Anti-TMPRSS6 antibody and its use

Abstract

Aspects of this disclosure provide an anti-TMPRSS6 antibody and methods of using it to promote hepcidin expression and to treat iron overload-related conditions, such as hemochromatosis, sickle cell disease, thalassemia, hemolysis, Diamond-Blackfan anemia, myelodysplastic syndrome (MDS), and blood transfusions.

Description

[Technical Field]

[0001] Cross-reference of related applications This application claims priority to U.S. Provisional Application No. 63 / 457,081, filed on April 4, 2023, the entire contents of which are incorporated herein by reference.

[0002] Electronic sequence list reference The contents of the electronic sequence listing (D084270011WO00-SEQ-LJG.xml; size: 95,921 bytes; and creation date: April 1, 2024) are incorporated herein by reference in their entirety. [Background technology]

[0003] Transmembrane serine protease 6 (TMPRSS6; also known as matryptase-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 coreceptors (e.g., hemoduvelin (HJV)), which leads to a decrease in hepcidin expression. This decrease in hepcidin expression levels leads to iron overload in various conditions. Improved compositions and methods are needed for the treatment of iron overload disorders. [Overview of the project]

[0004] Certain aspects of this disclosure relate to the recognition that iron overload occurs in various conditions (e.g., hemochromatosis, sickle cell disease (SCD), thalassemia, hemolysis, African iron overload, Diamond-Blackfan anemia, myelodysplastic syndrome (MDS), blood transfusion, etc.). In some embodiments, the methods and related compositions provided herein are useful for the treatment of various 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 are useful for inhibiting TMPRSS6 with the aim of reducing iron overload in the diseases and conditions described herein. In some embodiments, disease severity is remitted by reducing iron overload. Aspects of this disclosure provide anti-TMPRSS6 antibodies that have high binding affinity and specificity to TMPRSS6 and inhibit TMPRSS6 activity. Accordingly, in some embodiments, the 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 transfusion, etc. In some embodiments, the methods provided herein reduce iron overload in a subject. In some embodiments, the methods provided herein reduce systemic iron levels. In some embodiments, the methods provided herein reduce mean cellular hemoglobin concentration (MCHC) in a subject.

[0005] In some embodiments, the Disclosure provides a method for treating sickle cell disease (SCD) by administering an effective amount of anti-TMPRSS6 antibody to a subject. In some embodiments, the Disclosure provides a method and composition for treating iron overload in a subject having SCD by administering an effective amount of anti-TMPRSS6 antibody to a subject. In some embodiments, the method provided herein results in a reduction of painful attacks (i.e., occlusive attacks, VOCs), severity of VOCs (e.g., hospitalization and duration), systemic iron levels, mean cellular hemoglobin concentration (MCHC), hemoglobin S (HbS) polymerization, transfusion and / or hemolysis in a subject having SCD (e.g., hemoglobin SC disease). In some embodiments, the method provided herein reduces the frequency of painful attacks (VOCs) in hemoglobin SC disease.

[0006] In some aspects, the disclosure relates to a method for treating sickle cell disease (SCD), (a) HC CDR1, HC CDR2 and HC CDR3 heavy chain variable domains having the amino acid sequence of SEQ ID NO: 49, and LC CDR1, LC CDR2 and LC CDR3 light chain variable domains having the amino acid sequence of SEQ ID NO: 50, (b) HC CDR1, HC CDR2 and HC CDR3 heavy chain variable domains having the amino acid sequence of SEQ ID NO: 7, and LC CDR1, LC CDR2 and LC CDR3 light chain variable domains having the amino acid sequence of SEQ ID NO: 8, (c) HC CDR1, HC CDR2 and HC CDR3 heavy chain variable domains having the amino acid sequence of SEQ ID NO: 19 or 78, and LC CDR1, LC CDR2 and LC CDR3 light chain variable domains having the amino acid sequence of SEQ ID NO: 20, (d) HC CDR1, HC CDR2 and HC CDR3 of the heavy chain variable domain having the amino acid sequence of SEQ ID NO: 30, and LC CDR1, LC CDR2 and LC CDR3 of the light chain variable domain having the amino acid sequence of SEQ ID NO: 31, (e) The HC CDR1, HC CDR2, and HC CDR3 of the heavy chain variable domain having the amino acid sequence of SEQ ID NO: 37, and the LC CDR1, LC CDR2, and LC CDR3 of the light chain variable domain having the amino acid sequence of SEQ ID NO: 38, or (f) The HC CDR1, HC CDR2, and HC CDR3 of the heavy chain variable domain having the amino acid sequence of SEQ ID NO: 44, and the LC CDR1, LC CDR2, and LC CDR3 of the light chain variable domain having the amino acid sequence of SEQ ID NO: 45 Provided is a method comprising administering to a subject an effective amount of an anti-transmembrane serine protease 6 (TMPRSS6) antibody comprising

[0007] In some embodiments, the anti-TMPRSS6 antibody comprises the HC CDR1, HC CDR2, and HC CDR3 of the heavy chain variable domain having the amino acid sequence of SEQ ID NO: 49, and the LC CDR1, LC CDR2, and LC CDR3 of the light chain variable domain having the amino acid sequence of SEQ ID NO: 50.

[0008] In some embodiments, the anti-TMPRSS6 antibody is: (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 LC CDR3 having the amino acid sequence of SEQ ID NO: 29, (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 LC CDR3 having the amino acid sequence of SEQ ID NO: 6, (c) An HC CDR1 having the amino acid sequence of SEQ ID NO: 13, an HC CDR2 having the amino acid sequence of SEQ ID NO: 14, an HC CDR3 having the amino acid sequence of SEQ ID NO: 15, an LC CDR1 having the amino acid sequence of SEQ ID NO: 16, an LC CDR2 having the amino acid sequence of WAF, and an LC CDR3 having the amino acid sequence of SEQ ID NO: 18, (d) An HC CDR1 having the amino acid sequence of SEQ ID NO: 24, an HC CDR2 having the amino acid sequence of SEQ ID NO: 25, an HC CDR3 having the amino acid sequence of SEQ ID NO: 26, an LC CDR1 having the amino acid sequence of SEQ ID NO: 27, an LC CDR2 having the amino acid sequence of WAT, and an LC CDR3 having the amino acid sequence of SEQ ID NO: 29, (e) An HC CDR1 having the amino acid sequence of SEQ ID NO: 1, an HC CDR2 having the amino acid sequence of SEQ ID NO: 2, an HC CDR3 having the amino acid sequence of SEQ ID NO: 35, an LC CDR1 having the amino acid sequence of SEQ ID NO: 36, an LC CDR2 having the amino acid sequence of RAN, and an LC CDR3 having the amino acid sequence of SEQ ID NO: 6 or (f) An HC CDR1 having the amino acid sequence of SEQ ID NO: 13, an HC CDR2 having the amino acid sequence of SEQ ID NO: 43, an HC CDR3 having the amino acid sequence of SEQ ID NO: 15, an LC CDR1 having the amino acid sequence of SEQ ID NO: 16, an LC CDR2 having the amino acid sequence of WAF, and an LC CDR3 having the amino acid sequence of SEQ ID NO: 18 comprises.

[0009] In some embodiments, the anti-TMPRSS6 antibody comprises an HC CDR1 having the amino acid sequence of SEQ ID NO: 24, an HC CDR2 having the amino acid sequence of SEQ ID NO: 48, an HC CDR3 having the amino acid sequence of SEQ ID NO: 26, an LC CDR1 having the amino acid sequence of SEQ ID NO: 27, an LC CDR2 having the amino acid sequence of WAT, and an LC CDR3 having the amino acid sequence of SEQ ID NO: 29.

[0010] In some embodiments, the anti-TMPRSS June antibody is: (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) Heavy chain variable domain having the amino acid sequence of SEQ ID NO: 19 or 78 and light chain variable domain having the 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) Heavy chain variable domain having the amino acid sequence of SEQ ID NO: 44 and light chain variable domain having the amino acid sequence of SEQ ID NO: 45 Includes.

[0011] In some embodiments, the anti-TMPRSS6 antibody includes 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.

[0012] In some embodiments, the anti-TMPRSS6 antibody is: (a) 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, (b) A heavy chain containing the amino acid sequence of SEQ ID NO: 11 and a 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) A heavy chain containing the amino acid sequence of SEQ ID NO: 33 and a 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) Heavy chain containing the amino acid sequence of SEQ ID NO: 46 and light chain containing the amino acid sequence of SEQ ID NO: 47 Includes.

[0013] 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.

[0014] In some embodiments, antibody administration reduces iron overload in subjects. In some embodiments, antibody administration reduces hemolysis in subjects compared to subjects before administration. In some embodiments, subjects have recurrent moderate to severe vascular occlusive disorders (VOCs). In some embodiments, antibody administration reduces the frequency of VOCs in subjects compared to subjects before administration. In some embodiments, administration reduces the severity of VOCs compared to subjects before administration. In some embodiments, the severity of VOCs is measured by the frequency and / or duration of hospitalizations.

[0015] In some embodiments, antibody administration reduces systemic iron in the subject before administration. In some embodiments, antibody administration reduces mean cellular hemoglobin concentration (MCHC) compared to before administration. In some embodiments, administration reduces hemoglobin S (HbS) polymerization compared to the subject before administration. In some embodiments, administration reduces the frequency with which the subject requires blood transfusions compared to the subject before administration. In some embodiments, administration reduces inflammation in the subject compared to the subject before administration. In some embodiments, sickle cell disease is hemoglobin SS disease. In some embodiments, sickle cell disease is hemoglobin SC disease.

[0016] In some embodiments, the anti-TMPRSS6 antibody is used in conjunction with 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., narcotics, opioids, gabapentin, cannabis), blood transfusions, stem cell transplants, and Exagmglogene autotemcells. Autotemcel (exa-cel), lentiglobin, GMI-1070, VIT-2763 (vamifeport), ticagrelor, vitamin D, simvastatin, AG-348 (mitapivat sulfate), propranolol, epinephrine, regadenoson, atorvastatin, prasugrel, L-arginine, OTQ923, are administered as combination therapy in combination with oxygen therapy or gene therapy.

[0017] In some embodiments, the Disclosure provides a method for treating an iron-overload-related condition in subjects that require it, wherein the subjects do not have intermediate-type or type I hemochromatosis of β-thalassemia, and the method is (a) HC CDR1, HC CDR2 and HC CDR3 heavy chain variable domains having the amino acid sequence of SEQ ID NO: 49, and LC CDR1, LC CDR2 and LC CDR3 light chain variable domains having the amino acid sequence of SEQ ID NO: 50, (b) HC CDR1, HC CDR2 and HC CDR3 heavy chain variable domains having the amino acid sequence of SEQ ID NO: 7, and LC CDR1, LC CDR2 and LC CDR3 light chain variable domains having the amino acid sequence of SEQ ID NO: 8, (c) HC CDR1, HC CDR2 and HC CDR3 heavy chain variable domains having the amino acid sequence of SEQ ID NO: 19 or 78, and LC CDR1, LC CDR2 and LC CDR3 light chain variable domains having the amino acid sequence of SEQ ID NO: 20, (d) HC CDR1, HC CDR2 and HC CDR3 of the heavy chain variable domain having the amino acid sequence of SEQ ID NO: 30, and LC CDR1, LC CDR2 and LC CDR3 of the light chain variable domain having the amino acid sequence of SEQ ID NO: 31, (e) HC CDR1, HC CDR2 and HC CDR3 heavy chain variable domains having the amino acid sequence of SEQ ID NO: 37, and LC CDR1, LC CDR2 and LC CDR3 light chain variable domains having the amino acid sequence of SEQ ID NO: 38, or (f) HC CDR1, HC CDR2, and HC CDR3 heavy chain variable domains having the amino acid sequence of SEQ ID NO: 44, and LC CDR1, LC CDR2, and LC CDR3 light chain variable domains having the amino acid sequence of SEQ ID NO: 45 This includes administering an effective amount of anti-transmembrane serine protease 6 (TMPRSS6) antibody to the target.

[0018] In some embodiments, the anti-TMPRSS6 antibody comprises heavy chain variable domains HC CDR1, HC CDR2, and HC CDR3 having the amino acid sequence of SEQ ID NO: 49, and light chain variable domains LC CDR1, LC CDR2, and LC CDR3 having the amino acid sequence of SEQ ID NO: 50.

[0019] In some embodiments, the anti-TMPRSS6 antibody is (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 LC CDR3 having the amino acid sequence of SEQ ID NO: 29 (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 LC CDR3 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 LC CDR3 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 LC CDR3 having the amino acid sequence of SEQ ID NO: 29 (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 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 LC CDR3 having the amino acid sequence of SEQ ID NO: 18 Includes.

[0020] In some embodiments, the anti-TMPRSS6 antibody includes 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.

[0021] In some embodiments, the anti-TMPRSS6 antibody is (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) Heavy chain variable domain having the amino acid sequence of SEQ ID NO: 19 or 78 and light chain variable domain having the 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) Heavy chain variable domain having the amino acid sequence of SEQ ID NO: 44 and light chain variable domain having the amino acid sequence of SEQ ID NO: 45 Includes.

[0022] In some embodiments, the anti-TMPRSS6 antibody includes 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.

[0023] In some embodiments, the anti-TMPRSS6 antibody is (a) 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, (b) A heavy chain containing the amino acid sequence of SEQ ID NO: 11 and a 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) A heavy chain containing the amino acid sequence of SEQ ID NO: 33 and a 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) Heavy chain containing the amino acid sequence of SEQ ID NO: 46 and light chain containing the amino acid sequence of SEQ ID NO: 47 Includes.

[0024] 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.

[0025] In some embodiments, antibody administration reduces systemic iron levels in subjects compared to before administration. In some embodiments, antibody administration reduces mean cellular hemoglobin concentration (MCHC) compared to before administration. In some embodiments, antibody administration reduces iron overload in subjects compared to before administration.

[0026] In some embodiments, the subject has hereditary hemochromatosis. In some embodiments, the hereditary hemochromatosis is type 3 hereditary hemochromatosis.

[0027] In some embodiments, the subject has or is suspected of having thalassemia. In some embodiments, the thalassemia is α-thalassemia. In some embodiments, the α-thalassemia is severe, intermediate, or mild α-thalassemia. In some embodiments, the thalassemia is β-thalassemia. In some embodiments, the β-thalassemia is severe or mild β-thalassemia.

[0028] In some embodiments, the subject has or is suspected of having transfusion-associated iron overload. In some embodiments, the subject is receiving transfusions due to blood loss. In some embodiments, the subject is receiving repeated transfusions due to anemia.

[0029] In some embodiments, the subject has hemolytic anemia. In some embodiments, the subject has transfusion-dependent hemolytic anemia, pyruvate kinase deficiency hemolytic anemia, thalassemia-related hemolytic anemia, or sickle cell disease-related hemolytic anemia.

[0030] In some embodiments, the subject has African-type iron overload.

[0031] In some embodiments, the subject has Diamond-Blackfan anemia.

[0032] In some embodiments, the subject has myelodysplastic syndrome (MDS). In some embodiments, the MDS is refractory anemia with ring sideroblasts (RARS). In some embodiments, the subject has an SF3B1 mutation.

[0033] The aforementioned and other embodiments, implementations, actions, functions, features, and embodiments of this teaching can be better understood from the following description in conjunction with the accompanying drawings. [Brief explanation of the drawing]

[0034] [Figure 1] Figures 1A–1E demonstrate the dose-dependent effect of treating mice with inhibitory anti-TMPRSS6 antibodies. Mice were intraperitoneally injected with 0 (medium), 2, 5, or 10 mg / kg of anti-TMPRSS6 antibody and monitored for 10 days. Serum antibody (Figure 1A), iron (Figure 1B), and hepcidin (Figure 1C) concentrations were measured 10 days after injection. Figure 1D shows Hamp expression in the liver, and Figure 1E shows the TSAT% of mice over the course of the study. [Figure 2] Figures 2A–2D show the body, spleen, and liver weights of sickle cell (Townes model) mice administered with anti-TMPRSS6 antibody. Figure 2A shows the progression of total body weight over 8 weeks in mice administered with 0, 3, or 10 mg / kg of anti-TMPRSS6 antibody, stratified into male and female groups (referred to as M and F, respectively). Figure 2B shows the measured spleen weight / body weight of male and female mice 56 days after administration of 0, 3, or 10 mg / kg of anti-TMPRSS6 antibody. Figure 2C shows the measured liver weight / body weight of male and female mice 56 days after administration of 0, 3, or 10 mg / kg of anti-TMPRSS6 antibody. Figure 2D shows the liver weight / body weight of all mice (integrated male and female mice) 56 days after administration of 0 or 10 mg / kg of anti-TMPRSS6 antibody. [Figure 3]Figures 3A–3C show the reduction of hemolytic markers in sickle cell mice administered with anti-TMPRSS6 antibody. Lactate dehydrogenase (LDH) (Figure 3A), direct bilirubin (DBili) (Figure 3B), and total bilirubin (TBili) (Figure 3C) levels were measured in sickle cell mice 56 days after administration of 0, 3, or 10 mg / kg of anti-TMPRSS6 antibody. [Figure 4] Figures 4A–4C show the reduction of systemic inflammatory markers in sickle cell mice administered with anti-TMPRSS6 antibody. Levels of leukocytes (WBC) (Figure 4A), neutrophils (Figure 4B), and lymphocytes (Figure 4C), as measured by peroxidase, were measured in the blood of sickle cell mice 56 days after administration of 0, 3, or 10 mg / kg of anti-TMPRSS6 antibody. [Figure 5] Figures 5A–5C demonstrate the reduction in HbS polymerization in sickle cell mice treated with anti-TMPRSS6 antibody. Levels of HbS(CHCM) (Figure 5A), hypochromic erythrocytes (Figure 5B), and microcytic erythrocytes (Figure 5C) were measured in the blood of sickle cell mice 56 days after administration of 0, 3, or 10 mg / kg of anti-TMPRSS6 antibody. [Modes for carrying out the invention]

[0035] This disclosure relates, at least in part, to the recognition that iron overload occurs in various 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 are useful for treating various 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 are provided that are useful for inhibiting TMPRSS6 with the aim of reducing iron overload, thereby improving disease severity. Aspects of this disclosure provide anti-TMPRSS6 antibodies that have high binding affinity and specificity to TMPRSS6 and inhibit TMPRSS6 activity. Accordingly, in some embodiments, the present disclosure provides methods 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, and related anti-TMPRSS6 antibody compositions. In some embodiments, the methods provided herein reduce iron overload.

[0036] In some embodiments, the Disclosure provides a method for treating sickle cell disease (SCD) by administering an effective amount of anti-TMPRSS6 antibody to a subject. In some embodiments, the Disclosure provides a method and composition for treating iron overload in a subject having SCD by administering an effective amount of anti-TMPRSS6 antibody to a subject. In some embodiments, the method provided herein results in a reduction of the frequency of painful attacks (i.e., occlusive attacks, VOCs), the severity of VOCs (e.g., hospitalization and duration), hemoglobin S (HbS) polymerization, transfusion, and / or hemolysis in a subject having SCD (e.g., hemoglobin sickle cell disease). In some embodiments, the method provided herein reduces the frequency of painful attacks (VOCs) in hemoglobin sickle cell disease (SC).

[0037] The aforementioned and other aspects, implementations, actions, functions, features, and embodiments of this instruction can be better understood from the following description in conjunction with the accompanying drawings.

[0038] I. Definition And / or: As used herein, the term "and / or" should be interpreted as a specific disclosure of each of two or more designated features or components, whether or not the others are present. Thus, as used herein in phrases such as "A and / or B," the term "and / or" is intended to include "A and B," "A or B," "A" (alone) and "B" (alone). Similarly, as used in phrases such as "A, B and / or C," the term "and / or" is intended to include each of the following embodiments: 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).

[0039] Administering: As used herein, the terms “administer,” “administered,” “to administer,” or “to administer” mean to provide an antibody or a composition thereof to a subject in a manner that is physiologically and / or pharmacologically useful (for example, to treat a condition in the subject).

[0040] Affinity-matured antibody: The term "affinity-matured antibody" is used herein to refer to an antibody that has one or more modifications to one or more CDRs, resulting in an improved affinity (i.e., KD, kd, or ka) for the target antigen compared to a parent antibody that has no modifications. Exemplary affinity-matured antibodies may have nanomolar or even picomolar affinity for the target antigen. Various procedures for producing affinity-matured antibodies are known in the art, including screening of combinatory antibody libraries prepared using biodisplay. For example, Marks et al., BioTechnology, 10: 779-783 (1992) describes affinity maturation by VH and VL domain shuffling. Random mutagenesis of CDRs and / or framework residues is described by 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 and constant or hypermutation at activity-enhancing amino acid residue sites is described in U.S. Patent No. 6,914,128B1.

[0041] Antibody: As used herein, the term “antibody” means a polypeptide comprising at least one immunoglobulin variable domain or at least one site, e.g., a paratope, that specifically binds to an antigen. In some embodiments, the antibody comprises a paratope. In some embodiments, the paratope 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 an 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 bispecific antibody. 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 the group consisting of IgG, IgG1, IgG2, IgG2A, IgG2B, IgG2C, IgG3, IgG4, IgA1, IgA2, IgD, IgM, and IgE constant domains. In some embodiments, the antibody has a heavy (H) chain variable region (V in this specification). H (abbreviated as V), and / or light (L) chain variable region (V in this specification). L Includes (abbreviated as ). In some embodiments, the antibody includes a constant domain, e.g., an Fc region. The immunoglobulin constant domain refers to the heavy or light chain constant domain. The amino acid sequences of the human IgG heavy chain and light chain constant domains and their functional variations are known. With respect to 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 include a human alpha (α), delta (Δ), epsilon (ε), gamma (γ), or mu (μ) heavy chain. In certain embodiments, the antibody described herein includes the human gamma 1 CH1, CH2, and / or CH3 domains. In some embodiments, V HThe amino acid sequence of the domain includes the amino acid sequence of the human gamma (γ) heavy chain constant region, for example, any known in the art. Non-limiting examples of human constant region sequences are described in the art; see, for example, U.S. Patent No. 5,693,780 and Kabat EA et al., (1991) above. In some embodiments, V HThe domains include an amino acid sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 98%, or at least 99% identical to any of the variable chain constant regions provided herein. 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 by N-glycosylation, O-glycosylation, C-glycosylation, glyciation (GPI anchoring), 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 the present disclosure linked to a linker polypeptide or an immunoglobulin constant domain. The linker polypeptide comprises two or more amino acid residues joined by a peptide bond and is used to link one or more antigen-binding moieties. Examples of linker polypeptides 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, the antibody may be part of a larger immunoadhesion molecule formed by covalent or noncovalent bonding of the antibody or antibody moiety with one or more other proteins or peptides.Examples of such immunoadhesion molecules include the use of streptavidin core regions to create tetrameric scFv molecules [Kipriyanov, SM, et al. (1995) Human Antibodies and Hybridomas 6:93-101] and the use of cysteine ​​residues, marker peptides, and C-terminal polyhistidine tags to create divalent and biotinylated scFv molecules [Kipriyanov, SM, et al. (1994) Mol. Immunol. 31:1047-1058].

[0042] Approximately: As used herein, the terms “approximately” or “about” mean a value that is similar to the reference value stated, when applied to one or more of the values ​​of interest. In certain embodiments, unless otherwise stated or otherwise evident from the context, the terms “approximately” or “about” mean a range of values ​​that are within 15%, 14%, 13%, 12%, 11%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1%, or less in any direction of the reference value stated (except where such a number would exceed 100% of the possible value).

[0043] CDR: As used herein, the term “CDR” refers to the complementarity-determining region within the antibody variable sequence. A typical antibody molecule usually contains a heavy chain variable region (VH) and a light chain variable region (VL) that are involved in antigen binding. The VH and VL regions can be further subdivided into hypervariable regions, also known as “complementarity-determining regions” (“CDR”), which are interspersed with more conserved regions known as “framework regions” (“FR”). Each VH and VL typically consists of three CDRs and four FRs, arranged from amino-terminus to carboxy-terminus in the following order: FR1, CDR1, FR2, CDR2, FR3, CDR3, FR4. The extent of the framework regions and CDRs can be precisely identified using methods known in the art, for example, by the Kabat definition, IMGT definition, Chothia definition, AbM definition, and / or contact definition, all of which are well known in the art.Furthermore, 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(Manager), the International ImMunoGeneTics Information System(Manager)http: / / www.imgt.org, Lefranc, M.-P. http: / / dx.doi.org / 10.1037 / 0021-843X.112.2.213 Ruiz, M. et al., Nucleic Acids Res., 27:209-212 (1999); 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);Lefranc, M.-P. et al., Nucleic Acids 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. et al. (1987) J. Mol. Biol. 196:901–917, Al-Lazikani et al (1997) J. Molec. Biol. 273:927-948;and Almagro, J. Mol. Recognize. 17:132-143 (2004) at hgmp.mrc.ac.uk and at bioinf.org.uk / abs.As used herein, CDR may refer to a CDR as defined by any method known in the art. Two antibodies having the same CDR means that the two antibodies have the same amino acid sequence of the CDR, as determined by the same method, e.g., the IMGT definition.

[0044] In certain embodiments, there are three CDRs in the variable regions of the heavy and light chains, respectively, denoted as CDR1, CDR2, and CDR3 for each variable region. As used herein, the term “CDR set” refers to a group of three CDRs that occur in a single variable region capable of binding to an antigen. The exact boundaries of these CDRs are defined differently by different systems. The system described by Kabat [Kabat et al., Sequences of Proteins of Immunological Interest (National Institutes of Health, Bethesda, Md. (1987) and (1991))] provides not only a distinct residue numbering system applicable to any variable region of an antibody, but also the exact residue boundaries that define the three CDRs. These CDRs may be called Kabat CDRs. The sub-parts of the CDRs may be denoted as L1, L2, and L3 or H1, H2, and H3, where “L” and “H” indicate the light and heavy chain regions, respectively. These regions may also be called Chothia CDRs and have boundaries that overlap with Kabat CDRs. Other boundaries defining CDRs that overlap with Kabat CDRs have been described by Padlan [FASEB J. 9:133-139 (1995)] and MacCallum [J Mol Biol 262(5):732-45 (1996)]. Further other CDR boundary definitions do not have to strictly adhere to one of the above systems, but despite overlapping with Kabat CDRs, they may be shortened or extended to take into account predictions or experimental findings that do not significantly affect antigen binding for specific residues, groups of residues, or even the entire CDR. The methods used herein may utilize CDRs defined according to any of these systems, but preferred embodiments use CDRs defined by Kabat or Chothia.

[0045] In certain embodiments, the CDR of an antibody may have different amino acid sequences depending on the definition system used (e.g., IMGT definition, Kabat definition, or Chothia definition). The definition system annotates each amino acid of a given antibody sequence (e.g., VH or VL sequence) with a number, and the numbers corresponding to the heavy and light chain CDRs are provided in Table 2. The CDRs listed in Table 1 are defined by the Kabat definition. Those skilled in the art can obtain the CDR sequences of the anti-TMPRSS6 antibodies provided in Table 1 using different numbering systems.

[0046] [Table 1]

[0047] CDR transplant antibody: The term "CDR transplant antibody" refers to an antibody that contains heavy and light chain variable region sequences from a single species, but V H and / or V L This refers to antibodies in which one or more CDR region sequences are replaced with CDR sequences of another species, for example, antibodies having mouse heavy and light chain variable regions in which one or more mouse CDRs (e.g., CDR3) are replaced with human CDR sequences.

[0048] Chimeric antibody: The term "chimeric antibody" refers to an antibody that contains heavy chain and light chain variable region sequences from one species as well as a constant region sequence from another species, for example, an antibody having mouse heavy chain and light chain variable regions linked to a human constant region.

[0049] Complementarity: As used herein, the term “complementarity” refers to the ability of two nucleotides or two sets of nucleotides to pair precisely. In particular, complementarity is a term that characterizes the degree of hydrogen bond pairs that result in bonding between two nucleotides or two sets of nucleotides. For example, if a base at one position of an oligonucleotide can hydrogen bond with a base at the corresponding position of a target nucleic acid (e.g., mRNA), then the bases are considered complementary to each other at that position. Base pairs can include both classical Watson-Crick base pairs and non-Watson-Crick base pairs (e.g., fluctuation base pairs and Hoogsteen base pairs). For example, in some embodiments, due to complementary base pairing, an adenosine-type base (A) is complementary to a thymidine-type base (T) or a uracil-type base (U), a cytosine-type base (C) is complementary to a guanosine-type base (G), and a universal base, such as 3-nitropyrrole or 5-nitroindole, is considered complementary by hybridizing with any of A, C, U, or T. Inosine (I) is also considered a universal base in the art and is considered complementary to any of A, C, U, or T.

[0050] Conservative amino acid substitutions: As used herein, “conservative amino acid substitutions” refer to amino acid substitutions that do not alter the relative charge or size characteristics of the protein in which the substitution is made. Variants can be prepared according to methods of modifying polypeptide sequences known to those skilled in the art, for example, references that compile such methods, e.g., 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 amino acid substitutions include substitutions made in 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.

[0051] Tolerance Reactivity: As used herein, and in the context of targeted agents (e.g., antibodies), the term “tolerance reactivity” refers to the property of a drug to specifically bind to one or more antigens of similar types or classes (e.g., diverse homologous, paralogous, or orthologous antigens) with similar affinity or binding activity. For example, in some embodiments, an antibody that is tolerance reactive to similar types or classes of human and non-human primate antigens (e.g., human TMPRSS6 and non-human primate TMPRSS6) can bind to human and non-human primate antigens with similar affinity or binding activity. In some embodiments, the antibody is cross-reactive to similar types or classes of human and rodent antigens. In some embodiments, the antibody is cross-reactive to similar types or classes of rodent and non-human primate antigens. In some embodiments, the antibody is cross-reactive to similar types or classes of human, non-human primate, and rodent antigens.

[0052] Effective dose: As used herein, “effective dose” refers to the amount of each activator required to produce a therapeutic effect on a subject, either alone or in combination with one or more other activators. In some embodiments, the therapeutic effect is a decrease in TMPRSS6 levels or activity, an increase in hepcidin levels or activity, and / or a reduction in a disease condition (e.g., iron overload in sickle cell disease, thalassemia, hemochromatosis, etc.).

[0053] Framework: As used herein, the term “framework” or “framework sequence” refers to the sequence of the remaining variable region excluding the CDRs. Since the precise definition of a CDR sequence can be determined by different systems, the meaning of a framework sequence is assumed to be correspondingly subject to different interpretations. The six CDRs (CDR-L1, CDR-L2, and CDR-L3 of the light chain and CDR-H1, CDR-H2, and CDR-H3 of the heavy chain) also divide the framework regions of the light and heavy chains into four sub-regions (FR1, FR2, FR3, and FR4) of each chain, with CDR1 located between FR1 and FR2, CDR2 between FR2 and FR3, and CDR3 between FR3 and FR4. Without specifying a particular sub-region as FR1, FR2, FR3, or FR4, as otherwise mentioned, the framework region represents a combination of FRs within the variable region of a single, spontaneously occurring immunoglobulin chain. As used herein, FR represents one of four subregions, and FRs represents two or more of the four subregions that constitute the framework region. Human heavy and light chain acceptor sequences are known in the art. In one embodiment, acceptor sequences known in the art may be used in the antibodies disclosed herein. In some embodiments, the sequence of the disclosed framework region may vary by up to 20% (for example, the sequence may be approximately 80% (e.g., 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%) identical to the disclosed sequence).

[0054] 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. The human antibodies of the present disclosure may include amino acid residues not encoded by human germline immunoglobulin sequences [e.g., mutations introduced by random or site-directed mutagenesis in vitro or by somatic mutations in vivo], for example, in CDRs, particularly CDR3. However, as used herein, the term "human antibody" is not intended to include antibodies in which CDR sequences derived from the germline of another mammalian species, such as a mouse, are grafted onto human framework sequences.

[0055] Humanized Antibodies: The term "humanized antibody" includes heavy and light chain variable region sequences from non-human species (e.g., mouse), but at least a part of the V H and / or V L sequences are 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 human CDR sequences are introduced into non-human V H and V L sequences to replace the corresponding non-human CDR sequences. In one embodiment, humanized anti-TMPRSS6 antibodies and antigen-binding portions are provided. Such antibodies can be generated by obtaining mouse anti-TMPRSS6 monoclonal antibodies using humanization using in vitro genetic engineering, such as that disclosed in International Application PCT2005 / 123126A2 by Kasaian et al, after classical hybridoma technology.

[0056] A humanized antibody is a human immunoglobulin (recipient antibody) in which residues from the recipient's complementarity-determining region (CDR) are replaced with residues from a non-human species (donor antibody), such as mouse, rat, or rabbit, that possess the desired specificity, affinity, and capabilities. In some embodiments, Fv framework region (FR) residues of the human immunoglobulin are replaced with corresponding non-human residues. Furthermore, the humanized antibody may also include residues that are not found in the recipient antibody or in the transferred CDR or framework sequence, but are included to further refine and optimize antibody performance. Generally, a humanized antibody includes at least one, and typically two, substantially all, variable domains, where all or substantially all of the CDR region corresponds to that of a non-human immunoglobulin, and all or substantially all of the FR region corresponds to that of the human immunoglobulin consensus sequence. The humanized antibody also optimally and typically includes at least a portion of the immunoglobulin constant region or domain (Fc) of the human immunoglobulin. The antibody may have a modified Fc region as described in International Publication No. 99 / 58572. Other forms of the humanized antibody may have one or more CDRs (one, two, three, four, five, or six) that are modified with respect to the original antibody, which are also referred to as one or more CDRs derived from one or more CDRs from the original antibody. The humanized antibody may also undergo affinity maturation.

[0057] In some embodiments, humanization is achieved by transplanting a CDR (e.g., those shown in Table 1) into a human variable domain (e.g., IGKV1-NL1*01 and IGHV1-3*01 human variable domains). In some embodiments, the anti-TMPRSS6 antibody of this disclosure is a humanized variant comprising one or more amino acid substitutions compared to any one of the VHs listed in Table 1 (e.g., in the VH framework region) and / or one or more amino acid substitutions compared to any one of the VLs listed in Table 1 (e.g., in the VL framework region).

[0058] Isolated antibody: As used herein, “isolated antibody” is intended to mean an antibody that substantially does not contain other antibodies with different antigenic properties (for example, an isolated antibody that specifically binds to TMPRSS6 substantially does not contain antibodies that specifically bind to antigens other than TMPRSS6). An isolated antibody that specifically binds to TMPRSS6, however, may cross-react with other antigens. Furthermore, an isolated antibody may not substantially contain other cellular material and / or chemical substances.

[0059] Kabat Numbering: The terms “Kabat Numbering,” “Kabat Definition,” and “Kabat Labeling” are used interchangeably herein. As recognized in the Art, these terms refer to a system for numbering amino acid residues that are more variable (i.e., hypervariable) than other amino acid residues in the heavy and light chain variable regions of an antibody, or in its antigen-binding moiety [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 region, the hypervariable region is the range of amino acids 31-35 of CDR1, 50-65 of CDR2, and 95-102 of CDR3. Regarding the light chain variable region, the hypervariable region is located in the range of amino acids 24 to 34 of CDR1, 50 to 56 of CDR2, and 89 to 97 of CDR3.

[0060] Sequence Identity Percentage (%): With respect to a reference polypeptide (or nucleotide) sequence, the terms “Sequence Identity Percentage (%)”, “Identity Percentage (%)”, and “Identity Percentage to (%)” are defined as the percentage of amino acid residues (or nucleic acids) in a candidate sequence that are identical to an amino acid residue (or nucleic acid) in a reference polypeptide (or nucleotide) sequence, after the sequences have been aligned and gaps introduced as necessary to achieve the maximum sequence identity percentage, without considering any conservative substitutions as part of the sequence identity. Alignment aimed at determining the sequence identity percentage can be achieved in various ways within the scope of 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 aligning sequences, including any algorithm required to achieve the maximum alignment over the entire length of the sequences being compared. However, for the purposes of this specification, the amino acid (or nucleic acid) sequence identity % value is generated using the sequence comparison computer program ALIGN-2. The ALIGN-2 sequence comparison computer program was created by Genentech, Inc., and its source code, along with user documentation, has been filed with the U.S. Copyright Office, Washington, DC, 20559, and is registered under U.S. Copyright Registration Number 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 must be compiled for use on UNIX operating systems, including Digital UNIX V4.0D. All sequence comparison parameters are set by the ALIGN-2 program and do not change.

[0061] Recombinant Antibodies: As used herein, the term “recombinant antibodies” includes all antibodies (as described in detail herein) prepared, expressed, created, or isolated by recombinant means, such as antibodies expressed using recombinant expression vectors transfected into host cells, for example, antibodies isolated from recombinant, combinatorial 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) BioTechniques 29:128-145; Hoogenboom H., and Chames P. (2000) Immunology Today 21:371-378], antibodies isolated from animals (e.g., mice) that are transgenic of human immunoglobulin genes [e.g., Taylor, LD, et al. (1992) Nucl. Acids [See 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, created or isolated by any other means including splicing of human immunoglobulin gene sequences to other DNA sequences. In some embodiments, recombinant human antibodies are provided herein. In certain embodiments, such recombinant human antibodies have a variable region and a constant region derived from a human germline immunoglobulin sequence. However, in certain embodiments, such recombinant human antibodies are subjected to in vitro mutagenesis (or in vivo somatic mutagenesis, if a transgenic animal of a human Ig sequence is used), and thus the recombinant antibody is V H and V L The amino acid sequence of the region is human germline V H and V LThe sequence is sequence-derived or related, but does not necessarily have to be naturally present in the human antibody germline repertoire in vivo. One embodiment of the present disclosure provides a complete human antibody capable of binding to human TMPRSS6, which can be produced using techniques well known in the art, such as using a human Ig phage library, for example, disclosed in international application PCT2005 / 007699A2 by Jermutus et al.

[0062] Selective: As used herein, the terms “selective” or “selectively” refer to the ability of a molecule to produce an effect associated with its target molecule compared to a reference molecule (e.g., inhibit, antagonistize, agonize, etc.). For example, a molecule that selectively inhibits its target molecule means that this molecule can inhibit the target molecule to a degree that is distinguishable from the reference molecule in an inhibition assay or other inhibitory environment. For example, with respect to an inhibitor, the term “selectively inhibits” 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 not inhibited in an inhibition assay, to the extent that it enables selective inhibition of the target molecule, as described herein. After the reaction is complete, the signal produced by inhibiting the target molecule may be measured. The median inhibitory concentrations of the target molecule and the reference molecule may 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 antagonistize the target molecule.

[0063] Specific binding: As used herein, the term “specific binding” refers to the ability of a molecule to bind to a binding partner with a certain degree of affinity or binding strength so that the molecule can be used to distinguish the binding partner from a suitable control in a binding assay or other binding environment. With respect to an antibody, the term “specific binding” refers to the ability of an antibody to bind to a specific antigen with a certain degree of affinity or binding strength so that the antibody can be used to distinguish the specific antigen from others, as described herein, compared to a suitable reference antigen(s). In some embodiments, the antibody binds 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 K for binding to M or smaller targets D If present, the antibody binds specifically to the target. In some embodiments, the antibody binds specifically to TMPRSS6.

[0064] Subject: As used herein, the term “subject” refers to mammals. In some embodiments, the subject is a non-human primate or rodent. In some embodiments, the subject is a human. In some embodiments, the subject is a patient who has or is suspected of having a disease, e.g., a human patient. In some embodiments, the subject is a human patient who has or is suspected of having iron overload and / or one or more conditions resulting from iron overload.

[0065] Treatment: As used herein, the terms “treating,” “treated,” “treat,” or “treatment” refer to the application or administration of a composition comprising one or more active agents (e.g., anti-TMPRSS6 antibody) to a subject having a target disease or disorder, symptoms of a disease / disorder, or a predisposition to a disease / disorder, for the purpose of treating, curing, alleviating, reducing, altering, relieving, improving, or influencing a disorder, symptoms of a disease, or predisposition to a disease or disorder. Alleviating a target disease / disorder includes delaying or preventing the onset or progression of the disease, or reducing the severity of the disease. In some embodiments, target diseases or disorders of this disclosure include, but are not limited to, hemochromatosis, sickle cell disease (SCD), thalassemia, hemolysis, African iron overload, Diamond-Blackfan anemia, myelodysplastic syndrome (MDS), and blood transfusion.

[0066] II. Anti-TMPRSS6 antibody In some embodiments, the anti-TMPRSS6 antibody is an antibody that is specific to TMPRSS6. In some embodiments, antibodies that bind to human TMPRSS6 with high specificity and affinity are provided herein. 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 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.

[0067] TMPRSS6, also known as matryptase-2, is a serine protease within 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 Biol Chem. 2001; 276:857-60). The TTSP family of proteases comprises several subfamilies, including the matryptase subfamily, which includes TMPRSS2-5. TMPRSS6, to which antibodies in some embodiments of this disclosure specifically bind, 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 number: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 number:71753). In other embodiments, the antibody of this disclosure specifically binds to the TMPRSS6 protein encoded by the non-human primate TMPRSS6 gene (e.g., cynomolgus monkey (i.e., Macaca fascicularis), e.g., NCBI gene number:102139590). The TMPRSS6 gene spans 18 exons with 17 intervening introns, and the matryptase-2 protein domain boundary corresponds to the intron / exon junction of the coding gene across all species.Structurally, matryptase-2 contains the standard TTSP features, such as a short cytoplasmic amino-terminal tail, a transmembrane region, a stem region containing two complement protein subcomponents (C1r / C1s, sea urchin embryo growth factor, and bone morphogenetic protein 1 (CUB) domains), and three low-density lipoprotein receptor class A (LDLR) domains, as well as a carboxy-terminal trypsin-like serine protease domain. All of these structural features are robustly conserved across humans, macaques, dogs, cattle, mice, and rats, and human proteins share 95.6%, 91.1%, 85.6%, 80.1%, and 80.4% identity with matriptase-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).

[0068] The matriptase-2 proteolytic domain contains the serine protease triad of H, D, and S residues necessary for catalytic activity, as well as an SWG motif predicted to be located above the substrate S1 binding pocket, which positions the easily cleavable substrate bond in the correct orientation. Proteolytic activation of matriptase-2 is predicted to occur within the motif at the junction of the prodomain and catalytic domain (RIVGG (SEQ ID NO: 80)), which is characteristic 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 human and mouse tissues, the primary site of matriptase-2 mRNA expression is the liver. Specifically, matriptase-2 mRNA expression has been demonstrated to be restricted to hepatocytes.

[0069] Functionally, matryptase-2 acts to cleave hemoduvelin (HJV), a protein essential in the process of iron regulation via positive regulation of hepcidin expression. Hepcidin is a known negative regulator of iron absorption, release, and reuse. In the absence of matryptase-2, HJV levels increase, leading to increased hepcidin levels and consequently, inhibition of iron transport. The establishment of matryptase-2 as an essential component of iron homeostasis was 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 antibodies of this disclosure inhibit matryptase-2 activity to regulate iron homeostasis. In some embodiments, the antibodies of this disclosure inhibit matryptase-2 activity, thereby reducing iron transport through increased hepcidin activity.

[0070] In some embodiments, the anti-TMPRSS6 antibody described herein may be conjugated to a fragment of human TMPRSS6. The TMPRSS6 fragment may be about 5 to 425 amino acids, about 10 to 400 amino acids, about 50 to 350 amino acids, about 100 to 300 amino acids, about 150 to 250 amino acids, about 200 to 300 amino acids, or about 75 to 150 amino acids in length. In some embodiments, the fragment may contain a consecutive number of amino acids from TMPRSS6. In other embodiments, the fragment may contain a discontinuous number of amino acids derived from TMPRSS6.

[0071] 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 derived from crab-eating macaques, rhesus macaques, African green macaques, savanna monkeys, squirrel monkeys, night monkeys, pig-tailed macaques, or baboons). In some embodiments, the anti-TMPRSS6 antibody disclosed herein specifically binds to human TMPRSS6.

[0072] In some embodiments, the anti-TMPRSS6 antibody described herein is an affinity-mature clone. In some embodiments, the anti-TMPRSS6 antibody is at least about 10 times TMPRSS6 (e.g., human or non-human primate TMPRSS6). -4 M, 10 -5 M, 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 a smaller binding affinity (e.g., K DIt binds specifically to the TMPRSS6 protein (as shown by...). For example, the anti-TMPRSS6 antibody of this disclosure can bind to the TMPRSS6 protein (e.g., human TMPRSS6) with affinities between 5 pM and 750 nM, e.g., between 1 nM and 500 nM, e.g., between 10 nM and 450 nM, e.g., between 50 pM and 100 nM, e.g., between 500 pM and 50 nM. This disclosure also includes antibodies that compete with any of the antibodies described herein for binding to the TMPRSS6 protein (e.g., human TMPRSS6) and have affinities 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 embodiments, the anti-TMPRSS6 antibody described herein has a K content in the sub-nanomole range. D TMPRSS6 then binds.

[0073] 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 P2O] 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 given by the following formula: [[Bound]]=[[Free]]] / (Kd+[[[Free]]]) This relates to the concentration of free target protein ([free]).

[0074] K A It is not always necessary to make an accurate determination of K; in some cases, it is sufficient to obtain a quantitative indicator of affinity, which can be determined using methods such as ELISA or FACS analysis. A Because it is proportional to the affinity, it can be used for comparison, for example, to determine whether the affinity is higher, for example, twice as high, and a qualitative indicator of affinity can be obtained, or an affinity can be predicted, by functional assays, for example, activity in in vitro or in vivo assays.

[0075] Table 1 provides the heavy chain (HC) and light chain (LC) sequences, heavy chain variable domain (VH) and light chain variable domain (VL), CDR sequence, and heavy chain and light chain constant region sequences for non-limiting examples of anti-TMPRSS6 antibodies.

[0076] [Table 2-1] [Table 2-2] [Table 2-3] [Table 2-4] [Table 2-5] [Table 2-6] [Table 2-7] [Table 2-8] [Table 2-9] [Table 2-10]

[0077] In some embodiments, the anti-TMPRSS6 antibody of this disclosure comprises one or more HC CDR (e.g., HC CDR1, HC CDR2, or HC CDR3) amino acid sequences from any one of the anti-TMPRSS6 antibodies selected from Table 1. In some embodiments, the anti-TMPRSS6 antibody of this disclosure comprises HC CDR1, HC CDR2, and HC CDR3 provided for any one of the antibodies selected from Table 1. In some embodiments, the anti-TMPRSS6 antibody of this disclosure comprises one or more LC CDR (e.g., LC CDR1, LC CDR2, or LC CDR3) amino acid sequences from any one of the anti-TMPRSS6 antibodies selected from Table 1. In some embodiments, the anti-TMPRSS6 antibody of this disclosure comprises LC CDR1, LC CDR2, and LC CDR3 provided for any one of the anti-TMPRSS6 antibodies selected from Table 1.

[0078] In some embodiments, the anti-TMPRSS6 antibody of this disclosure comprises HC CDR1, HC CDR2, HC CDR3, LC CDR1, LC CDR2, and LC CDR3 according to any one of the anti-TMPRSS6 antibodies selected from Table 1. In some embodiments, the antibody heavy chain and light chain CDR3 domains 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 comprise at least the heavy chain and / or light chain CDR3 of any one of the anti-TMPRSS6 antibodies selected from Table 1.

[0079] 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).

[0080] Also within the scope of this disclosure are any exemplary functional variants of anti-TMPRSS6 antibodies disclosed herein. Functional variants retain substantially similar binding and biological activity (e.g., substantially similar binding affinity, binding specificity, inhibitory activity, or combination thereof) to the reference antibody, while exhibiting V (Varying) compared to the reference antibody. H and / or V L It may contain one or more amino acid residue variations in one or more medium or HC CDRs and / or one or more LC CDRs.

[0081] 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 of the HC CDR1, HC CDR2, HC CDR3, LC CDR1, LC CDR2, and / or LC CDR3 sequences from one of the anti-TMPRSS6 antibodies selected from Table 1. In some embodiments, the positions of one or more CDRs in the VH (e.g., HC CDR1, HC CDR2, or HC CDR3) and / or VL (e.g., LC CDR1, LC CDR2, or LC CDR3) regions of the antibodies described herein may vary by one, two, three, four, five, or six amino acid positions, as long as immunospecific binding to TMPRSS6 (e.g., human TMPRSS6) is maintained (e.g., substantially maintained at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, or at least 95% of the binding of the original antibody from which it is derived). For example, in some embodiments, the position defining the CDR of any antibody described herein may be altered by shifting the N-terminal and / or C-terminal boundary of the CDR by one, two, three, four, five, or six amino acids compared to any one CDR position of any antibody described herein, as long as immunospecific binding to TMPRSS6 (e.g., human TMPRSS6) is maintained (e.g., substantially maintaining at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, or at least 95% of the binding of the original antibody from which it originates). In another embodiment, the length of one or more CDRs in the VH (e.g., HC CDR1, HC CDR2, or HC CDR3) and / or VL (e.g., LC CDR1, LC CDR2, or LC CDR3) regions of the antibodies described herein may vary by one, two, three, four, five, or more amino acids (e.g., short or long), as long as immunospecific binding to TMPRSS6 (e.g., human TMPRSS6) is maintained (e.g., substantially maintained at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, or at least 95% of the binding of the original antibody from which it originates).

[0082] Therefore, in some embodiments, the HC CDR1, HC CDR2, HC CDR3, LC CDR1, LC CDR2, and / or LC CDR3 described herein may have one, two, three, four, five, or more amino acids shorter than one or more of the CDRs described herein (e.g., CDRs from any of the anti-HJV antibodies selected from Table 1), as long as immunospecific binding to TMPRSS6 (e.g., human TMPRSS6) is maintained (e.g., substantially maintained by at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, or at least 95% compared to binding 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 have one, two, three, four, five, or more amino acids longer than one or more of the CDRs described herein (e.g., CDRs from any of the anti-TMPRSS6 antibodies selected from Table 1), provided that immunospecific binding to TMPRSS6 (e.g., human TMPRSS6) is maintained (e.g., substantially maintained by at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, or at least 95% compared to the binding of the original antibody from which it is derived). In some embodiments, the amino portions of HC CDR1, HC CDR2, HC CDR3, LC CDR1, LC CDR2, and / or LC CDR3 described herein may be extended by one, two, three, four, five, or more amino acids compared to one or more CDRs described herein (e.g., CDRs from any of the anti-TMPRSS6 antibodies selected from Table 1), provided that immunospecific binding to TMPRSS6 (e.g., human TMPRSS6) is maintained (e.g., substantially maintained by at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, or at least 95% compared to binding to the original antibody from which it is derived).In some embodiments, the carboxyl portions of HC CDR1, HC CDR2, HC CDR3, LC CDR1, LC CDR2, and / or LC CDR3 described herein may be elongated by one, two, three, four, five, or more amino acids compared to one or more CDRs described herein (e.g., CDRs from any of the anti-TMPRSS6 antibodies selected from Table 1), provided that immunospecific binding to TMPRSS6 (e.g., human TMPRSS6) is maintained (e.g., substantially maintained by at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, or at least 95% compared to binding to the original antibody from which it is derived). In some embodiments, the amino portions 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 CDRs described herein (e.g., CDRs from any of the anti-TMPRSS6 antibodies selected from Table 1), provided that immunospecific binding to TMPRSS6 (e.g., human TMPRSS6) is maintained (e.g., substantially maintained by at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, or at least 95% compared to binding to the original antibody from which it is derived). In some embodiments, the carboxyl portions 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 CDRs described herein (e.g., CDRs from any of the anti-TMPRSS6 antibodies selected from Table 1), provided that immunospecific binding to TMPRSS6 (e.g., human TMPRSS6) is maintained (e.g., substantially maintained by at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, or at least 95% compared to binding to the original antibody from which it is derived).Any method can be used, for example, using the binding assays and conditions described in the Art, to determine whether immunospecific binding to TMPRSS6 (e.g., human TMPRSS6) is maintained.

[0083] In some examples, any anti-TMPRSS6 antibody of this disclosure has one or more CDR (e.g., HC CDR or LC CDR) sequences substantially similar to any one of the anti-TMPRSS6 antibodies selected from Table 1. For example, an antibody may contain one or more CDR sequences from any anti-TMPRSS6 antibody selected from Table 1, containing up to five, four, three, two, or one amino acid residue variation compared to the corresponding CDR region of any one of the CDRs provided herein (e.g., a CDR from any of the anti-TMPRSS6 antibodies selected from Table 1), provided that immunospecific binding to TMPRSS6 (e.g., human TMPRSS6) is maintained (e.g., substantially maintained by at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, or at least 95% compared to the binding of 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. Conservation variations can be introduced into the CDR at positions where the residues would not be involved in interaction with the TMPRSS6 protein (e.g., human TMPRSS6 protein) if determined, for example, based on the crystal structure. Some aspects of this disclosure provide anti-TMPRSS6 antibodies comprising one or more heavy-chain variable (VH) and / or light-chain variable (VL) domains provided herein. In some embodiments, any VH domain provided herein comprises one or more HC CDR sequences provided herein (e.g., HC CDR1, HC CDR2, and HC CDR3), for example, any CDR-H sequence provided in any one of the anti-TMPRSS6 antibodies selected from Table 1. In some embodiments, any VL domain provided herein comprises one or more CDR-L sequences provided herein (e.g., LC CDR1, LC CDR2, and LC CDR3), for example, any LC CDR sequence provided in any one of the anti-TMPRSS6 antibodies selected from Table 1.

[0084] In some embodiments, the anti-TMPRSS6 antibody of this disclosure comprises any antibody comprising the heavy chain variable domain and / or light chain variable domain of any one anti-TMPRSS6 antibody selected from Table 1, as well as any variants thereof. In some embodiments, the anti-TMPRSS6 antibody of this disclosure comprises any antibody comprising the heavy chain variable and light chain variable pairs of any anti-TMPRSS6 antibody selected from Table 1.

[0085] Aspects of this disclosure provide anti-TMPRSS6 antibodies having heavy-chain variable (VH) and / or light-chain variable (VL) domain amino acid sequences homologous to any of those described herein. In some embodiments, the anti-TMPRSS6 antibody includes a heavy-chain variable sequence or a light-chain variable sequence that is 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 99%) identical to any one of the heavy-chain variable sequences and / or light-chain variable sequences of an anti-TMPRSS6 antibody selected from Table 1. In some embodiments, the heavy-chain variable and / or light-chain variable amino acid sequences are unchanged within any CDR sequence 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, excluding any CDR sequences provided herein. In some embodiments, any anti-TMPRSS6 antibody provided herein includes a heavy-chain variable sequence and a light-chain variable sequence that include a framework sequence which is 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% identical to the framework sequence of any anti-TMPRSS6 antibody selected from Table 1.

[0086] In some embodiments, the anti-TMPRSS6 antibody of this disclosure is an antibody comprising a VH containing 20 or fewer amino acid variations (e.g., 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1 or fewer amino acid variations) compared to any VH of any of the anti-TMPRSS6 antibodies listed in Table 1. Alternatively or additionally, the anti-TMPRSS6 antibody of this disclosure is a humanized antibody comprising a VL containing 20 or fewer amino acid variations (e.g., 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 9, 8, 7, 6, 5, 4, 3, 2, or 1 or fewer amino acid variations) compared to any one VL of any of the anti-TMPRSS6 antibodies listed in Table 1. In some embodiments, the heavy-chain variable and / or light-chain variable amino acid sequence remains unchanged within any CDR sequence provided herein. For example, in some embodiments, the number of amino acid variations (e.g., 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1 or fewer amino acid variations) can occur within the heavy-chain variable and / or light-chain variable sequence, excluding any CDR sequence provided herein. In some embodiments, any anti-TMPRSS6 antibody provided herein includes a heavy chain variable sequence comprising a framework sequence containing 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1 or fewer amino acid variations for any one VH framework sequence of an anti-TMPRSS6 antibody selected from Table 1, and / or a light chain variable sequence comprising a framework sequence containing 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1 or fewer amino acid variations for any one VL framework sequence of an anti-TMPRSS6 antibody selected from Table 1.

[0087] In some embodiments, the anti-TMPRSS6 antibody of this disclosure is a humanized antibody (e.g., a humanized variant containing one or more CDRs from Table 1). In some embodiments, the anti-TMPRSS6 antibody of this disclosure comprises HC CDR1, HC CDR2, HC CDR3, LC CDR1, LC CDR2, and LC CDR3, which are identical to HC CDR1, HC CDR2, HC CDR3, LC CDR1, LC CDR2, and LC CDR3 shown in Table 1, and includes a humanized heavy chain variable region and / or a humanized light chain variable region. In some embodiments, the humanized heavy chain variable and / or humanized light chain variable amino acid sequence is unchanged within any CDR sequence 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, excluding any CDR sequences provided herein. In some embodiments, the humanized anti-TMPRSS6 antibodies provided herein include heavy-chain variable sequences and light-chain variable sequences that include a framework sequence which is 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% identical to the framework sequence of any anti-TMPRSS6 antibody selected from Table 1. In some embodiments, the humanized heavy-chain variable and / or humanized light-chain variable amino acid sequences are unchanged within any CDR sequences provided herein. For example, in some embodiments, the number of amino acid variations (e.g., 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1 or fewer amino acid variations) can occur within heavy-chain variable and / or light-chain variable sequences, excluding any CDR sequences provided herein.In some embodiments, the humanized anti-TMPRSS6 antibody provided herein includes a heavy chain variable sequence comprising a framework sequence containing 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1 or fewer amino acid variations for any one VH framework sequence of an anti-TMPRSS6 antibody selected from Table 1, and / or a light chain variable sequence comprising a framework sequence containing 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1 or fewer amino acid variations for any one VL framework sequence of an anti-TMPRSS6 antibody selected from Table 1.

[0088] In some embodiments, the anti-TMPRSS6 antibody of this disclosure comprises heavy chain variable domains HC CDR1, HC CDR2, and HC CDR3 having the amino acid sequence of SEQ ID NO: 7. Alternatively or additionally, the anti-TMPRSS6 antibody of this disclosure comprises light chain variable domains LC CDR1, LC CDR2, and LC CDR3 having the amino acid sequence of SEQ ID NO: 8.

[0089] In some embodiments, the anti-TMPRSS6 antibody of the present disclosure is defined by the Kabat definition system as comprising 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.

[0090] In some embodiments, the anti-TMPRSS6 antibody of this disclosure comprises HC CDR1, HC CDR2, and HC CDR3, each containing a total of five or fewer amino acid variations (e.g., five, four, three, two, or one or fewer amino acid variations) compared to 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. Where used in any part of this disclosure, "total" means that the total number of amino acid variations in all three heavy chain CDRs is within a defined range. Alternatively or additionally, the anti-TMPRSS6 antibody of this disclosure comprises LC CDR1, LC CDR2, and LC CDR3, each containing a total of five or fewer amino acid variations (e.g., five, four, three, two, or one or fewer amino acid variations) 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.

[0091] In some embodiments, the anti-TMPRSS6 antibody of the present 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, and HC CDR3 having the amino acid sequence of SEQ ID NO: 3, with a total of 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%) identical HC CDR1, HC CDR2, and HC CDR3 having the amino acid sequence of SEQ ID NO: 3. Alternatively or additionally, the anti-TMPRSS6 antibody of this disclosure comprises LC CDR1, LC CDR2, and LC CDR3 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, which together comprise 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%) identical LC CDR1, LC CDR2, and LC CDR3 having the amino acid sequence of SEQ ID NO: 6.

[0092] In some embodiments, the anti-TMPRSS6 antibody of the present disclosure includes HC CDR1 having three or fewer amino acid variations (e.g., three, two, or one or fewer amino acid variations) compared to HC CDR1 having the amino acid sequence of SEQ ID NO: 1; HC CDR2 having three or fewer amino acid variations (e.g., three, two, or one or fewer amino acid variations) compared to HC CDR2 having the amino acid sequence of SEQ ID NO: 2; and / or HC CDR3 having three or fewer amino acid variations (e.g., three, two, or one or fewer amino acid variations) compared to HC CDR3 having the amino acid sequence of SEQ ID NO: 3. Alternatively or additionally, the anti-TMPRSS6 antibodies of this disclosure include LC CDR1 having three or fewer amino acid variations (e.g., three, two, or one or fewer amino acid variations) compared to LC CDR1 having the amino acid sequence of SEQ ID NO. 4; LC CDR2 having three or fewer amino acid variations (e.g., three, two, or one or fewer amino acid variations) compared to LC CDR2 having the amino acid sequence of RAN; and / or LC CDR3 having three or fewer amino acid variations (e.g., three, two, or one or fewer amino acid variations) compared to LC CDR3 having the amino acid sequence of SEQ ID NO. 6.

[0093] In some embodiments, the anti-TMPRSS6 antibody of this disclosure is HC CDR1 which is 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%) identical to HC CDR1 which has the amino acid sequence of SEQ ID NO: 1; HC CDR2 which is 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%) identical to HC CDR2 which has the amino acid sequence of SEQ ID NO: 2; and / or HC CDR2 which has the amino acid sequence of SEQ ID NO: 3 Includes HC CDR3 which is at least 80% identical to CDR3 (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%).Alternatively or additionally, the anti-TMPRSS6 antibody of this disclosure is 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%) identical to LC CDR1 having the amino acid sequence of SEQ ID NO: 4; LC CDR2 having the amino acid sequence of RAN is 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%) identical to LC CDR1 having the amino acid sequence of SEQ ID NO: 6 Includes LC CDR3 which is at least 80% identical to CDR3 (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%).

[0094] In some embodiments, the anti-TMPRSS6 antibody of the present disclosure comprises VH, which contains the amino acid sequence of SEQ ID NO: 7. Alternatively or additionally, the anti-TMPRSS6 antibody of the present disclosure comprises VL, which contains the amino acid sequence of SEQ ID NO: 8.

[0095] In some embodiments, the anti-TMPRSS6 antibody of the present disclosure includes a VH containing 20 or fewer amino acid variations compared to the VH described in SEQ ID NO: 7 (e.g., 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1 or fewer amino acid variations). Alternatively, the anti-TMPRSS6 antibody of the present disclosure includes a VL containing 20 or fewer amino acid variations compared to the VL described in SEQ ID NO: 8 (e.g., 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 9, 8, 7, 6, 5, 4, 3, 2, or 1 or fewer amino acid variations). In some embodiments, the number of amino acid variations (e.g., 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1 or fewer amino acid variations) can occur within the VH of SEQ ID NO: 7 and / or the VL of SEQ ID NO: 8, excluding any CDR sequence within them. In some embodiments, the anti-TMPRSS6 antibody provided herein includes a heavy chain variable sequence comprising a framework sequence containing 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1 or fewer amino acid variations relative to the VH framework sequence of SEQ ID NO: 7, and / or a light chain variable sequence comprising a framework sequence containing 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1 or fewer amino acid variations relative to the VL framework sequence of SEQ ID NO: 8.

[0096] In some embodiments, the anti-TMPRSS6 antibody of the present disclosure comprises a VH having an amino acid sequence that is 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%) identical to the VH described in SEQ ID NO: 7. Alternatively, the anti-TMPRSS6 antibody of the present disclosure comprises a VL having an amino acid sequence that is 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 99%) identical to the VL described in SEQ ID NO: 8. 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 VH of sequence number 7 and / or the VL of sequence number 8, excluding any CDR sequences within them. In some embodiments, the anti-TMPRSS6 antibody provided herein includes a heavy chain variable sequence that is 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% identical to the framework sequence of VH of SEQ ID NO: 7, and / or a light chain variable sequence that includes a framework sequence that is 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% identical to the framework sequence of VL of SEQ ID NO: 8.

[0097] In some embodiments, the anti-TMPRSS6 antibody of the present disclosure comprises heavy chain variable domains HC CDR1, HC CDR2, and HC CDR3 having the amino acid sequence of SEQ ID NO: 19 or 78. Alternatively, the anti-TMPRSS6 antibody of the present disclosure comprises light chain variable domains LC CDR1, LC CDR2, and LC CDR3 having the amino acid sequence of SEQ ID NO: 20.

[0098] In some embodiments, the anti-TMPRSS6 antibody of the present disclosure is defined by the Kabat definition system as comprising 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.

[0099] In some embodiments, the anti-TMPRSS6 antibody of the Disclosure comprises HC CDR1, HC CDR2, and HC CDR3, each containing a total of five or fewer amino acid variations (e.g., five, four, three, two, or one or fewer amino acid variations) 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. Alternatively or additionally, the anti-TMPRSS6 antibody of the Disclosure comprises LC CDR1, LC CDR2, and LC CDR3, each containing a total of five or fewer amino acid variations (e.g., five, four, three, two, or one or fewer amino acid variations) compared to 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.

[0100] In some embodiments, the anti-TMPRSS6 antibody of the present 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, and HC CDR3 having the amino acid sequence of SEQ ID NO: 15, with 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 99%) identical HC CDR1, HC CDR2, and HC CDR3 having the amino acid sequence of SEQ ID NO: 15. Alternatively or additionally, the anti-TMPRSS6 antibody of this disclosure comprises 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, together comprising LC CDR1, LC CDR2, and LC CDR3 that are 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%) identical.

[0101] In some embodiments, the anti-TMPRSS6 antibody of the present disclosure includes HC CDR1 having three or fewer amino acid variations (e.g., three, two, or one or fewer amino acid variations) compared to HC CDR1 having the amino acid sequence of SEQ ID NO. 13; HC CDR2 having three or fewer amino acid variations (e.g., three, two, or one or fewer amino acid variations) compared to HC CDR2 having the amino acid sequence of SEQ ID NO. 14; and / or HC CDR3 having three or fewer amino acid variations (e.g., three, two, or one or fewer amino acid variations) compared to HC CDR3 having the amino acid sequence of SEQ ID NO. 15. Alternatively or additionally, the anti-TMPRSS6 antibodies of this disclosure include LC CDR1 having three or fewer amino acid variations (e.g., three, two, or one or fewer amino acid variations) compared to LC CDR1 having the amino acid sequence of SEQ ID NO. 16; LC CDR2 having three or fewer amino acid variations (e.g., three, two, or one or fewer amino acid variations) compared to LC CDR2 having the amino acid sequence of WAF; and / or LC CDR3 having three or fewer amino acid variations (e.g., three, two, or one or fewer amino acid variations) compared to LC CDR3 having the amino acid sequence of SEQ ID NO. 18.

[0102] In some embodiments, the anti-TMPRSS6 antibody of this disclosure is HC CDR1 which is 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%) identical to HC CDR1 having the amino acid sequence of SEQ ID NO: 13; HC CDR2 which is 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%) identical to HC CDR2 having the amino acid sequence of SEQ ID NO: 14; and / or HC CDR2 having the amino acid sequence of SEQ ID NO: 15 Includes HC CDR3 which is at least 80% identical to CDR3 (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%).Alternatively or additionally, the anti-TMPRSS6 antibody of this disclosure is 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%) identical to LC CDR1 having the amino acid sequence of SEQ ID NO: 16; LC CDR2 having the amino acid sequence of WAF is 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%) identical to LC CDR1 having the amino acid sequence of SEQ ID NO: 18; and / or LC CDR2 having the amino acid sequence of SEQ ID NO: 18 Includes LC CDR3 which is at least 80% identical to CDR3 (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%).

[0103] In some embodiments, the anti-TMPRSS6 antibody of this disclosure comprises VH, which includes the amino acid sequence of SEQ ID NO: 19 or 78. Alternatively or additionally, the anti-TMPRSS6 antibody of this disclosure comprises VL, which includes the amino acid sequence of SEQ ID NO: 20.

[0104] In some embodiments, the anti-TMPRSS6 antibody of this disclosure comprises a VH containing 20 or fewer amino acid variations (e.g., 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1 or fewer amino acid variations) compared to the VH described in SEQ ID NO: 19 or 78. Alternatively, the anti-TMPRSS6 antibody of this disclosure comprises a VL containing 20 or fewer amino acid variations (e.g., 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 9, 8, 7, 6, 5, 4, 3, 2, or 1 or fewer amino acid variations) compared to the VL described in SEQ ID NO: 20. In some embodiments, the number of amino acid variations (e.g., 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1 or fewer amino acid variations) can occur within the VH of SEQ ID NO: 19 or 78 and / or the VL of SEQ ID NO: 20, excluding any CDR sequence among them. In some embodiments, the anti-TMPRSS6 antibody provided herein includes a heavy chain variable sequence comprising a framework sequence containing 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1 or fewer amino acid variations relative to the VH framework sequence of SEQ ID NO: 19 or 78, and / or a light chain variable sequence comprising a framework sequence containing 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1 or fewer amino acid variations relative to the VL framework sequence of SEQ ID NO: 20.

[0105] In some embodiments, the anti-TMPRSS6 antibody of the present disclosure comprises a VH having an amino acid sequence that is 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%) identical to the VH described in SEQ ID NO: 19 or 78. Alternatively or additionally, the anti-TMPRSS6 antibody of the present disclosure comprises a VL having an amino acid sequence that is 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%) identical to the VL described 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 occur within the VH of sequence number 19 or 78 and / or the VL of sequence number 20, excluding any CDR sequences within them. In some embodiments, the anti-TMPRSS6 antibody provided herein includes a heavy chain variable sequence that is 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% identical to the VH framework sequence of SEQ ID NO: 19 or 78, and / or a light chain variable sequence that includes a framework sequence that is 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% identical to the VL framework sequence of SEQ ID NO: 20.

[0106] In some embodiments, the anti-TMPRSS6 antibody of the present disclosure comprises heavy chain variable domains HC CDR1, HC CDR2, and HC CDR3 having the amino acid sequence of SEQ ID NO: 30. Alternatively, the anti-TMPRSS6 antibody of the present disclosure comprises light chain variable domains LC CDR1, LC CDR2, and LC CDR3 having the amino acid sequence of SEQ ID NO: 31.

[0107] In some embodiments, the anti-TMPRSS6 antibody of the present disclosure is defined by the Kabat definition system as comprising 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.

[0108] In some embodiments, the anti-TMPRSS6 antibody of the Disclosure comprises HC CDR1, HC CDR2, and HC CDR3, each containing a total of five or fewer amino acid variations (e.g., five, four, three, two, or one or fewer amino acid variations) 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. Alternatively or additionally, the anti-TMPRSS6 antibody of the Disclosure comprises LC CDR1, LC CDR2, and LC CDR3, each containing a total of five or fewer amino acid variations (e.g., five, four, three, two, or one or fewer amino acid variations) 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.

[0109] In some embodiments, the anti-TMPRSS6 antibody of the present 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, and HC CDR3 having the amino acid sequence of SEQ ID NO: 26, together comprising 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 99%) identical HC CDR1, HC CDR2, and HC CDR3. Alternatively or additionally, the anti-TMPRSS6 antibody of this disclosure comprises 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, together comprising LC CDR1, LC CDR2, and LC CDR3 that are 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 99%) identical.

[0110] In some embodiments, the anti-TMPRSS6 antibody of the present disclosure includes HC CDR1 having three or fewer amino acid variations (e.g., three, two, or one or fewer amino acid variations) compared to HC CDR1 having the amino acid sequence of SEQ ID NO. 24; HC CDR2 having three or fewer amino acid variations (e.g., three, two, or one or fewer amino acid variations) compared to HC CDR2 having the amino acid sequence of SEQ ID NO. 25; and / or HC CDR3 having three or fewer amino acid variations (e.g., three, two, or one or fewer amino acid variations) compared to HC CDR3 having the amino acid sequence of SEQ ID NO. 26. Alternatively or additionally, the anti-TMPRSS6 antibodies of this disclosure include LC CDR1 having three or fewer amino acid variations (e.g., three, two, or one or fewer amino acid variations) compared to LC CDR1 having the amino acid sequence of SEQ ID NO. 27; LC CDR2 having three or fewer amino acid variations (e.g., three, two, or one or fewer amino acid variations) compared to LC CDR2 having the amino acid sequence of WAT; and / or LC CDR3 having three or fewer amino acid variations (e.g., three, two, or one or fewer amino acid variations) compared to LC CDR3 having the amino acid sequence of SEQ ID NO. 29.

[0111] In some embodiments, the anti-TMPRSS6 antibody of this disclosure is HC CDR1 which is 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%) identical to HC CDR1 having the amino acid sequence of SEQ ID NO: 24; HC CDR2 which is 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%) identical to 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 Includes HC CDR3 which is at least 80% identical to CDR3 (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%).Alternatively or additionally, the anti-TMPRSS6 antibody of this disclosure is 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%) identical to LC CDR1 having the amino acid sequence of SEQ ID NO: 27; LC CDR2 having the amino acid sequence of WAT is 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%) identical to LC CDR1 having the amino acid sequence of SEQ ID NO: 29 Includes LC CDR3 which is at least 80% identical to CDR3 (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%).

[0112] In some embodiments, the anti-TMPRSS6 antibody of this disclosure comprises VH, which contains the amino acid sequence of SEQ ID NO: 30. Alternatively or additionally, the anti-TMPRSS6 antibody of this disclosure comprises VL, which contains the amino acid sequence of SEQ ID NO: 31.

[0113] In some embodiments, the anti-TMPRSS6 antibody of the present disclosure includes a VH containing 20 or fewer amino acid variations compared to the VH described in SEQ ID NO: 30 (e.g., 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1 or fewer amino acid variations). Alternatively, the anti-TMPRSS6 antibody of the present disclosure includes a VL containing 20 or fewer amino acid variations compared to the VL described in SEQ ID NO: 31 (e.g., 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 9, 8, 7, 6, 5, 4, 3, 2, or 1 or fewer amino acid variations). In some embodiments, the number of amino acid variations (e.g., 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1 or fewer amino acid variations) can occur within the VH of SEQ ID NO: 30 and / or the VL of SEQ ID NO: 31, excluding any CDR sequence among them. In some embodiments, the anti-TMPRSS6 antibody provided herein includes a heavy chain variable sequence comprising a framework sequence containing 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1 or fewer amino acid variations relative to the VH framework sequence of SEQ ID NO: 30, and / or a light chain variable sequence comprising a framework sequence containing 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1 or fewer amino acid variations relative to the VL framework sequence of SEQ ID NO: 31.

[0114] In some embodiments, the anti-TMPRSS6 antibody of the present disclosure comprises a VH having an amino acid sequence that is 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%) identical to the VH described in SEQ ID NO: 30. Alternatively, or further, the anti-TMPRSS6 antibody of the present disclosure comprises a VL having an amino acid sequence that is 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%) identical to the VL described in SEQ ID NO: 31. 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 VH of sequence number 30 and / or the VL of sequence number 31, excluding any CDR sequences within it. In some embodiments, the anti-TMPRSS6 antibody provided herein includes a heavy chain variable sequence that is 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% identical to the VH framework sequence of SEQ ID NO: 30, and / or a light chain variable sequence that includes a framework sequence that is 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% identical to the VL framework sequence of SEQ ID NO: 31.

[0115] In some embodiments, the anti-TMPRSS6 antibody of this disclosure comprises HC CDR1, HC CDR2, and HC CDR3 heavy chain variable domains having the amino acid sequence of SEQ ID NO: 37. Alternatively or additionally, the anti-TMPRSS6 antibody of this disclosure comprises LC CDR1, LC CDR2, and LC CDR3 light chain variable domains having the amino acid sequence of SEQ ID NO: 38.

[0116] In some embodiments, the anti-TMPRSS6 antibody of the present disclosure is defined by the Kabat definition system as comprising 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.

[0117] In some embodiments, the anti-TMPRSS6 antibody of the present disclosure comprises HC CDR1, HC CDR2, and HC CDR3, each containing a total of five or fewer amino acid variations (e.g., five, four, three, two, or one or fewer amino acid variations) compared to 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. Alternatively or additionally, the anti-TMPRSS6 antibody of the present disclosure comprises LC CDR1, LC CDR2, and LC CDR3, each containing a total of five or fewer amino acid variations (e.g., five, four, three, two, or one or fewer amino acid variations) 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.

[0118] In some embodiments, the anti-TMPRSS6 antibody of the present 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, and HC CDR3 having the amino acid sequence of SEQ ID NO: 35, with 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%) identical HC CDR1, HC CDR2, and HC CDR3 having the amino acid sequence of SEQ ID NO: 35. Alternatively or additionally, the anti-TMPRSS6 antibody of this disclosure comprises 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, together comprising LC CDR1, LC CDR2, and LC CDR3 that are 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%) identical.

[0119] In some embodiments, the anti-TMPRSS6 antibody of the present disclosure includes HC CDR1 having three or fewer amino acid variations (e.g., three, two, or one or fewer amino acid variations) compared to HC CDR1 having the amino acid sequence of SEQ ID NO. 1; HC CDR2 having three or fewer amino acid variations (e.g., three, two, or one or fewer amino acid variations) compared to HC CDR2 having the amino acid sequence of SEQ ID NO. 2; and / or HC CDR3 having three or fewer amino acid variations (e.g., three, two, or one or fewer amino acid variations) compared to HC CDR3 having the amino acid sequence of SEQ ID NO. 35. Alternatively or additionally, the anti-TMPRSS6 antibodies of this disclosure include LC CDR1 having three or fewer amino acid variations (e.g., three, two, or one or fewer amino acid variations) compared to LC CDR1 having the amino acid sequence of SEQ ID NO. 36; LC CDR2 having three or fewer amino acid variations (e.g., three, two, or one or fewer amino acid variations) compared to LC CDR2 having the amino acid sequence of RAN; and / or LC CDR3 having three or fewer amino acid variations (e.g., three, two, or one or fewer amino acid variations) compared to LC CDR3 having the amino acid sequence of SEQ ID NO. 6.

[0120] In some embodiments, the anti-TMPRSS6 antibody of this disclosure is HC CDR1 which is 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%) identical to HC CDR1 which has the amino acid sequence of SEQ ID NO: 1; HC CDR2 which is 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%) identical to HC CDR2 which has the amino acid sequence of SEQ ID NO: 2; and / or HC CDR2 which has the amino acid sequence of SEQ ID NO: 35 Includes HC CDR3 which is at least 80% identical to CDR3 (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%).Alternatively or additionally, the anti-TMPRSS6 antibody of this disclosure is 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%) identical to LC CDR1 having the amino acid sequence of SEQ ID NO: 36; LC CDR2 having the amino acid sequence of RAN is 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%) identical to LC CDR1 having the amino acid sequence of SEQ ID NO: 6; and / or LC CDR2 having the amino acid sequence of SEQ ID NO: 6 Includes LC CDR3 which is at least 80% identical to CDR3 (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%).

[0121] In some embodiments, the anti-TMPRSS6 antibody of the present disclosure comprises VH, which contains the amino acid sequence of SEQ ID NO: 37. Alternatively or additionally, the anti-TMPRSS6 antibody of the present disclosure comprises VL, which contains the amino acid sequence of SEQ ID NO: 38.

[0122] In some embodiments, the anti-TMPRSS6 antibody of this disclosure includes a VH containing 20 or fewer amino acid variations compared to the VH described in SEQ ID NO: 37 (e.g., 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1 or fewer amino acid variations). Alternatively or additionally, the anti-TMPRSS6 antibody of this disclosure includes a VL containing 20 or fewer amino acid variations compared to the VL described in SEQ ID NO: 38 (e.g., 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 9, 8, 7, 6, 5, 4, 3, 2, or 1 or fewer amino acid variations). In some embodiments, the number of amino acid variations (e.g., 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1 or fewer amino acid variations) can occur within the VH of SEQ ID NO: 37 and / or the VL of SEQ ID NO: 38, excluding any CDR sequence among them. In some embodiments, the anti-TMPRSS6 antibody provided herein includes a heavy chain variable sequence comprising a framework sequence containing 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1 or fewer amino acid variations relative to the VH framework sequence of SEQ ID NO: 37, and / or a light chain variable sequence comprising a framework sequence containing 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1 or fewer amino acid variations relative to the VL framework sequence of SEQ ID NO: 38.

[0123] In some embodiments, the anti-TMPRSS6 antibody of the present 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%) the same amino acid sequence as the VH described in SEQ ID NO: 37. Alternatively or additionally, the anti-TMPRSS6 antibody of the present 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%) the same amino acid sequence as the VL described in SEQ ID NO: 38. 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 VH of sequence number 37 and / or the VL of sequence number 38, excluding any CDR sequences within it. In some embodiments, the anti-TMPRSS6 antibody provided herein includes a heavy chain variable sequence that is 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% identical to the VH framework sequence of SEQ ID NO: 37, and / or a light chain variable sequence that includes a framework sequence that is 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% identical to the VL framework sequence of SEQ ID NO: 38.

[0124] In some embodiments, the anti-TMPRSS6 antibody of this disclosure comprises HC CDR1, HC CDR2, and HC CDR3 heavy chain variable domains having the amino acid sequence of SEQ ID NO: 44. Alternatively or additionally, the anti-TMPRSS6 antibody of this disclosure comprises LC CDR1, LC CDR2, and LC CDR3 light chain variable domains having the amino acid sequence of SEQ ID NO: 45.

[0125] In some embodiments, the anti-TMPRSS6 antibody of the present disclosure is defined by the Kabat definition system as comprising 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.

[0126] In some embodiments, the anti-TMPRSS6 antibody of the Disclosure comprises HC CDR1, HC CDR2, and HC CDR3, each containing a total of five or fewer amino acid variations (e.g., five, four, three, two, or one or fewer amino acid variations) 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. Alternatively or additionally, the anti-TMPRSS6 antibody of the Disclosure comprises LC CDR1, LC CDR2, and LC CDR3, each containing a total of five or fewer amino acid variations (e.g., five, four, three, two, or one or fewer amino acid variations) compared to 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.

[0127] In some embodiments, the anti-TMPRSS6 antibody of the present 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, and HC CDR3 having the amino acid sequence of SEQ ID NO: 15, with a total of 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%) identical HC CDR1, HC CDR2, and HC CDR3 having the amino acid sequence of SEQ ID NO: 15. Alternatively or additionally, the anti-TMPRSS6 antibody of this disclosure comprises 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, together comprising LC CDR1, LC CDR2, and LC CDR3 that are 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%) identical.

[0128] In some embodiments, the anti-TMPRSS6 antibody of the present disclosure includes HC CDR1 having three or fewer amino acid variations (e.g., three, two, or one or fewer amino acid variations) compared to HC CDR1 having the amino acid sequence of SEQ ID NO. 13; HC CDR2 having three or fewer amino acid variations (e.g., three, two, or one or fewer amino acid variations) compared to HC CDR2 having the amino acid sequence of SEQ ID NO. 43; and / or HC CDR3 having three or fewer amino acid variations (e.g., three, two, or one or fewer amino acid variations) compared to HC CDR3 having the amino acid sequence of SEQ ID NO. 15. Alternatively or additionally, the anti-TMPRSS6 antibodies of this disclosure include LC CDR1 having three or fewer amino acid variations (e.g., three, two, or one or fewer amino acid variations) compared to LC CDR1 having the amino acid sequence of SEQ ID NO. 16; LC CDR2 having three or fewer amino acid variations (e.g., three, two, or one or fewer amino acid variations) compared to LC CDR2 having the amino acid sequence of WAF; and / or LC CDR3 having three or fewer amino acid variations (e.g., three, two, or one or fewer amino acid variations) compared to LC CDR3 having the amino acid sequence of SEQ ID NO. 18.

[0129] In some embodiments, the anti-TMPRSS6 antibody of this disclosure is HC CDR1 which is 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%) identical to HC CDR1 having the amino acid sequence of SEQ ID NO: 13; HC CDR2 which is 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%) identical to 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 Includes HC CDR3 which is at least 80% identical to CDR3 (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%).Alternatively or additionally, the anti-TMPRSS6 antibody of this disclosure is 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%) identical to LC CDR1 having the amino acid sequence of SEQ ID NO: 16; LC CDR2 having the amino acid sequence of WAF is 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%) identical to LC CDR1 having the amino acid sequence of SEQ ID NO: 18; and / or LC CDR2 having the amino acid sequence of SEQ ID NO: 18 Includes LC CDR3 which is at least 80% identical to CDR3 (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%).

[0130] In some embodiments, the anti-TMPRSS6 antibody of the present disclosure comprises VH, which contains the amino acid sequence of SEQ ID NO: 44. Alternatively or additionally, the anti-TMPRSS6 antibody of the present disclosure comprises VL, which contains the amino acid sequence of SEQ ID NO: 45.

[0131] In some embodiments, the anti-TMPRSS6 antibody of this disclosure includes a VH containing 20 or fewer amino acid variations compared to the VH described in SEQ ID NO: 44 (e.g., 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1 or fewer amino acid variations). Alternatively or additionally, the anti-TMPRSS6 antibody of this disclosure includes a VL containing 20 or fewer amino acid variations compared to the VL described in SEQ ID NO: 45 (e.g., 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 9, 8, 7, 6, 5, 4, 3, 2, or 1 or fewer amino acid variations). In some embodiments, the number of amino acid variations (e.g., 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1 or fewer amino acid variations) can occur within the VH of SEQ ID NO: 44 and / or the VL of SEQ ID NO: 45, excluding any CDR sequence among them. In some embodiments, the anti-TMPRSS6 antibody provided herein includes a heavy chain variable sequence comprising a framework sequence containing 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1 or fewer amino acid variations relative to the VH framework sequence of SEQ ID NO: 44, and / or a light chain variable sequence comprising a framework sequence containing 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1 or fewer amino acid variations relative to the VL framework sequence of SEQ ID NO: 45.

[0132] In some embodiments, the anti-TMPRSS6 antibody of the present disclosure comprises a VH having at least 80% (e.g., at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or 99%) of the amino acid sequence of the VH described in SEQ ID NO: 44. Alternatively or additionally, the anti-TMPRSS6 antibody of the present 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%) the same amino acid sequence as the VL described in SEQ ID NO: 45. 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 VH of sequence number 44 and / or the VL of sequence number 45, excluding any CDR sequences within them. In some embodiments, the anti-TMPRSS6 antibody provided herein includes a heavy chain variable sequence that is 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% identical to the VH framework sequence of SEQ ID NO: 44, and / or a light chain variable sequence that includes a framework sequence that is 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% identical to the VL framework sequence of SEQ ID NO: 45.

[0133] In some embodiments, the anti-TMPRSS6 antibody of this disclosure comprises HC CDR1, HC CDR2, and HC CDR3 heavy chain variable domains having the amino acid sequence of SEQ ID NO: 49. Alternatively or additionally, the anti-TMPRSS6 antibody of this disclosure comprises LC CDR1, LC CDR2, and LC CDR3 light chain variable domains having the amino acid sequence of SEQ ID NO: 50.

[0134] In some embodiments, the anti-TMPRSS6 antibody of the present disclosure is defined by the Kabat definition system as comprising 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.

[0135] In some embodiments, the anti-TMPRSS6 antibody of the Disclosure comprises HC CDR1, HC CDR2, and HC CDR3, each containing a total of five or fewer amino acid variations (e.g., five, four, three, two, or one or fewer amino acid variations) 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. Alternatively or additionally, the anti-TMPRSS6 antibody of the Disclosure comprises LC CDR1, LC CDR2, and LC CDR3, each containing a total of five or fewer amino acid variations (e.g., five, four, three, two, or one or fewer amino acid variations) 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.

[0136] In some embodiments, the anti-TMPRSS6 antibody of the present disclosure comprises at least 80% (e.g., at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%) of HC CDR1, HC CDR2, and HC CDR3 having the amino acid sequence of SEQ ID NO: 24, HC CDR1 having the amino acid sequence of SEQ ID NO: 48, and HC CDR3 having the amino acid sequence of SEQ ID NO: 26. Alternatively or additionally, the anti-TMPRSS6 antibody of this disclosure comprises 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, together comprising LC CDR1, LC CDR2, and LC CDR3 that are 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%) identical.

[0137] In some embodiments, the anti-TMPRSS6 antibody of the present disclosure includes HC CDR1 having three or fewer amino acid variations (e.g., three, two, or one or fewer amino acid variations) compared to HC CDR1 having the amino acid sequence of SEQ ID NO. 24; HC CDR2 having three or fewer amino acid variations (e.g., three, two, or one or fewer amino acid variations) compared to HC CDR2 having the amino acid sequence of SEQ ID NO. 48; and / or HC CDR3 having three or fewer amino acid variations (e.g., three, two, or one or fewer amino acid variations) compared to HC CDR3 having the amino acid sequence of SEQ ID NO. 26. Alternatively or additionally, the anti-TMPRSS6 antibodies of this disclosure include LC CDR1 having three or fewer amino acid variations (e.g., three, two, or one or fewer amino acid variations) compared to LC CDR1 having the amino acid sequence of SEQ ID NO. 27; LC CDR2 having three or fewer amino acid variations (e.g., three, two, or one or fewer amino acid variations) compared to LC CDR2 having the amino acid sequence of WAT; and / or LC CDR3 having three or fewer amino acid variations (e.g., three, two, or one or fewer amino acid variations) compared to LC CDR3 having the amino acid sequence of SEQ ID NO. 29.

[0138] In some embodiments, the anti-TMPRSS6 antibody of this disclosure is HC CDR1 which is 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%) identical to HC CDR1 having the amino acid sequence of SEQ ID NO: 24; HC CDR2 which is 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%) identical to 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 Includes HC CDR3 which is at least 80% identical to CDR3 (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%).Alternatively or additionally, the anti-TMPRSS6 antibody of this disclosure is 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%) identical to LC CDR1 having the amino acid sequence of SEQ ID NO: 27; LC CDR2 having the amino acid sequence of WAT is 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%) identical to LC CDR1 having the amino acid sequence of SEQ ID NO: 29 Includes LC CDR3 which is at least 80% identical to CDR3 (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%).

[0139] In some embodiments, the anti-TMPRSS6 antibody of this disclosure comprises VH, which contains the amino acid sequence of SEQ ID NO: 49. Alternatively or additionally, the anti-TMPRSS6 antibody of this disclosure comprises VL, which contains the amino acid sequence of SEQ ID NO: 50.

[0140] In some embodiments, the anti-TMPRSS6 antibody of this disclosure includes a VH containing 20 or fewer amino acid variations compared to the VH described in SEQ ID NO: 49 (e.g., 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1 or fewer amino acid variations). Alternatively or additionally, the anti-TMPRSS6 antibody of this disclosure includes a VL containing 20 or fewer amino acid variations compared to the VL described in SEQ ID NO: 50 (e.g., 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 9, 8, 7, 6, 5, 4, 3, 2, or 1 or fewer amino acid variations). In some embodiments, the number of amino acid variations (e.g., 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1 or fewer amino acid variations) can occur within the VH of SEQ ID NO: 49 and / or the VL of SEQ ID NO: 50, excluding any CDR sequence among them. In some embodiments, the anti-TMPRSS6 antibody provided herein includes a heavy chain variable sequence comprising a framework sequence containing 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1 or fewer amino acid variations relative to the VH framework sequence of SEQ ID NO: 49, and / or a light chain variable sequence comprising a framework sequence containing 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1 or fewer amino acid variations relative to the VL framework sequence of SEQ ID NO: 50.

[0141] In some embodiments, the anti-TMPRSS6 antibody of the present 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%) the same amino acid sequence as the VH described in SEQ ID NO: 49. Alternatively or additionally, the anti-TMPRSS6 antibody of the present 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%) the same amino acid sequence as the VL described in SEQ ID NO: 50. 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 VH of sequence number 49 and / or the VL of sequence number 50, excluding any CDR sequences within them. In some embodiments, the anti-TMPRSS6 antibody provided herein includes a heavy chain variable sequence that is 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% identical to the VH framework sequence of SEQ ID NO: 49, and / or a light chain variable sequence that includes a framework sequence that is 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% identical to the VL framework sequence of SEQ ID NO: 50.

[0142] In some embodiments, the anti-TMPRSS6 antibodies described herein are chimeric antibodies that may contain a heavy chain constant region and a light chain constant region from a human antibody. A chimeric antibody refers to an antibody having a variable region or a portion of a variable region from a first species and a constant region from a second species. Typically, in these chimeric antibodies, both the light chain and heavy chain variable regions mimic the variable region of an antibody derived from one species of mammal (e.g., a non-human mammal, e.g., mouse, rabbit, and rat), and the constant region is homologous to a sequence in an antibody derived from another mammal, e.g., human. In some embodiments, amino acid modifications may be made in the variable region and / or the constant region.

[0143] In some embodiments, the anti-TMPRSS6 antibody of this disclosure comprises the VH domain and / or VL domain of any one of the anti-TMPRSS6 antibodies selected from Table 1, and comprises a constant region containing the amino acid sequence of the constant region of an IgG, IgE, IgM, IgD, IgA, or IgY immunoglobulin molecule, any class of immunoglobulin molecule (e.g., IgG1, IgG2, IgG3, IgG4, IgA1, and IgA2), or any subclass (e.g., IgG2a and IgG2b). Non-limiting examples of human constant regions are described in the Art, see, for example, Kabat EA et al., (1991) cited above.

[0144] In some embodiments, any light chain of the anti-TMPRSS6 antibody described herein may further include a CL, which may be any light chain constant region (CL) known in the art. In some examples, the CL is a kappa light chain. In other examples, the CL is a lambda light chain. In some embodiments, the CL is a kappa light chain.

[0145] Other antibody heavy and light chain constant regions are well known in the art and are available, for example, in the IMGT database (www.imgt.org) or at www.vbase2.org / vbstat.php, both of which are incorporated herein by reference.

[0146] In some embodiments, the anti-TMPRSS6 antibody described herein comprises a heavy chain comprising a heavy chain constant region that is 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% compared to any one of the VHs listed in Table 1 or any variant thereof and SEQ ID NO: 9. In some embodiments, the anti-TMPRSS6 antibody described herein comprises a heavy chain comprising a heavy chain constant region containing 20 or fewer amino acid variations (e.g., 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1 or fewer amino acid variations) compared to any one of the VHs listed in Table 1 or any variant thereof and SEQ ID NO: 9. In some embodiments, the anti-TMPRSS6 antibodies described herein include a heavy chain comprising one of the VHs listed in Table 1 or any variant thereof, and the heavy chain constant region described in SEQ ID NO: 9.

[0147] In some embodiments, the anti-TMPRSS6 antibodies described herein include a heavy chain containing a heavy chain constant region that is 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% identical to any of the VHs listed in Table 1 or any variant thereof and SEQ ID NO: 21. In some embodiments, the anti-TMPRSS6 antibodies described herein include a heavy chain containing a heavy chain constant region that has 20 or fewer amino acid variations (e.g., 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1 or fewer amino acid variations) compared to any of the VHs listed in Table 1 or any variant thereof and SEQ ID NO: 21. In some embodiments, the anti-TMPRSS6 antibodies described herein include a heavy chain comprising one of the VHs listed in Table 1 or any variant thereof, and the heavy chain constant region described in SEQ ID NO: 21.

[0148] In some embodiments, the anti-TMPRSS6 antibodies described herein include a heavy chain comprising a heavy chain constant region that is 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% identical to any of the VHs listed in Table 1 or any variant thereof and SEQ ID NO: 39. In some embodiments, the anti-TMPRSS6 antibodies described herein include a heavy chain comprising a heavy chain constant region containing 20 or fewer amino acid variations (e.g., 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1 or fewer amino acid variations) compared to any of the VHs listed in Table 1 or any variant thereof and SEQ ID NO: 39. In some embodiments, the anti-TMPRSS6 antibodies described herein include a heavy chain comprising one of the VHs listed in Table 1 or any variant thereof, and the heavy chain constant region described in SEQ ID NO: 39.

[0149] In some embodiments, the anti-TMPRSS6 antibodies described herein include a heavy chain comprising a heavy chain constant region that is 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% identical to any of the VHs listed in Table 1 or any variant thereof and SEQ ID NO: 51. In some embodiments, the anti-TMPRSS6 antibodies described herein include a heavy chain comprising a heavy chain constant region containing 20 or fewer amino acid variations (e.g., 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1 or fewer amino acid variations) compared to any of the VHs listed in Table 1 or any variant thereof and SEQ ID NO: 51. In some embodiments, the anti-TMPRSS6 antibodies described herein include a heavy chain comprising one of the VHs listed in Table 1 or any variant thereof, and the heavy chain constant region described in SEQ ID NO: 51.

[0150] In some embodiments, the anti-TMPRSS6 antibody described herein includes a light chain comprising a light chain constant region that is 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% compared to any one of the VLs listed in Table 1 or any variant thereof and SEQ ID NO: 10. In some embodiments, the anti-TMPRSS6 antibody described herein includes a light chain comprising a light chain constant region containing 20 or fewer amino acid variations (e.g., 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1 or fewer amino acid variations) compared to any one of the VLs listed in Table 1 or any variant thereof and SEQ ID NO: 10. In some embodiments, the anti-TMPRSS6 antibodies described herein include a light chain comprising one of the VLs listed in Table 1 or any variant thereof, and the light chain constant region described in SEQ ID NO: 10.

[0151] In some embodiments, the anti-TMPRSS6 antibody described herein includes a light chain comprising a light chain constant region that is 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% compared to any one of the VLs listed in Table 1 or any variant thereof and SEQ ID NO: 40. In some embodiments, the anti-TMPRSS6 antibody described herein includes a light chain comprising a light chain constant region containing 20 or fewer amino acid variations (e.g., 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1 or fewer amino acid variations) compared to any one of the VLs listed in Table 1 or any variant thereof and SEQ ID NO: 40. In some embodiments, the anti-TMPRSS6 antibodies described herein include a light chain comprising one of the VLs listed in Table 1 or any variant thereof, and the light chain constant region described in SEQ ID NO: 40. Examples of IgG heavy chain and light chain amino acid sequences of the described anti-TMPRSS6 antibodies are provided in Table 1 above.

[0152] In some embodiments, the anti-TMPRSS6 antibody of this disclosure comprises a heavy chain containing 20 or fewer amino acid variations compared to the heavy chain described in SEQ ID NO: 11 (e.g., 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1 or fewer amino acid variations). Alternatively or additionally, the anti-TMPRSS6 antibody of this disclosure comprises a light chain containing 20 or fewer amino acid variations compared to the light chain described in SEQ ID NO: 12 (e.g., 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 9, 8, 7, 6, 5, 4, 3, 2, or 1 or fewer amino acid variations). In some embodiments, the anti-TMPRSS6 antibody described herein comprises a heavy chain containing an amino acid sequence that is 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% identical to SEQ ID NO: 11. Alternatively or additionally, the anti-TMPRSS6 antibody described herein comprises a light chain containing an amino acid sequence that is 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% identical to SEQ ID NO: 12.

[0153] In some embodiments, the anti-TMPRSS6 antibody of the present disclosure comprises a heavy chain containing 20 or fewer amino acid variations (e.g., 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1 or fewer amino acid variations) compared to the heavy chain described in SEQ ID NO: 17 or 22. Alternatively or additionally, the anti-TMPRSS6 antibody of the present disclosure comprises a light chain containing 20 or fewer amino acid variations (e.g., 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 9, 8, 7, 6, 5, 4, 3, 2, or 1 or fewer amino acid variations) compared to the light chain described in SEQ ID NO: 23. In some embodiments, the anti-TMPRSS6 antibody described herein comprises a heavy chain containing an amino acid sequence that is at least 80% (e.g., 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%) identical to SEQ ID NO: 17 or 22. Alternatively or additionally, the anti-TMPRSS6 antibody described herein comprises a light chain containing an amino acid sequence that is at least 80% (e.g., 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%) identical to 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 additionally, the anti-TMPRSS6 antibody described herein comprises a light chain containing the amino acid sequence of SEQ ID NO: 23.

[0154] In some embodiments, the anti-TMPRSS6 antibody of this disclosure comprises a heavy chain containing 20 or fewer amino acid variations (e.g., 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1 or fewer amino acid variations) compared to the heavy chain described in SEQ ID NO: 33. Alternatively or additionally, the anti-TMPRSS6 antibody of this disclosure comprises a light chain containing 20 or fewer amino acid variations (e.g., 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 9, 8, 7, 6, 5, 4, 3, 2, or 1 or fewer amino acid variations) compared to the light chain described in SEQ ID NO: 34. In some embodiments, the anti-TMPRSS6 antibody described herein comprises a heavy chain containing an amino acid sequence that is at least 80% (e.g., 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%) identical to SEQ ID NO: 33. Alternatively or additionally, the anti-TMPRSS6 antibody described herein comprises a light chain containing an amino acid sequence that is at least 80% (e.g., 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%) identical to 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 additionally, the anti-TMPRSS6 antibody described herein comprises a light chain containing the amino acid sequence of SEQ ID NO: 34.

[0155] In some embodiments, the anti-TMPRSS6 antibody of this disclosure comprises a heavy chain containing 20 or fewer amino acid variations (e.g., 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1 or fewer amino acid variations) compared to the heavy chain described in SEQ ID NO: 41. Alternatively or additionally, the anti-TMPRSS6 antibody of this disclosure comprises a light chain containing 20 or fewer amino acid variations (e.g., 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 9, 8, 7, 6, 5, 4, 3, 2, or 1 or fewer amino acid variations) compared to the light chain described in SEQ ID NO: 42. In some embodiments, the anti-TMPRSS6 antibody described herein comprises a heavy chain containing an amino acid sequence that is at least 80% (e.g., 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%) identical to SEQ ID NO: 41. Alternatively or additionally, the anti-TMPRSS6 antibody described herein comprises a light chain containing an amino acid sequence that is at least 80% (e.g., 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%) identical to 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 additionally, the anti-TMPRSS6 antibody described herein comprises a light chain containing the amino acid sequence of SEQ ID NO: 42.

[0156] In some embodiments, the anti-TMPRSS6 antibody of this disclosure comprises a heavy chain containing 20 or fewer amino acid variations (e.g., 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1 or fewer amino acid variations) compared to the heavy chain described in SEQ ID NO: 46. Alternatively or additionally, the anti-TMPRSS6 antibody of this disclosure comprises a light chain containing 20 or fewer amino acid variations (e.g., 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 9, 8, 7, 6, 5, 4, 3, 2, or 1 or fewer amino acid variations) compared to the light chain described in SEQ ID NO: 47. In some embodiments, the anti-TMPRSS6 antibody described herein comprises a heavy chain containing an amino acid sequence that is at least 80% (e.g., 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%) identical to SEQ ID NO: 46. Alternatively or additionally, the anti-TMPRSS6 antibody described herein comprises a light chain containing an amino acid sequence that is at least 80% (e.g., 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%) identical to 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 additionally, the anti-TMPRSS6 antibody described herein comprises a light chain containing the amino acid sequence of SEQ ID NO: 47.

[0157] In some embodiments, the anti-TMPRSS6 antibody of this disclosure comprises a heavy chain containing 20 or fewer amino acid variations (e.g., 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1 or fewer amino acid variations) compared to the heavy chain described in SEQ ID NO: 52. Alternatively or additionally, the anti-TMPRSS6 antibody of this disclosure comprises a light chain containing 20 or fewer amino acid variations (e.g., 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 9, 8, 7, 6, 5, 4, 3, 2, or 1 or fewer amino acid variations) compared to the light chain described in SEQ ID NO: 53. In some embodiments, the anti-TMPRSS6 antibody described herein comprises a heavy chain containing at least 80% (e.g., 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%) the same amino acid sequence as SEQ ID NO: 52. Alternatively or additionally, the anti-TMPRSS6 antibody described herein comprises a light chain containing at least 80% (e.g., 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%) the same amino acid sequence as 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 additionally, the anti-TMPRSS6 antibody described herein comprises a light chain containing the amino acid sequence of SEQ ID NO: 53.

[0158] The anti-TMPRSS6 antibodies described herein may be in any antibody form, including but not limited to intact (i.e., full-length) 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 antibodies described herein are scFv. In some embodiments, the anti-TMPRSS6 antibodies described herein are scFv-Fab (e.g., scFv fused to a portion of the constant region).

[0159] In some embodiments, conservative mutations may be introduced into the antibody sequence (e.g., CDR or framework sequence) at a position where the residue is unlikely to be involved in interaction with the target antigen (e.g., TMPRSS6), for example, when determined based on the crystal structure. In some embodiments, one, two or more mutations (e.g., amino acid substitutions) are introduced into the Fc region of the anti-TMPRSS6 antibody described herein [e.g., the CH2 domain (residues 231-340 of human IgG1) and / or the CH3 domain (residues 341-447 of human IgG1) and / or the hinge region] 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.

[0160] In some embodiments, for example, as described in U.S. Patent No. 5,677,425, one, two, or more mutations (e.g., amino acid substitutions) are introduced into the hinge region of the Fc region (CH1 domain), resulting in a change (e.g., an increase or decrease) in the number of cysteine ​​residues in the hinge region. The number of cysteine ​​residues in the hinge region of the CH1 domain may be changed, for example, to facilitate the assembly of the light and heavy chains, or to alter (e.g., increase or decrease) the stability of the antibody, or to facilitate linker conjugation.

[0161] In some embodiments, one, two, or more mutations (e.g., amino acid substitutions) are introduced into the Fc region of the antibodies described herein [e.g., in the CH2 domain (residues 231-340 of human IgG1) and / or the CH3 domain (residues 341-447 of human IgG1) and / or the hinge region, using numbering by the Kabat numbering system (e.g., the EU index in Kabat)] to increase or decrease the affinity of the antibody to the Fc receptor. Mutations in the Fc region of antibodies that decrease or increase the affinity of the antibody to the Fc receptor and techniques for introducing such mutations into the Fc receptor or fragments thereof are known to those skilled in the art. Examples of mutations in the Fc receptor of an antibody that can be made to alter the affinity of the antibody to the Fc receptor are described, for example, in Smith P et al., (2012) PNAS 109: 6181-6186, U.S. Patent No. 6,737,056, and International Publication No. 97 / 34631, which are incorporated herein by reference.

[0162] In some embodiments, one, two, or more amino acid mutations (i.e., substitutions, insertions, or deletions) are introduced into the IgG constant domain 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. For example, see International Publication No. 97 / 34631; and U.S. Patents No. 5,869,046, No. 6,121,022, No. 6,277,375, and No. 6,165,745 for examples of mutations that alter (e.g., decrease or increase) the half-life of an antibody in vivo.

[0163] In some embodiments, one, two, or more amino acid mutations (i.e., substitutions, insertions, or deletions) are introduced into the IgG constant domain or its FcRn binding fragment (preferably an Fc or hinge-Fc domain fragment) to decrease 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 IgG constant domain 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 231-340 of human IgG1) and / or the third constant (CH3) domain (residues 341-447 of human IgG1) using numbering by the EU index in Kabat [Kabat EA et al., (1991) cited above]. In some embodiments, the constant region of IgG1 of the antibodies described herein includes a substitution of methionine (M) to tyrosine (Y) at position 252, serine (S) to threonine (T) at position 254, and a substitution of threonine (T) to glutamic acid (E) at position 256, as numbered by the EU index in Kabat. See U.S. Patent No. 7,658,921, incorporated herein by reference. This type of mutant IgG, referred to as the “YTE mutant,” has been shown to exhibit a four-fold increase in half-life compared to the wild-type version of the same antibody [see Dall'Acqua WF et al., (2006) J Biol Chem 281: 23514-24].

[0164] In some embodiments, one, two, or more amino acid substitutions are introduced into the Fc region of the IgG constant domain to alter the effector function of an anti-TMPRSS6 antibody. The effector ligand whose affinity to it is altered may be, for example, the Fc receptor or the C1 component of complement. This approach is described in more detail in U.S. Patents No. 5,624,821 and No. 5,648,260. In some embodiments, one or more amino acid substitutions may be introduced into the Fc region of the antibodies described herein to remove a potential glycosylation site on the Fc region that may reduce Fc receptor binding [see, for example, Shields RL et al., (2001) J Biol Chem 276: 6591-604].

[0165] In some embodiments, one or more amino acids in the constant region of the anti-TMPRSS6 antibody described herein may be replaced with different amino acid residues so that the antibody has modified Clq binding and / or reduced or absent complement-dependent cytotoxicity (CDC). This approach 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 ability to immobilize complement. This approach is described in International Publication No. 94 / 29351. In some embodiments, the Fc region of the antibody described herein is modified to increase the antibody's ability to mediate antibody-dependent cytotoxicity (ADCC) and / or increase its affinity for the Fcγ receptor. This approach is described in International Publication No. 00 / 42072.

[0166] In some embodiments, the heavy and / or light chain variable domain sequences of the antibodies provided herein may be used, for example, to generate CDR-implanted, chimeric, humanized, or compound human antibodies or antigen-binding fragments as described elsewhere herein. As will be understood by those skilled in the art, any variant, CDR-implanted, chimeric, humanized, or compound antibody derived from any antibody provided herein may be useful in the compositions and methods described herein, maintaining the ability to specifically bind to TMPRSS6, and as a result, the variant, CDR-implanted, chimeric, humanized, or compound 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 compared to the original antibody from which it is derived.

[0167] In some embodiments, the antibodies provided herein include mutations that confer desired properties to the antibody. For example, to avoid potential complications resulting from Fab-arm exchange, which are known to occur with native IgG4 mAbs, the antibodies provided herein may include a stabilizing "Adair" mutation [Angal S., et al., "A single amino acid substitution abolishes the heterogeneity of chimeric mouse / human (IgG4) antibody," Mol Immunol 30, 105-108; 1993], where serine 228 (EU numbering; residue 241, Kabat numbering) is converted to proline, resulting in an IgG1-like hinge sequence. Thus, any of the antibodies may include a stabilizing "Adair" mutation.

[0168] 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 by N-glycosylation, O-glycosylation, C-glycosylation, glycieation (GPI anchoring), 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, N-acetylgalactosamine units, galactose units, fucose units, or phospholipid units. In some embodiments, there are 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. In some embodiments, the glycosylated antibody is completely or partially glycosylated. In some embodiments, the antibody is glycosylated by chemical reaction or enzymatic means. In some embodiments, the antibody is glycosylated in vitro or inside a cell, and the cell may lack an enzyme in the N- or O-glycosylation pathway, such as glycosyltransferase. In some embodiments, the antibody is functionalized with a sugar or carbohydrate molecule, as described in International Patent Application Publication No. 2014065661, published on May 1, 2014, titled "Modified Antibody, Antibody-Conjugate and Method for Preparation thereof".

[0169] In some embodiments, any one of the anti-TMPRSS6 antibodies described herein may include a signal peptide (e.g., an N-terminal signal peptide) in the heavy chain and / or light chain sequence. In some embodiments, the anti-TMPRSS6 antibody described herein includes any one of the VH and VL sequences described herein, any one of the IgG heavy chain and light chain sequences, or any one of the F(ab') heavy chain and light chain sequences, and further includes a signal peptide (e.g., an N-terminal signal peptide).

[0170] III. Preparation of anti-TMPRSS6 antibody The antibodies capable of binding to TMPRSS6 described herein can be prepared by any method known in the art. See, for example, Harlow and Lane, (1998) Antibodies: A Laboratory Manual, Cold Spring Harbor Laboratory, New York.

[0171] In some embodiments, antibodies specific to a target antigen (e.g., TMPRSS6) may be produced by conventional hybridoma techniques. A full-length target antigen or a fragment thereof, which may be linked to a carrier protein, e.g., KLH, may be used to immunize a host animal to produce antibodies that bind to that antigen. The routes and schedules for immunization of the host animal are generally carried out according to established prior art for antibody stimulation and production, as further described herein. General techniques for mouse, humanization, and human antibody production are known in the art and are described herein. Any mammalian subject, including humans, or antibody-producing cells therefrom, are assumed to be engineered to serve as a basis for the production of mammalian hybridoma cell lines, including human hybridoma cell lines. Typically, the host animal is inoculated intraperitoneally, intramuscularly, orally, subcutaneously, in the sole of the foot, and / or intradermally, in amounts of immunogen, including those described herein.

[0172] If desired, the antibody of interest (monoclonal or polyclonal) (e.g., produced by a hybridoma) may be sequenced, and the polynucleotide sequence may then be cloned into a vector for expression or reproduction. The sequence encoding the antibody of interest may be maintained in the vector in host cells, which may then be expanded and frozen for future use. Alternatively, the polynucleotide sequence may be used for genetic engineering to “humanize” the antibody or to improve the antibody’s affinity (affinity maturation) or other characteristics. For example, the constant region may be engineered to more closely resemble the human constant region so that the antibody avoids an immune response when used in clinical trials and treatments in humans. It may be desirable to genetically engineer the antibody sequence to obtain higher affinity and higher efficacy against the target antigen. It is evident to those skilled in the art that one or more polynucleotide changes can be made to an antibody while still maintaining its binding specificity to the target antigen.

[0173] In other embodiments, fully human antibodies may be obtained by using commercially available mice engineered to express specific human immunoglobulin proteins. Transgenic animals designed to produce a more desirable (e.g., fully human antibodies) or more robust immune response may also be used for humanization or the production of human antibodies. Examples of such techniques include XenomouseRTM from Amgen, Inc. (Fremont, CA) and HuMAb-MouseRTM and TC Mouse™ from Medarex, Inc. (Princeton, NJ) or H2L2 mice from Harbour Antibodies BV (Holland). Alternatively, antibodies may be produced by recombination using phage display or yeast techniques. 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 produce human antibodies and antibody fragments in vitro from the immunoglobulin variable (V) domain gene repertoire from non-immunized donors.

[0174] Antigen-binding fragments of intact antibodies (full-length antibodies) can be prepared by routine methods. For example, F(ab')2 fragments can be produced by pepsin digestion of the antibody molecule, and Fab fragments can be generated by reducing the disulfide crosslinks of the F(ab')2 fragment. Genetically engineered antibodies, such as humanized antibodies, chimeric antibodies, single-chain antibodies, and bispecific antibodies, can be produced, for example, by conventional recombination techniques. In one example, 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 that have the ability to specifically bind to the 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 may be placed in one or more expression vectors, which are then transfected into host cells, such as Escherichia coli (E. coli) cells, Simian COS cells, Chinese hamster ovary (CHO) cells, human HEK293 cells, or myeloma cells that do not otherwise produce immunoglobulin proteins, in order to synthesize monoclonal antibodies in recombinant host cells. See, for example, PCT International Publication 87 / 04462. The DNA can then be modified, for example, by substituting coding sequences for human heavy and light chain constant domains at the location of homologous mouse sequences [Morrison et al., (1984) Proc. Nat. Acad. Sci. 81:6851], or by covalently joining all or part of a coding sequence for a non-immunoglobulin polypeptide to an immunoglobulin coding sequence. In this manner, genetically modified antibodies with binding specificity to target antigens, such as “chimeric” or “hybrid” antibodies, can be prepared.

[0175] Single-chain antibodies can be prepared via recombinant techniques by linking nucleotide sequences encoding a heavy-chain variable region and a light-chain variable region. Preferably, a flexible linker is incorporated between the two variable regions.

[0176] Alternatively, phage or yeast scFv libraries can be produced by applying techniques described for the production of single-chain antibodies (U.S. Patents 4,946,778 and 4,704,692), and TMPRSS6-specific scFv clones can be identified from the library using routine procedures. Positive clones can then be subjected to further screening to identify clones with high TMPRSS6 binding affinity.

[0177] Antibodies obtained according to methods known in the art and described herein can be characterized using methods known in the art. For example, one method is to identify the epitopes to which the antigen binds, or "epitope mapping." There are many methods known in the art for mapping and characterizing the location of epitopes on proteins, including, for example, elucidation of the crystalline structure of antibody-antigen complexes, competitive assays, gene fragment expression assays, and synthetic peptide-based assays, as described in Chapter 11 of Harlow and Lane, Using Antibodies, a Laboratory Manual, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY, 1999. In one example, epitope mapping can be achieved using H / D-Ex (hydrogen-deuterium exchange) coupled with protein hydrolysis and mass spectrometry. In an additional example, epitope mapping can be used to determine the sequences to which the antibody binds. Epitopes can be linear epitopes, i.e., those contained within a single stretch of amino acids, or conformational epitopes formed by three-dimensional interactions of amino acids, which do not necessarily have to be contained within a single stretch (linear primary structure sequence). Peptides of variable length (e.g., at least 4-6 amino acids) may be isolated or synthesized (e.g., by recombination) and used for antibody-binding assays. In another example, the epitope to which the antibody binds may be determined in a systematic screening by using overlap peptides derived from the target antigen sequence and determining antibody binding. According to a gene fragment expression assay, an open reading frame encoding the target antigen is fragmented randomly or by a specific genetic composition, and the reactivity of the expressed fragments of the antigen with the antibody to be tested is determined. The gene fragments may be produced, for example, by PCR, then transcribed in vitro and translated into proteins in the presence of radioactive amino acids. Antibody binding to the radiolabeled antigen fragments is then determined by immunoprecipitation and gel electrophoresis.A particular epitope can also be identified by using a large library of random peptide sequences presented on the surface of phage particles (a phage library). Alternatively, a defined library of overlapping peptide fragments can be tested for binding to a test antibody in a simple binding assay. In additional examples, antigen-binding domain mutagenesis, domain swapping experiments, and alanine scanning mutagenesis may be performed to identify the sufficient and / or required residues for epitope binding. Alternatively, a competitive assay may be performed using other antibodies known to bind to the same antigen to determine whether an antibody binds to the same epitope as another antibody. Competitive assays are well known to those skilled in the art.

[0178] In some cases, anti-TMPRSS6 antibodies are prepared by recombinant techniques as illustrated below. The nucleic acids encoding the heavy and light chains of the anti-TMPRSS6 antibodies described herein can be cloned into a single expression vector, with each nucleotide sequence operably ligated to a suitable promoter. In one example, each of the nucleotide sequences encoding the heavy and light chains is operably ligated to a separate promoter. Alternatively, the nucleotide sequences encoding the heavy and light chains can be operably ligated to a single promoter, resulting in both the heavy and light chains being expressed from the same promoter. If necessary, an internal ribosome entry site (IRES) may be inserted between the heavy chain and light chain sequences.

[0179] In some cases, the nucleotide sequences encoding the two chains of an antibody are cloned into two vectors, and the two vectors can be introduced into the same or different cells. If the two chains are expressed in different cells, each of them can be isolated from the host cell expressing it, and the isolated heavy and light chains can be mixed and incubated under suitable conditions to enable antibody formation.

[0180] Generally, nucleic acid sequences encoding one or all of an antibody chain can be cloned into a suitable expression vector operably linked to a suitable promoter using methods known in the art. For example, the nucleotide sequence and vector can be contacted with restriction enzymes under suitable conditions, thereby creating complementary ends on each molecule that can pair with each other and ligate together with the ligase. Alternatively, synthetic nucleic acid linkers can be ligated to the ends of the gene. These synthetic linkers contain nucleic acid sequences corresponding to specific restriction sites in the vector. The choice of expression vector / promoter depends on the type of host cell for use in antibody production.

[0181] Various promoters are available for the expression of the antibodies described herein, including, but not limited to, the cytomegalovirus (CMV) initial promoter, viral LTRs such as Rous sarcoma virus LTR, HIV-LTR, HTLV-1 LTR, Simian virus 40 (SV40) initial promoter, Escherichia coli lac UV promoter, and herpes simplex virus tk promoter.

[0182] Adjustable promoters may also be used. Such adjustable promoters include those that use a lac repressor from E. coli as a transcription modulator to regulate transcription from mammalian cell promoters containing a lac operator (Brown, M. et al., Cell, 49:603-612 (1987)), and those that use a tetracycline repressor (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 estradiol, RU486, diphenol murislerone, or FK506 dimer, VP16, or p65 using rapamycin. Inducible systems are available from several manufacturers, including Invitrogen, Clontech, and Ariad.

[0183] A moduloable promoter containing a repressor may be used with the operon. In one embodiment, a lac repressor from E. coli can function as a transcription modulator to regulate transcription from a mammalian cell promoter possessing a lac operator (M. Brown et al., Cell, 49:603-612 (1987)); Gossen and Bujard (1992); (M. Gossen et al., Natl. Acad. Sci. USA, 89:5547-5551 (1992)). A tetracycline repressor (tetR) is combined with a transcription activator (VP 16) to create a tetR-mammalian cell transcription activator fusion protein, tTa(tetR-VP 16), tetO, which has a minimal promoter derived from a human cytomegalovirus (hCMV) promoter to create a tetR-tet operator system for controlling gene expression in mammalian cells. In one embodiment, a tetracycline-inducible switch is used. Rather than a tetR-mammalian cell transcription factor fusion derivative, the tetracycline repressor (tetR) alone can function as a potent transmodulator for regulating gene expression in mammalian cells when the tetracycline operator is appropriately positioned downstream of the TATA element of the CMVIE promoter (Yao et al., Human Gene Therapy). One particular advantage of this tetracycline-inducible switch is that, in order to achieve its tunable effect, it does not require the use of a tetracycline repressor-mammalian cell transactivator or repressor fusion protein, which can be toxic to cells 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)].

[0184] Additionally, the vector may contain some or all of the following, for example: a selection marker gene, e.g., the neomycin gene for selection of stable or transient transfectants in mammalian cells; an enhancer / promoter sequence from the earliest gene of human CMV for high levels of transcription; transcription termination and RNA processing signals from SV40 for mRNA stability; the SV40 polyomatous replication origin and ColE1 for proper episomal replication; an internal ribosome binding site (IRES), a versatile multiple cloning site; and T7 and SP6 RNA promoters for in vitro transcription of sense and antisense RNA. Suitable vectors and methods for producing vectors containing the transgene are well known and available in the art. Examples of polyadenylation signals useful for carrying out the methods described herein include, but are not limited to, the human collagen I polyadenylation signal, the human collagen II polyadenylation signal, and the SV40 polyadenylation signal.

[0185] One or more vectors (e.g., expression vectors) containing nucleic acids encoding any of the antibodies (e.g., nucleic acid coding sequences listed in Table 3) may be introduced into host cells suitable for antibody production. 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 SV40-transformed monkey kidney CV1 lineage (COS) cells. In some embodiments, the host cells expressing the anti-TMPRSS6 antibody are CHO cells. The host cells may be cultured under conditions suitable for the expression of the antibody or any polypeptide chain thereof. Such antibodies or their polypeptide chains can be recovered by cultured cells (e.g., from cells or culture supernatant) via conventional methods, such as affinity purification. If necessary, the polypeptide chain of the antibody can be incubated under suitable conditions for a suitable period of time that allows for antibody production. In some embodiments, the host cell contains a nucleic acid encoding the heavy chain of the anti-TMPRSS6 antibody. In some embodiments, the host cell contains a nucleic acid encoding the light chain of the anti-TMPRSS6 antibody. In some embodiments, the host cell contains a nucleic acid encoding both the heavy chain and the light chain.

[0186] In some embodiments, the antibody preparation methods described herein involve a recombinant expression vector encoding both the heavy and light chains of the 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. Positively transformed host cells are selected and cultured under suitable conditions that allow expression of the two polypeptide chains forming the antibody, and the two chains can be recovered from the cells or culture medium. If necessary, the two chains recovered from the host cells can be incubated under suitable conditions that allow antibody formation.

[0187] In one example, two recombinant expression vectors are provided, one encoding the heavy chain of the anti-TMPRSS6 antibody and the other encoding the light chain of the anti-TMPRSS6 antibody. Both recombinant expression vectors can be introduced into suitable host cells (e.g., dhfr-CHO cells) by conventional methods, such as calcium phosphate-mediated transfection.

[0188] Alternatively, each expression vector can be introduced into a suitable host cell. A positive transformant can be selected and cultured under suitable conditions that allow expression of the antibody polypeptide chain. If two expression vectors are introduced into the same host cell, the antibody produced in the cell can be recovered from the host cell or culture medium. If necessary, the polypeptide chain can be recovered from the host cell or culture medium and then incubated under suitable conditions that allow antibody formation. If two expression vectors are introduced into different host cells, each of them can be recovered from the corresponding host cell or corresponding culture medium. The two polypeptide chains can then be incubated under suitable conditions for antibody formation.

[0189] Standard molecular biology techniques are used to prepare recombinant expression vectors, transfect host cells, select transformants, culture the host cells, and recover antibodies from the culture medium. For example, some antibodies can be isolated by affinity chromatography using a protein A or protein G linked matrix.

[0190] Any of the nucleic acids encoding the heavy chain, light chain, or both of the anti-TMPRSS6 antibody described herein (for example, provided in Table 3), vectors containing the same (for example, expression vectors), or host cells containing the vector are within the scope of this disclosure.

[0191] [Table 3-1] [Table 3-2] Table 3-3 Table 3-4 Table 3-5 Table 3-6 Table 3-7 Table 3-8 Table 3-9 Table 3-10 Table 3-11 Table 3-12 Table 3-13

[0192] In some embodiments, the Disclosure provides an isolated nucleic acid containing a sequence identical to at least 60% (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) of sequence SEQ ID NO: 54 and / or an isolated nucleic acid containing a sequence identical to at least 60% (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) of sequence SEQ ID NO: 55. In some embodiments, the Disclosure provides an expression vector comprising an isolated nucleic acid containing a sequence identical to at least 60% (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) of sequence SEQ ID NO: 54 and / or an isolated nucleic acid containing a sequence identical to at least 60% (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) of sequence SEQ ID NO: 55.

[0193] In some embodiments, the Disclosure provides an isolated nucleic acid containing a sequence identical to at least 60% (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) of sequence SEQ ID NO: 56 and / or an isolated nucleic acid containing a sequence identical to at least 60% (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) of sequence SEQ ID NO: 57. In some embodiments, the Disclosure provides an expression vector comprising an isolated nucleic acid containing a sequence identical to at least 60% (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) of sequence SEQ ID NO: 56 and / or an isolated nucleic acid containing a sequence identical to at least 60% (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) of sequence SEQ ID NO: 57.

[0194] In some embodiments, the Disclosure provides an isolated nucleic acid containing a sequence identical to at least 60% (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) of sequence SEQ ID NO: 58 and / or an isolated nucleic acid containing a sequence identical to at least 60% (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) of sequence SEQ ID NO: 59. In some embodiments, the Disclosure provides an expression vector comprising an isolated nucleic acid containing a sequence identical to at least 60% (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) of sequence SEQ ID NO: 58 and / or an isolated nucleic acid containing a sequence identical to at least 60% (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) of sequence SEQ ID NO: 59.

[0195] In some embodiments, the Disclosure provides an isolated nucleic acid containing a sequence identical to at least 60% (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) of sequence SEQ ID NO: 60 and / or an isolated nucleic acid containing a sequence identical to at least 60% (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) of sequence SEQ ID NO: 61. In some embodiments, the Disclosure provides an expression vector comprising an isolated nucleic acid containing a sequence identical to at least 60% (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) of sequence SEQ ID NO: 60 and / or an isolated nucleic acid containing a sequence identical to at least 60% (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) of sequence SEQ ID NO: 61.

[0196] In some embodiments, the Disclosure provides an isolated nucleic acid containing a sequence identical to at least 60% (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) of sequence SEQ ID NO: 62 and / or an isolated nucleic acid containing a sequence identical to at least 60% (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) of sequence SEQ ID NO: 63. In some embodiments, the Disclosure provides an expression vector comprising an isolated nucleic acid containing a sequence identical to at least 60% (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) of SEQ ID NO: 62 and / or an isolated nucleic acid containing a sequence identical to at least 60% (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) of SEQ ID NO: 63.

[0197] In some embodiments, the Disclosure provides an isolated nucleic acid containing a sequence identical to at least 60% (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) of sequence SEQ ID NO: 5 and / or an isolated nucleic acid containing a sequence identical to at least 60% (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) of sequence SEQ ID NO: 32. In some embodiments, the Disclosure provides an expression vector comprising an isolated nucleic acid containing a sequence identical to at least 60% (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) of sequence SEQ ID NO: 5 and / or an isolated nucleic acid containing a sequence identical to at least 60% (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) of sequence SEQ ID NO: 32.

[0198] In some embodiments, the Disclosure provides an isolated nucleic acid containing a sequence identical to at least 60% (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) of sequence SEQ ID NO: 79 and / or an isolated nucleic acid containing a sequence identical to at least 60% (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) of sequence SEQ ID NO: 28. In some embodiments, the Disclosure provides an expression vector comprising an isolated nucleic acid containing at least 60% (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) the same sequence as SEQ ID NO: 79, and / or an isolated nucleic acid containing at least 60% (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) the same sequence as SEQ ID NO: 28.

[0199] In some embodiments, the Disclosure provides isolated nucleic acids containing sequences identical to sequence number 64 by at least 60% (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%), and / or isolated nucleic acids containing sequences identical to sequence number 65 by at least 60% (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%). In some embodiments, the Disclosure provides an expression vector comprising an isolated nucleic acid containing at least 60% (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) the same sequence as SEQ ID NO: 64, and / or an isolated nucleic acid containing at least 60% (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) the same sequence as SEQ ID NO: 65.

[0200] In some embodiments, the Disclosure provides isolated nucleic acids containing sequences identical to sequence number 66 by at least 60% (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%), and / or isolated nucleic acids containing sequences identical to sequence number 67 by at least 60% (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%). In some embodiments, the Disclosure provides an expression vector comprising an isolated nucleic acid containing at least 60% (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) the same sequence as SEQ ID NO: 66, and / or an isolated nucleic acid containing at least 60% (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) the same sequence as SEQ ID NO: 67.

[0201] In some embodiments, the Disclosure provides an isolated nucleic acid containing a sequence identical to at least 60% (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) of sequence SEQ ID NO: 68 and / or an isolated nucleic acid containing a sequence identical to at least 60% (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) of sequence SEQ ID NO: 69. In some embodiments, the Disclosure provides an expression vector comprising an isolated nucleic acid containing a sequence identical to at least 60% (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) of sequence SEQ ID NO: 68 and / or an isolated nucleic acid containing a sequence identical to at least 60% (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) of sequence SEQ ID NO: 69.

[0202] In some embodiments, the Disclosure provides an isolated nucleic acid containing a sequence identical to at least 60% (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) of sequence SEQ ID NO: 70 and / or an isolated nucleic acid containing a sequence identical to at least 60% (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) of sequence SEQ ID NO: 71. In some embodiments, the Disclosure provides an expression vector comprising an isolated 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%) the same sequence as SEQ ID NO: 70, and / or an isolated 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%) the same sequence as SEQ ID NO: 71.

[0203] In some embodiments, the Disclosure provides isolated nucleic acids containing sequences identical to sequence number 72 by at least 60% (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) and / or isolated nucleic acids containing sequences identical to sequence number 73 by at least 60% (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%). In some embodiments, the Disclosure provides an expression vector comprising an isolated nucleic acid containing a sequence identical to at least 60% (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) of sequence SEQ ID NO: 72 and / or an isolated nucleic acid containing a sequence identical to at least 60% (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) of sequence SEQ ID NO: 73.

[0204] In some embodiments, the Disclosure provides isolated nucleic acids containing sequences identical to sequence number 74 by at least 60% (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%), and / or isolated nucleic acids containing sequences identical to sequence number 75 by at least 60% (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%). In some embodiments, the Disclosure provides an expression vector comprising an isolated 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%) the same sequence as SEQ ID NO: 74, and / or an isolated 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%) the same sequence as SEQ ID NO: 75.

[0205] In some embodiments, the Disclosure provides isolated nucleic acids containing sequences identical to sequence number 76 by at least 60% (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) and / or isolated nucleic acids containing sequences identical to sequence number 77 by at least 60% (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%). In some embodiments, the Disclosure provides an expression vector comprising an isolated nucleic acid containing at least 60% (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) the same sequence as SEQ ID NO: 76, and / or an isolated nucleic acid containing at least 60% (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) the same sequence as SEQ ID NO: 77.

[0206] In some embodiments, the anti-TMPRSS6 described herein is produced by (i) expressing nucleic acid identical to at least 60% (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) of SEQ ID NO: 54, and / or (ii) expressing nucleic acid identical to at least 60% (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) of SEQ ID NO: 55.

[0207] In some embodiments, the anti-TMPRSS6 antibody described herein is produced by (i) expressing nucleic acid identical to at least 60% (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) of SEQ ID NO: 56, and / or (ii) expressing nucleic acid identical to at least 60% (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) of SEQ ID NO: 57.

[0208] In some embodiments, the anti-TMPRSS6 described herein is produced by (i) expressing nucleic acid identical to at least 60% (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) of SEQ ID NO: 58 and / or (ii) expressing nucleic acid identical to at least 60% (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) of SEQ ID NO: 59.

[0209] In some embodiments, the anti-TMPRSS6 described herein is produced by expressing (i) nucleic acid identical to at least 60% (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) of SEQ ID NO: 60 and / or (ii) nucleic acid identical to at least 60% (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) of SEQ ID NO: 61.

[0210] In some embodiments, the anti-TMPRSS6 antibody described herein is produced by (i) expressing nucleic acid identical to at least 60% (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) of SEQ ID NO: 62, and / or (ii) expressing nucleic acid identical to at least 60% (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) of SEQ ID NO: 63.

[0211] In some embodiments, the anti-TMPRSS6 antibodies described herein are produced by (i) expressing nucleic acid identical to at least 60% (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) of SEQ ID NO: 5, and / or (ii) expressing nucleic acid identical to at least 60% (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) of SEQ ID NO: 32.

[0212] In some embodiments, the anti-TMPRSS6 antibody described herein is produced by (i) expressing nucleic acid identical to at least 60% (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) of SEQ ID NO: 79 and / or (ii) expressing nucleic acid identical to at least 60% (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) of SEQ ID NO: 80.

[0213] In some embodiments, the anti-TMPRSS6 antibody described herein is produced by (i) expressing nucleic acid identical to at least 60% (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) of SEQ ID NO: 64 and / or (ii) expressing nucleic acid identical to at least 60% (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) of SEQ ID NO: 65.

[0214] In some embodiments, the anti-TMPRSS6 antibody described herein is produced by (i) expressing nucleic acid identical to at least 60% (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) of SEQ ID NO: 66 and / or (ii) expressing nucleic acid identical to at least 60% (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) of SEQ ID NO: 67.

[0215] In some embodiments, the anti-TMPRSS6 antibody described herein is produced by (i) expressing nucleic acid identical to at least 60% (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) of SEQ ID NO: 68 and / or (ii) expressing nucleic acid identical to at least 60% (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) of SEQ ID NO: 69.

[0216] In some embodiments, the anti-TMPRSS6 antibody described herein is produced by (i) expressing nucleic acid identical to at least 60% (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) of SEQ ID NO: 70 and / or (ii) expressing nucleic acid identical to at least 60% (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) of SEQ ID NO: 71.

[0217] In some embodiments, the anti-TMPRSS6 antibody described herein is produced by (i) expressing nucleic acid identical to at least 60% (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) of SEQ ID NO: 72 and / or (ii) expressing nucleic acid identical to at least 60% (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) of SEQ ID NO: 73.

[0218] In some embodiments, the anti-TMPRSS6 antibody described herein is produced by (i) expressing nucleic acid identical to at least 60% (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) of SEQ ID NO: 74, and / or (ii) expressing nucleic acid identical to at least 60% (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) of SEQ ID NO: 75.

[0219] In some embodiments, the anti-TMPRSS6 described herein is produced by (i) expressing nucleic acid identical to at least 60% (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) of SEQ ID NO: 76 and / or (ii) expressing nucleic acid identical to at least 60% (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) of SEQ ID NO: 77.

[0220] In some embodiments, this disclosure provides recombinant cells expressing the anti-TMPRSS6 antibody described herein (e.g., recombinant cells for antibody production).

[0221] Accordingly, the present disclosure provides a method for producing an antibody, the method comprising culturing recombinant cells under conditions suitable for the expression of an antibody from an expression vector by recombinant cells. Recombinant cells expressing the antibody can be cultured under any preferred conditions known in the art. In some embodiments, the method further comprises isolating the antibody from the culture medium in which the cells(s) were cultured using any preferred known method in the art.

[0222] IV. Pharmaceutical Compositions In addition to the antibodies described herein, the encoding nucleic acid or nucleic acid set, vectors containing the same, or host cells containing the vectors can be mixed with pharmaceutically acceptable carriers (excipients) to form pharmaceutical compositions for use in the treatment of target diseases. "Acceptable" means that the carrier must be compatible with the active ingredient of the composition (and preferably have the ability to stabilize the active ingredient) and must not be harmful to the subject being treated. Pharmaceutically acceptable excipients (carriers) include buffers well known in the art. See, for example, Remington: The Science and Practice of Pharmacy 20th Ed. (2000) Lippincott Williams and Wilkins, Ed. K. E. Hoover.

[0223] The anti-TMPRSS6 antibody-containing pharmaceutical compositions disclosed herein may further comprise a suitable buffer. A buffer is a weak acid or base used to maintain the pH of a solution near a selected value after the addition of another acid or base. In some examples, the buffers disclosed herein can be buffers having the ability to maintain physiological pH regardless of changes in carbon dioxide concentration (produced by cell respiration). Exemplary buffers include, but are not limited to, HEPES [4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid] buffer, Dulbecco's phosphate buffered saline (DPBS) buffer, or phosphate buffered saline (PBS) buffer. Such buffers may include disodium hydrogen phosphate and sodium chloride, or potassium dihydrogen phosphate and potassium chloride.

[0224] In some embodiments, the buffer in the pharmaceutical compositions described herein may maintain a pH value of about 5-8. For example, the pH of the pharmaceutical composition can 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 examples, the pharmaceutical composition may have a pH value lower than 7, such as about 7, 6.8, 6.5, 6.3, 6, 5.8, 5.5, 5.3, or 5.

[0225] The pharmaceutical compositions described herein include one or more suitable salts. A salt is an ionic compound that can be formed by the neutralization reaction of an acid and a base [Skoog, D.A; West, D.M.; Holler, J.F.; Crouch, S.R. (2004). "chapters 14-16". Fundamentals of Analytical Chemistry (8th ed.)]. Since a salt is composed of an associated number of cations (positively charged ions) and anions (negatively charged ions), the product is electrically neutral (has no net charge).

[0226] In some embodiments, the pharmaceutical composition can include 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. K. E. Hoover]. In some embodiments, the pharmaceutical composition can be formulated for intravenous injection. In some embodiments, the pharmaceutical composition can be formulated for subcutaneous injection.

[0227] Pharmaceutical compositions used for in vivo administration must be sterile. This can be readily achieved, for example, by filtration through a sterile filtration membrane. Therapeutic antibody compositions are generally placed in containers having a sterile access port, such as an intravenous or subcutaneous solution bag or vial having a stopper penetrable by a hypodermic needle.

[0228] V. Method of Use In certain embodiments, the Disclosure provides methods and related compositions (e.g., anti-TMPRSS6 antibody) for treating iron overload and iron overload-related conditions, including, for example, sickle cell disease (SCD), thalassemia (e.g., α-thalassemia, severe β-thalassemia, mild β-thalassemia), hemochromatosis (e.g., type 1 hemochromatosis, type 2 hemochromatosis, type 3 hemochromatosis, type 4 hemochromatosis), transfusion-associated iron overload (e.g., repeated transfusions for anemia, large-volume transfusions for blood loss due to trauma), hemolytic anemia (e.g., transfusion-dependent hemolytic anemia, pyruvate kinase deficiency hemolytic anemia), African iron overload, Blackfan-diamond anemia, refractory anemia with ring sideroblasts (RARS), and myelodysplastic syndromes (MDS) (e.g., SF3B1-associated MDS). In some embodiments, the disclosure provides a method for treating sickle cell disease (SCD). In some embodiments, the disclosure provides a method for treating sickle cell disease in subjects with iron overload.

[0229] Aspects of this disclosure relate to methods and compositions (e.g., anti-TMPRSS6 antibodies) useful for treating iron overload. Iron overload occurs when there is an excess of iron storage in the body. In some embodiments, iron overload is primary iron overload caused by a genetic disorder (e.g., hemochromatosis). In some embodiments, iron overload is secondary iron overload caused by a condition that increases iron storage in the body (e.g., blood transfusion or hemolysis) or a disorder that impairs red blood cell production (i.e., ineffective erythropoiesis) (e.g., sickle cell disease, thalassemia, refractory anemia with ring sideroblasts (RARS), MDS). Excess iron is deposited in organs throughout the body, which can lead to organ damage. Organs commonly affected by iron deposition include the liver, heart, and endocrine glands.

[0230] In some embodiments, matriptase-2 regulates iron homeostasis by regulating hepcidin levels in the subject. Iron is an essential component for almost all living cells and organisms. Excess iron is harmful (for example, due to its redox reactivity which promotes oxidative stress). Therefore, dysregulation of iron metabolism leads to disease. In mammals, the majority of iron in the body (e.g., >70%) is distributed in red blood cells and mediates oxygen transport within hemoglobin. Aged red blood cells are removed by tissue macrophages, and the iron from aged red blood cells is recycled to erythroblasts for reuse. Iron release into plasma involves ferrous iron (Fe). 2+ Ferrous ferrous 3+ It undergoes oxidation and is captured by iron carrier transferrin. The main function of transferrin is iron delivery to tissues via transferrin receptor 1 (TfR1). Duodenal intestinal cells move iron from the intestinal lumen via divalent metal transporter 1 (DMT1) and excrete it into the plasma via ferroportin.

[0231] The inclusion of iron in the bloodstream is crucial for systemic iron homeostasis and is negatively regulated by the iron-regulating hormone hepcidin (see, e.g., Ganz T. Systemic iron homeostasis. Physiol Rev. (2013) 93:1721-41). Hepcidin is expressed as a prepropeptide in hepatocytes and undergoes proteolytic processing. Mature hepcidin is a cysteine-rich peptide of 25 amino acids. In some situations, the binding of hepcidin to ferroportin in tissue macrophages, duodenal intestinal cells, and other target cells triggers ubiquitination, internalization, and degradation of ferroportin in lysosomes, which leads to iron sequestration within macrophages, inhibition of dietary iron absorption, and, consequently, a decrease in plasma iron levels. Therefore, iron levels can be regulated by manipulating hepcidin levels in the target. Hepcidin expression is regulated by the BMP signaling pathway and the IL-6-JAK-STAT signaling pathway. The binding of BMPs (e.g., BMP6 or BMP2) to BMP receptors on the cell membrane of hepatocytes (e.g., type I (ALK2 and ALK3), type II (ActRIIA and BMPR2) BMP receptors) via the BMP pathway promotes hepcidin expression. BMP coreceptors, human homeostatic iron regulatory protein (HFE), hemoduvelin (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 matriptase-2 (TMPRSS6), which cleaves and inactivates type I and II BMP receptors as well as HJV (Wahedi et al. Matriptase-2 suppresses hepcidin expression by cleaving multiple components of the hepcidin induction pathway. J Biol Chem. (2017) 292:18354-71).In some embodiments, cleavage of HJV by matriptase-2 reduces the amount of cell surface HJV available for BMP signaling, and consequently reduces hepcidin expression. In some embodiments, loss of hemomodoverin function may be associated with iron overload (e.g., type 2 hemochromatosis). 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 amplify hepcidin synthesis in response to IL-6 and are unable to initiate an effective hypoirhemic response to acute inflammation. Therefore, in some embodiments, inhibition of matriptase-2 (e.g., by an anti-TMPRSS6 antibody) positively modulates hepcidin expression by maintaining / increasing HJV activity. In some embodiments, increased hepcidin levels due to inhibition of matriptase-2 (e.g., by an anti-TMPRSS6 antibody) reduce iron levels in the subject, thereby addressing iron overload.

[0232] 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. An increase in hepcidin levels 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, a decrease in systemic iron leads to a decrease in mean cellular 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, by administering anti-TMPRSS6 antibody, one or more of the following symptoms are treated and / or improved: arthralgia, fatigue, weight loss, skin discoloration, abdominal pain, hair loss, decreased libido, blurred memory, or arrhythmia. In some embodiments, the methods provided herein include targeting an effective dose of anti-TMPRSS6 antibody for the purpose of improving symptoms and conditions associated with secondary iron overload, such as sickle cell disease, thalassemia, refractory anemia with ring sideroblasts (RARS), and mesenteric dysplasia (MDS), as described elsewhere herein. In some embodiments, the methods provided herein involve administering an effective dose of anti-TMPRSS6 antibody to target a patient to treat sickle cell disease (SCD), thalassemia (e.g., α-thalassemia, severe β-thalassemia, mild β-thalassemia), hemochromatosis (e.g., type 1 hemochromatosis, type 2 hemochromatosis, type 3 hemochromatosis, type 4 hemochromatosis), transfusion-associated iron overload (e.g., repeated transfusions for anemia, large-volume transfusions for blood loss due to trauma), hemolytic anemia (e.g., transfusion-dependent hemolytic anemia, pyruvate kinase deficiency hemolytic anemia), African iron overload, Blackfan-diamond anemia, refractory anemia with ring sideroblasts (RARS), or myelodysplastic syndrome (MDS) (e.g., SF3B1-associated MDS).

[0233] Further embodiments of this disclosure relate to methods and compositions (e.g., anti-TMPRSS6 antibodies) useful for the treatment of sickle cell disease (SCD). In some embodiments, the methods provided herein include administering an effective amount of anti-TMPRSS6 antibody to a subject in order to treat SCD in the subject. In some embodiments, the methods provided herein include administering an effective amount of anti-TMPRSS6 antibody to a subject for the purpose of treating SCD in a subject having iron overload. In some embodiments, the methods provided herein reduce iron overload in a subject having SCD. In some embodiments, the methods described herein reduce systemic iron levels in a subject having SCD. In some embodiments, the methods provided herein reduce mean cellular hemoglobin concentration (MCHC) in a subject having SCD. 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 SCD (e.g., occlusive epidemic). Sickle cell disease (SCD) is a comprehensive term for a group of genetic disorders affecting the hemoglobin gene. Sickle cell disease (SCD) is usually caused by an autosomal recessive mutation (e.g., single nucleotide substitution) in the β-globin gene, which leads to the synthesis of sickle hemoglobin (HbS) and sickle-shaped 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+ (beta) thalassemia, hemoglobin SB0 (beta-zero) thalassemia, hemoglobin SD disease, hemoglobin SE disease, and hemoglobin SO disease. In hemoglobin S syndrome (i.e., sickle cell anemia), the target group is homozygous for a mutation in the hemoglobin S (HbS) gene and produces only hemoglobin S.In hemoglobin SSS disease, the typical disc shape of red blood cells (RBCs) is altered to a sickle shape, and this deformation disrupts their primary function. Other symptoms of hemoglobin SSS disease include fatigue, recurrent infections, periodic pain, and visceral injury. Iron supplements are not considered effective in increasing the level of hemoglobin present in the blood. In hemoglobin SC disease, the subject has one allele of the HbS gene and one allele of the hemoglobin C (HbC) gene. Generally, anemia in hemoglobin SC disease is less severe than in hemoglobin SSS disease. This is likely because the hemoglobin C gene does not polymerize as rapidly as hemoglobin S. Therefore, only a small number of sickle cells are formed. The symptoms of hemoglobin SSS disease and hemoglobin SC disease are similar, but differ slightly in severity.

[0234] In SCD (e.g., hemoglobin SS disease and / or hemoglobin SC disease), during deoxygenation in capillaries, HbS polymerizes into stiff fibers, causing strain on the red blood cell membrane, which can lead to intravascular hemolysis and inflammation. In some embodiments, the degree of HbS polymerization is related to the hemoglobin concentration in red blood cells. The hemoglobin concentration in red blood cells can be measured by mean cytoplasmic hemoglobin concentration (MCHC), which is a measure of the average amount of hemoglobin in a single red blood cell. In some embodiments, HbS polymerization in SCD patients can be reduced by decreasing the hemoglobin concentration in red blood cells (see, for example, Sunshine et al., Requirements for therapeutic inhibition of sickle hemoglobin gelation, Nature volume 275, pages 238-240 (1978)). In some embodiments, reducing systemic iron levels results in a reduction of HbS polymerization.In some embodiments, reducing systemic iron leads to a decrease in MCHC, which in turn leads to a decrease in HbS polymerization (e.g., 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 Clin Invest. 1984 Jan; 73(1): 116-123; Brewin et al., The pleiotropic effects of α-thalassemia on HbSS and HbSC sickle cell disease: Reduced erythrocyte cation co-transport activity, serum erythropoietin, and transfusion) (See Am J Hematol. 2022 Oct;97(10):1275-1285; burden, do not translate into increased survival.) Intravascular hemolysis in SCD results in the release of cell-free hemoglobin into circulation, which oxidizes and releases reactive heme into the vascular structure. In some embodiments, the release of cell-free hemoglobin into circulation causes iron overload in SCD patients. In some embodiments, extracellular heme activates endothelial cell adhesion molecules, inducing 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).As used herein, the terms vascular occlusive attack (VOC), vascular occlusion, painful attack, and vascular occlusive painful attack are interchangeable. Vascular occlusion is a fundamental pathological process in SCD (Frenette PS. Sickle cell vaso-occlusion: multistep and multicellular paradigm. Curr Opin Hematol. (2002) 9:101-6; Pathare et al., Cytokines in sickle cell disease. Hematology. (2003) 8:329-37). In VOCs, in some embodiments, the vascular lumen is blocked by cells that obstruct capillary blood flow to various organs and other parts of the body. This triggers an inflammatory process that leads to painful attacks and damage to various tissues (e.g., brain, liver, kidneys, lungs, spleen, etc.). Painful attacks affect virtually all patients with SCD, often beginning in late infancy and recurring throughout life. In some embodiments, patients with SCD are treated with blood transfusions (e.g., periodic or chronic transfusions on demand). In some embodiments, transfusions accelerate hemolysis and / or iron overload (e.g., Raghunath et al., Iron Overload in Sickle Cell Disease, Advances in Hematology, Volume 2010 | Article ID 272940).

[0235] In some embodiments, subjects with SCD have splenic hematopoietic retention. The primary function of the spleen is the removal of defective RBCs, including sickle-shaped RBCs. In patients with SCD, blood flow through the spleen is slow, leading to decreased oxygen pressure and, consequently, increased polymerization of HbS. Due to the narrow capillaries in the splenic vascular bed, the spleen then becomes filled with blood cells. Splenic hematopoietic retention (also known as splenic retention) is characterized by acutely dilated spleen (splenomegaly) and hemoglobin levels more than 2 g / dL below baseline levels in affected individuals (Bender and Carlberg, September 15, 2003 [updated December 28, 2023]. In: Adam MP, Feldman J, Mirzaa GM, Pagon RA, Wallace SE, Bean LJH, Gripp KW, Amemiya A, editors. GeneReviews® [Internet]. Seattle (WA): University of Washington, Seattle; 1993-2024). In some embodiments, subjects with splenic hematopoietic retention require hospitalization, splenectomy, and / or blood transfusion. In some embodiments, administration of anti-TMPRSS6 antibody reduces splenic hematopoietic retention (e.g., frequency and / or severity) in subjects with SCD compared to subjects before administration or compared to untreated subjects with SCD. In some embodiments, administration of anti-TMPRSS6 antibodies reduces hospitalizations associated with splenic hematopoietic retention (e.g., hospitalizations associated with splenectomy) in subjects with SCD compared to subjects before administration or to untreated subjects with SCD. In some embodiments, administration of anti-TMPRSS6 antibodies reduces the frequency of transfusions required in subjects with SCD compared to subjects before administration or to untreated subjects with SCD. In some embodiments, administration of anti-TMPRSS6 antibodies reduces the size of the dilated spleen (e.g., subjects with splenomegaly) in subjects with SCD compared to subjects before administration or to untreated subjects with SCD.

[0236] In some embodiments, subjects with SCD have extramedullary hematopoiesis (EMH) (see, e.g., 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 may occur due to chronic anemia and the body's compensatory response to maintain adequate blood cell production. In some embodiments, EMH occurs in subjects with SCD due to chronic hemolytic anemia, splenomegaly, ineffective erythropoiesis, VOCs, and / or organ damage and dysfunction, all of which contribute to an increased need for blood cell production. In some embodiments, administration of anti-TMPRSS6 antibody reduces EMH in subjects with SCD compared to subjects before administration or compared to untreated subjects with SCD.

[0237] 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 Jun 16;13(6):e15680). In some embodiments, hepatomegaly in subjects with SCD is caused by congestion in blood vessels caused by sickle cells, which in some embodiments leads to hepatic infarction of the liver. In some embodiments, the liver becomes dilated due to congestion caused by blocked blood flow. In some embodiments, hepatomegaly in subjects with SCD is caused by sickle cell episodes (e.g., hepatic hematuria). In some embodiments, during splenic hematuria episodes, blood also becomes trapped in the liver, leading to its dilation. In some embodiments, hepatomegaly in subjects with SCD is caused by hepatic dysfunction. In some embodiments, hepatic 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 may be caused by gallbladder complications (e.g., gallstones and cholecystitis), leading to bile duct obstruction and hepatomegaly. In some embodiments, hepatomegaly in subjects with SCD may be caused by hepatic iron overload (e.g., from chronic transfusion therapy) (e.g., secondary hemochromatosis). Excess iron deposition in the liver may lead to hepatomegaly and hepatic dysfunction. In some embodiments, hepatomegaly in subjects with SCD may be caused at least in part by hepatotoxicity associated with drug therapies (e.g., hydroxyurea) taken to treat SCD, leading to drug-induced liver injury. In some embodiments, administration of anti-TMPRSS6 antibodies 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 antibodies reduces hepatic vascular retention 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 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 reduces 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 reduces hepatic iron overload in subjects with SCD compared to subjects before administration or to untreated subjects with SCD. In some embodiments, administration of anti-TMPRSS6 antibody reduces hepatic dysfunction in subjects with SCD compared to subjects before administration or to untreated subjects with SCD. In some embodiments, administration of anti-TMPRSS6 antibody reduces hepatic infarction in subjects with SCD compared to subjects before administration or to untreated subjects with SCD. In some embodiments, administration of an anti-TMPRSS6 antibody reduces hepatic congestion (e.g., biliary obstruction) in subjects with SCD compared to subjects before administration or compared to untreated subjects with SCD.

[0238] In some embodiments, the disclosure provides methods and compositions (e.g., anti-TMPRSS6 antibodies) useful for reducing volatile organic compounds (VOCs) in subjects having SCD (e.g., SCD subjects with or without blood transfusions) via iron restriction. In some embodiments, administration of an anti-TMPRSS6 antibody reduces the frequency of VOCs in subjects having SCD (e.g., SCD subjects with or without blood transfusions). In some embodiments, administration of an anti-TMPRSS6 antibody reduces the severity of VOCs in subjects having SCD (e.g., SCD subjects with or without blood transfusions). In some embodiments, administration of an anti-TMPRSS6 antibody results in an increase in hepcidin levels in subjects having SCD (e.g., SCD subjects with or without blood transfusions) compared to subjects before receiving the anti-TMPRSS6 antibody, or subjects having SCD (e.g., SCD subjects with or without blood transfusions) but not receiving the anti-TMPRSS6 antibody, thereby reducing circulating iron (e.g., circulating iron available for HbS synthesis).

[0239] In some embodiments, administration of an anti-TMPRSS6 antibody results in a decrease in circulating iron (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%) in subjects with SCD (e.g., SCD subjects with or without blood transfusions) compared to subjects without receiving the anti-TMPRSS6 antibody. In some embodiments, administration of an anti-TMPRSS6 antibody results in a decrease in circulating iron (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%) in subjects with SCD compared to subjects before receiving the anti-TMPRSS6 antibody or subjects with SCD (e.g., hemoglobin SC disease) but not receiving the anti-TMPRSS6 antibody. Circulating iron levels can be assessed by conventional clinical trials, such as measuring serum iron, transferrin saturation (TSAT), or total iron-binding capacity (TIBC).

[0240] In some embodiments, administration of an anti-TMPRSS6 antibody results in a subject with SCD (e.g., a subject with or without blood transfusions) having an increased circulating hepcidin-25 level (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%, 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 the circulating hepcidin-25 level) compared to a subject with SCD before receiving the anti-TMPRSS6 antibody or a subject with SCD (e.g., a subject with or without blood transfusions). In some embodiments, administration of an anti-TMPRSS6 antibody results in an increase in circulating hepcidin-25 levels (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%, 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 subjects with SCD (e.g., hemoglobin SC disease) compared to subjects before receiving the anti-TMPRSS6 antibody or subjects with SCD (e.g., hemoglobin SC disease) but without receiving the anti-TMPRSS6 antibody.

[0241] In some embodiments, administration of an anti-TMPRSS6 antibody results in an increase in Hamp expression. In some embodiments, administration of an anti-TMPRSS6 antibody results in an increase in Hamp expression (e.g., an increase 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 Hamp expression) in subjects with SCD (e.g., SCD subjects with or without blood transfusions) compared to subjects without receiving the anti-TMPRSS6 antibody. In some embodiments, administration of an anti-TMPRSS6 antibody results in increased Hamp expression (e.g., a decrease 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%) in subjects with SCD (e.g., hemoglobin SC disease) compared to subjects before receiving the anti-TMPRSS6 antibody or subjects with SCD (e.g., hemoglobin SC disease) but without receiving the anti-TMPRSS6 antibody.

[0242] In some embodiments, administration of an anti-TMPRSS6 antibody results in a decrease in transferrin saturation (TSAT). In some embodiments, administration of an 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% of TSAT) in subjects with SCD (e.g., SCD subjects with or without blood transfusions) compared to subjects without receiving the anti-TMPRSS6 antibody. In some embodiments, administration of an anti-TMPRSS6 antibody results in a reduction in TSAT (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% of TSAT) in subjects with SCD (e.g., hemoglobin SC disease) compared to subjects before receiving the anti-TMPRSS6 antibody or subjects with SCD (e.g., hemoglobin SC disease) but without receiving the anti-TMPRSS6 antibody.

[0243] In some embodiments, administration of anti-TMPRSS6 antibody results in a decrease in extramedullary hematopoiesis. In some embodiments, administration of anti-TMPRSS6 antibody results in a decrease in extramedullary hematopoiesis (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% of extramedullary hematopoiesis) in subjects with SCD (e.g., SCD subjects with or without blood transfusions) compared to subjects without anti-TMPRSS6 antibody. In some embodiments, administration of an anti-TMPRSS6 antibody results in a reduction in extramyelopoeia (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% of extramyelopoeia) in subjects with SCD (e.g., hemoglobin SC disease) compared to subjects before receiving the anti-TMPRSS6 antibody or subjects with SCD (e.g., hemoglobin SC disease) but without receiving the anti-TMPRSS6 antibody. Extramedullary hematopoiesis can be assessed by preferred known methods, such as the ratio of spleen weight to body weight, imaging studies (e.g., X-ray, CT, MRI, ultrasound), biopsy, blood tests (e.g., hematological cytomorphology or blast cell levels), bone marrow examinations (e.g., changes in bone marrow cell type, composition and structure), or erythropoietin levels and reticulocyte count.

[0244] 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 (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% of hepatomegaly) in subjects with SCD (e.g., SCD subjects with or without blood transfusions) compared to subjects without anti-TMPRSS6 antibody treatment. In some embodiments, administration of an anti-TMPRSS6 antibody results in a reduction in hepatomegaly (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% of hepatomegaly) in subjects with SCD (e.g., hemoglobin SC disease) compared to subjects before receiving the anti-TMPRSS6 antibody or subjects with SCD (e.g., hemoglobin SC disease) but without receiving the anti-TMPRSS6 antibody. Hepatomegaly can be assessed by preferred known methods, such as the ratio of liver weight to body weight, imaging studies (e.g., CT, MRI, ultrasound, hepatic elastography), blood tests (e.g., liver function tests, markers of liver injury [alanine transaminase and aspartate transaminase]), and biopsy.

[0245] In some embodiments, administration of an anti-TMPRSS6 antibody results in a decrease in the frequency of vaso-occlusive crises (VOCs) (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% decrease in VOC frequency) in a subject with SCD (e.g., a SCD subject with or without transfusion) compared to a subject having SCD (e.g., a SCD subject with or without transfusion) prior to receiving the anti-TMPRSS6 antibody but who has not received the anti-TMPRSS6 antibody. In some embodiments, administration of an anti-TMPRSS6 antibody results in a decrease in the frequency of VOCs (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% decrease in VOC frequency) in a subject with SCD (e.g., hemoglobin SC disease) compared to a subject having SCD (e.g., hemoglobin SC disease) prior to receiving the anti-TMPRSS6 antibody but who has not received the anti-TMPRSS6 antibody.

[0246] In some embodiments, administration of an anti-TMPRSS6 antibody results in a reduction in the severity of vascular occlusive disease (VOC) (e.g., hospitalization and / or duration) in subjects with SCD (e.g., SCD subjects with or without blood transfusions) compared to subjects before receiving the anti-TMPRSS6 antibody or subjects with SCD (e.g., SCD subjects with or without blood transfusions) (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% of VOC severity (e.g., hospitalization and / or duration)). In some embodiments, administration of an anti-TMPRSS6 antibody results in a reduction in the frequency of vascular occlusive disorders (VOCs) in subjects with SCD (e.g., hemoglobin-mediated sclerosis) compared to subjects before receiving the anti-TMPRSS6 antibody or subjects with SCD (e.g., hemoglobin-mediated sclerosis) but without receiving the 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% of VOCs).In some embodiments, administration of an anti-TMPRSS6 antibody results in a reduction in the severity of vascular occlusive disorders (e.g., hospitalization and / or duration) (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% of VOC) in subjects with SCD (e.g., hemoglobin SC disease) compared to subjects before receiving the anti-TMPRSS6 antibody or subjects with SCD (e.g., hemoglobin SC disease) but without receiving the anti-TMPRSS6 antibody. VOCs (e.g., frequency and / or severity) can be assessed by preferred known methods, such as a comprehensive metabolic panel including complete hematology with leukocyte differential count, platelet count, reticulocyte count, and liver and kidney function tests. Typical laboratory findings include a sharp decrease in hemoglobin concentration, an increase in platelet count, an increase in reticulocyte count, and elevated serum urea.

[0247] In some embodiments, a decrease in the administration of anti-TMPRSS6 antibody results in a decrease in systemic iron. In some embodiments, the administration of anti-TMPRSS6 antibody results in a decrease in MCHC. In some embodiments, a decrease in MCHC results in a decrease in HbS polymerization. In some embodiments, the administration of anti-TMPRSS6 antibody results in a decrease in MCHC (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% of MCHC) in subjects with SCD (e.g., SCD subjects with or without blood transfusions) compared to subjects without anti-TMPRSS6 antibody. In some embodiments, administration of an 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% of MCHC) in subjects with SCD (e.g., hemoglobin SC disease) compared to subjects before receiving the anti-TMPRSS6 antibody or subjects with SCD (e.g., hemoglobin SC disease) but without receiving the anti-TMPRSS6 antibody.In some embodiments, administration of an anti-TMPRSS6 antibody results in a reduction in HbS polymerization (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% of HbS polymerization) in subjects with SCD (e.g., SCD subjects with or without blood transfusions) compared to subjects without receiving the anti-TMPRSS6 antibody. In some embodiments, administration of an anti-TMPRSS6 antibody results in a reduction in HbS polymerization (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% of HbS polymerization) in subjects with SCD (e.g., hemoglobin SC disease) compared to subjects before receiving the anti-TMPRSS6 antibody or subjects with SCD (e.g., hemoglobin SC disease) but without receiving the anti-TMPRSS6 antibody. HbS polymerization can be evaluated by suitable known methods, such as solubility sickling tests, 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).

[0248] In some embodiments, administration of an anti-TMPRSS6 antibody results in a reduction in sickle cyst formation of RBCs (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% of RBC sickle cyst formation in subjects with SCD (e.g., SCD subjects with or without blood transfusions) compared to subjects without receiving the anti-TMPRSS6 antibody. In some embodiments, administration of an anti-TMPRSS6 antibody results in a reduction in RBC sickling (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% of RBC sickling compared to subjects with SCD (e.g., hemoglobin SC disease) before receiving the anti-TMPRSS6 antibody or subjects with SCD (e.g., hemoglobin SC disease) but without receiving the anti-TMPRSS6 antibody. RBC sickling can be evaluated by a preferred known method, such as a blood smear.

[0249] In some embodiments, administration of an anti-TMPRSS6 antibody results in a reduction in hypochromic RBC levels (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% of hypochromic RBC levels) in subjects with SCD (e.g., SCD subjects with or without blood transfusions) compared to subjects without receiving the anti-TMPRSS6 antibody. In some embodiments, administration of an anti-TMPRSS6 antibody results in a reduction in hypochromic RBC levels (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% hypochromic RBC levels) in subjects with SCD (e.g., hemoglobin SC disease) compared to subjects before receiving the anti-TMPRSS6 antibody or subjects with SCD (e.g., hemoglobin SC disease) but without receiving the anti-TMPRSS6 antibody.

[0250] In some embodiments, administration of an anti-TMPRSS6 antibody results in a reduction in microcytic RBC levels (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% of microcytic RBC levels) in subjects with SCD (e.g., SCD subjects with or without blood transfusions) compared to subjects without receiving the anti-TMPRSS6 antibody. In some embodiments, administration of an anti-TMPRSS6 antibody results in a reduction in microcytic RBC levels (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% microcytic RBC levels) in subjects with SCD (e.g., hemoglobinic sclerosis) compared to subjects before receiving the anti-TMPRSS6 antibody or subjects with SCD (e.g., hemoglobinic sclerosis) but without receiving the anti-TMPRSS6 antibody.

[0251] In some embodiments, administration of an anti-TMPRSS6 antibody results in a reduction in hemolysis (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% of hemolysis) in subjects with SCD (e.g., SCD subjects with or without blood transfusions) compared to subjects without receiving the anti-TMPRSS6 antibody. In some embodiments, administration of an anti-TMPRSS6 antibody results in a reduction in hemolysis (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% of hemolysis) in subjects with SCD (e.g., hemoglobin SC disease) compared to subjects before receiving the anti-TMPRSS6 antibody or subjects with SCD (e.g., hemoglobin SC disease) but without receiving the anti-TMPRSS6 antibody. Hemolysis can be assessed by preferred known methods, such as lactate dehydrogenase (LDH), unconjugated bilirubin, total bilirubin, direct bilirubin, and haptoglobin tests, reticulocyte count, or total heme and hemopexin (see, e.g., Hemolytic Anemia; ARUP Consult®; obtained March 15, 2023; see arupconsult.com / content / hemolytic-anemias).

[0252] In some embodiments, administration of an anti-TMPRSS6 antibody results in a decrease in lactate dehydrogenase (LDH). In some embodiments, administration of an anti-TMPRSS6 antibody results in a decrease in LDH (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% of LDH) in subjects with SCD (e.g., SCD subjects with or without blood transfusions) compared to subjects without receiving the anti-TMPRSS6 antibody. In some embodiments, administration of an anti-TMPRSS6 antibody results in a decrease in LDH (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% of LDH) in subjects with SCD (e.g., hemoglobin SC disease) compared to subjects before receiving the anti-TMPRSS6 antibody or subjects with SCD (e.g., hemoglobin SC disease) but without receiving the anti-TMPRSS6 antibody.

[0253] In some embodiments, administration of an anti-TMPRSS6 antibody results in a decrease in bilirubin (e.g., direct bilirubin and / or total bilirubin). In some embodiments, administration of an anti-TMPRSS6 antibody results in a decrease in bilirubin (e.g., direct bilirubin and / or total bilirubin) (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% of bilirubin) in subjects with SCD (e.g., SCD subjects with or without blood transfusions) compared to subjects without receiving the anti-TMPRSS6 antibody. In some embodiments, administration of an anti-TMPRSS6 antibody results in a reduction in bilirubin (e.g., direct bilirubin and / or total bilirubin) (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% of bilirubin) in subjects with SCD (e.g., hemoglobin SC disease) compared to subjects before receiving the anti-TMPRSS6 antibody or subjects with SCD (e.g., hemoglobin SC disease) but not receiving the anti-TMPRSS6 antibody.

[0254] In some embodiments, administration of an anti-TMPRSS6 antibody results in a reduction of inflammation (e.g., systemic inflammation). In some embodiments, administration of an 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% of inflammation) in subjects with SCD (e.g., SCD subjects with or without blood transfusions) before receiving the anti-TMPRSS6 antibody compared to subjects who have not received the anti-TMPRSS6 antibody. In some embodiments, administration of an 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% of inflammation) in subjects with SCD (e.g., hemoglobin SC disease) compared to subjects before receiving the anti-TMPRSS6 antibody or subjects with SCD (e.g., hemoglobin SC disease) but without receiving the anti-TMPRSS6 antibody. Inflammation can be assessed by preferred 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.

[0255] In some embodiments, administration of anti-TMPRSS6 antibody results in a decrease in white blood cell (WBC) count. In some embodiments, administration of anti-TMPRSS6 antibody results in a decrease in WBC count (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 WBC count) in subjects with SCD (e.g., SCD subjects with or without blood transfusions) compared to subjects without anti-TMPRSS6 antibody. In some embodiments, administration of an anti-TMPRSS6 antibody results in a reduction in WBC 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% of WBC count) in subjects with SCD (e.g., hemoglobin SC disease) compared to subjects before receiving the anti-TMPRSS6 antibody or subjects with SCD (e.g., hemoglobin SC disease) but without receiving the anti-TMPRSS6 antibody.

[0256] In some embodiments, administration of an anti-TMPRSS6 antibody results in a decrease in neutrophil count. In some embodiments, administration of an anti-TMPRSS6 antibody results in a decrease in neutrophil count (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 neutrophil count (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., SCD subjects with or without blood transfusions) compared to subjects without receiving the anti-TMPRSS6 antibody. In some embodiments, administration of an anti-TMPRSS6 antibody results in a decrease in neutrophil count (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., hemoglobin SC disease) compared to subjects before receiving the anti-TMPRSS6 antibody or subjects with SCD (e.g., hemoglobin SC disease) but without receiving the anti-TMPRSS6 antibody.

[0257] In some embodiments, administration of an anti-TMPRSS6 antibody results in a decrease in lymphocyte count. In some embodiments, administration of an anti-TMPRSS6 antibody results in a decrease in lymphocyte count (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., SCD subjects with or without blood transfusions) compared to subjects without receiving the anti-TMPRSS6 antibody. In some embodiments, administration of an 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., hemoglobin SC disease) compared to subjects before receiving the anti-TMPRSS6 antibody or subjects with SCD (e.g., hemoglobin SC disease) but without receiving the anti-TMPRSS6 antibody.

[0258] In some embodiments, administration of an anti-TMPRSS6 antibody results in a reduction in the frequency of blood transfusions (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 the frequency of blood transfusions) in subjects with SCD (e.g., SCD subjects with or without blood transfusions) compared to subjects without receiving the anti-TMPRSS6 antibody. In some embodiments, administration of an anti-TMPRSS6 antibody results in a reduction in transfusion sessions (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% of transfusion sessions) in subjects with SCD (e.g., hemoglobin SC disease) before receiving the anti-TMPRSS6 antibody or in subjects with SCD (e.g., hemoglobin SC disease) but who have not received the anti-TMPRSS6 antibody.

[0259] In some embodiments, the Disclosure relates to any known therapeutic agent for treating SCD, e.g., hemoglobin S polymerization inhibitors (e.g., voxelotol), therapeutic agents for reducing VOCs (e.g., hydroxyurea, L-glutamine oral powder, chryzanlizumab, selective pyruvate kinase-R (PKR) activators), analgesics (e.g., narcotics, opioids, gabapentin, cannabis), blood transfusions, stem cell transplants, and exagamglosine autotemcells (exa-ce). The present invention provides a method for treating subjects with SCD by administering an effective dose of anti-TMPRSS6 antibody in combination with lenticulin, GMI-1070, VIT-2763 (vamifeport), ticagrelor, vitamin D, simvastatin, AG-348 (mitapivat sulfate), propranolol, epinephrine, regadenoson, atorvastatin, prasugrel, L-arginine, OTQ923, oxygen therapy, and gene therapy.

[0260] Further aspects of this disclosure relate to methods and compositions (e.g., anti-TMPRSS6 antibodies) useful for treating thalassemia (e.g., α-thalassemia, severe β-thalassemia, or mild β-thalassemia) and / or one or more conditions resulting from thalassemia in subjects. In some embodiments, the methods provided herein involve administering an effective amount of anti-TMPRSS6 antibody to a subject having iron overload for the purpose of treating thalassemia (e.g., α-thalassemia, severe β-thalassemia, or mild β-thalassemia). In some embodiments, the methods provided herein reduce iron overload in a subject having thalassemia (e.g., α-thalassemia, severe β-thalassemia, or mild β-thalassemia). "Thalassemia" includes a group of hereditary blood disorders caused by defects in the synthesis of one or more hemoglobin chains. In some embodiments, α-thalassemia is caused by a decrease or absence of alpha-globin chain synthesis. In some embodiments, β-thalassemia is caused by a decrease or absence of betaglobin chain synthesis. In some embodiments, an imbalance in hemoglobin chains leads to hemolysis and impaired erythrogenesis.

[0261] Alpha-thalassemia is the result of a defect or absence of alpha-globin chain synthesis, leading to an excess of beta-globin chains. Alpha-globin chain production is controlled by two genes (i.e., four alpha-globin genes) each on chromosome 16, and defective production is usually caused by the deletion of one or more of these genes (Farashi et al., Molecular basis of α-thalassemia. Blood Cells Mol Dis 2018; 70:43-53). A single gene deletion results in an α-thalassemia silent carrier state, which is asymptomatic and presents with normal hematological findings. A 2-gene deletion causes a mild form of α-thalassemia with microcytosis, usually without anemia. A 3-gene deletion results in marked production of hemoglobin H (HbH) with four beta-chains. An intermediate form of α-thalassemia, or HbH disorder, causes microcytic anemia, hemolysis, and splenomegaly. The deletion of four genes results in the marked production of hemoglobin Bart (Hb Bart), which has four gamma chains. Severe α-thalassemia with Hb Bart usually leads to lethal fetal hydrops (Harteveld and Higgs, α-thalassemia. Orphanet J Rare Dis 2010; 5:13).

[0262] β-thalassemia is the result of a defect or absence of beta-taglobin chain synthesis, leading to an excess of alpha chains and, consequently, impaired erythropoiesis. Beta-taglobin chain production is controlled by the beta-taglobin gene on chromosome 11. Beta-taglobin chain production can range from near-normal to completely absent, leading to varying degrees of excess alpha-globin versus beta-taglobin 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 either two missing or defective beta-taglobin genes, resulting in the absence of beta-taglobin chains. Subjects with severe β-thalassemia typically present with severe anemia (e.g., within the first two years of life) and require regular red blood cell (RBC) transfusions. Findings in untreated or poorly transfused individuals with severe β-thalassemia may include hemolysis, growth retardation, pallor, jaundice, insufficient muscular system, hepatosplenomegaly, lower leg ulcers, clot formation from extramedullary hematopoiesis, and skeletal changes resulting from bone marrow expansion and proliferation. In some embodiments, treatment options for severe β-thalassemia include regular transfusions, iron chelation, and management of secondary complications of iron overload. In some conditions, splenectomy may be necessary, and bone marrow transplantation remains the only curative treatment currently available. Regular transfusion therapy can lead to endocrine complications (growth retardation, impaired sexual maturation, diabetes, and insufficient parathyroid, thyroid, pituitary, and, less commonly, adrenal), iron overload-related complications including dilated cardiomyopathy, hepatic fibrosis, and cirrhosis.

[0263] Patients with intermediate β-thalassemia, caused by the loss or defect of two betaglobin genes, exhibit significant genetic heterogeneity and clinical polymorphisms, present with moderate anemia later in life, and do not require regular blood transfusions. Key clinical features in these patients include erythrocyte myelometra with medullary and extramedullary hematopoiesis and its complications (osteoporosis, mainly erythropoietic tissue masses affecting the spleen, liver, lymph nodes, chest and spine, as well as bone deformities and typical facial changes), hemolytic gallstones, iron accumulation, painful lower leg ulcers, and increased predisposition to thrombosis. In some embodiments, patients with intermediate-type β-thalassemia may receive blood transfusions if they have severe, persistent symptoms (e.g., persistent leg ulcers and associated pain, pulmonary hypertension, severe anemia, growth retardation) (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).

[0264] In mild β-thalassemia, only one betaglobin gene is damaged or lost. Subjects with mild β-thalassemia are mildly anemia, hypochromic, and microcytic (i.e., elevated levels of hypochromic and / or microcytic erythrocytes, respectively). In some embodiments, iron overload occurs in subjects with mild β-thalassemia who are supplemented with iron due to their anemia.

[0265] 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 increased morbidity 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 transfusion (e.g., in severe β-thalassemia and intermediate β-thalassemia). In some embodiments, iron overload in patients with beta-thalassemia (e.g., primarily severe and intermediate forms of β-thalassemia) is secondary to ineffective erythropoiesis characterized by increased 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, suppression of hepcidin expression enhances iron absorption from the intestines, enabling iron release from macrophages, thereby further increasing iron levels. Due to the underlying iron loading mechanisms and / or rates of iron accumulation, different organs are affected differently by iron overload in different forms of thalassemia (see, e.g., Taher and Saliba, Iron overload in thalassemia: different organs at different rates. Hematology Am Soc Hematol Educ Program 2017; 2017(1):265-271). In some embodiments, the methods described herein treat thalassemia (e.g., α-thalassemia, severe β-thalassemia, or mild β-thalassemia) in the subject. In some embodiments, the thalassemia is not intermediate β-thalassemia.In some embodiments, this disclosure provides a method for treating iron overload in subjects having thalassemia (e.g., α-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 matriptase-2 activity, subsequently increasing hemoduvelin (HJV) levels, which leads to an increase in hepcidin levels. Subsequently, in some embodiments, hepcidin acts to inhibit iron absorption, release, and reuse, thereby reducing iron levels. In some embodiments, the method herein reduces or improves one or more symptoms associated with thalassemia. In some embodiments, the method herein reduces symptoms associated with iron overload in subjects having thalassemia (e.g., arthralgia, abdominal pain, fatigue, weakness, diabetes, heart failure, and / or hepatic failure).

[0266] In some embodiments, administration of anti-TMPRSS6 antibody in subjects with thalassemia (e.g., α-thalassemia, severe β-thalassemia, or mild β-thalassemia) results in an increase in circulating hepcidin-25 levels (e.g., at least 5%, at least 10%, at least 15%, at least 20%, at least 25%) compared to subjects before receiving TMPRSS6 antibody or subjects with thalassemia (e.g., α-thalassemia, severe β-thalassemia, or mild β-thalassemia) but who have not received anti-TMPRSS6 antibody. This results in an increase of 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 circulating hepcidin-25 levels.

[0267] In some embodiments, administration of an anti-TMPRSS6 antibody results in a reduction of hemolysis (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% reduction in hemolysis) in subjects having thalassemia (e.g., α-thalassemia, severe β-thalassemia, or mild β-thalassemia) compared to subjects before receiving the anti-TMPRSS6 antibody or subjects having thalassemia (e.g., α-thalassemia, severe β-thalassemia, or mild β-thalassemia) but not receiving the anti-TMPRSS6 antibody. Hemolysis can be evaluated by preferred known methods described elsewhere herein.

[0268] In some embodiments, administration of an anti-TMPRSS6 antibody results in a reduction in transfusion sessions (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% of transfusion sessions) in subjects with thalassemia (e.g., severe α-thalassemia, intermediate α-thalassemia, severe β-thalassemia) compared to subjects before receiving the anti-TMPRSS6 antibody or subjects with thalassemia (e.g., severe α-thalassemia, intermediate α-thalassemia, severe β-thalassemia) but without receiving the anti-TMPRSS6 antibody.

[0269] In some embodiments, the Disclosure provides a method for treating a subject having thalassemia by administering an effective dose of anti-TMPRSS6 antibody to the subject in combination with any known therapeutic agent for treating thalassemia, such as blood transfusion, iron chelation, folic acid supplements, bow marrow transplantation, stem cell transplantation, deferipron, ruspatercept, splenectomy, or gene therapy.

[0270] Further embodiments of this disclosure relate to methods and compositions (e.g., anti-TMPRSS6 antibodies) useful for treating hereditary hemochromatosis (HH). In some embodiments, the methods provided herein involve administering an effective amount of anti-TMPRSS6 antibody to a subject for treating HH in that subject. In some embodiments, the methods provided herein involve administering an effective amount of anti-TMPRSS6 antibody to a subject for the purpose of treating HH in a subject having iron overload. In some embodiments, the methods provided herein reduce iron overload in a subject having HH. In some embodiments, the methods provided herein involve administering an effective amount of anti-TMPRSS6 antibody to a subject for the purpose of treating and / or improving symptoms and conditions associated with HH. Hereditary hemochromatosis (HH) includes several genetic disorders that can lead to progressive iron overload. There are four main types of HH, classified based on the proteins involved in iron homeostasis that 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 frequent hereditary form of iron overload. In some embodiments, type 1 HH results from a G to A transition at nucleotide 845 of the HFE gene, resulting in a cysteine-to-tyrosine substitution at amino acid 282 (C282Y), also known as type 1a HH. In other embodiments, type 1 HH results from an H63D mutation. In some embodiments, 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 H63D hemochromatosis mutation, Gastroenterology. 2002 Mar;122(3):646-51).In some embodiments, type 1 HH is the result of a C282Y / H63D substitution and is classified as type HH 1b. In some embodiments, subjects with type HH 1b rarely develop clinically significant iron overload 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 embodiments, type 1 HH is the result of an S65C mutation in the HFE gene and is called type 1c HH. Generally, S65C mutations are considered to be polymorphisms of no clinical significance.

[0271] Type 2 HH, also known as juvenile hemochromatosis, is associated with mutations in the HJV gene (type 2A) or the hepatic antimicrobial protein (HAMP) gene (type 2B), respectively, 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 embodiments, type 2 HH tends to lead to the most severe form of primary iron overload. In some embodiments, type 2 HH occurs primarily in young individuals.

[0272] Type 3 HH is associated with mutations in the transferrin receptor 2 (TFR2) gene, leading 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 a decrease in hepcidin transcription (see, for example, Christal et al., The role of hepatic transferrin receptor 2 in the regulation of iron homeostasis in the body, Front Pharmacol. 2014; 5: 34). In some embodiments, iron overload is present in type 3 HH, but is considerably less severe than in type 2 HH.

[0273] Type 4 HH is the result of a mutation in the ferroportin 1 (FPN1 gene). Type 4A HH, also known as FPN disease, is an autosomal dominant form of hemochromatosis caused by a mutation in the FPN1 gene (SLC40A1) (Abboud et al., novel mammalian iron-regulated protein involved in intracellular iron metabolism, J Biol Chem. 2000 Jun 30;275(26):19906-12). In type 4A HH, hepcidin production is normal, but FPN1 transport function is impaired, leading to intracellular iron retention, low plasma iron levels, and normal or low transferrin saturation, but elevated serum ferritin levels (33). The spleen is the organ most affected in type 4A HH due to high FPN1 activity at the macrophage level. Type 4B HH is a form of iron overload due to FPN1's resistance to hepcidin.

[0274] In some embodiments, the method provided herein involves administering an effective amount of anti-TMPRSS6 antibody to a subject to treat HH in the subject. In some embodiments, the method provided herein involves administering an effective amount of anti-TMPRSS6 antibody to a subject for the purpose of treating a subject having 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 matriptase-2 activity, subsequently increasing hemoduvelin (HJV) levels, which leads to an increase in hepcidin levels. Subsequently, in some embodiments, hepcidin acts to inhibit iron absorption, release, and reuse, thereby decreasing iron levels, and thereby reducing iron overload in a subject having 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 hereditary hemochromatosis (e.g., type 1 HH, type 2, type 3 HH, or type 4 HH). In some embodiments, the methods described herein reduce symptoms associated with iron overload in subjects having hereditary hemochromatosis (e.g., type 1 HH, type 2, type 3 HH, or type 4 HH), such as arthralgia, abdominal pain, fatigue, weakness, diabetes, decreased libido, impotence, heart failure, liver failure, bronze or gray skin color, and memory fog.

[0275] In some embodiments, the Disclosure provides a method for treating iron overload in subjects having hereditary hemochromatosis (e.g., type I HH, type II HH, or type IV HH) by administering an effective dose of anti-TMPRSS6 antibody to the subject in combination with any known treatment for hereditary hemochromatosis (e.g., type I HH, type II HH, type III HH, or type IV HH), such as venotomy, iron chelation (e.g., deferoxamine or deferasirox), or supportive care for complications.

[0276] Further aspects of this disclosure relate to methods and compositions (e.g., anti-TMPRSS6 antibodies) useful for treating myelodysplastic syndromes (MDS). In some embodiments, the methods provided herein involve administering an effective amount of anti-TMPRSS6 antibody to a subject for treating MDS in that subject. In some embodiments, the methods provided herein involve administering an effective amount of anti-TMPRSS6 antibody to a subject for the purpose of treating MDS in a subject having iron overload. In some embodiments, the methods provided herein reduce iron overload in a subject having MDS. In some embodiments, the methods provided herein involve administering an effective amount of anti-TMPRSS6 antibody to a subject for the purpose of treating and / or improving symptoms and conditions associated with MDS. Myelodysplastic syndromes (MDS) are a heterogeneous group of hematological neoplasms of varying severity affecting one or more hematopoietic systems. In some embodiments, ineffective erythropoiesis leads to dysregulation of iron metabolism. In some embodiments, ineffective erythropoiesis suppresses hepcidin production in the liver, and therefore leads to unrestricted intestinal iron uptake, so iron overload begins to develop in MDS patients before they become transfusion-dependent. In some embodiments, ineffective erythropoiesis, a characteristic of MDS, causes massive proliferation of myeloerythroblasts 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 creates a high demand for iron, leading to suppression of hepcidin. In some embodiments, hepcidin levels are heterogeneous across different MDS subtypes. In some embodiments, the lowest hepcidin levels have been observed in refractory anemia with ring sideroblasts (RARS). In some embodiments, RARS patients have high levels of toxic non-transferrin-bound iron compared to patients with other types of MDS that have high hepcidin levels.Furthermore, in some embodiments, tissue hypoxia resulting from ineffective erythropoiesis leads to increased erythropoietin production, followed by low hepcidin levels, and then iron overload (Cui et al., Serum iron metabolism and erythropoiesis in patients with myelodysplastic syndrome not receiving RBC transfusions. Leukemia research. 2014;38:545-550). Therefore, in some embodiments, the present disclosure provides methods and compositions for treating refractory anemia with ring sideroblasts (RARS). In some embodiments, the methods provided herein involve administering an effective amount of anti-TMPRSS6 antibody to a subject for the purpose of treating and / or improving symptoms and conditions associated with RARS. In some embodiments, the methods provided herein involve administering an effective amount of anti-TMPRSS6 antibody to a subject for the purpose of reducing iron overload associated with RARS. In some embodiments, administration of anti-TMPRSS6 antibody results in increased hepcidin levels in a subject having MDS (e.g., RARS). In some embodiments, administration of an anti-TMPRSS6 antibody results in a reduction of iron overload in subjects with MDS (e.g., RARS).

[0277] In some embodiments, subjects with MDS have a mutation in the SF3B1 gene. SF3B1 is a gene that encodes a component of the RNA splicing mechanism, and mutations in this gene have been found in various types of MDS, including RARS. In some embodiments, MDS patients with SF3B1 mutations have a significantly lower hepcidin-to-ferritin ratio compared to those 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 erythrocyte neovitability in MDS patients with SF3B1 mutations leads to low hepcidin levels in MDS-RARS patients with SF3B1 mutations, ultimately leading to excessive iron release from RES and iron overload in the parenchyma. Therefore, in some embodiments, this disclosure provides methods and compositions for treating MDS with SF3B1 mutations. 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 MDS having an SF3B1 mutation. In some embodiments, the methods provided herein include administering an effective amount of anti-TMPRSS6 antibody to a subject for the purpose of reducing iron excess associated with MDS having an SF3B1 mutation. In some embodiments, administration of anti-TMPRSS6 antibody results in an increase in hepcidin levels in a subject having MDS (e.g., MDS having an SF3B1 mutation). In some embodiments, administration of anti-TMPRSS6 antibody results in a decrease in iron excess in a subject having MDS (e.g., MDS having an SF3B1 mutation).

[0278] In some embodiments, RARS patients have a high prevalence 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. Leukemia research. 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. Accordingly, in some embodiments, this disclosure provides methods and compositions for treating RARS with SF3B1 mutations. In some embodiments, the methods provided herein involve administering an effective amount of anti-TMPRSS6 antibody to a subject for the purpose of treating and / or improving symptoms and conditions associated with RARS with SF3B1 mutations. In some embodiments, the methods provided herein involve administering an effective amount of anti-TMPRSS6 antibody to a subject for the purpose of reducing iron overload associated with RARS having an SF3B1 mutation. In some embodiments, administration of anti-TMPRSS6 antibody results in an increase in hepcidin levels in a subject having MDS (e.g., RARS having an SF3B1 mutation). In some embodiments, administration of anti-TMPRSS6 antibody results in a decrease in iron overload in a subject having MDS (e.g., RARS having an SF3B1 mutation).

[0279] Furthermore, in some embodiments, MDS patients have anemia, and some require regular red blood cell transfusions. Chronic transfusion therapy is also a major cause of iron overload in patients with MDS. In some embodiments, MDS patients with iron overload had reduced overall survival and poor outcomes after allogeneic stem cell transplantation compared to those without iron overload (see, for example, 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 embodiments, the method provided herein involves targeting an effective dose of anti-TMPRSS6 antibody for the purpose of reducing transfusion-associated iron overload in MDS patients.

[0280] In some embodiments, the methods provided herein improve symptoms and / or clinical outcomes associated with iron overload, such as survival, cardiovascular function, hepatic function, and immune function, in subjects having MDS (e.g., RARS, MDS with SF3B1, RARS with SF3B1).

[0281] In some embodiments, the Disclosure provides a method for treating subjects having MDS (e.g., RARS, MDS with SF3B1, RARS with SF3B1) by administering an effective dose of anti-TMPRSS6 antibody to the subject in combination with any known therapeutic agent 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, ruspatercept, granulocyte colony-stimulating factor (GCSF), anti-thymocyte globulin (ATG), cyclosporine (CSA), hypomethylating agent (HMA), lenalidomide (LEN), thrombopoietin receptor agonist (TPO-RA), etc.

[0282] Further embodiments of this disclosure relate to methods and compositions (e.g., anti-TMPRSS6 antibodies) useful for treating hemolytic anemia (e.g., transfusion-dependent hemolytic anemia, pyruvate kinase deficiency hemolytic anemia). In some embodiments, hemolysis causes a massive outflow of hemoglobin into the bloodstream, subsequently leading to iron overload. In some embodiments, hemolysis induces EPO production, thereby activating erythrocyte synthesis (e.g., without ineffective erythrocyte synthesis). In some embodiments, the increase in EPO and erythrocyte synthesis suppresses hepcidin expression, thereby leading to iron overload in subjects with hemolytic anemia. Accordingly, 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 anti-TMPRSS6 antibody to a subject for the purpose of 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 for the purpose of treating hemolytic anemia in a subject having iron overload. In some embodiments, the methods provided herein reduce iron overload in a subject having hemolytic anemia. 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 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 for the purpose of reducing iron overload associated with hemolytic anemia (e.g., transfusion-dependent hemolytic anemia, pyruvate kinase deficiency hemolytic anemia). In some embodiments, administration of an anti-TMPRSS6 antibody results in an increase in hepcidin levels in subjects with hemolytic anemia (e.g., transfusion-dependent hemolytic anemia, pyruvate kinase deficiency hemolytic anemia). In some embodiments, administration of an anti-TMPRSS6 antibody results in a decrease in iron overload in subjects with hemolytic anemia (e.g., transfusion-dependent hemolytic anemia, pyruvate kinase deficiency hemolytic anemia).

[0283] Further embodiments of this disclosure relate to methods and compositions (e.g., anti-TMPRSS6 antibodies) useful for treating transfusion-associated iron overload. Transfusions have been widely used in medical practice since the early 20th century to treat various conditions (e.g., anemia and / or bleeding), and have sometimes been overused. Transfused red blood cells (RBCs) provide beneficial effects to the recipient, e.g., increasing blood volume, decreasing blood viscosity, and increasing oxygen carrying capacity. In some embodiments, transfusions (e.g., large volume transfusions) are used in subjects with blood loss (e.g., from trauma or surgery). In some embodiments, transfusions (e.g., repeated transfusions) are used in subjects with anemia (e.g., SCD, thalassemia, hemochromatosis, MDS, iron deficiency anemia, Diamond-Blackfan anemia, etc.). In some embodiments, when transfused, the subject receives excess iron. In some embodiments, excess iron from transfusions is stored in various tissues, causing iron overload. Furthermore, in some embodiments, blood transfusion causes hemolysis, thereby releasing iron into circulation and further causing iron overload. In some embodiments, therefore, in some embodiments, the present disclosure provides methods and compositions for treating iron overload associated with blood transfusion (e.g., transfusion for anemia and / or bleeding). In some embodiments, the methods provided herein involve administering an effective amount of anti-TMPRSS6 antibody to a subject to treat iron overload associated with blood transfusion (e.g., transfusion for anemia and / or bleeding). In some embodiments, the methods provided herein reduce iron overload in a subject having hemolytic anemia. In some embodiments, the methods provided herein involve administering an effective amount of anti-TMPRSS6 antibody to a subject for the purpose of reducing iron overload associated with blood transfusion (e.g., transfusion for anemia and / or bleeding). In some embodiments, administration of anti-TMPRSS6 antibody results in an increase in hepcidin levels in a subject having iron overload associated with blood transfusion (e.g., transfusion for anemia and / or bleeding). In some embodiments, administration of an 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).

[0284] Further embodiments of this disclosure relate to methods and compositions (e.g., anti-TMPRSS6 antibodies) useful for treating African iron overload. In some embodiments, the methods provided herein involve administering an effective amount of anti-TMPRSS6 antibody to a subject for African iron overload. In some embodiments, the methods provided herein reduce iron overload in a subject having African iron overload. African iron overload is iron overload resulting from an iron-rich diet and genetic factors. In some embodiments, administration of anti-TMPRSS6 antibody results in an increase in hepcidin levels in a subject having African iron overload. In some embodiments, administration of anti-TMPRSS6 antibody results in a decrease in iron overload in a subject having African iron overload.

[0285] Further embodiments of this disclosure relate to methods and compositions (e.g., anti-TMPRSS6 antibodies) useful for treating Diamond-Blackfan anemia. In some embodiments, the methods provided herein involve administering an effective amount of anti-TMPRSS6 antibody to a subject for treating Diamond-Blackfan anemia. In some embodiments, the methods provided herein involve administering an effective amount of anti-TMPRSS6 antibody to a subject for the purpose of treating Diamond-Blackfan anemia in a subject having iron overload. In some embodiments, the methods provided herein reduce iron overload in a subject. In some embodiments, the methods provided herein involve administering an effective amount of anti-TMPRSS6 antibody to a subject for the purpose of treating symptoms and conditions associated with Diamond-Blackfan anemia. Diamond-Blackfan anemia (DBA) is a hereditary blood disorder that affects the ability of the bone marrow to produce red blood cells. In some embodiments, Diamond-Blackfan anemia is caused by mutations 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, e.g., 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 anaemia with iron overload: A serious issue, 2022 Oct;199(2):171-172). Repeated transfusions, as described elsewhere herein, result in iron overload. Accordingly, in some embodiments, the methods provided herein include administering an effective amount of anti-TMPRSS6 antibody to a subject with Diamond-Blackfan anemia for the purpose of reducing transfusion-related iron overload.In some embodiments, administration of anti-TMPRSS6 antibodies results in an increase in hepcidin levels in Diamond-Blackfan anemia patients receiving blood transfusions. In some embodiments, administration of anti-TMPRSS6 antibodies results in a decrease in iron overload in Diamond-Blackfan anemia patients receiving blood transfusions.

[0286] Determining whether the amount of antibody (e.g., anti-TMPRSS6 antibody) antagonist achieved a therapeutic effect is obvious to those skilled in the art based on the teachings provided herein. The effective dose will vary, as recognized by those skilled in the art, depending on the specific condition being treated, the severity of the condition, individual patient parameters including age, health status, size, sex, and weight, the duration of treatment, the nature of any concomitant therapy, the specific route of administration, and factors within the knowledge and expertise of the healthcare professional. The specific administration regimen used in the methods described herein, i.e., dose, timing, and frequency, will depend on the specific subject and the medical history of that subject, as described herein.

[0287] Empirical considerations, such as time to maximum effect, half-life, and / or time above a certain concentration, will usually contribute to dose determination. For example, antibodies compatible with the human immune system, e.g., humanized or fully human antibodies, may be used to extend the half-life of the antibody and to prevent the antibody from being attacked by the host immune system. Other reasons for dose adjustment include differences in pharmacokinetic or pharmacodynamic responses driven by sex, age, individual responses, and polymorphisms related to antibody targets and / or receptors involved in antibody clearance. The frequency of administration may generally be determined and adjusted over the course of therapy, based on the treatment and / or suppression and / or recovery and / or delay of the target disease / impairment. Alternatively, a sustained-release formulation of the antibody may be appropriate. Various formulations and devices for achieving sustained release are known in the art.

[0288] The frequency of administration may vary according to the claimed method. In some embodiments, the composition may be administered once. In some embodiments, the composition may be administered on multiple occasions. In some embodiments, the frequency of administration may be weekly, every two weeks, every three weeks, every four weeks, every five weeks, every six weeks, every seven weeks, every eight weeks, every nine weeks, or every ten weeks; or once a month, every two months, or every three months, or more. In some embodiments, the composition may be administered daily, twice a week, weekly, twice a month, monthly, or at any time interval that provides suitable (e.g., maximum) efficacy while minimizing safety risks to the subject. Generally, efficacy and treatment and safety risks may be monitored throughout the course of treatment.

[0289] In some embodiments, a subject may be administered a composition provided herein (e.g., an anti-TMPRSS6 antibody antagonist) at intervals of one or more times over a period of time. In some cases, the periods during which the subject is administered the composition at intervals of one or more times may be divided into periods during which the subject is not administered the composition. In some embodiments, the relative duration of each period may be determined based on the subject's response to the treatment or the severity of the disease or both, and / or on the judgment of the treating physician.

[0290] In some embodiments, antibodies may be administered parenterally. For example, parenterally administered compositions may be administered topically, transmucosally, subcutaneously, intradermally, intravenously, intraperitoneally, intramuscularly, intra-arterially, intra-arterially, or by infusion techniques.

[0291] In some embodiments, the antibody (e.g., anti-TMPRSS6 antibody) is administered intravenously. In some embodiments, the antibody (e.g., anti-TMPRSS6 antibody) is administered subcutaneously.

[0292] For intravenous injection, water-soluble antibodies can be administered by drip infusion, in which a pharmaceutical formulation containing the antibody and physiologically acceptable excipients is injected. Physiologically acceptable excipients may include, for example, 5% dextrose, 0.9% saline, Ringer's solution, or other suitable excipients. Other injectable compositions may contain various carriers, such as vegetable oils, dimet...

Claims

1. 1. A method for treating sickle cell disease (SCD), (a) HC CDR1, HC CDR2, and HC CDR3 heavy chain variable domains having the amino acid sequence of SEQ ID NO: 49, and LC CDR1, LC CDR2, and LC CDR3 light chain variable domains having the amino acid sequence of SEQ ID NO: 50, (b) HC CDR1, HC CDR2, and HC CDR3 heavy chain variable domains having the amino acid sequence of SEQ ID NO: 7, and LC CDR1, LC CDR2, and LC CDR3 light chain variable domains having the amino acid sequence of SEQ ID NO: 8, (c) HC CDR1, HC CDR2, and HC CDR3 heavy chain variable domains having the amino acid sequence of SEQ ID NO: 19 or 78, and LC CDR1, LC CDR2, and LC CDR3 light chain variable domains having the amino acid sequence of SEQ ID NO: 20, (d) HC CDR1, HC CDR2, and HC CDR3 heavy chain variable domains having the amino acid sequence of SEQ ID NO: 30, and LC CDR1, LC CDR2, and LC CDR3 light chain variable domains having the amino acid sequence of SEQ ID NO: 31, (e) HC CDR1, HC CDR2, and HC CDR3 heavy chain variable domains having the amino acid sequence of SEQ ID NO: 37, and LC CDR1, LC CDR2, and LC CDR3 light chain variable domains having the amino acid sequence of SEQ ID NO: 38, or (f) HC CDR1, HC CDR2, and HC CDR3 heavy chain variable domains having the amino acid sequence of SEQ ID NO: 44, and LC CDR1, LC CDR2, and LC CDR3 light chain variable domains having the amino acid sequence of SEQ ID NO: 45 A method comprising administering an effective amount of anti-transmembrane serine protease 6 (TMPRSS6) antibody to a target.

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

50.

3. Anti-TMPRSS6 antibody, (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 LC CDR3 having the amino acid sequence of SEQ ID NO: 29 (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 LC CDR3 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 LC CDR3 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 LC CDR3 having the amino acid sequence of SEQ ID NO: 29 (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 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 LC CDR3 having the amino acid sequence of SEQ ID NO: 18 The method according to claim 1 or 2, including the method according to claim 1 or 2.

4. The method according to 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. Anti-TMPRSS6 antibody, (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 the amino acid sequence of SEQ ID NO: 19 or 78 and a light chain variable domain having the 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) Heavy chain variable domain having the amino acid sequence of SEQ ID NO: 44 and light chain variable domain having the amino acid sequence of SEQ ID NO: 45 The method according to any one of claims 1 to 4, including the method described in any one of claims 1 to 4.

6. The method according to 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. Anti-TMPRSS6 antibody, (a) 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 (b) A heavy chain containing the amino acid sequence of SEQ ID NO: 11 and a 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) A heavy chain containing the amino acid sequence of SEQ ID NO: 33 and a 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) Heavy chain containing the amino acid sequence of SEQ ID NO: 46 and light chain containing the amino acid sequence of SEQ ID NO: 47 The method according to any one of claims 1 to 6, including

8. The method according to 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 according to any one of claims 1 to 8, wherein the administration of an antibody reduces iron excess in a subject.

10. The method according to any one of claims 1 to 9, wherein the administration of an antibody reduces hemolysis in the subject compared to the subject before administration.

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

12. The method according to any one of claims 1 to 11, wherein the administration of an antibody reduces the frequency of VOCs in the subject compared to the subject before administration.

13. The method according to any one of claims 1 to 12, wherein the administration reduces the severity of VOCs compared to the subject before administration.

14. The method according to claim 13, wherein the severity of VOCs is measured by the frequency and / or duration of hospitalizations.

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

16. The method according to any one of claims 1 to 15, wherein the administration of an antibody reduces the mean cellular hemoglobin concentration (MCHC) compared to before administration.

17. The method according to any one of claims 1 to 16, wherein the administration reduces hemoglobin S (HbS) polymerization compared to the subject before administration.

18. The method according to any one of claims 1 to 17, wherein the administration reduces the frequency with which the subject requires blood transfusions compared to the subject before administration.

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

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

21. The method according to any one of claims 1 to 20, wherein an anti-TMPRSS6 antibody is administered as a combination therapy in combination with a hemoglobin S polymerization inhibitor (e.g., voxelotol), a therapeutic agent for reducing VOCs (e.g., hydroxyurea, L-glutamine oral powder, chryzanlizumab, selective pyruvate kinase-R (PKR) activator), analgesics (e.g., narcotics, opioids, gabapentin, cannabis), blood transfusion, stem cell transplantation, exagumgrogline autotemcell (exa-cel), lenticulin, GMI-1070, VIT-2763 (bamifeport), ticagrelor, vitamin D, simvastatin, AG-348 (mitapivat sulfate), propranolol, epinephrine, regadenoson, atorvastatin, prasugrel, L-arginine, OTQ923, oxygen therapy, or gene therapy.

Images