) a nucleic acid having at least 60% (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) identity with SEQ ID NO: 80.

[0299] In some embodiments, the anti-TMPRSS6 described herein is generated by expressing: (i) a nucleic acid having at least 60% (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) identity with SEQ ID NO: 64, and / or (ii) a nucleic acid having at least 60% (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) identity with SEQ ID NO: 65.

[0300] In some embodiments, the anti-TMPRSS6 described herein is generated by expressing: (i) a nucleic acid having at least 60% (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) identity with SEQ ID NO: 66, and / or (ii) a nucleic acid having at least 60% (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) identity with SEQ ID NO: 67.

[0301] In some embodiments, the anti-TMPRSS6 described herein is generated by expressing: (i) a nucleic acid having at least 60% (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) identity with SEQ ID NO: 68, and / or (ii) a nucleic acid having at least 60% (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) identity with SEQ ID NO: 69.

[0302] In some embodiments, the anti-TMPRSS6 described herein is generated by expressing: (i) a nucleic acid having at least 60% (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) identity with SEQ ID NO: 70, and / or (ii) a nucleic acid having at least 60% (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) identity with SEQ ID NO: 71.

[0303] In some embodiments, the anti-TMPRSS6 described herein is generated by expressing: (i) a nucleic acid having at least 60% (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) identity with SEQ ID NO: 73, and / or (ii) a nucleic acid having at least 60% (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) identity with SEQ ID NO: 73.

[0304] In some embodiments, the anti-TMPRSS6 described herein is generated by expressing: (i) a nucleic acid having at least 60% (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) identity with SEQ ID NO: 74, and / or (ii) a nucleic acid having at least 60% (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) identity with SEQ ID NO: 75.

[0305] In some embodiments, the anti-TMPRSS6 described herein is generated by expressing: (i) a nucleic acid having at least 60% (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) identity with SEQ ID NO: 77, and / or (ii) a nucleic acid having at least 60% (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) identity with SEQ ID NO: 77.

[0306] In some embodiments, this disclosure provides recombinant cells expressing the anti-TMPRSS6 antibody described herein (e.g., recombinant cells for antibody production).

[0307] Therefore, this disclosure provides a method for generating antibodies, the method comprising culturing recombinant cells under conditions suitable for expressing antibodies from an expression vector via recombinant cells. The recombinant cells expressing antibodies can be cultured under any suitable conditions known in the art. In some embodiments, the method further includes isolating antibodies from the culture medium in which the cells are cultured using any suitable method known in the art.

[0308] IV. Pharmaceutical Compositions

[0309] The antibodies described herein, as well as those encoding nucleic acids or nucleic acid sequences, carriers containing these, or host cells containing said carriers, can be mixed with pharmaceutically acceptable excipients to form pharmaceutical compositions for treating a target disease. "Acceptable" means that the carrier must be compatible with (and preferably able to stabilize) the active ingredient of the composition and harmless to the subject of treatment. Pharmaceutically acceptable excipients (carriers) include buffers known in the art. See, for example, Remington: The Science and Practice of Pharmacy, 20th edition. (2000) Lippincott Williams and Wilkins, Ed. KE Hoover.

[0310] The pharmaceutical compositions containing anti-TMPRSS6 antibodies disclosed herein may also include suitable buffers. Buffers are weak acids or weak bases used to maintain the pH of the solution near a selected value after the addition of another acid or base. In some instances, the buffers disclosed herein may be buffers that maintain physiological pH even if the concentration of carbon dioxide (produced by cellular respiration) changes. Exemplary buffers include, but are not limited to, HEPES (4-(2-hydroxyethyl)-1-piperazine ethanesulfonic acid) buffer, Dulbecco's phosphate-buffered saline (DPBS) buffer, or phosphate-buffered saline (PBS) buffer. Such buffers may contain disodium hydrogen phosphate and sodium chloride, or potassium dihydrogen phosphate and potassium chloride.

[0311] In some embodiments, the buffer in the pharmaceutical composition described herein may maintain a pH of about 5 to 8. For example, the pH of the pharmaceutical composition may be about 7.1, 7.2, 7.3, 7.4, 7.5, 7.6, 7.7, 7.8, 7.9, or 8.0. In other instances, the pH of the pharmaceutical composition may be below 7, for example, about 7, 6.8, 6.5, 6.3, 6, 5.8, 5.5, 5.3, or 5.

[0312] The pharmaceutical compositions described herein comprise one or more suitable salts. Salts are ionic compounds that can be formed by the neutralization reaction of acids and bases. (Skoog, DA; West, DM; Holler, JF; Crouch, SR (2004). “chapters 14–16”. Fundamentals of Analytical Chemistry (8th ed.)). Salts consist of a relevant number of cations (positively charged ions) and anions (negative ions) such that the product is electrically neutral (without net charge).

[0313] In some embodiments, the pharmaceutical composition may comprise a pharmaceutically acceptable carrier, excipient, or stabilizer in the form of a lyophilized formulation or an aqueous solution. (Remington: The Science and Practice of Pharmacy 20th Ed. (2000) Lippincott Williams and Wilkins, Ed. KE Hoover). In some embodiments, the pharmaceutical composition may be formulated for intravenous injection. In some embodiments, the pharmaceutical composition may be formulated for subcutaneous injection.

[0314] Pharmaceutical compositions intended for internal administration must be sterile. This can be readily achieved, for example, by filtration through a sterile filter membrane. Therapeutic antibody compositions are typically placed in containers with sterile access ports, such as intravenous or subcutaneous solution bags or vials (with stoppers that can be punctured by a subcutaneous injection needle).

[0315] V. Usage Method

[0316] In some aspects, this disclosure provides methods and related compositions (e.g., anti-TMPRSS6 antibodies) for treating iron overload and iron overload-related conditions, including, for example, sickle cell disease (SCD), thalassemia (e.g., alpha-thalassemia, severe beta-thalassemia, mild beta-thalassemia), hemochromatosis (e.g., hemochromatosis type 1, hemochromatosis type 2, hemochromatosis type 3, hemochromatosis type 4), transfusion-related iron overload (e.g., repeated transfusions due to anemia, massive transfusions due to traumatic blood loss), hemolytic anemia (e.g., transfusion-dependent hemolytic anemia, pyruvate kinase deficiency hemolytic anemia), African iron overload, Blackfan-Diamond anemia, ringed sideroblastic refractory anemia (RARS), and myelodysplastic syndromes (MDS) (e.g., SF3B1-related MDS). In some embodiments, this disclosure provides methods for treating sickle cell disease (SCD). In some implementations, this disclosure provides a method for treating sickle cell disease in subjects suffering from iron overload.

[0317] Some aspects of this disclosure relate to methods and compositions (e.g., anti-TMPRSS6 antibodies) that can be used to treat iron overload. Iron overload occurs when there is an excess of iron stored in the body. In some embodiments, iron overload is primary iron overload caused by a hereditary condition (e.g., hemochromatosis). In some embodiments, iron overload is secondary iron overload caused by conditions that increase iron storage in the body (e.g., transfusion or hemolysis) or by diseases that impair erythrocyte production (i.e., ineffective erythropoiesis) (e.g., sickle cell disease, thalassemia, refractory anemia of ringed sideroblasts (RARS), MDS). Excess iron deposits in organs throughout the body, which can lead to organ damage. Organs commonly damaged due to iron deposition are the liver, heart, and endocrine glands.

[0318] In some respects, proteolytic enzyme-2 regulates iron homeostasis by modulating hepcidin levels in the organism. Iron is an essential component of almost all living cells and organisms. Excess iron is harmful (e.g., it promotes oxidative stress due to its redox reactivity). Therefore, iron metabolism disorders can lead to disease. In mammals, most of the body's iron (e.g., >70%) is distributed in red blood cells and mediates oxygen transport within hemoglobin. Senescent red blood cells are cleared by tissue macrophages, while iron from senescent red blood cells is recycled to erythroblasts for reuse. The release of iron into the plasma involves transferrin, specifically ferrous iron (Fe2+). 2+ The output product. After its output, ferrous ions are oxidized to ferric iron (Fe3+) by ceruloplasmin (a circulating ferrous oxidase). 3+ Iron is captured by transferrin, a siderophore. The main function of transferrin is to deliver iron to tissues via transferrin receptor 1 (TfR1). Duodenal intestinal epithelial cells convert iron from the intestinal lumen via divalent metal transporter 1 (DMT1) and then export it to the plasma via transferrin.

[0319] Iron entry into the bloodstream is crucial for systemic iron homeostasis and is negatively regulated by hepcidin (i.e., the iron-regulating hormone) (see, e.g., Ganz T. Systemic iron homeostasis. Physiol Rev. (2013) 93:1721-41). Hepcidin is expressed as a pre-propeptide in hepatocytes and undergoes proteolysis. Mature hepcidin is a 25-amino acid cysteine-rich peptide. In some cases, the binding of hepcidin to transferrin in tissue macrophages, duodenal intestinal epithelial cells, and other target cells triggers the ubiquitination, internalization, and degradation of transferrin in lysosomes. This leads to iron sequestration within macrophages, inhibiting dietary iron absorption and consequently reducing plasma iron levels. Therefore, iron levels can be regulated by manipulating hepcidin levels in the target organism. Hepcidin expression is regulated by the BMP signaling pathway and the IL-6-JAK-STAT signaling pathway. Hepcidin expression is promoted by the binding of BMPs (e.g., BMP6 or BMP2) to BMP receptors (e.g., type I (ALK2 and ALK3), type II (ActRIIA and BMPR2) BMP receptors) on the hepatocyte plasma membrane via the BMP pathway. BMP co-receptors, human iron homeostasis regulator (HFE), hemoblastin (HJV), and transferrin receptor 2 (TFR2) are each required for hepcidin expression via the BMP signaling pathway. The BMP-HJV signaling pathway is negatively regulated by serine protease cleavage enzyme 2 (TMPRSS6), which cleaves type I and type II BMP receptors and HJV, thereby inactivating them (Wahedi et al. Matriptase-2 suppresses hepcidin expression by cleaving multiple components of the hepcidin induction pathway. J BiolChem. (2017) 292:18354-71). In some embodiments, cleavage of HJV by proteolytic enzyme 2 reduces the amount of cell surface HJV available for participation in BMP signaling, thereby decreasing hepcidin expression. In some embodiments, loss of hemoblast function may be associated with iron overload (e.g., hemochromatosis type 2). In some embodiments, inhibition of HJV reduces hepcidin expression induced by the IL-6-JAK-STAT signaling pathway. For example, in some embodiments, homozygous HJV knockdown animals are unable to enhance hepcidin synthesis in response to IL-6 and cannot produce an effective hypohemoemic response to acute inflammation.Therefore, in some embodiments, inhibition of proteolytic enzyme 2 (e.g., via an anti-TMPRSS6 antibody) positively regulates hepcidin expression by maintaining / enhancing HJV activity. In some embodiments, iron overload is treated by reducing iron levels in a subject by increasing hepcidin levels through inhibition of proteolytic enzyme 2 (e.g., via an anti-TMPRSS6 antibody).

[0320] In some embodiments, the methods provided herein include administering an effective amount of anti-TMPRSS6 antibody to a subject for the purpose of increasing hepcidin expression. The increased hepcidin level in the subject reduces iron levels. In some embodiments, the methods provided herein treat iron overload in the subject by increasing hepcidin levels. In some embodiments, the methods provided herein reduce systemic iron. In some embodiments, the reduced systemic iron results in a decrease in mean corpuscular hemoglobin concentration (MCHC). In some embodiments, the methods provided herein include administering an effective amount of anti-TMPRSS6 antibody to a subject for the purpose of treating and / or improving symptoms and conditions associated with iron overload. For example, administering anti-TMPRSS6 antibody treats and / or improves one or more of the following symptoms: arthralgia, fatigue, weight loss, skin discoloration, abdominal pain, hair loss, loss of libido, memory impairment, or cardiac arrhythmia. In some implementations, the methods provided herein include administering an effective amount of anti-TMPRSS6 antibody to a subject for the purpose of improving symptoms and conditions associated with secondary iron overload (e.g., sickle cell disease, thalassemia, refractory anemia of ringed sideroblasts (RARS), MDS), as described elsewhere herein. In some implementations, the methods provided herein include administering an effective amount of anti-TMPRSS6 antibody to a subject for the treatment of sickle cell disease (SCD), thalassemia (e.g., alpha-thalassemia, severe beta-thalassemia, mild beta-thalassemia), hemochromatosis (e.g., hemochromatosis type 1, hemochromatosis type 2, hemochromatosis type 3, hemochromatosis type 4), transfusion-related iron overload (e.g., repeated transfusions due to anemia, massive transfusions due to traumatic blood loss), hemolytic anemia (e.g., transfusion-dependent hemolytic anemia, pyruvate kinase deficiency hemolytic anemia), African iron overload, Blackfan Diamond anemia, ring sideroblast-refractory anemia (RARS), and myelodysplastic syndromes (MDS) (e.g., SF3B1-associated MDS).

[0321] Other aspects of this disclosure relate to methods and compositions (e.g., anti-TMPRSS6 antibodies) that can be used to treat sickle cell disease (SCD). In some embodiments, the methods provided herein include administering an effective amount of an anti-TMPRSS6 antibody to a subject for the treatment of SCD in the subject. In some embodiments, the methods provided herein include administering an effective amount of an anti-TMPRSS6 antibody to a subject for the purpose of treating SCD in a subject with iron overload. In some embodiments, the methods provided herein reduce iron overload in a subject with SCD. In some embodiments, the methods provided herein reduce systemic iron levels in a subject with SCD. In some embodiments, the methods provided herein reduce mean corpuscular hemoglobin concentration (MCHC) in a subject with SCD. In some embodiments, the methods provided herein include administering an effective amount of an anti-TMPRSS6 antibody to a subject for the purpose of treating and / or improving symptoms and conditions associated with SCD (e.g., occlusive crisis). Sickle cell disease (SCD) is a collective term for a group of genetic diseases affecting hemoglobin genes. Sickle cell disease (SCD) is typically caused by an autosomal recessive mutation (e.g., a single nucleotide substitution) in the β-globin gene, which leads to the synthesis of sickle hemoglobin (HbS) and sickle red blood cells (RBCs) (Sundd et al., Pathophysiology of sickle cell disease. Annu Rev Pathol. 2019;14(1):263-292; Kato GJ, et al. Sickle cell disease. Nat Rev Dis Primers. 2018;4(1):18010). There are several types of sickle cell disease: hemoglobin SS disease, hemoglobin SC disease, hemoglobin SB+ (β) thalassemia, hemoglobin SB 0 (β 0) thalassemia, hemoglobin SD, hemoglobin SE, and hemoglobin SO. In hemoglobin SS disease (i.e., sickle cell anemia), the individual is homozygous for a mutation in the hemoglobin S (HbS) gene and produces only hemoglobin S. In hemoglobin SS disease, the typical disc shape of red blood cells (RBCs) changes to a sickle shape; this deformation disrupts their primary function. Other symptoms of hemoglobin SS disease include fatigue, recurrent infections, periodic pain, and damage to internal organs. Iron supplementation is not considered effective in increasing the level of hemoglobin present in the blood. In hemoglobin SC disease, the subject possesses one allele of the HbS gene and one allele of the hemoglobin C (HbC) gene. Typically, anemia is less severe in hemoglobin SC disease compared to hemoglobin SS disease. This may be because the hemoglobin C gene does not polymerize as rapidly as hemoglobin S. Therefore, this results in the formation of fewer sickle cells.The symptoms of hemoglobin SS disease and hemoglobin SC disease are similar, but the severity differs slightly.

[0322] In SCD (e.g., hemoglobin SS and / or hemoglobin SC), HbS polymerizes in rigid fibers during capillary deoxygenation, causing RBC membrane deformation, which can lead to intravascular hemolysis and inflammation. In some embodiments, the degree of HbS polymerization is correlated with the hemoglobin concentration in erythrocytes. The hemoglobin concentration in erythrocytes can be measured by mean erythrocyte hemoglobin concentration (MCHC). MCHC is a measure of the average amount of hemoglobin in a single erythrocyte. In some embodiments, reducing the hemoglobin concentration in erythrocytes can reduce HbS polymerization in patients with SCD (see, for example, Sunshine et al., Requirements for therapeutic inhibition of sickle haemoglobingelation, Nature volume 275, pages 238–240 (1978)). In some embodiments, reducing systemic iron leads to a reduction in HbS polymerization.In some implementations, reduced systemic iron leads to a decrease in MCHC, which in turn leads to a reduction in HbS aggregation (see, for example, Castro et al., Iron restriction in sickle cell anemia: Time for controlled clinical studies, American Journal of Hematology, Vol. 90, No. 12, December 2015; Brugnara, Less (Fe) is more (Hb) in SCA, Blood (2021) 137(11): 1446–1447; Embury et al., Concurrent sickle cell anemia and alpha-thalassemia. Effect on pathological properties of sickle erythrocytes, J ClinInvest. 1984 Jan; 73(1): 116–123; Brewin et al., The pleiotropic effects of alpha-thalassemia on HbSS and HbSC sickle cell disease: Reduced erythrocytecation co-transport activity, serum erythropoietin, and transfusion burden, do not translate). Into increased survival, Am J Hematol. 2022 Oct;97(10):1275-1285). Intravascular hemolysis in SCD results in the release of cell-free hemoglobin into circulation, which is oxidized and releases reactive heme into the vascular system. In some embodiments, the release of cell-free hemoglobin into circulation causes iron overload in patients with SCD. In some embodiments, cell-free heme activates endothelial cell adhesion molecules and induces leukocyte activation and migration, as well as the release of reactive oxygen species, cytokines, and chemokines. In some embodiments, HbS polymerization, hemolysis, and the release of hemoglobin and / or cell-free heme lead to vascular occlusive crisis (VOC). The terms vascular occlusive crisis (VOC), vascular occlusion, pain crisis, and vascular occlusive pain crisis used herein are interchangeable.Vascular occlusion is a major pathological process in SCD (Frenette PS. Sickle cell vaso-occlusion: multistep and multicellular paradigm. Curr Opin Hematol. (2002) 9:101-6; Pathra et al., Cytokines insickle cell disease. Hematology. (2003) 8:329-37). During VOC, in some implementations, the lumen of a blood vessel is blocked by cells, interrupting the flow of capillary blood to various organs and other parts of the body. This accelerates the inflammatory process, leading to a painful crisis and damage to multiple tissues, such as the brain, liver, kidneys, lungs, spleen, etc. Painful crises affect almost all patients with SCD, typically beginning in late infancy and recurring throughout life. In some implementations, patients with SCD are treated with blood transfusions (e.g., periodic on-demand transfusions or long-term transfusions). In some implementations, blood transfusion accelerates hemolysis and / or iron overload (e.g., Raghunath et al., Iron Overload in Sickle Cell Disease, Advances in Hematology, Volume 2010 | Article ID 272940).

[0323] In some implementations, subjects with SCD exhibit splenic sequestration. The spleen's primary function is to remove defective red blood cells, including sickle-shaped red blood cells. In patients with SCD, blood flow through the spleen is slow, leading to reduced oxygen tension and thus increased HbS aggregation. Due to the narrow capillaries in the splenic vascular bed, the spleen subsequently becomes filled with blood cells. Splenic sequestration (also known as splenic pooling) is characterized by acute splenomegaly (splenomegaly) accompanied by hemoglobin levels more than 2 g / dL below the affected individual's baseline (Bender and Carlberg, 2003 Sep 15 [updated 2023 Dec 28]. In: Adam MP, Feldman J, Mirzaa GM, Pagon RA, Wallace SE, BeanLJH, Gripp KW, Amemiya A, editors. GeneReviews® [Internet]. Seattle (WA): University of Washington, Seattle; 1993–2024.). In some implementations, subjects with splenic sequestration require hospitalization, splenectomy, and / or blood transfusions. In some implementations, administration of anti-TMPRSS6 antibody reduces splenic sequestration (e.g., frequency and / or severity) in subjects with SCD compared to subjects before administration or to untreated subjects with SCD. In some implementations, administration of anti-TMPRSS6 antibody reduces hospitalizations associated with splenic sequestration in subjects with SCD (e.g., hospitalizations associated with splenectomy) compared to subjects before administration or to untreated subjects with SCD. In some implementations, administration of anti-TMPRSS6 antibody reduces the frequency of blood transfusions required by subjects with SCD compared to subjects before administration or to untreated subjects with SCD. In some implementations, administration of anti-TMPRSS6 antibody reduces the size of the enlarged spleen in subjects with SCD (e.g., subjects with splenomegaly) compared to subjects before administration or to untreated subjects with SCD.

[0324] In some implementations, subjects with SCD experience extramedullary hematopoiesis (EMH) (see, for example, Gupta, et al., Clinicopathological characteristics and management of extramedullary hematopoiesis: A review. Pediatric Hematology Oncology Journal 7.4 (2022): 182-186). EMH refers to the production of blood cells outside the bone marrow, typically occurring in organs such as the liver, spleen, and lymph nodes. In subjects with SCD, EMH can occur due to chronic anemia and compensatory bodily responses to maintain adequate blood cell production. In some implementations, EMH in subjects with SCD occurs due to chronic hemolytic anemia, splenomegaly, ineffective erythropoiesis, VOCs and / or organ damage and organ dysfunction, all of which contribute to an increased demand for blood cell production. In some implementations, administration of anti-TMPRSS6 antibody reduces EMH in subjects with SCD compared to subjects prior to administration or compared to untreated subjects with SCD.

[0325] In some embodiments, subjects with SCD have hepatomegaly (see, for example, Burley et al., Acute Liver Failure in Sickle Cell Disease: A Perfect Storm. Cureus. 2021 Jun16;13(6):e15680). In some embodiments, the hepatomegaly in subjects with SCD is caused by intravascular congestion due to sickle cells, which in some embodiments leads to hepatic infarction. In some embodiments, the liver is enlarged due to congestion caused by obstructed blood flow. In some embodiments, the hepatomegaly in subjects with SCD is caused by sickle cell crisis (e.g., hepatic sequestration). In some embodiments, during a splenic sequestration crisis, blood also stagnates in the liver, leading to hepatomegaly. In some embodiments, the hepatomegaly in subjects with SCD is caused by liver dysfunction. In some embodiments, liver dysfunction in subjects with SCD is caused by chronic hemolysis, iron overload (e.g., from repeated transfusions), and / or hepatic congestion. In some embodiments, hepatomegaly in subjects with SCD is caused by gallbladder complications (e.g., gallstones and cholecystitis), which can lead to bile duct obstruction and hepatomegaly. In some embodiments, hepatomegaly in subjects with SCD is caused by iron overload of the liver (e.g., from chronic transfusion therapy) (e.g., secondary hemochromatosis). Excessive iron deposition in the liver can lead to hepatomegaly and liver dysfunction. In some embodiments, hepatomegaly in subjects with SCD is at least partially caused by hepatotoxicity associated with drugs used to treat SCD (e.g., hydroxyurea), leading to drug-induced liver injury. In some embodiments, administration of anti-TMPRSS6 antibody reduces hepatomegaly in subjects with SCD compared to subjects before administration or compared to untreated subjects with SCD. In some embodiments, administration of anti-TMPRSS6 antibody reduces liver sequestration in subjects with SCD compared to subjects before administration or compared to untreated subjects with SCD. In some embodiments, administration of anti-TMPRSS6 antibody reduced hepatic congestion in subjects with SCD compared to subjects before administration or to untreated subjects with SCD. In some embodiments, administration of anti-TMPRSS6 antibody reduced gallbladder complications (e.g., gallstones and / or cholecystitis) in subjects with SCD compared to subjects before administration or to untreated subjects with SCD. In some embodiments, administration of anti-TMPRSS6 antibody reduced hepatic iron overload in subjects with SCD compared to subjects before administration or to untreated subjects with SCD.In some embodiments, administration of the anti-TMPRSS6 antibody reduced liver dysfunction in subjects with SCD compared to subjects before administration or to untreated subjects with SCD. In some embodiments, administration of the anti-TMPRSS6 antibody reduced hepatic infarction in subjects with SCD compared to subjects before administration or to untreated subjects with SCD. In some embodiments, administration of the anti-TMPRSS6 antibody reduced hepatic congestion (e.g., bile duct obstruction) in subjects with SCD compared to subjects before administration or to untreated subjects with SCD.

[0326] In some embodiments, this disclosure provides methods and compositions (e.g., anti-TMPRSS6 antibody) for reducing VOCs in subjects with SCD (e.g., those receiving blood transfusions or not) by iron restriction. In some embodiments, administration of the anti-TMPRSS6 antibody reduces the frequency of VOCs in subjects with SCD (e.g., those receiving blood transfusions or not). In some embodiments, administration of the anti-TMPRSS6 antibody reduces the severity of VOCs in subjects with SCD (e.g., those receiving blood transfusions or not). In some embodiments, administration of the anti-TMPRSS6 antibody results in increased hepcidin levels in subjects with SCD (e.g., those receiving blood transfusions or not) relative to subjects prior to receiving the anti-TMPRSS6 antibody or subjects with SCD (e.g., those receiving blood transfusions or not) but not receiving the anti-TMPRSS6 antibody, thereby reducing circulating iron (e.g., circulating iron available for HbS synthesis).

[0327] In some implementations, the administration of anti-TMPRSS6 antibody results in a reduction in circulating iron in subjects with SCD (e.g., those receiving blood transfusions or those without blood transfusions) compared to subjects who received anti-TMPRSS6 antibody before or subjects with SCD (e.g., those receiving blood transfusions or those without blood transfusions) who did not receive anti-TMPRSS6 antibody (e.g., a reduction in circulating iron of at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%). In some implementations, administration of the anti-TMPRSS6 antibody results in a decrease in circulating iron in subjects with SCD (e.g., hemoglobinic SC) compared to subjects who had received the anti-TMPRSS6 antibody prior to treatment or subjects with SCD (e.g., hemoglobinic SC) who did not receive the anti-TMPRSS6 antibody (e.g., a decrease in circulating iron of at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%). Circulating iron levels can be evaluated by routine laboratory tests such as measurements of serum iron, transferrin saturation (TSAT), or total iron-binding capacity (TIBC).

[0328] In some implementations, the administration of anti-TMPRSS6 antibody results in an increase in circulating hepcidin-25 levels in subjects with SCD (e.g., those receiving blood transfusions or those without blood transfusions) relative to subjects who received anti-TMPRSS6 antibody before or subjects with SCD (e.g., those receiving blood transfusions or those without blood transfusions) who did not receive anti-TMPRSS6 antibody (e.g., an increase in circulating hepcidin-25 levels of at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 100%, at least 1.5 times, at least 2 times, at least 3 times, at least 4 times, at least 5 times, at least 10 times, at least 20 times, at least 50 times, or at least 100 times). In some implementations, the administration of anti-TMPRSS6 antibody results in an increase in circulating hepcidin-25 levels in subjects with SCD (e.g., hemoglobin SC disease) relative to subjects who received anti-TMPRSS6 antibody before or subjects with SCD (e.g., hemoglobin SC disease) who did not receive anti-TMPRSS6 antibody (e.g., an increase in circulating hepcidin-25 levels of at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 100%, at least 1.5 times, at least 2 times, at least 3 times, at least 4 times, at least 5 times, at least 10 times, at least 20 times, at least 50 times, or at least 100 times).

[0329] In some embodiments, administration of anti-TMPRSS6 antibody results in increased Hamp expression. In some embodiments, administration of anti-TMPRSS6 antibody results in increased Hamp expression in subjects with SCD (e.g., those receiving blood transfusions or not receiving blood transfusions) relative to subjects who previously received anti-TMPRSS6 antibody or subjects with SCD (e.g., those receiving blood transfusions or not receiving blood transfusions) who did not receive anti-TMPRSS6 antibody (e.g., an increase in Hamp expression of at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or 100%). In some implementations, compared to subjects who had received anti-TMPRSS6 antibodies before or subjects with SCD (e.g., hemoglobin SC disease) who did not receive anti-TMPRSS6 antibodies, administration of anti-TMPRSS6 antibodies resulted in increased Hamp expression in subjects with SCD (e.g., hemoglobin SC disease) (e.g., a reduction in bilirubin of at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or 100%).

[0330] In some embodiments, administration of anti-TMPRSS6 antibody results in a decrease in transferrin saturation (TSAT). In some embodiments, administration of anti-TMPRSS6 antibody results in a decrease in TSAT (e.g., a decrease of at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or 100%) in subjects with SCD (e.g., subjects with SCD who received blood transfusions or did not receive blood transfusions) compared to subjects who received anti-TMPRSS6 antibody prior to administration or subjects with SCD who received blood transfusions but did not receive anti-TMPRSS6 antibody. In some implementations, the administration of anti-TMPRSS6 antibody results in a reduction in TSAT in subjects with SCD (e.g., hemoglobin SC disease) compared to subjects who received anti-TMPRSS6 antibody before or subjects with SCD (e.g., hemoglobin SC disease) who did not receive anti-TMPRSS6 antibody (e.g., a reduction in TSAT of at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or 100%).

[0331] In some embodiments, administration of anti-TMPRSS6 antibody results in a reduction in extramedullary hematopoiesis. In some embodiments, administration of anti-TMPRSS6 antibody results in a reduction in extramedullary hematopoiesis (e.g., a reduction of at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or 100%) in subjects with SCD (e.g., subjects with SCD who received blood transfusions or did not receive blood transfusions) compared to subjects who received anti-TMPRSS6 antibody prior to administration or subjects with SCD who received blood transfusions but did not receive anti-TMPRSS6 antibody. In some implementations, administration of anti-TMPRSS6 antibody results in a reduction in extramedullary hematopoiesis in subjects with SCD (e.g., hemoglobinic SC disease) compared to subjects who did not receive anti-TMPRSS6 antibody prior to treatment or subjects with SCD (e.g., hemoglobinic SC disease) who did not receive anti-TMPRSS6 antibody (e.g., a reduction in extramedullary hematopoiesis of at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or 100%). Extramedullary hematopoiesis can be evaluated by suitable known methods, such as spleen weight to body weight ratio, imaging studies (e.g., X-ray, CT, MRI, ultrasound), biopsy, blood tests (e.g., blood cell morphology or blast level), bone marrow examination (e.g., bone marrow cellularity, changes in composition and structure), or erythropoietin levels and reticulocyte counts.

[0332] In some embodiments, administration of anti-TMPRSS6 antibody results in a reduction in hepatomegaly. In some embodiments, administration of anti-TMPRSS6 antibody results in a reduction in hepatomegaly in subjects with SCD (e.g., those receiving blood transfusions or not receiving blood transfusions) compared to subjects who previously received anti-TMPRSS6 antibody or subjects with SCD (e.g., those receiving blood transfusions or not receiving blood transfusions) who did not receive anti-TMPRSS6 antibody (e.g., a reduction in hepatomegaly of at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or 100%). In some implementations, administration of anti-TMPRSS6 antibody results in a reduction in hepatomegaly in subjects with SCD (e.g., hemoglobinocardial SC disease) compared to subjects who had previously received anti-TMPRSS6 antibody or subjects with SCD (e.g., hemoglobinocardial SC disease) who did not receive anti-TMPRSS6 antibody (e.g., a reduction in hepatomegaly of at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or 100%). Hepatomegaly can be evaluated by suitable known methods, such as liver weight to body weight ratio, imaging studies (e.g., CT, MRI, ultrasound, liver elastography), blood tests (e.g., liver function tests, liver injury markers [alanine aminotransferase and aspartate aminotransferase]), and biopsy.

[0333] In some implementations, the administration of anti-TMPRSS6 antibody results in a reduction in the frequency of vascular occlusive crisis (VOC) in subjects with SCD (e.g., those receiving blood transfusions or those without blood transfusions) compared to subjects who received anti-TMPRSS6 antibody before or subjects with SCD (e.g., those receiving blood transfusions or those without blood transfusions) who did not receive anti-TMPRSS6 antibody (e.g., a reduction in VOC frequency of at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or 100%). In some implementations, the administration of anti-TMPRSS6 antibody results in a reduction in VOC frequency in subjects with SCD (e.g., hemoglobin SC disease) compared to subjects who received anti-TMPRSS6 antibody before or subjects with SCD (e.g., hemoglobin SC disease) who did not receive anti-TMPRSS6 antibody (e.g., a reduction in VOC frequency of at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or 100%).

[0334] In some implementations, the administration of anti-TMPRSS6 antibody results in a reduction in the severity (e.g., hospitalization and / or duration) of vascular occlusive crisis (VOC) in subjects with SCD (e.g., those receiving blood transfusions or those without) compared to subjects who received anti-TMPRSS6 antibody prior to receiving it or subjects with SCD (e.g., those receiving blood transfusions or those without) without receiving anti-TMPRSS6 antibody (e.g., a reduction in VOC severity (e.g., hospitalization and / or duration) of at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or 100%). In some implementations, compared to subjects who received anti-TMPRSS6 antibodies before or subjects with SCD (e.g., hemoglobin SC disease) who did not receive anti-TMPRSS6 antibodies, administration of anti-TMPRSS6 antibodies resulted in a reduction in the frequency of vascular occlusive crisis (VOC) in subjects with SCD (e.g., hemoglobin SC disease) (e.g., a reduction in VOC of at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or 100%). In some implementations, administration of anti-TMPRSS6 antibody results in a reduction in the severity (e.g., hospitalization and / or duration) of vascular occlusive crisis (VOC) in subjects with SCD (e.g., hemoglobin SC disease) compared to subjects prior to receiving anti-TMPRSS6 antibody or subjects with SCD (e.g., hemoglobin SC disease) who did not receive anti-TMPRSS6 antibody (e.g., a reduction in VOC of at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or 100%). VOC (e.g., frequency and / or severity) can be evaluated using suitable known methods, such as complete blood count with white blood cell differential, platelet count, reticulocyte count, and comprehensive metabolomics including liver and kidney function tests. Typical laboratory results include an acute decrease in hemoglobin concentration, an increase in platelet count, an increase in reticulocyte count, and an increase in serum urea.

[0335] In some embodiments, the administration of anti-TMPRSS6 antibody results in a reduction in systemic iron. In some embodiments, the administration of anti-TMPRSS6 antibody results in a reduction in MCHC. In some embodiments, the reduced MCHC leads to a reduction in HbS polymerization. In some embodiments, the administration of anti-TMPRSS6 antibody results in a reduction in MCHC (e.g., a reduction of at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or 100%) in subjects with SCD (e.g., subjects with SCD who received a blood transfusion or did not receive a blood transfusion) compared to subjects who received anti-TMPRSS6 antibody prior to receiving it or subjects with SCD who did not receive anti-TMPRSS6 antibody. In some implementations, the administration of anti-TMPRSS6 antibody results in a reduction of MCHC in subjects with SCD (e.g., hemoglobin SC disease) by at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or 100%, compared to subjects who received anti-TMPRSS6 antibody before or subjects with SCD (e.g., hemoglobin SC disease) who did not receive anti-TMPRSS6 antibody. In some implementations, the administration of anti-TMPRSS6 antibody results in a reduction in HbS polymerization in subjects with SCD (e.g., those receiving blood transfusions or those without blood transfusions) compared to subjects who received anti-TMPRSS6 antibody before or subjects with SCD (e.g., those receiving blood transfusions or those without blood transfusions) who did not receive anti-TMPRSS6 antibody (e.g., a reduction in HbS polymerization of at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or 100%).In some implementations, administration of the anti-TMPRSS6 antibody results in a reduction in HbS polymerization in subjects with SCD (e.g., hemoglobinic SC disease) compared to those who received the anti-TMPRSS6 antibody prior to treatment or those who had SCD (e.g., hemoglobinic SC disease) but did not receive the anti-TMPRSS6 antibody (e.g., a reduction in HbS polymerization of at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or 100%). HbS polymerization can be evaluated by suitable known methods, such as solubility sickle assay, hemoglobin electrophoresis, isoelectric focusing, or high-performance liquid chromatography (see, for example, Arishi et al., Techniques for the Detection of Sickle Cell Disease: A Review, Micromachines 2021, 12(5), 519).

[0336] In some implementations, the administration of anti-TMPRSS6 antibody results in a reduction in RBC sickling in subjects with SCD (e.g., those who received blood transfusions or did not receive blood transfusions) compared to subjects who received anti-TMPRSS6 antibody before or subjects with SCD (e.g., those who received blood transfusions or did not receive blood transfusions) (e.g., a reduction in RBC sickling of at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or 100%). In some implementations, compared to subjects who had received anti-TMPRSS6 antibodies before or subjects with SCD (e.g., hemoglobinic SC disease) who did not receive anti-TMPRSS6 antibodies, administration of anti-TMPRSS6 antibodies resulted in a reduction in RBC sickling in subjects with SCD (e.g., hemoglobinic SC disease) (e.g., a reduction in RBC sickling of at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or 100%). RBC sickling can be evaluated by suitable known methods such as blood smears.

[0337] In some implementations, the administration of anti-TMPRSS6 antibody results in a reduction in hypochromic RBC levels in subjects with SCD (e.g., those receiving blood transfusions or those without) compared to subjects who received anti-TMPRSS6 antibody prior to receiving it or subjects with SCD (e.g., those receiving blood transfusions or those without) but who did not receive anti-TMPRSS6 antibody (e.g., a reduction in hypochromic RBC levels of at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or 100%). In some implementations, the administration of anti-TMPRSS6 antibody results in a reduction in hypochromic RBC levels in subjects with SCD (e.g., hemoglobin SC disease) compared to subjects who received anti-TMPRSS6 antibody prior to treatment or subjects with SCD (e.g., hemoglobin SC disease) who did not receive anti-TMPRSS6 antibody (e.g., a reduction in hypochromic RBC levels of at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or 100%).

[0338] In some implementations, the administration of anti-TMPRSS6 antibody results in a reduction in microcytic RBC levels in subjects with SCD (e.g., those receiving blood transfusions or not receiving blood transfusions) compared to subjects who received anti-TMPRSS6 antibody before or subjects with SCD (e.g., those receiving blood transfusions or not receiving blood transfusions) who did not receive anti-TMPRSS6 antibody (e.g., a reduction in microcytic RBC levels of at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or 100%). In some implementations, compared to subjects who received anti-TMPRSS6 antibody before or subjects with SCD (e.g., hemoglobin SC disease) who did not receive anti-TMPRSS6 antibody, administration of anti-TMPRSS6 antibody resulted in a reduction in microcytic RBC levels in subjects with SCD (e.g., hemoglobin SC disease) (e.g., a reduction in microcytic RBC levels of at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or 100%).

[0339] In some implementations, the administration of anti-TMPRSS6 antibody results in a reduction in hemolysis in subjects with SCD (e.g., those who received blood transfusions or did not receive blood transfusions) compared to subjects who received anti-TMPRSS6 antibody before or subjects with SCD (e.g., those who received blood transfusions or did not receive blood transfusions) (e.g., a reduction in hemolysis of at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or 100%). In some implementations, the administration of anti-TMPRSS6 antibody results in a reduction in hemolysis in subjects with SCD (e.g., hemoglobin SC disease) compared to subjects who received anti-TMPRSS6 antibody before or subjects with SCD (e.g., hemoglobin SC disease) who did not receive anti-TMPRSS6 antibody (e.g., a reduction in hemolysis of at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or 100%). Hemolysis can be evaluated by appropriate known methods, such as lactate dehydrogenase (LDH), unbound bilirubin, total bilirubin, direct bilirubin and haptoglobin tests, reticulocyte count, or total heme and heme-binding protein (see, for example, Hemolytic Anemias. ARUPConsult®. Retrieved March 15, 2023, arupconsult.com / content / hemolytic-anemias).

[0340] In some embodiments, administration of anti-TMPRSS6 antibody results in a reduction of lactate dehydrogenase (LDH). In some embodiments, administration of anti-TMPRSS6 antibody results in a reduction of LDH in subjects with SCD (e.g., those receiving blood transfusions or not receiving blood transfusions) relative to subjects who received anti-TMPRSS6 antibody or subjects with SCD (e.g., those receiving blood transfusions or not receiving blood transfusions) who did not receive anti-TMPRSS6 antibody (e.g., a reduction of LDH of at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or 100%). In some implementations, the administration of anti-TMPRSS6 antibody results in a reduction of LDH in subjects with SCD (e.g., hemoglobin SC disease) by at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or 100%, compared to subjects who received anti-TMPRSS6 antibody before or subjects with SCD (e.g., hemoglobin SC disease) who did not receive anti-TMPRSS6 antibody.

[0341] In some embodiments, administration of anti-TMPRSS6 antibody results in a reduction of bilirubin (e.g., direct bilirubin and / or total bilirubin). In some embodiments, administration of anti-TMPRSS6 antibody results in a reduction of bilirubin (e.g., direct bilirubin and / or total bilirubin) in subjects with SCD (e.g., those receiving blood transfusions or those without blood transfusions) relative to subjects who received anti-TMPRSS6 antibody or subjects with SCD who did not receive anti-TMPRSS6 antibody (e.g., those receiving blood transfusions or those without blood transfusions). The reduction is described as at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or 100%. In some implementations, the administration of anti-TMPRSS6 antibody results in a reduction in bilirubin (e.g., direct bilirubin and / or total bilirubin) in subjects with SCD (e.g., hemoglobin SC disease) compared to subjects who received anti-TMPRSS6 antibody before or subjects with SCD (e.g., hemoglobin SC disease) who did not receive anti-TMPRSS6 antibody (e.g., a reduction of at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or 100%).

[0342] In some embodiments, administration of anti-TMPRSS6 antibody results in a reduction of inflammation (e.g., systemic inflammation). In some embodiments, administration of anti-TMPRSS6 antibody results in a reduction of inflammation (e.g., a reduction of at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or 100%) in subjects with SCD (e.g., subjects with SCD who received a blood transfusion or did not receive a blood transfusion) compared to subjects who received anti-TMPRSS6 antibody before or subjects with SCD (e.g., subjects with SCD who received a blood transfusion or did not receive a blood transfusion). In some implementations, administration of the anti-TMPRSS6 antibody results in a reduction in inflammation (e.g., a reduction of at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or 100%) in subjects with SCD (e.g., hemoglobin SC disease) compared to subjects who had not received the anti-TMPRSS6 antibody prior to administration. Inflammation can be assessed by suitable known methods, such as blood tests (e.g., C-reactive protein (CRP) levels, erythrocyte sedimentation rate (ESR), white blood cell (WBC) count, neutrophil count, lymphocyte count, and levels of pro-inflammatory cytokines), imaging studies (e.g., CT, MRI, PET scans), and biopsies.

[0343] In some embodiments, administration of anti-TMPRSS6 antibody results in a reduction in white blood cell (WBC) count. In some embodiments, administration of anti-TMPRSS6 antibody results in a reduction in WBC count (e.g., at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or 100%) in subjects with SCD (e.g., those receiving blood transfusions or not) compared to subjects who previously received anti-TMPRSS6 antibody or subjects with SCD who did not receive anti-TMPRSS6 antibody. In some implementations, the administration of anti-TMPRSS6 antibody results in a reduction in WBC count in subjects with SCD (e.g., hemoglobin SC disease) by at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or 100%, compared to subjects who received anti-TMPRSS6 antibody before or subjects with SCD (e.g., hemoglobin SC disease) who did not receive anti-TMPRSS6 antibody.

[0344] In some embodiments, administration of anti-TMPRSS6 antibody results in a reduction in neutrophil count. In some embodiments, administration of anti-TMPRSS6 antibody results in a reduction in neutrophil count (e.g., a reduction of at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or 100%) in subjects with SCD (e.g., subjects with SCD who received a blood transfusion or did not receive a blood transfusion) compared to subjects who received anti-TMPRSS6 antibody prior to administration or subjects with SCD who received anti-TMPRSS6 antibody but did not receive anti-TMPRSS6 antibody. In some implementations, the administration of anti-TMPRSS6 antibody results in a reduction in neutrophil count in subjects with SCD (e.g., hemoglobin SC disease) compared to subjects who received anti-TMPRSS6 antibody prior to treatment or subjects with SCD (e.g., hemoglobin SC disease) who did not receive anti-TMPRSS6 antibody (e.g., a reduction in neutrophil count of at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or 100%).

[0345] In some embodiments, administration of anti-TMPRSS6 antibody results in a reduction in lymphocyte count. In some embodiments, administration of anti-TMPRSS6 antibody results in a reduction in lymphocyte count (e.g., a reduction of at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or 100%) in subjects with SCD (e.g., subjects with SCD who received blood transfusions or did not receive blood transfusions) compared to subjects who received anti-TMPRSS6 antibody prior to administration or subjects with SCD who received blood transfusions but did not receive anti-TMPRSS6 antibody. In some implementations, the administration of anti-TMPRSS6 antibody results in a reduction in lymphocyte count in subjects with SCD (e.g., hemoglobin SC disease) by at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or 100%, compared to subjects who received anti-TMPRSS6 antibody before or subjects with SCD (e.g., hemoglobin SC disease) who did not receive anti-TMPRSS6 antibody.

[0346] In some implementations, the administration of anti-TMPRSS6 antibody results in a reduction in transfusion frequency (e.g., at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or 100%) in subjects with SCD (e.g., those receiving transfusions or those not receiving transfusions) compared to subjects who did not receive anti-TMPRSS6 antibody prior to receiving it or subjects with SCD (e.g., those receiving transfusions or not receiving transfusions). In some implementations, the administration of anti-TMPRSS6 antibody results in a reduction in transfusion duration for subjects with SCD (e.g., hemoglobin SC disease) compared to subjects who received anti-TMPRSS6 antibody prior to receiving it or subjects with SCD (e.g., hemoglobin SC disease) who did not receive anti-TMPRSS6 antibody (e.g., a reduction in transfusion duration of at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or 100%).

[0347] In some embodiments, this disclosure provides a method for treating a subject with SCD by administering an effective amount of an anti-TMPRSS6 antibody in combination with any known treatment for SCD, such as hemoglobin S polymerization inhibitors (e.g., vorselotor), VOC-reducing agents (e.g., hydroxyurea, L-glutamine in oral powder form, lizapril, selective pyruvate kinase R (PKR) activators), pain relievers (e.g., anesthetics, opioids, gabapentin, cannabis), blood transfusions, stem cell transplantation, Exagmglogene autotemcel (exa-cel), LentiGlobin, GMI-1070, VIT-2763 (vamifeport), ticagrelor, vitamin D, simvastatin, AG-348 (mitapiva sulfate), propranolol, epinephrine, regardsone, atorvastatin, prasugrel, L-arginine, OTQ923, oxygen therapy, and gene therapy.

[0348] Other aspects of this disclosure relate to methods and compositions (e.g., anti-TMPRSS6 antibodies) that can be used to treat thalassemia (e.g., alpha-thalassemia, severe beta-thalassemia, or mild beta-thalassemia) and / or one or more conditions caused by thalassemia in subjects. In some embodiments, the methods provided herein include administering an effective amount of an anti-TMPRSS6 antibody to a subject for the purpose of treating thalassemia (e.g., alpha-thalassemia, severe beta-thalassemia, or mild beta-thalassemia) in subjects with iron overload. In some embodiments, the methods provided herein reduce iron overload in subjects with thalassemia (e.g., alpha-thalassemia, severe beta-thalassemia, or mild beta-thalassemia). “Thalassemia” includes the group of inherited blood disorders caused by defects in the synthesis of one or more hemoglobin chains. In some embodiments, alpha-thalassemia is caused by a reduction or lack of alpha-globin chain synthesis. In some embodiments, beta-thalassemia is caused by a reduction or lack of beta-globin chain synthesis. In some implementations, an imbalance in hemoglobin chains leads to hemolysis and impairs erythrocyte production.

[0349] Alpha-thalassemia results from insufficient or absent synthesis of alpha-globin chains, leading to an excess of beta-globin chains. Alpha-globin chain production is controlled by two genes on each chromosome 16 (i.e., four alpha-globin genes), and a deficiency is usually caused by the deletion of one or more of these genes (Farashi et al., Molecular basis of alpha-thalassemia. Blood Cells Mol Dis 2018; 70:43-53). A single gene deletion results in a silent carrier state of alpha-thalassemia, characterized by asymptomatic and normal hematological results. Double gene deletion causes mild alpha-thalassemia with microcytosis and usually without anemia. Triple gene deletion leads to an overproduction of hemoglobin H (HbH), which has four beta chains. Intermediate alpha-thalassemia or HbH disease causes microcytic anemia, hemolysis, and splenomegaly. Quadruple gene deletion leads to an overproduction of hemoglobin Bart's (HbBart's), which has four gamma chains. Severe alpha-thalassemia with HB Bart syndrome often leads to fatal fetal hydrops (Harteveld and Higgs, alpha-thalassemia. Orphanet J Rare Dis 2010; 5:13).

[0350] β-thalassemia results from insufficient or absent synthesis of β-globin chains, leading to an excess of α-globin chains and consequently impaired erythropoiesis. β-globin chain production is controlled by β-globin genes on chromosome 11. β-globin chain production can range from near-normal to completely absent, resulting in varying degrees of α-globin excess relative to β-globin chain production. Three main forms have been described: severe β-thalassemia (also known as transfusion-dependent β-thalassemia or Cooley's anemia), intermediate β-thalassemia, and mild β-thalassemia (Origa, β-Thalassemia. Genet Med 2017; 19(6):609-619). Individuals with severe β-thalassemia have two deleted or defective β-globin genes, resulting in the absence of β-globin chains. Subjects with severe β-thalassemia typically (e.g., in the first two years of life) exhibit severe anemia and require regular red blood cell (RBC) transfusions. Findings in untreated or poorly transfused patients with severe β-thalassemia may include hemolysis, growth retardation, pallor, jaundice, poor muscle tissue, hepatosplenomegaly, leg ulcers, masses due to extramedullary hematopoiesis, and skeletal changes due to bone marrow amplification. In some implementations, treatment options for severe β-thalassemia include regular transfusions, iron chelation, and management of secondary complications of iron overload. In some cases, spleen removal may be necessary, and bone marrow transplantation remains the only definitive cure currently available. Routine infusion therapy can lead to iron overload-related complications, including endocrine complications (growth retardation, impaired sexual maturation, diabetes, and parathyroid, thyroid, pituitary, and less commonly adrenal insufficiency), dilated cardiomyopathy, liver fibrosis, and cirrhosis.

[0351] Patients with intermediate β-thalassemia are caused by two missing or defective β-globin genes, exhibiting significant genetic heterogeneity and clinical polymorphism. They present with moderate anemia late in life and do not require regular transfusions. The main clinical features of these patients are erythroid bone marrow hypertrophy with intramedullary and extramedullary hematopoiesis and its complications (osteoporosis, erythropoietic tissue masses mainly affecting the spleen, liver, lymph nodes, chest and spine, as well as bone deformities and typical facial changes), hemolytic gallstones, iron deposition, painful leg ulcers, and an increased tendency to thrombosis. In some implementations, if a subject with intermediate β-thalassemia has severe and persistent symptoms (e.g., persistent leg ulcers and pain associated with leg ulcers, pulmonary hypertension, severe anemia, growth retardation), the subject may be able to tolerate a blood transfusion (see, for example, Cappellini et al., Guidelines for the Clinical Management of Thalassaemia, 2nd revised edition, Chapter 11, Thalassaemia Intermedia and HbE, Nicosia (CY), Thalassaemia International Federation; 2008, ISBN-13: 978-9963-623-70-9).

[0352] In mild β-thalassemia, only one β-globin gene is damaged or deleted. Subjects with mild β-thalassemia are mildly anemic, hypochromic, and microcytic (i.e., have elevated hypochromic and / or microcytic erythrocyte levels, respectively). In some embodiments, subjects with mild β-thalassemia who are receiving iron supplementation due to their anemia experience iron overload.

[0353] In some embodiments, iron overload is present in subjects with thalassemia (e.g., all forms of α-thalassemia and β-thalassemia). In some embodiments, iron overload is associated with an increased incidence of thalassemia in subjects with thalassemia (e.g., all forms of α-thalassemia and β-thalassemia). In some embodiments, iron overload in thalassemia subjects is secondary to blood transfusion (e.g., in severe β-thalassemia and intermediate β-thalassemia). In some embodiments, iron overload in β-thalassemia subjects (e.g., primarily severe β-thalassemia and intermediate β-thalassemia) is secondary to ineffective erythropoiesis, characterized by enhanced intestinal iron absorption mediated by hepcidin inhibition (see, e.g., Gardenghi et al., Ineffective erythropoiesis in β-thalassemia is characterized by increased iron absorption mediated by down-regulation of hepcidin and up-regulation of ferroportin, Blood. 2007 Jun 1; 109(11): 5027-5035). In some embodiments, inhibition of hepcidin expression enhances iron absorption from the gut and allows iron release from macrophages, thereby further increasing iron levels. In different forms of thalassemia, iron overload has different effects on different organs due to underlying iron loading mechanisms and / or iron deposition rates (see, e.g., Taher and Saliba, Ironoverload in thalassemia: different organs at different rates. Hematology AmSoc Hematol Educ Program 2017; 2017(1):265-271). In some embodiments, the methods described herein treat thalassemia (e.g., alpha-thalassemia, severe β-thalassemia, or mild β-thalassemia) in subjects. In some embodiments, the thalassemia is not intermediate β-thalassemia. In some embodiments, this disclosure provides a method for treating iron overload in subjects with thalassemia (e.g., alpha-thalassemia, severe β-thalassemia, or mild β-thalassemia) by administering an anti-TMPRSS6 antibody to the subject. In some embodiments, the anti-TMPRSS6 antibody disclosed herein inhibits protein lyase 2 activity, subsequently increasing hemojuglin (HJV) levels, leading to increased hepcidin levels. Subsequently, in some embodiments, hepcidin further acts to inhibit iron uptake, release, and recycling, thereby reducing iron levels.In some implementations, the methods described herein reduce or improve one or more symptoms associated with thalassemia. In some implementations, the methods described herein reduce symptoms associated with iron overload (e.g., joint pain, abdominal pain, fatigue, weakness, diabetes, heart failure, and / or liver failure) in subjects with thalassemia.

[0354] In some implementations, the administration of anti-TMPRSS6 antibody results in an increase in circulating hepcidin-25 levels in subjects with thalassemia (e.g., alpha-thalassemia, severe beta-thalassemia, or mild beta-thalassemia) compared to subjects who received the anti-TMPRSS6 antibody before or subjects with thalassemia (e.g., alpha-thalassemia, severe beta-thalassemia, or mild beta-thalassemia) who did not receive the anti-TMPRSS6 antibody (e.g., an increase in circulating hepcidin-25 levels of at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 100%, at least 1.5 times, at least 2 times, at least 3 times, at least 4 times, at least 5 times, at least 10 times, at least 20 times, at least 50 times, or at least 100 times).

[0355] In some implementations, administration of the anti-TMPRSS6 antibody results in a reduction in hemolysis in subjects with thalassemia (e.g., alpha-thalassemia, severe beta-thalassemia, or mild beta-thalassemia) compared to subjects who received the anti-TMPRSS6 antibody prior to treatment or subjects with thalassemia (e.g., alpha-thalassemia, severe beta-thalassemia, or mild beta-thalassemia) who did not receive the anti-TMPRSS6 antibody (e.g., a reduction in hemolysis of at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or 100%). Hemolysis can be evaluated using suitable known methods described elsewhere herein.

[0356] In some implementations, the administration of anti-TMPRSS6 antibody results in a reduction in transfusion duration (e.g., at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or 100%) in subjects with thalassemia (e.g., severe alpha-thalassemia, intermediate alpha-thalassemia, severe beta-thalassemia) compared to subjects who received anti-TMPRSS6 antibody before or subjects with thalassemia (e.g., severe alpha-thalassemia, intermediate alpha-thalassemia, severe beta-thalassemia) but did not receive anti-TMPRSS6 antibody, compared to subjects who did not receive anti-TMPRSS6 antibody before.

[0357] In some embodiments, this disclosure provides a method for treating a subject with thalassemia by administering an effective amount of anti-TMPRSS6 antibody to the subject in combination with any known treatment for thalassemia, such as blood transfusion, iron chelation, folic acid supplementation, bone marrow transplantation, stem cell transplantation, deferiprone, luspatercept, splenectomy, or gene therapy.

[0358] Other aspects of this disclosure relate to methods and compositions (e.g., anti-TMPRSS6 antibodies) for treating hereditary hemochromatosis (HH). In some embodiments, the methods provided herein include administering an effective amount of an anti-TMPRSS6 antibody to a subject for treating HH in the subject. In some embodiments, the methods provided herein include administering an effective amount of an anti-TMPRSS6 antibody to a subject for treating HH in a subject suffering from iron overload. In some embodiments, the methods provided herein reduce iron overload in a subject suffering from HH. In some embodiments, the methods provided herein include administering an effective amount of an anti-TMPRSS6 antibody to a subject to treat and / or improve symptoms and conditions associated with HH. Hereditary hemochromatosis (HH) includes many inherited conditions that can lead to progressive iron overload. There are four main types of hemochromatosis (HH), classified based on which proteins involved in iron homeostasis are affected (see, for example, Kowdley et al., ACG Clinical Guideline: Hereditary Hemochromatosis, The American Journal of Gastroenterology 114(8):p 1202-1218, August 2019). Type 1 HH is the most common genetic form of iron overload. In some embodiments, type 1 HH results from a G-to-A shift at nucleotide 845 of the HFE gene, leading to a cysteine ​​substitution at amino acid 282 for a tyrosine residue (C282Y), referred to as type 1a HH. In other embodiments, type 1 HH results from an H63D mutation. In some implementations, subjects with the H63D mutation in the HFE gene are not at risk of developing clinically significant iron overload (see, for example, Gochee et al., A population-based study of the biochemical and clinical expression of the H63Dhemochromatosis mutation, Gastroenterology. 2002 Mar;122(3):646-51). In some implementations, type 1 HH is the result of C282Y / H63D substitution and is classified as type 1b HH.In some implementations, clinically significant iron overload is rare in individuals with type 1b HH unless cofactors such as alcohol or hepatitis C virus (HCV) are involved (see, for example, Alissa et al., The Clinical Relevance of Compound Heterozygosity for the C282Y and H63D Substitutions in Hemochromatosis, Clinical Gastroenterology & Hepatology, November 2006, Volume 4 (11), 1403–1410). In some implementations, type 1 HH results from an S65C mutation in the HFE gene and is referred to as type 1c HH. Typically, the S65C mutation is considered a polymorphism of no clinical significance.

[0359] Type 2 hemochromatosis, also known as juvenile hemochromatosis, is associated with mutations in either the HJV gene (type 2A) or the hepatic antimicrobial protein (HAMP) gene (type 2B), leading to hepcidin deficiency (Papanikolaou et al., Mutations in HFE2 cause iron overload in chromosome 1q-linked juvenile hemochromatosis, Nat Genet. 2004 Jan;36(1):77-82). In some implementations, type 2 hemochromatosis tends to result in the most severe form of primary iron overload. In some implementations, type 2 hemochromatosis primarily occurs in young individuals.

[0360] Type 3 hemochromatosis (HH) is associated with mutations in the transferrin receptor 2 (TFR2) gene, which also lead to hepcidin deficiency (Camaschella et al., The gene TFR2 is mutated in a new type of haemochromatosis mapping to 7q22, Nat Genet. 2000 May;25(1):14-5). In some embodiments, mutations in the TFR2 gene cause HH through reduced hepcidin transcription (see, for example, Christal et al., The role of hepatic transferrin receptor 2 in the regulation of ironhomeostasis in the body, Front Pharmacol. 2014; 5: 34). In some embodiments, iron overload is present in type 3 HH but is far less severe than in type 2 HH.

[0361] Type 4 hemochromatosis (HH) results from mutations in the membrane iron transporter 1 (FPN1 gene). Type 4A HH, also known as FPN disease, is an autosomal dominant form of hemochromatosis caused by mutations in the FPN1 gene (SLC40A1) (Abboud et al., novel mammalian iron-regulated protein involved in intracellular ironmetabolism, J Biol Chem. 2000 Jun 30;275(26):19906-12). In type 4A HH, hepcidin production is normal, but the export function of FPN1 is weakened, leading to intracellular iron retention, accompanied by low levels of plasma iron and normal or low levels of transferrin saturation, but elevated serum ferritin levels (33). The spleen is the most affected organ in type 4A HH due to its high FPN1 activity at the macrophage level. Type 4B HH is a form of iron overload caused by resistance of FPN1 to hepcidin.

[0362] In some embodiments, the methods provided herein include administering an effective amount of an anti-TMPRSS6 antibody to a subject for the treatment of hemochromatosis (HH) in the subject. In some embodiments, the methods provided herein include administering an effective amount of an anti-TMPRSS6 antibody to a subject for the treatment of a patient with a hereditary hemochromatosis with iron overload (e.g., type 1 HH, type 2, type 3 HH, or type 4 HH). In some embodiments, the anti-TMPRSS6 antibody disclosed herein inhibits proteolytic enzyme-2 activity, subsequently increasing hemojuglin (HJV) levels, leading to elevated hepcidin levels. Subsequently, in some embodiments, hepcidin further inhibits iron absorption, release, and recycling, thereby reducing iron levels and thus reducing iron overload in patients with a hereditary hemochromatosis (e.g., type 1 HH, type 2, type 3 HH, or type 4 HH). In some embodiments, the methods described herein reduce or improve one or more symptoms associated with a hereditary hemochromatosis (e.g., type 1 HH, type 2, type 3 HH, or type 4 HH). In some implementations, the methods described herein reduce iron overload-related symptoms in patients with hereditary hemochromatosis (e.g., type 1 HH, type 2, type 3 HH, or type 4 HH), such as joint pain, abdominal pain, fatigue, weakness, diabetes, loss of libido, impotence, heart failure, liver failure, bronze or gray skin, and memory impairment.

[0363] In some embodiments, this disclosure provides a method for treating iron overload in subjects suffering from hereditary hemochromatosis (e.g., type I HH, or type 3 HH), the method being performed by administering an effective amount of anti-TMPRSS6 antibody to the subject in combination with any known treatment for hereditary hemochromatosis (e.g., type I HH, type 2, type 3 HH, or type 4 HH) (e.g., venipuncture, iron chelation (e.g., deferoxamine or deferasirox), supportive care for complications, etc.).

[0364] Other aspects of this disclosure relate to methods and compositions (e.g., anti-TMPRSS6 antibodies) for treating myelodysplastic syndromes (MDS). In some embodiments, the methods provided herein include administering an effective amount of an anti-TMPRSS6 antibody to a subject for treating MDS in the subject. In some embodiments, the methods provided herein include administering an effective amount of an anti-TMPRSS6 antibody to a subject for treating MDS in a subject suffering from iron overload. In some embodiments, the methods provided herein reduce iron overload in a subject suffering from MDS. In some embodiments, the methods provided herein include administering an effective amount of an anti-TMPRSS6 antibody to a subject to treat and / or improve symptoms and conditions associated with MDS. Myelodysplastic syndromes (MDS) are a heterogeneous group of hematologic malignancies of varying severity that affect one or more hematopoietic lineages. In some embodiments, ineffective erythropoiesis leads to iron metabolism disorders. In some embodiments, iron overload begins to occur in an MDS patient before the patient becomes transfusion dependent because ineffective erythropoiesis inhibits hepcidin production in the liver, resulting in unrestricted intestinal iron uptake. In some implementations, ineffective erythropoiesis (a hallmark of MDS) leads to a massive expansion of erythroblasts in the bone marrow due to reduced production of mature RBCs (Sebastiani G et al., Pharmacological targeting of the hepcidin / ferroportin axis. Frontiers in Pharmacology. 2016;7:160.). This results in a high demand for iron, leading to inhibition of hepcidin. In some implementations, hepcidin levels vary across different MDS subtypes. In some implementations, the lowest hepcidin levels are observed in ringed sideroblastic refractory anemia (RARS). In some implementations, RARS patients have high levels of toxic nontransferrin-bound iron compared to patients with other types of MDS who have higher hepcidin levels. Furthermore, and in some embodiments, tissue hypoxia caused by ineffective erythropoiesis triggers increased erythropoietin production, which then leads to low hepcidin levels and consequently iron overload (Cui et al., Serum iron metabolism and erythropoiesis in patients with myelodysplastic syndrome not receiving RBCtransfusions. Leukemia research. 2014;38:545–550). Therefore, in some embodiments, this disclosure provides methods and compositions for treating refractory anemia of ringed sideroblasts (RARS).In some embodiments, the methods provided herein include administering an effective amount of anti-TMPRSS6 antibody to a subject to treat and / or improve symptoms and conditions associated with RARS. In some embodiments, the methods provided herein include administering an effective amount of anti-TMPRSS6 antibody to a subject to reduce iron overload associated with RARS. In some embodiments, administration of anti-TMPRSS6 antibody results in increased hepcidin levels in subjects with MDS (e.g., RARS). In some embodiments, administration of anti-TMPRSS6 antibody results in reduced iron overload in subjects with MDS (e.g., RARS).

[0365] In some embodiments, the subject of MDS has a mutation in the SF3B1 gene. SF3B1 is a gene encoding a component of the RNA splicing mechanism, and mutations in this gene have been found in a variety of types of MDS, including RARS. In some embodiments, MDS patients with the SF3B1 mutation have significantly lower hepcidin to ferritin ratios compared to patients without the mutation (Ilaria et al., Inappropriately low hepcidin levels in patients with myelodysplastic syndrome carrying a somatic mutation of SF3B1. Haematologica. 2013;98:420–423). In some embodiments, ineffective erythropoiesis in MDS patients with the SF3B1 mutation leads to low hepcidin levels in MDS-RARS patients with the SF3B1 mutation, ultimately resulting in excessive release of iron from the RES and substantial iron overload. Therefore, in some embodiments, this disclosure provides methods and compositions for treating MDS with the SF3B1 mutation. In some embodiments, the methods provided herein include administering an effective amount of anti-TMPRSS6 antibody to a subject to treat and / or improve symptoms and conditions associated with MDS having an SF3B1 mutation. In some embodiments, the methods provided herein include administering an effective amount of anti-TMPRSS6 antibody to a subject to reduce iron overload associated with MDS having an SF3B1 mutation. In some embodiments, administration of the anti-TMPRSS6 antibody results in increased hepcidin levels in subjects with MDS (e.g., MDS with an SF3B1 mutation). In some embodiments, administration of the anti-TMPRSS6 antibody results in reduced iron overload in subjects with MDS (e.g., MDS with an SF3B1 mutation).

[0366] In some embodiments, RARS patients have a high incidence of SF3B1 mutations (see, for example, Zhu et al., SF3B1-mutated myelodysplastic syndrome with ring sideroblasts harbors more severe iron overload and corresponding over-erythropoiesis. Leukemiaresearch. 2016;44:8–16). In some embodiments, hepcidin levels are lower in RARS subjects with SF3B1 mutations compared to wild-type RARS patients without SF3B1 mutations. In some embodiments, iron overload is more severe in RARS subjects with SF3B1 mutations compared to RARS patients without SF3B1 mutations. Therefore, in some embodiments, this disclosure provides methods and compositions for treating RARS with SF3B1 mutations. In some embodiments, the methods provided herein include administering an effective amount of an anti-TMPRSS6 antibody to a subject to treat and / or improve symptoms and conditions associated with RARS with SF3B1 mutations. In some embodiments, the methods provided herein include administering an effective amount of anti-TMPRSS6 antibody to a subject to reduce iron overload associated with RARS having an SF3B1 mutation. In some embodiments, administration of the anti-TMPRSS6 antibody results in increased hepcidin levels in subjects with MDS (e.g., RARS with an SF3B1 mutation). In some embodiments, administration of the anti-TMPRSS6 antibody results in reduced iron overload in subjects with MDS (e.g., RARS with an SF3B1 mutation).

[0367] Furthermore, in some implementations, MDS patients have anemia, and some require regular red blood cell transfusions. Long-term transfusion therapy is also a major cause of iron overload in patients with MDS. In some implementations, MDS patients with iron overload have lower overall survival and worse outcomes after allogeneic stem cell transplantation compared to patients without iron overload (see, e.g., Gattermann, Iron overload in myelodysplastic syndromes (MDS), International Journal of Hematology volume 107, pages 55–63 (2018); Lyle et al., Iron Overload in Myelodysplastic Syndromes: Pathophysiology, Consequences, Diagnosis, and Treatment, J Adv Pract Oncol. 2018;9(4):392-405). Therefore, in some implementations, the methods described herein include administering an effective amount of anti-TMPRSS6 antibody to the subject to reduce transfusion-related iron overload in MDS patients.

[0368] In some implementations, the methods provided herein improve symptoms and / or clinical outcomes associated with iron overload in subjects with MDS (e.g., RARS, MDS with SF3B1, RARS with SF3B1), such as survival, cardiovascular function, liver function, and immune function.

[0369] In some embodiments, this disclosure provides a method for treating a subject with MDS (e.g., RARS, MDS with SF3B1, RARS with SF3B1) by administering an effective amount of an anti-TMPRSS6 antibody to the subject in combination with any known treatment for treating MDS (e.g., RARS, MDS with SF3B1, RARS with SF3B1), such as iron chelation, bone marrow transplantation, stem cell transplantation, blood transfusion, EPO, rotezip, granulocyte colony-stimulating factor (GCSF), antithymocyte globulin (ATG), cyclosporine (CSA), hypomethylating agent (HMA), lenalidomide (LEN), thrombopoietin receptor agonist (TPO-RA), etc.

[0370] Other aspects of this disclosure relate to methods and compositions (e.g., anti-TMPRSS6 antibodies) for treating hemolytic anemia (e.g., transfusion-dependent hemolytic anemia, pyruvate kinase deficiency hemolytic anemia). In some embodiments, hemolysis results in a large outflow of hemoglobin into the bloodstream, leading to iron overload. In some embodiments, hemolysis induces EPO production to activate erythropoiesis (e.g., in the absence of ineffective erythropoiesis). In some embodiments, increased EPO and erythropoiesis inhibit hepcidin expression, thereby inducing iron overload in subjects with hemolytic anemia. Therefore, in some embodiments, this disclosure provides methods and compositions for treating hemolytic anemia (e.g., transfusion-dependent hemolytic anemia, pyruvate kinase deficiency hemolytic anemia). In some embodiments, the methods provided herein include administering an effective amount of an anti-TMPRSS6 antibody to a subject for treating hemolytic anemia (e.g., transfusion-dependent hemolytic anemia, pyruvate kinase deficiency hemolytic anemia). In some embodiments, the methods provided herein include administering an effective amount of anti-TMPRSS6 antibody to a subject to treat hemolytic anemia in a subject with iron overload. In some embodiments, the methods provided herein reduce iron overload in a subject suffering from hemolytic anemia. In some embodiments, the methods provided herein include administering an effective amount of anti-TMPRSS6 antibody to a subject to treat and / or improve symptoms and conditions associated with hemolytic anemia (e.g., transfusion-dependent hemolytic anemia, pyruvate kinase deficiency hemolytic anemia). In some embodiments, the methods provided herein include administering an effective amount of anti-TMPRSS6 antibody to a subject to reduce iron overload associated with hemolytic anemia (e.g., transfusion-dependent hemolytic anemia, pyruvate kinase deficiency hemolytic anemia). In some embodiments, administration of anti-TMPRSS6 antibody results in increased hepcidin levels in a subject suffering from hemolytic anemia (e.g., transfusion-dependent hemolytic anemia, pyruvate kinase deficiency hemolytic anemia). In some implementations, administration of anti-TMPRSS6 antibody results in a reduction of iron overload in subjects with hemolytic anemia (e.g., transfusion-dependent hemolytic anemia, pyruvate kinase deficiency hemolytic anemia).

[0371] Other aspects of this disclosure relate to methods and compositions (e.g., anti-TMPRSS6 antibodies) for treating transfusion-related iron overload. Since the early 20th century, blood transfusions have been widely and sometimes overused in medical practice to treat a variety of conditions (e.g., anemia and / or bleeding). Transfused red blood cells (RBCs) provide beneficial effects to the recipient, such as increasing blood volume, reducing blood viscosity, and improving oxygen-carrying capacity. In some embodiments, blood transfusions (e.g., massive transfusions) are used for recipients of blood loss (e.g., blood loss due to trauma or surgery). In some embodiments, blood transfusions (e.g., repeated transfusions) are used for recipients of anemia (e.g., SCD, thalassemia, hemochromatosis, MDS, iron deficiency anemia, Diamond-Blackfan anemia, etc.). In some embodiments, the recipient receives an excess of iron through transfusion. In some embodiments, the excess iron from the transfusion is stored in various tissues, leading to iron overload. Furthermore, in some embodiments, the transfusion causes hemolysis, thereby releasing iron into the circulation, further causing iron overload. In some embodiments, this disclosure provides methods and compositions for treating iron overload associated with blood transfusions (e.g., transfusions for anemia and / or bleeding). In some embodiments, the methods provided herein include administering an effective amount of an anti-TMPRSS6 antibody to a subject for treating iron overload associated with blood transfusions (e.g., transfusions for anemia and / or bleeding). In some embodiments, the methods provided herein reduce iron overload in subjects suffering from hemolytic anemia. In some embodiments, the methods provided herein include administering an effective amount of an anti-TMPRSS6 antibody to a subject to reduce iron overload associated with blood transfusions (e.g., transfusions for anemia and / or bleeding). In some embodiments, administration of the anti-TMPRSS6 antibody results in an increase in hepcidin levels in subjects with iron overload associated with blood transfusions (e.g., transfusions for anemia and / or bleeding). In some embodiments, administration of the anti-TMPRSS6 antibody results in a reduction of iron overload in subjects with iron overload associated with blood transfusions (e.g., transfusions for anemia and / or bleeding).

[0372] Other aspects of this disclosure relate to methods and compositions (e.g., anti-TMPRSS6 antibodies) for treating African iron overload. In some embodiments, the methods provided herein include administering an effective amount of an anti-TMPRSS6 antibody to a subject for treating African iron overload. In some embodiments, the methods provided herein reduce iron overload in subjects suffering from African iron overload. African iron overload is iron overload caused by a high-iron diet and genetic factors. In some embodiments, administration of the anti-TMPRSS6 antibody results in an increase in hepcidin levels in subjects suffering from African iron overload. In some embodiments, administration of the anti-TMPRSS6 antibody results in a decrease in iron overload in subjects suffering from African iron overload.

[0373] Other aspects of this disclosure relate to methods and compositions (e.g., anti-TMPRSS6 antibodies) for treating Diamond-Blackfan anemia. In some embodiments, the methods provided herein include administering an effective amount of an anti-TMPRSS6 antibody to a subject for treating Diamond-Blackfan anemia. In some embodiments, the methods provided herein include administering an effective amount of an anti-TMPRSS6 antibody to a subject for treating Diamond-Blackfan anemia in a subject with iron overload. In some embodiments, the methods provided herein reduce iron overload in a subject. In some embodiments, the methods provided herein include administering an effective amount of an anti-TMPRSS6 antibody to a subject for treating symptoms and conditions associated with Diamond-Blackfan anemia. Diamond-Blackfan anemia (DBA) is a hereditary blood disorder that affects the bone marrow's ability to produce red blood cells. In some embodiments, Diamond-Blackfan anemia is caused by genetic changes in several genes, including but not limited to: RPS19, RPL5, RPS10, RPL11, RPL35A, RPS7, RPS17, RPS24, RPS26, and GATA1 genes. In some embodiments, patients with DBA are dependent on blood transfusions because they cannot produce red blood cells (see, for example, Roggero et al., Severe iron overload in Blackfan-Diamond anemia: a case-control study, Am J Hematol. 2009 Nov;84(11):729-32; Quarello et al., Diamond-Blackfan anemia with iron overload: A serious issue, 2022 Oct;199(2):171-172). Repeated transfusions lead to iron overload, as described elsewhere herein. Therefore, in some embodiments, the methods provided herein include administering an effective amount of anti-TMPRSS6 antibody to the subject to reduce transfusion-related iron overload in subjects with Diamond-Blackfan anemia. In some embodiments, the administration of anti-TMPRSS6 antibody results in an increase in hepcidin levels in Diamond-Blackfan anemia subjects receiving transfusions. In some embodiments, the administration of anti-TMPRSS6 antibody results in a reduction in iron overload in Diamond-Blackfan anemia subjects receiving transfusions.

[0374] Based on the teachings provided herein, it will be apparent to those skilled in the art that determining whether an antibody (e.g., an anti-TMPRSS6 antibody) is effective will be appropriate. As will be recognized by those skilled in the art, the effective amount varies depending on: the specific condition being treated, the severity of the condition, individual patient parameters (including age, physical condition, body size, sex, and weight), the duration of treatment, the nature of concurrent treatment (if any), the specific route of administration, and other factors within the knowledge and expertise of the healthcare practitioner. As discussed herein, the specific dosing regimens used in the methods described herein—namely, the dosage, timing, and repetition—will depend on the specific subject and that subject's medical history.

[0375] Empirical considerations, such as the time to reach maximum effect, half-life, and / or time above a specific concentration, often help determine the dosage. For example, antibodies compatible with the human immune system, such as humanized or fully human antibodies, can be used to prolong the antibody's half-life and prevent it from being attacked by the host immune system. Other reasons for dosage adjustment include differences in pharmacokinetic or pharmacodynamic responses driven by sex, age, individual response, antibody target, and / or receptor polymorphisms involved in antibody clearance. The frequency of administration can be determined and adjusted during the course of treatment and is typically (but not necessarily) based on the treatment and / or inhibition and / or improvement and / or delay of the target disease / symptom. Alternatively, a sustained-release formulation of the ant...

Claims

1. A method for treating sickle cell disease (SCD), the method comprising administering to a subject an effective amount of an anti-transmembrane serine protease 6 (TMPRSS6) antibody, said antibody comprising: (a) HC CDR1, HC CDR2 and HCCDR3 having the amino acid sequence of SEQ ID NO: 49, and LC CDR1, LC CDR2 and LCCDR3 having the amino acid sequence of SEQ ID NO: 50; (b) HC CDR1, HC CDR2 and HCCDR3 having the heavy chain variable domain of the amino acid sequence of SEQ ID NO: 7, and LC CDR1, LC CDR2 and LCCDR3 having the light chain variable domain of the amino acid sequence of SEQ ID NO: 8; (c) HC CDR1, HC CDR2 and HCCDR3 having the amino acid sequence of SEQ ID NO: 19 or 78, and LC CDR1, LC CDR2 and LCCDR3 having the amino acid sequence of SEQ ID NO: 20; (d) HC CDR1, HC CDR2 and HCCDR3 having the amino acid sequence of SEQ ID NO: 30, and LC CDR1, LC CDR2 and LCCDR3 having the amino acid sequence of SEQ ID NO: 31; (e) HC CDR1, HC CDR2, and HCCDR3 having the heavy chain variable domain of the amino acid sequence of SEQ ID NO: 37, and LC CDR1, LC CDR2, and LCCDR3 having the light chain variable domain of the amino acid sequence of SEQ ID NO: 38; or (f) HC CDR1, HC CDR2 and HCCDR3 having the amino acid sequence of SEQ ID NO: 44 and the light chain variable domain having the amino acid sequence of SEQ ID NO:

45.

2. The method of claim 1, wherein the anti-TMPRSS6 antibody comprises HC CDR1, HC CDR2 and HC CDR3 having the amino acid sequence of SEQ ID NO: 49, and LC CDR1, LC CDR2 and LC CDR3 having the amino acid sequence of SEQ ID NO:

50.

3. The method of claim 1 or 2, wherein the anti-TMPRSS6 antibody comprises: (a) HC CDR1 having the amino acid sequence of SEQ ID NO: 24, HC CDR2 having the amino acid sequence of SEQ ID NO: 48, HC CDR3 having the amino acid sequence of SEQ ID NO: 26, LC CDR1 having the amino acid sequence of SEQ ID NO: 27, LC CDR2 having the amino acid sequence of WAT, and LCCDR3 having the amino acid sequence of SEQ ID NO: 29; (b) HC CDR1 having the amino acid sequence of SEQ ID NO: 1, HCCDR2 having the amino acid sequence of SEQ ID NO: 2, HC CDR3 having the amino acid sequence of SEQ ID NO: 3, LCCDR1 having the amino acid sequence of SEQ ID NO: 4, LCCDR2 having the amino acid sequence of RAN, and LCCDR3 having the amino acid sequence of SEQ ID NO: 6; (c) HC CDR1 having the amino acid sequence of SEQ ID NO: 13, HC CDR2 having the amino acid sequence of SEQ ID NO: 14, HC CDR3 having the amino acid sequence of SEQ ID NO: 15, LC CDR1 having the amino acid sequence of SEQ ID NO: 16, LC CDR2 having the amino acid sequence of WAF, and LCCDR3 having the amino acid sequence of SEQ ID NO: 18; (d) HC CDR1 having the amino acid sequence of SEQ ID NO: 24, HC CDR2 having the amino acid sequence of SEQ ID NO: 25, HC CDR3 having the amino acid sequence of SEQ ID NO: 26, LC CDR1 having the amino acid sequence of SEQ ID NO: 27, LC CDR2 having the amino acid sequence of WAT, and LCCDR3 having the amino acid sequence of SEQ ID NO: 29; (e) HC CDR1 having the amino acid sequence of SEQ ID NO: 1, HCCDR2 having the amino acid sequence of SEQ ID NO: 2, HC CDR3 having the amino acid sequence of SEQ ID NO: 35, LCCDR1 having the amino acid sequence of SEQ ID NO: 36, LC CDR2 having the amino acid sequence of RAN, and LC CDR3 having the amino acid sequence of SEQ ID NO: 6; or (f) HC CDR1 having the amino acid sequence of SEQ ID NO: 13, HC CDR2 having the amino acid sequence of SEQ ID NO: 43, HC CDR3 having the amino acid sequence of SEQ ID NO: 15, LC CDR1 having the amino acid sequence of SEQ ID NO: 16, LC CDR2 having the amino acid sequence of WAF, and LCCDR3 having the amino acid sequence of SEQ ID NO:

18.

4. The method of any one of claims 1 to 3, wherein the anti-TMPRSS6 antibody comprises HC CDR1 having the amino acid sequence of SEQ ID NO: 24, HC CDR2 having the amino acid sequence of SEQ ID NO: 48, HC CDR3 having the amino acid sequence of SEQ ID NO: 26, LC CDR1 having the amino acid sequence of SEQ ID NO: 27, LC CDR2 having the amino acid sequence of WAT, and LC CDR3 having the amino acid sequence of SEQ ID NO:

29.

5. The method of any one of claims 1 to 4, wherein the anti-TMPRSS6 antibody comprises: (a) A heavy chain variable domain having the amino acid sequence of SEQ ID NO: 49 and a light chain variable domain having the amino acid sequence of SEQ ID NO: 50; (b) A heavy chain variable domain having the amino acid sequence of SEQ ID NO: 7 and a light chain variable domain having the amino acid sequence of SEQ ID NO: 8; (c) A heavy chain variable domain having an amino acid sequence of SEQ ID NO: 19 or 78 and a light chain variable domain having an amino acid sequence of SEQ ID NO: 20; (d) A heavy chain variable domain having the amino acid sequence of SEQ ID NO: 30 and a light chain variable domain having the amino acid sequence of SEQ ID NO: 31; (e) A heavy chain variable domain having the amino acid sequence of SEQ ID NO: 37 and a light chain variable domain having the amino acid sequence of SEQ ID NO: 38; or (f) A heavy chain variable domain having the amino acid sequence of SEQ ID NO: 44 and a light chain variable domain having the amino acid sequence of SEQ ID NO:

45.

6. The method of any one of claims 1 to 5, wherein the anti-TMPRSS6 antibody comprises a heavy chain variable domain having the amino acid sequence of SEQ ID NO: 49 and a light chain variable domain having the amino acid sequence of SEQ ID NO:

50.

7. The method of any one of claims 1 to 6, wherein the anti-TMPRSS6 antibody comprises: (a) The heavy chain containing the amino acid sequence of SEQ ID NO: 52 and the light chain containing the amino acid sequence of SEQ ID NO: 53; (b) The heavy chain containing the amino acid sequence of SEQ ID NO: 11 and the light chain containing the amino acid sequence of SEQ ID NO: 12; (c) A heavy chain containing the amino acid sequence of SEQ ID NO: 17 or 22, and a light chain containing the amino acid sequence of SEQ ID NO: 23; (d) The heavy chain containing the amino acid sequence of SEQ ID NO: 33 and the light chain containing the amino acid sequence of SEQ ID NO: 34; (e) A heavy chain containing the amino acid sequence of SEQ ID NO: 41 and a light chain containing the amino acid sequence of SEQ ID NO: 42; or (f) A heavy chain containing the amino acid sequence of SEQ ID NO: 46 and a light chain containing the amino acid sequence of SEQ ID NO:

47.

8. The method of any one of claims 1 to 7, wherein the anti-TMPRSS6 antibody comprises a heavy chain containing the amino acid sequence of SEQ ID NO: 52 and a light chain containing the amino acid sequence of SEQ ID NO:

53.

9. The method of any one of claims 1 to 8, wherein the administration of the antibody reduces iron overload in the subject.

10. The method of any one of claims 1 to 9, wherein the application of the antibody reduces hemolysis in the subject relative to the subject prior to the application.

11. The method of any one of claims 1 to 10, wherein the subject has recurrent moderate to severe vascular occlusive crisis (VOC).

12. The method of any one of claims 1 to 11, wherein the application of the antibody reduces the frequency of VOCs in the object relative to the object prior to the application.

13. The method of any one of claims 1 to 12, wherein the application reduces the severity of VOC relative to the object prior to the application.

14. The method of claim 13, wherein the severity of VOC is measured by hospitalization frequency and / or hospitalization duration.

15. The method of any one of claims 1 to 14, wherein the administration of the antibody reduces systemic iron in the subject prior to administration.

16. The method of any one of claims 1 to 15, wherein the administration of the antibody reduces the mean erythrocyte hemoglobin concentration (MCHC) relative to the period prior to the administration.

17. The method of any one of claims 1 to 16, wherein the application reduces hemoglobin S (HbS) polymerization relative to the object prior to the application.

18. The method of any one of claims 1 to 17, wherein the application reduces the frequency at which the subject requires blood transfusions relative to the subject prior to the application.

19. The method of any one of claims 1 to 18, wherein the sickle cell disease is hemoglobin SS disease.

20. The method of any one of claims 1 to 18, wherein the sickle cell disease is hemoglobin SC disease.

21. The method of any one of claims 1 to 20, wherein the anti-TMPRSS6 antibody is administered as a co-treatment in combination with the following: a hemoglobin S-polymerization inhibitor (e.g., vorcelothorax), a therapeutic agent for reducing VOCs (e.g., hydroxyurea, L-glutamine oral powder, lizapril, selective pyruvate kinase-R (PKR) activator), analgesics (e.g., anesthetics, opioids, gabapentin, cannabis), blood transfusion, stem cell transplantation, Exagmglogene autotemcel (exa-cel), LentiGlobin, GMI-1070, VIT-2763 (vamifeport), ticagrelor, vitamin D, simvastatin, AG-348 (mitapiva sulfate), propranolol, adrenaline, regardsone, atorvastatin, prasugrel, L-arginine, OTQ923, oxygen therapy, or gene therapy.

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