ALK2 antibodies and methods of use thereof

Abstract

To provide pharmaceutical compositions and methods for bone diseases, anemia, HO, Sjogren's syndrome, DIPG, MO, PCO, and cardiac hypertrophy and / or cardiac fibrosis.SOLUTION: The invention features ALK2 antibodies and antigen binding fragments thereof (e.g., ALK2 binding fragments). The invention also features pharmaceutical compositions and methods of using the ALK2 antibodies or antigen binding fragments thereof to treat bone disease or damage, low red blood cell levels (e.g., anemia or blood loss), heterotopic ossification (e.g., heterotopic ossification resulting from fibrodysplasia ossificans progressiva), Sjogren's syndrome (e.g., dry eye associated with Sjogren's syndrome), multiple osteochondroma, diffuse intrinsic pontine glioma, posterior capsule opacification, or cardiac hypertrophy and / or cardiac fibrosis.SELECTED DRAWING: Figure 5

Description

[Background technology]

[0001] Healthy bone undergoes constant remodeling involving both bone destruction and bone growth. Bone growth is mediated by osteoblasts, whereas osteoclasts resorb bone. Pathologies arise when these systems are imbalanced, either by downregulation of the anabolic program, upregulation of the catabolic system, or a combination of both, resulting in net bone loss. Thus, controlling the balance of bone remodeling may be useful not only for promoting healing of injuries to bone, but also for treating disorders such as osteoporosis, which are associated with bone loss and bone demineralization.

[0002] Bone damage can result from a variety of underlying causes, including bone loss associated with aging or cancer, genetic conditions, and adverse side effects of drug treatments. The World Health Organization estimates that osteoporosis alone affects 75 million people in the United States, Europe, and Japan and is a significant risk factor for bone damage. In general, the totality of bone loss represents a pathological condition with few effective treatments. Instead, treatment focuses on immobilization, exercise, and dietary changes rather than drugs that directly promote bone growth and increase bone density. For osteoporosis, estrogen, calcitonin, osteocalcin in combination with vitamin K, or high doses of calcium in the diet are all used as therapeutic interventions. Other therapeutic approaches to osteoporosis include bisphosphonates, parathyroid hormone, parathyroid hormone-related protein (PTHrP), calcimimetics, statins, anabolic steroids, lanthanum and strontium salts, and sodium fluoride. However, such therapeutic agents are often associated with undesirable side effects.

[0003] Anemia is a global health problem with health consequences that impact both morbidity and mortality. In the United States alone, the prevalence of anemia nearly doubled from 2003 to 2012. Symptoms of anemia include fatigue, weakness, shortness of breath, palpitations, and decreased cognitive ability, with children, pregnant women, women of reproductive age, and the elderly being found to be at highest risk of developing anemia. The most common form of anemia is iron deficiency anemia, but anemia can also be caused by chronic disease, blood loss, and the destruction of red blood cells. Iron deficiency anemia can be treated with iron supplements, but many other forms of anemia, such as aplastic anemia, anemia of chronic disease, and hemolytic anemia, may require blood transfusions.

[0004] Heterotopic ossification (HO) is the pathological formation of extraskeletal bone in soft tissues. This process occurs in patient populations with severe trauma, such as large-area burns, musculoskeletal injuries, orthopedic surgery, spinal cord injuries, and in those with a genetic disorder known as fibrodysplasia ossificans progressiva (FOP). FOP is caused by hyperactivating mutations in the type I bone morphogenetic protein (BMP) receptor activin A receptor type I (ACVR1), and patients with FOP can develop heterotopic bone lesions in muscles, tendons, ligaments, and other connective tissues without substantial trauma. The clinical sequelae of these pathological heterotopic bone formations, whether in the context of trauma or genetic mutations, include non-healing wounds, chronic pain, and joint immobility. In FOP, the formation of heterotopic bone can result in bony bridges that develop across joints, gradually restricting movement, and progressive ossification can result in death due to loss of compliance of the rib cage.

[0005] Treatment options for HO are limited, as bone often recurs after surgical resection, and attempts to surgically remove excess bone may result in further bone growth. Additionally, some patients may have HO in a vulnerable location that is unresectable. The risks of surgery may outweigh the benefits of resection, especially in the face of recurrence. Treatment options are needed that can prevent HO before it first occurs in at-risk patients and / or reduce the amount of HO, reduce the recurrence of HO, or prevent further ectopic bone formation in patients with FOP. Currently, there is no approved treatment for FOP.

[0006] Sjögren's syndrome is a systemic autoimmune disease identified by its two most common symptoms: dry eyes and dry mouth. It may also be characterized by joint pain, swelling, and stiffness, swollen salivary glands, skin rash or dry skin, vaginal dryness, persistent dry cough, and prolonged fatigue. Sjögren's syndrome can also cause dysfunction of organs such as the kidneys, gastrointestinal system, blood vessels, lungs, liver, pancreas, and central nervous system. It is one of the most common autoimmune diseases, with over 4 million Americans suffering from the disease, 90% of whom are women. Dry eyes associated with Sjögren's syndrome can be treated with immunosuppressants and steroid eye drops. However, steroid eye drops should not be used for long periods of time due to side effects such as increased intraocular pressure and increased susceptibility to infection.

[0007] Diffuse intrinsic pontine gliomas (DIPGs) are pediatric brain tumors originating from the pons and accounting for approximately 20% of all pediatric brain tumors. DIPGs remain incurable with a median survival of 10-11 months. The inability to identify successful treatments for DIPGs may be due to a lack of understanding of the biology of the disease, due in part to the vulnerable location of the tumors, and because biopsies were not commonly performed until recently. As a result, many trials to date have been based on genetic alterations found in adult glioblastomas, which have recently been found to be molecularly distinct from DIPGs.

[0008] Multiple osteochondromas (MO), also called congenital multiple exostoses, is a rare genetic disorder characterized by the development of multiple benign (non-cancerous) bone tumors called osteochondromas, often in the growing ends (diaphysis) of the long bones of the legs, arms, and fingers, and in flat bones such as the hips and scapula. The number of osteochondros and the bones in which they are located vary widely among affected individuals. Osteochondromas are not present at birth, but approximately 96% of affected individuals develop multiple osteochondromas by the age of 12. These osteochondromas usually continue to grow until just after puberty and can cause bone deformities, skeletal abnormalities, limb length discrepancies, short stature, pain, reduced range of motion, and pressure on nerves, blood vessels, spinal cord, and tissues surrounding the osteochondromas. Congenital multiple osteochondromas are inherited as an autosomal dominant disorder and are associated with mutations in the exostosin-1 (EXT1) or exostosin-2 (EXT2) genes. Current treatment options include a "watch and wait" approach (if there are no symptoms), surgical removal of the osteochondroma, corrective osteotomy, and epiphyseal closure or limb lengthening.

[0009] Posterior capsule opacification (PCO) is the most frequent complication associated with decreased vision after cataract surgery and intraocular lens implantation. During cataract surgery, the eye's natural lens is replaced with an artificial intraocular lens, which is placed inside the lens capsule. PCO occurs when lens epithelial cells migrate toward the posterior capsule, proliferate and differentiate, and grow to the back of the capsule, thickening it and making it slightly opaque. This reduces the amount of light that can reach the retina, which can result in blurred or cloudy vision, or problems with bright light and glare. PCO can be treated using Nd:YAG laser capsulotomy, but the potential complications and the considerable cost of treatment underscore the importance of identifying new PCO therapies.

[0010] Cardiac hypertrophy is an abnormal enlargement or thickening of the myocardium due to an increase in the size of cardiomyocytes and changes in other myocardial components such as the extracellular matrix. Cardiac hypertrophy can be physiological, occurring in response to exercise or pregnancy, or pathological, occurring in response to stress (e.g., hemodynamic stress), such as hypertension or valvular heart disease. Cardiac fibrosis may also be characterized by abnormal thickening of the heart valves due to inappropriate proliferation of cardiac fibroblasts or excessive deposition of extracellular matrix in the myocardium. This results in stiffer, less compliant muscle and may occur during the progression to heart failure. There is a lack of effective therapies to reduce or reverse cardiac fibrosis, and existing therapies for cardiac hypertrophy, such as adrenergic antagonists, renin-angiotensin system modulators such as angiotensin-converting enzyme (ACE) inhibitors, and angiotensin receptor blockers, may result in a ceiling effect characterized by lack of efficacy or even regression in some patients.

[0011] There is a need for new and effective treatments for bone disease, anemia, HO (e.g., HO due to FOP), Sjogren's syndrome (e.g., dry eye associated with Sjogren's syndrome), DIPG, MO, PCO, and cardiac hypertrophy and / or cardiac fibrosis. Summary of the Invention

[0012] Described herein are activin receptor-like kinase-2 (ALK2, also known as activin A receptor type I (ACVR1)) antibodies, such as ALK2 neutralizing antibodies and ALK2-binding fragments thereof. The ALK2 antibodies and antigen-binding fragments thereof described herein can be formulated into pharmaceutical compositions for administration to a subject, such as a human subject. The ALK2 antibodies or antigen-binding fragments thereof and compositions described herein can be administered to a subject, such as a human subject, to treat or prevent a bone disease (e.g., a disease or condition involving bone damage, such as primary osteoporosis, secondary osteoporosis, osteopenia, osteopetrosis, bone fracture, bone loss associated with bone cancer or cancer metastasis, Paget's disease, renal osteodystrophy, treatment-related bone loss, diet-related bone loss, bone loss associated with the treatment of obesity, low gravity-related bone loss, or bone loss associated with immobility), low red blood cell levels (e.g., low hemoglobin levels or low red blood cell count, e.g., anemia), heterotopic ossification (HO), e.g., heterotopic ossification due to fibrodysplasia ossificans progressiva (FOP), Sjogren's syndrome (e.g., dry eye associated with Sjogren's syndrome), diffuse intrinsic pontine glioma (DIPG), multiple osteochondromas (MO), posterior capsule opacification (PCO), or cardiac hypertrophy and / or cardiac fibrosis.

[0013] Exemplary embodiments of the present invention are described in the paragraphs listed below. E1. An isolated antibody, or an ALK2-binding fragment thereof, comprising: (1) a light chain variable domain, the light chain complementarity determining region (CDR) 1 comprising or consisting of an amino acid sequence selected from SGSSSNIGSNYVS (SEQ ID NO:1) and SGDX1X2X3X4X5X6X7X8 (SEQ ID NO:2, wherein X1 is S or N, X2 is I or L, X3 is P, G, or R, X4 is S, T, or K, X5 is F, K, or Y, X6 is F, Y, or S, X7 is A or V, and X8 is S, Y, or H); a light chain CDR2 comprising or consisting of the amino acid sequence X1X2IYX3X4X5X6RPS (SEQ ID NO:3, where X1 is V or L, X2 is V or L, X3 is K, R, G or Y, X4 is N or D, X5 is N or S, and X6 is H, N, D, or K); and a light chain CDR2 comprising or consisting of an amino acid sequence selected from ASWDHSDRFYV (SEQ ID NO:4), YVTAPWKSIW (SEQ ID NO:5), YSADAQQMKA (SEQ ID NO:6), QVYASVHRM (SEQ ID NO:7), and QTYDWSHFGW (SEQ ID NO:8). and (2) a heavy chain CDR1 comprising or consisting of the amino acid sequence GX1TFX2SX3X4X5X6 (SEQ ID NO: 9, where X1 is G or F, X2 is S or N, X3 is Y, H, S, or A, X4 is G or A, X5 is V, M, or I, and X6 is S or H); No. 11, where Xi is A or R, X2 is S or G, and X3 is G or Y), and a heavy chain CDR2 comprising or consisting of an amino acid sequence selected from WVSVISSDGGSTYYADSVKG (SEQ ID NO: 12); and a heavy chain CDR3 comprising or consisting of an amino acid sequence selected from EIGSLDI (SEQ ID NO: 13), DYGVAFAY (SEQ ID NO: 14), DYGGLKFDY (SEQ ID NO: 15), GPTQAIHYFAY (SEQ ID NO: 16), and AGFILGSLGVAWMDV (SEQ ID NO: 17).

[0014] E2. The isolated antibody of E1, or an ALK2-binding fragment thereof, wherein the light chain CDR1 comprises or consists of the sequence of SEQ ID NO:2, and X1 is S. E3. The isolated antibody of E1, or an ALK2-binding fragment thereof, wherein the light chain CDR1 comprises or consists of the sequence of SEQ ID NO:2 and X1 is N.

[0015] E4. An isolated antibody of any one of E1-E3, or an ALK2-binding fragment thereof, wherein the light chain CDR1 comprises or consists of the sequence of SEQ ID NO:2 and X2 is I. E5. An isolated antibody of any one of E1-E3, or an ALK2-binding fragment thereof, wherein the light chain CDR1 comprises or consists of the sequence of SEQ ID NO:2 and X2 is L.

[0016] E6. An isolated antibody of any one of E1-E5, or an ALK2-binding fragment thereof, wherein the light chain CDR1 comprises or consists of the sequence of SEQ ID NO:2 and X3 is P. E7. An isolated antibody of any one of E1-E5, or an ALK2-binding fragment thereof, wherein the light chain CDR1 comprises or consists of the sequence of SEQ ID NO:2 and X3 is G.

[0017] E8. An isolated antibody of any one of E1-E5, or an ALK2-binding fragment thereof, wherein the light chain CDR1 comprises or consists of the sequence of SEQ ID NO:2, and X3 is R. E9. An isolated antibody of any one of E1-E8, or an ALK2-binding fragment thereof, wherein the light chain CDR1 comprises or consists of the sequence of SEQ ID NO:2 and X4 is S.

[0018] E10. An isolated antibody of any one of E1-E8, or an ALK2-binding fragment thereof, wherein the light chain CDR1 comprises or consists of the sequence of SEQ ID NO:2, and X4 is T. E11. An isolated antibody of any one of E1-E8, or an ALK2-binding fragment thereof, wherein the light chain CDR1 comprises or consists of the sequence of SEQ ID NO:2 and X4 is K.

[0019] E12. An isolated antibody of any one of E1-E11, or an ALK2-binding fragment thereof, wherein the light chain CDR1 comprises or consists of the sequence of SEQ ID NO:2, and X5 is F. E13. An isolated antibody of any one of E1-E11, or an ALK2-binding fragment thereof, wherein the light chain CDR1 comprises or consists of the sequence of SEQ ID NO:2 and X5 is K.

[0020] E14. The isolated antibody of any one of E1-E11, or an ALK2-binding fragment thereof, wherein the light chain CDR1 comprises or consists of the sequence of SEQ ID NO:2, and X5 is Y. E15. An isolated antibody of any one of E1-E14, or an ALK2-binding fragment thereof, wherein the light chain CDR1 comprises or consists of the sequence of SEQ ID NO:2, and X6 is F.

[0021] E16. An isolated antibody of any one of E1-E14, or an ALK2-binding fragment thereof, wherein the light chain CDR1 comprises or consists of the sequence of SEQ ID NO:2, and X6 is Y. E17. An isolated antibody of any one of E1-E14, or an ALK2-binding fragment thereof, wherein the light chain CDR1 comprises or consists of the sequence of SEQ ID NO:2, and X6 is S.

[0022] E18. An isolated antibody of any one of E1-E17, or an ALK2-binding fragment thereof, wherein the light chain CDR1 comprises or consists of the sequence of SEQ ID NO:2, and X7 is A. E19. An isolated antibody of any one of E1-E17, or an ALK2-binding fragment thereof, wherein the light chain CDR1 comprises or consists of the sequence of SEQ ID NO:2 and X7 is V.

[0023] E20. An isolated antibody of any one of E1-E19, or an ALK2-binding fragment thereof, wherein the light chain CDR1 comprises or consists of the sequence of SEQ ID NO:2 and X8 is S. E21. An isolated antibody of any one of E1-E19, or an ALK2-binding fragment thereof, wherein the light chain CDR1 comprises or consists of the sequence of SEQ ID NO:2, and X8 is Y.

[0024] E22. An isolated antibody of any one of E1-E19, or an ALK2-binding fragment thereof, wherein the light chain CDR1 comprises or consists of the sequence of SEQ ID NO:2, X7 is A or V, and X8 is H.

[0025] E23. An isolated antibody of any one of E1-E22, or an ALK2-binding fragment thereof, wherein the light chain CDR2 comprises or consists of the sequence of SEQ ID NO:3 and X1 is V. E24. The isolated antibody of any one of E1-E22, or an ALK2-binding fragment thereof, wherein the light chain CDR2 comprises or consists of the sequence of SEQ ID NO:3, and X1 is L.

[0026] E25. The isolated antibody of any one of E1-E24, or an ALK2-binding fragment thereof, wherein the light chain CDR2 comprises or consists of the sequence of SEQ ID NO:3 and X2 is V. E26. The isolated antibody of any one of E1-E24, or an ALK2-binding fragment thereof, wherein the light chain CDR2 comprises or consists of the sequence of SEQ ID NO:3, and X2 is L.

[0027] E27. The isolated antibody of any one of E1-E26, or an ALK2-binding fragment thereof, wherein the light chain CDR2 comprises or consists of the sequence of SEQ ID NO:3, and X3 is K. E28. The isolated antibody of any one of E1-E26, or an ALK2-binding fragment thereof, wherein the light chain CDR2 comprises or consists of the sequence of SEQ ID NO:3, and X3 is R.

[0028] E29. The isolated antibody of any one of E1-E26, or an ALK2-binding fragment thereof, wherein the light chain CDR2 comprises or consists of the sequence of SEQ ID NO:3, and X3 is G. E30. The isolated antibody of any one of E1-E26, or an ALK2-binding fragment thereof, wherein the light chain CDR2 comprises or consists of the sequence of SEQ ID NO:3, and X3 is Y.

[0029] E31. An isolated antibody of any one of E1-E30, or an ALK2-binding fragment thereof, wherein the light chain CDR2 comprises or consists of the sequence of SEQ ID NO:3 and X4 is N. E32. An isolated antibody of any one of E1-E30, or an ALK2-binding fragment thereof, wherein the light chain CDR2 comprises or consists of the sequence of SEQ ID NO:3 and X4 is D.

[0030] E33. An isolated antibody of any one of E1-E32, or an ALK2-binding fragment thereof, wherein the light chain CDR2 comprises or consists of the sequence of SEQ ID NO:3, and X5 is N. E34. An isolated antibody of any one of E1-E32, or an ALK2-binding fragment thereof, wherein the light chain CDR2 comprises or consists of the sequence of SEQ ID NO:3, and X5 is S.

[0031] E35. An isolated antibody of any one of E1-E34, or an ALK2-binding fragment thereof, wherein the light chain CDR2 comprises or consists of the sequence of SEQ ID NO:3 and X6 is H. E36. An isolated antibody of any one of E1-E34, or an ALK2-binding fragment thereof, wherein the light chain CDR2 comprises or consists of the sequence of SEQ ID NO:3 and X6 is N.

[0032] E37. An isolated antibody of any one of E1-E34, or an ALK2-binding fragment thereof, wherein the light chain CDR2 comprises or consists of the sequence of SEQ ID NO:3, and X6 is D. E38. An isolated antibody of any one of E1-E34, or an ALK2-binding fragment thereof, wherein the light chain CDR2 comprises or consists of the sequence of SEQ ID NO:3 and X6 is K.

[0033] E39. An isolated antibody of any one of E1-E38, or an ALK2-binding fragment thereof, wherein the heavy chain CDR1 comprises or consists of the sequence of SEQ ID NO: 9 and X1 is G. E40. An isolated antibody of any one of E1-E38, or an ALK2-binding fragment thereof, wherein the heavy chain CDR1 comprises or consists of the sequence of SEQ ID NO: 9, and X1 is F.

[0034] E41. An isolated antibody of any one of E1-E40, or an ALK2-binding fragment thereof, wherein the heavy chain CDR1 comprises or consists of the sequence of SEQ ID NO: 9, and X2 is S. E42. An isolated antibody of any one of E1-E40, or an ALK2-binding fragment thereof, wherein the heavy chain CDR1 comprises or consists of the sequence of SEQ ID NO: 9 and X2 is N.

[0035] E43. An isolated antibody of any one of E1-E42, or an ALK2-binding fragment thereof, wherein the heavy chain CDR1 comprises or consists of the sequence of SEQ ID NO: 9, and X3 is Y. E44. An isolated antibody of any one of E1-E42, or an ALK2-binding fragment thereof, wherein the heavy chain CDR1 comprises or consists of the sequence of SEQ ID NO: 9 and X3 is H.

[0036] E45. An isolated antibody of any one of E1-E42, or an ALK2-binding fragment thereof, wherein the heavy chain CDR1 comprises or consists of the sequence of SEQ ID NO: 9, and X3 is S. E46. The isolated antibody of any one of E1-E42, or an ALK2-binding fragment thereof, wherein the heavy chain CDR1 comprises or consists of the sequence of SEQ ID NO: 9 and X3 is A.

[0037] E47. The isolated antibody of any one of E1-E46, or an ALK2-binding fragment thereof, wherein the heavy chain CDR1 comprises or consists of the sequence of SEQ ID NO: 9 and X4 is G. E48. The isolated antibody of any one of E1-E46, or an ALK2-binding fragment thereof, wherein the heavy chain CDR1 comprises or consists of the sequence of SEQ ID NO: 9 and X4 is A.

[0038] E49. An isolated antibody of any one of E1-E48, or an ALK2-binding fragment thereof, wherein the heavy chain CDR1 comprises or consists of the sequence of SEQ ID NO: 9 and X5 is V. E50. An isolated antibody of any one of E1-E48, or an ALK2-binding fragment thereof, wherein the heavy chain CDR1 comprises or consists of the sequence of SEQ ID NO: 9, and X5 is M.

[0039] E51. An isolated antibody of any one of E1 to E48, or an ALK2-binding fragment thereof, wherein the heavy chain CDR1 comprises or consists of the sequence of SEQ ID NO: 9, and X5 is I. E52. An isolated antibody of any one of E1-E51, or an ALK2-binding fragment thereof, wherein the heavy chain CDR1 comprises or consists of the sequence of SEQ ID NO: 9 and X6 is S.

[0040] E53. An isolated antibody of any one of E1-E51, or an ALK2-binding fragment thereof, wherein the heavy chain CDR1 comprises or consists of the sequence of SEQ ID NO: 9 and X6 is H. E54. An isolated antibody of any one of E1-E53, or an ALK2-binding fragment thereof, wherein the heavy chain CDR2 comprises or consists of the sequence of SEQ ID NO: 10, and X1 is G.

[0041] E55. An isolated antibody of any one of E1-E53, or an ALK2-binding fragment thereof, wherein the heavy chain CDR2 comprises or consists of the sequence of SEQ ID NO: 10, and X1 is R.

[0042] E56. An isolated antibody of any one of E1-E55, or an ALK2-binding fragment thereof, wherein the heavy chain CDR2 comprises or consists of the sequence of SEQ ID NO: 10, and X2 is H.

[0043] E57. An isolated antibody of any one of E1-E55, or an ALK2-binding fragment thereof, wherein the heavy chain CDR2 comprises or consists of the sequence of SEQ ID NO: 10, and X2 is D.

[0044] E58. An isolated antibody of any one of E1-E57, or an ALK2-binding fragment thereof, wherein the heavy chain CDR2 comprises or consists of the sequence of SEQ ID NO: 10, and X3 is I.

[0045] E59. An isolated antibody of any one of E1 to E57, or an ALK2-binding fragment thereof, wherein the heavy chain CDR2 comprises or consists of the sequence of SEQ ID NO: 10, and X3 is T.

[0046] E60. The isolated antibody of any one of E1-E53, or an ALK2-binding fragment thereof, wherein the heavy chain CDR2 comprises or consists of the sequence of SEQ ID NO: 11, and X1 is A.

[0047] E61. The isolated antibody of any one of E1-E53, or an ALK2-binding fragment thereof, wherein the heavy chain CDR2 comprises or consists of the sequence of SEQ ID NO: 11, and X1 is R.

[0048] E62. An isolated antibody of any one of E1-E53, E60, and E61, or an ALK2-binding fragment thereof, wherein the heavy chain CDR2 comprises or consists of the sequence of SEQ ID NO:11, and X2 is S.

[0049] E63. An isolated antibody of any one of E1-E53, E60, and E61, or an ALK2-binding fragment thereof, wherein the heavy chain CDR2 comprises or consists of the sequence of SEQ ID NO: 11, and X2 is G.

[0050] E64. An isolated antibody of any one of E1 to E53, and E60 to E63, or an ALK2-binding fragment thereof, wherein the heavy chain CDR2 comprises or consists of the sequence of SEQ ID NO: 11, and X3 is G.

[0051] E65. An isolated antibody of any one of E1 to E53, and E60 to E63, or an ALK2-binding fragment thereof, wherein the heavy chain CDR2 comprises or consists of the sequence of SEQ ID NO: 11, and X3 is Y.

[0052] E66. The isolated antibody of E1, or an ALK2-binding fragment thereof, wherein the light chain CDR1 comprises or consists of the sequence SGSSSNIGSNYVS (SEQ ID NO:1). E67. The isolated antibody of E1, or an ALK2-binding fragment thereof, wherein the light chain CDR1 comprises or consists of the sequence SGDSIPSFFAS (SEQ ID NO: 18).

[0053] E68. The isolated antibody of E1, or an ALK2-binding fragment thereof, wherein the light chain CDR1 comprises or consists of the sequence SGDNIGTKYAY (SEQ ID NO: 19). E69. The isolated antibody of E1, or an ALK2-binding fragment thereof, wherein the light chain CDR1 comprises or consists of the sequence SGDNLRKYSAH (SEQ ID NO: 20).

[0054] E70. The isolated antibody of E1, or an ALK2-binding fragment thereof, wherein the light chain CDR1 comprises or consists of the sequence SGDSLGSKSVH (SEQ ID NO:21). E71. An isolated antibody of any one of E1 and E66 to E70, or an ALK2-binding fragment thereof, wherein the light chain CDR2 comprises or consists of the sequence VLIYKNNHRPS (SEQ ID NO: 24).

[0055] E72. An isolated antibody of any one of E1 and E66 to E70, or an ALK2-binding fragment thereof, wherein the light chain CDR2 comprises or consists of the sequence LVIYRDSNRPS (SEQ ID NO: 25).

[0056] E73. An isolated antibody of any one of E1 and E66 to E70, or an ALK2-binding fragment thereof, wherein the light chain CDR2 comprises or consists of the sequence LVIYGDSDRPS (SEQ ID NO: 26).

[0057] E74. An isolated antibody of any one of E1 and E66 to E70, or an ALK2-binding fragment thereof, wherein the light chain CDR2 comprises or consists of the sequence LVIYYDNKRPS (SEQ ID NO:27).

[0058] E75. An isolated antibody of any one of E1 and E66 to E70, or an ALK2-binding fragment thereof, wherein the light chain CDR2 comprises or consists of the sequence LVIYRDSKRPS (SEQ ID NO:28).

[0059] E76. An isolated antibody of any one of E1 and E66 to E75, or an ALK2-binding fragment thereof, wherein the light chain CDR3 comprises or consists of the sequence ASWDHSDRFYV (SEQ ID NO: 4).

[0060] E77. An isolated antibody of any one of E1 and E66 to E75, or an ALK2-binding fragment thereof, wherein the light chain CDR3 comprises or consists of the sequence YVTAPWKSIW (SEQ ID NO:5).

[0061] E78. An isolated antibody of any one of E1 and E66 to E75, or an ALK2-binding fragment thereof, wherein the light chain CDR3 comprises or consists of the sequence YSADAQQMKA (SEQ ID NO: 6).

[0062] E79. An isolated antibody of any one of E1 and E66 to E75, or an ALK2-binding fragment thereof, wherein the light chain CDR3 comprises or consists of the sequence QVYASVHRM (SEQ ID NO: 7).

[0063] E80. An isolated antibody of any one of E1 and E66 to E75, or an ALK2-binding fragment thereof, wherein the light chain CDR3 comprises or consists of QTYDWSHFGW (SEQ ID NO: 8).

[0064] E81. An isolated antibody of any one of E1 and E66 to E80, or an ALK2-binding fragment thereof, wherein the heavy chain CDR1 comprises or consists of the sequence GGTFSSYGVS (SEQ ID NO: 31).

[0065] E82. An isolated antibody of any one of E1 and E66 to E80, or an ALK2-binding fragment thereof, wherein the heavy chain CDR1 comprises or consists of the sequence GFTFSSHAMS (SEQ ID NO: 32).

[0066] E83. An isolated antibody of any one of E1 and E66 to E80, or an ALK2-binding fragment thereof, wherein the heavy chain CDR1 comprises or consists of the sequence GFTNFSSAMS (SEQ ID NO: 33).

[0067] E84. An isolated antibody of any one of E1 and E66 to E80, or an ALK2-binding fragment thereof, wherein the heavy chain CDR1 comprises or consists of the sequence GGTFSSYAIH (SEQ ID NO: 34).

[0068] E85. An isolated antibody of any one of E1 and E66 to E80, or an ALK2-binding fragment thereof, wherein the heavy chain CDR1 comprises or consists of the sequence GFTFSSAAMH (SEQ ID NO: 35).

[0069] E86. An isolated antibody of any one of E1 and E66 to E85, or an ALK2-binding fragment thereof, wherein the heavy chain CDR2 comprises or consists of the sequence WMGGIIPHFGIANYAQKFQG (SEQ ID NO: 36).

[0070] E87. An isolated antibody of any one of E1 and E66 to E85, or an ALK2-binding fragment thereof, wherein the heavy chain CDR2 comprises or consists of the sequence WVGRIKSKADSGTTDYAAPVKG (SEQ ID NO: 37).

[0071] E88. An isolated antibody of any one of E1 and E66 to E85, or an ALK2-binding fragment thereof, wherein the heavy chain CDR2 comprises or consists of the sequence WVGRIKSKRDGYTTDYAAPVKG (SEQ ID NO: 38).

[0072] E89. An isolated antibody of any one of E1 and E66 to E85, or an ALK2-binding fragment thereof, wherein the heavy chain CDR2 comprises or consists of the sequence WMGRIIPDFGTANYAQKFQG (SEQ ID NO: 39).

[0073] E90. An isolated antibody of any one of E1 and E66 to E85, or an ALK2-binding fragment thereof, wherein the heavy chain CDR2 comprises or consists of the sequence WVSVISSDGGSTYYADSVKG (SEQ ID NO: 12).

[0074] E91. An isolated antibody of any one of E1 and E66 to E90, or an ALK2-binding fragment thereof, wherein the heavy chain CDR3 comprises or consists of the sequence EIGSLDI (SEQ ID NO: 13).

[0075] E92. An isolated antibody of any one of E1 and E66 to E90, or an ALK2-binding fragment thereof, wherein the heavy chain CDR3 comprises or consists of the sequence DYGVAFAY (SEQ ID NO: 14).

[0076] E93. An isolated antibody of any one of E1 and E66 to E90, or an ALK2-binding fragment thereof, wherein the heavy chain CDR3 comprises or consists of the sequence DYGGLKFDY (SEQ ID NO: 15).

[0077] E94. An isolated antibody of any one of E1 and E66 to E90, or an ALK2-binding fragment thereof, wherein the heavy chain CDR3 comprises or consists of the sequence GPTQAIHYFAY (SEQ ID NO: 16).

[0078] E95. An isolated antibody of any one of E1 and E66 to E90, or an ALK2-binding fragment thereof, wherein the heavy chain CDR3 comprises or consists of the sequence AGFILGSLGVAWMDV (SEQ ID NO: 17).

[0079] E96. The isolated antibody of E1, or an ALK2-binding fragment thereof, wherein the light chain CDR2 comprises or consists of the sequence LVIYX1DX2X3RPS (sequence number 22, where X1 is R, G, or Y, X2 is S or N, and X3 is N, D, or K).

[0080] E97. The isolated antibody of E96, or an ALK2-binding fragment thereof, wherein the light chain CDR2 comprises or consists of the sequence of SEQ ID NO: 22, and X1 is R. E98. The isolated antibody of E96, or an ALK2-binding fragment thereof, wherein the light chain CDR2 comprises or consists of the sequence of SEQ ID NO: 22 and X1 is G.

[0081] E99. The isolated antibody of E96, or an ALK2-binding fragment thereof, wherein the light chain CDR2 comprises or consists of the sequence of SEQ ID NO: 22 and X1 is Y. E100. The isolated antibody of any one of E96 to E99, or an ALK2-binding fragment thereof, wherein the light chain CDR2 comprises or consists of the sequence of SEQ ID NO: 22 and X2 is S.

[0082] E101. The isolated antibody of any one of E96 to E99, or an ALK2-binding fragment thereof, wherein the light chain CDR2 comprises or consists of the sequence of SEQ ID NO: 22 and X2 is N.

[0083] E102. An isolated antibody of any one of E96 to E101, or an ALK2-binding fragment thereof, wherein the light chain CDR2 comprises or consists of the sequence of SEQ ID NO: 22 and X3 is N.

[0084] E103. An isolated antibody of any one of E96 to E101, or an ALK2-binding fragment thereof, wherein the light chain CDR2 comprises or consists of the sequence of SEQ ID NO: 22 and X3 is D.

[0085] E104. An isolated antibody of any one of E96 to E101, or an ALK2-binding fragment thereof, wherein the light chain CDR2 comprises or consists of the sequence of SEQ ID NO: 22 and X3 is K.

[0086] E105. The isolated antibody of E1, or an ALK2-binding fragment thereof, wherein the light chain CDR2 comprises or consists of the sequence LVIYRDSX1RPS (SEQ ID NO: 23, where X1 is N or K).

[0087] E106. The isolated antibody of E105, or an ALK2-binding fragment thereof, wherein the light chain CDR2 comprises or consists of the sequence of SEQ ID NO: 23, and X1 is N. E107. The isolated antibody of E105, or an ALK2-binding fragment thereof, wherein the light chain CDR2 comprises or consists of the sequence of SEQ ID NO: 23, and X1 is K.

[0088] E108. The isolated antibody of E1, or an ALK2-binding fragment thereof, wherein the heavy chain CDR1 comprises or consists of the sequence GFTFSSX1AMX2 (SEQ ID NO: 29, where X1 is H or A and X2 is S or H).

[0089] E109. The isolated antibody of E108, or an ALK2-binding fragment thereof, wherein the heavy chain CDR1 comprises or consists of the sequence of SEQ ID NO: 29, and X1 is H. E110. The isolated antibody of E108, or an ALK2-binding fragment thereof, wherein the heavy chain CDR1 comprises or consists of the sequence of SEQ ID NO: 29, and X1 is A.

[0090] E111. The isolated antibody of any one of E108 to E110, or an ALK2-binding fragment thereof, wherein the heavy chain CDR1 comprises or consists of the sequence of SEQ ID NO: 29, and X2 is S.

[0091] E112. The isolated antibody of any one of E108 to E110, or an ALK2-binding fragment thereof, wherein the heavy chain CDR1 comprises or consists of the sequence of SEQ ID NO: 29 and X2 is H.

[0092] E113. The isolated antibody of E1, or an ALK2-binding fragment thereof, wherein the heavy chain CDR1 comprises or consists of the sequence GFTFX1SX2AMS (SEQ ID NO: 30, where X1 is S or N and X2 is H or S).

[0093] E114. The isolated antibody of E113, or an ALK2-binding fragment thereof, wherein the heavy chain CDR1 comprises or consists of the sequence of SEQ ID NO: 30, and X1 is S. E115. The isolated antibody of E113, or an ALK2-binding fragment thereof, wherein the heavy chain CDR1 comprises or consists of the sequence of SEQ ID NO: 30, and X1 is N.

[0094] E116. The isolated antibody of any one of E113 to E115, or an ALK2-binding fragment thereof, wherein the heavy chain CDR1 comprises or consists of the sequence of SEQ ID NO: 30, and X2 is H.

[0095] E117. The isolated antibody of any one of E113 to E115, or an ALK2-binding fragment thereof, wherein the heavy chain CDR1 comprises or consists of the sequence of SEQ ID NO: 30, and X2 is S.

[0096] E118. The isolated antibody of E1, or an ALK2-binding fragment thereof, wherein the light chain CDR1 comprises or consists of the amino acid sequence SGSSSNIGSNYVS (SEQ ID NO:1), the light chain CDR2 comprises or consists of the amino acid sequence VLIYKNNHRPS (SEQ ID NO:24), and the light chain CDR3 comprises or consists of the amino acid sequence ASWDHSDRFYV (SEQ ID NO:4).

[0097] E119. The isolated antibody of E1, or an ALK2-binding fragment thereof, wherein the heavy chain CDR1 comprises or consists of the amino acid sequence GGTFSSYGVS (SEQ ID NO: 31), the heavy chain CDR2 comprises or consists of the amino acid sequence WMGGIIPHFGIANYAQKFQG (SEQ ID NO: 36), and the heavy chain CDR3 comprises or consists of the amino acid sequence EIGSLDI (SEQ ID NO: 13).

[0098] E120. The isolated antibody of E1, or an ALK2-binding fragment thereof, wherein the light chain CDR1 comprises or consists of the amino acid sequence SGSSSNIGSNYVS (SEQ ID NO:1), the light chain CDR2 comprises or consists of the amino acid sequence VLIYKNNHRPS (SEQ ID NO:24), the light chain CDR3 comprises or consists of the amino acid sequence ASWDHSDRFYV (SEQ ID NO:4), the heavy chain CDR1 comprises or consists of the amino acid sequence GGTFSSYGVS (SEQ ID NO:31), the heavy chain CDR2 comprises or consists of the amino acid sequence WMGGIIPHFGIANYAQKFQG (SEQ ID NO:36), and the heavy chain CDR3 comprises or consists of the amino acid sequence EIGSLDI (SEQ ID NO:13).

[0099] E121. The isolated antibody of E1, or an ALK2-binding fragment thereof, wherein the light chain CDR1 consists of the amino acid sequence SGSSSNIGSNYVS (SEQ ID NO: 1), the light chain CDR2 consists of the amino acid sequence VLIYKNNHRPS (SEQ ID NO: 24), the light chain CDR3 consists of the amino acid sequence ASWDHSDRFYV (SEQ ID NO: 4), the heavy chain CDR1 consists of the amino acid sequence GGTFSSYGVS (SEQ ID NO: 31), the heavy chain CDR2 consists of the amino acid sequence WMGGIIPHFGIANYAQKFQG (SEQ ID NO: 36), and the heavy chain CDR3 consists of the amino acid sequence EIGSLDI (SEQ ID NO: 13).

[0100] E122. The isolated antibody of E1, or an ALK2-binding fragment thereof, wherein the light chain CDR1 comprises or consists of the amino acid sequence SGDSIPSFFAS (SEQ ID NO: 18), the light chain CDR2 comprises or consists of the amino acid sequence LVIYRDSNRPS (SEQ ID NO: 25), and the light chain CDR3 comprises or consists of the amino acid sequence YVTAPWKSIW (SEQ ID NO: 5).

[0101] E123. The isolated antibody of E1, or an ALK2-binding fragment thereof, wherein the heavy chain CDR1 comprises or consists of the amino acid sequence GFTFSSHAMS (SEQ ID NO: 32), the heavy chain CDR2 comprises or consists of the amino acid sequence WVGRIKSKADSGTTDYAAPVKG (SEQ ID NO: 37), and the heavy chain CDR3 comprises or consists of the amino acid sequence DYGVAFAY (SEQ ID NO: 14).

[0102] E124. The isolated antibody of E1, or an ALK2-binding fragment thereof, wherein the light chain CDR1 comprises or consists of the amino acid sequence SGDSIPSFFAS (SEQ ID NO: 18), the light chain CDR2 comprises or consists of the amino acid sequence LVIYRDSNRPS (SEQ ID NO: 25), the light chain CDR3 comprises or consists of the amino acid sequence YVTAPWKSIW (SEQ ID NO: 5), the heavy chain CDR1 comprises or consists of the amino acid sequence GFTFSSHAMS (SEQ ID NO: 32), the heavy chain CDR2 comprises or consists of the amino acid sequence WVGRIKSKADSGTTDYAAPVKG (SEQ ID NO: 37), and the heavy chain CDR3 comprises or consists of the amino acid sequence DYGVAFAY (SEQ ID NO: 14).

[0103] E125. The isolated antibody of E1, or an ALK2-binding fragment thereof, wherein the light chain CDR1 consists of the amino acid sequence SGDSIPSFFAS (SEQ ID NO: 18), the light chain CDR2 consists of the amino acid sequence LVIYRDSNRPS (SEQ ID NO: 25), the light chain CDR3 consists of the amino acid sequence YVTAPWKSIW (SEQ ID NO: 5), the heavy chain CDR1 consists of the amino acid sequence GFTFSSHAMS (SEQ ID NO: 32), the heavy chain CDR2 consists of the amino acid sequence WVGRIKSKADSGTTDYAAPVKG (SEQ ID NO: 37), and the heavy chain CDR3 consists of the amino acid sequence DYGVAFAY (SEQ ID NO: 14).

[0104] E126. The isolated antibody of E1, or an ALK2-binding fragment thereof, wherein the light chain CDR1 comprises or consists of the amino acid sequence SGDNIGTKYAY (SEQ ID NO: 19), the light chain CDR2 comprises or consists of the amino acid sequence LVIYGDSDRPS (SEQ ID NO: 26), and the light chain CDR3 comprises or consists of the amino acid sequence YSADAQQMKA (SEQ ID NO: 6).

[0105] E127. The isolated antibody of E1, or an ALK2-binding fragment thereof, wherein the heavy chain CDR1 comprises or consists of the amino acid sequence GFTNFSSAMS (SEQ ID NO: 33), the heavy chain CDR2 comprises or consists of the amino acid sequence WVGRIKSKRDGYTTDYAAPVKG (SEQ ID NO: 38), and the heavy chain CDR3 comprises or consists of the amino acid sequence DYGGLKFDY (SEQ ID NO: 15).

[0106] E128. The isolated antibody of E1, or an ALK2-binding fragment thereof, wherein the light chain CDR1 comprises or consists of the amino acid sequence SGDNIGTKYAY (SEQ ID NO: 19), the light chain CDR2 comprises or consists of the amino acid sequence LVIYGDSDRPS (SEQ ID NO: 26), the light chain CDR3 comprises or consists of the amino acid sequence YSADAQQMKA (SEQ ID NO: 6), the heavy chain CDR1 comprises or consists of the amino acid sequence GFTNFSSAMS (SEQ ID NO: 33), the heavy chain CDR2 comprises or consists of the amino acid sequence WVGRIKSKRDGYTTDYAAPVKG (SEQ ID NO: 38), and the heavy chain CDR3 comprises or consists of the amino acid sequence DYGGLKFDY (SEQ ID NO: 15).

[0107] E129. The isolated antibody of E1, or an ALK2-binding fragment thereof, wherein the light chain CDR1 consists of the amino acid sequence SGDNIGTKYAY (SEQ ID NO: 19), the light chain CDR2 consists of the amino acid sequence LVIYGDSDRPS (SEQ ID NO: 26), the light chain CDR3 consists of the amino acid sequence YSADAQQMKA (SEQ ID NO: 6), the heavy chain CDR1 consists of the amino acid sequence GFTFNSSAMS (SEQ ID NO: 33), the heavy chain CDR2 consists of the amino acid sequence WVGRIKSKRDGYTTDYAAPVKG (SEQ ID NO: 38), and the heavy chain CDR3 consists of the amino acid sequence DYGGLKFDY (SEQ ID NO: 15).

[0108] E130. The isolated antibody of E1, or an ALK2-binding fragment thereof, wherein the light chain CDR1 comprises or consists of the amino acid sequence SGDNLRKYSAH (SEQ ID NO: 20), the light chain CDR2 comprises or consists of the amino acid sequence LVIYYDNKRPS (SEQ ID NO: 27), and the light chain CDR3 comprises or consists of the amino acid sequence QVYASVHRM (SEQ ID NO: 7).

[0109] E131. The isolated antibody of E1, or an ALK2-binding fragment thereof, wherein the heavy chain CDR1 comprises or consists of the amino acid sequence GGTFSSYAIH (SEQ ID NO: 34), the heavy chain CDR2 comprises or consists of the amino acid sequence WMGRIIPDFGTANYAQKFQG (SEQ ID NO: 39), and the heavy chain CDR3 comprises or consists of the amino acid sequence GPTQAIHYFAY (SEQ ID NO: 16).

[0110] E132. The isolated antibody of E1, or an ALK2-binding fragment thereof, wherein the light chain CDR1 comprises or consists of the amino acid sequence SGDNLRKYSAH (SEQ ID NO:20), the light chain CDR2 comprises or consists of the amino acid sequence LVIYYDNKRPS (SEQ ID NO:27), the light chain CDR3 comprises or consists of the amino acid sequence QVYASVHRM (SEQ ID NO:7), the heavy chain CDR1 comprises or consists of the amino acid sequence GGTFSSYAIH (SEQ ID NO:34), the heavy chain CDR2 comprises or consists of the amino acid sequence WMGRIIPDFGTANYAQKFQG (SEQ ID NO:39), and the heavy chain CDR3 comprises or consists of the amino acid sequence GPTQAIHYFAY (SEQ ID NO:16).

[0111] E133. The isolated antibody of E1, or an ALK2-binding fragment thereof, wherein the light chain CDR1 consists of the amino acid sequence SGDNLRKYSAH (SEQ ID NO: 20), the light chain CDR2 consists of the amino acid sequence LVIYYDNKRPS (SEQ ID NO: 27), the light chain CDR3 consists of the amino acid sequence QVYASVHRM (SEQ ID NO: 7), the heavy chain CDR1 consists of the amino acid sequence GGTFSSYAIH (SEQ ID NO: 34), the heavy chain CDR2 consists of the amino acid sequence WMGRIIPDFGTANYAQKFQG (SEQ ID NO: 39), and the heavy chain CDR3 consists of the amino acid sequence GPTQAIHYFAY (SEQ ID NO: 16).

[0112] E134. The isolated antibody of E1, or an ALK2-binding fragment thereof, wherein the light chain CDR1 comprises or consists of the amino acid sequence SGDSLGSKSVH (SEQ ID NO: 21), the light chain CDR2 comprises or consists of the amino acid sequence LVIYRDSKRPS (SEQ ID NO: 28), and the light chain CDR3 comprises or consists of the amino acid sequence QTYDWSHFGW (SEQ ID NO: 8).

[0113] E135. The isolated antibody of E1, or an ALK2-binding fragment thereof, wherein the heavy chain CDR1 comprises or consists of the amino acid sequence of GFTFSSAAMH (SEQ ID NO: 35), the heavy chain CDR2 comprises or consists of the amino acid sequence of WVSVISSDGGSTYYADSVKG (SEQ ID NO: 12), and the heavy chain CDR3 comprises or consists of the amino acid sequence of AGFILGSLGVAWMDV (SEQ ID NO: 17).

[0114] E136. The isolated antibody of E1, or an ALK2-binding fragment thereof, wherein the light chain CDR1 comprises or consists of the amino acid sequence SGDSLGSKSVH (SEQ ID NO:21), the light chain CDR2 comprises or consists of the amino acid sequence LVIYRDSKRPS (SEQ ID NO:28), the light chain CDR3 comprises or consists of the amino acid sequence QTYDWSHFGW (SEQ ID NO:8), the heavy chain CDR1 comprises or consists of the amino acid sequence GFTFSSAAMH (SEQ ID NO:35), the heavy chain CDR2 comprises or consists of the amino acid sequence WVSVISSDGGSTYYADSVKG (SEQ ID NO:12), and the heavy chain CDR3 comprises or consists of the amino acid sequence AGFILGSLGVAWMDV (SEQ ID NO:17).

[0115] E137. The isolated antibody of E1, or an ALK2-binding fragment thereof, wherein the light chain CDR1 consists of the amino acid sequence SGDSLGSKSVH (SEQ ID NO: 21), the light chain CDR2 consists of the amino acid sequence LVIYRDSKRPS (SEQ ID NO: 28), the light chain CDR3 consists of the amino acid sequence QTYDWSHFGW (SEQ ID NO: 8), the heavy chain CDR1 consists of the amino acid sequence GFTFSSAAMH (SEQ ID NO: 35), the heavy chain CDR2 consists of the amino acid sequence WVSVISSDGGSTYYADSVKG (SEQ ID NO: 12), and the heavy chain CDR3 consists of the amino acid sequence AGFILGSLGVAWMDV (SEQ ID NO: 17).

[0116] E138. The isolated antibody of E1, or an ALK2-binding fragment thereof, wherein the antibody has at least 90% (e.g., 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more) sequence identity to amino acids 1 to 331 of sequence SEQ ID NO:67, or at least 95% sequence identity to amino acids 1 to 331 of sequence SEQ ID NO:67, excluding light chain CDR1, CDR2, and CDR3 and heavy chain CDR1, CDR2, and CDR3.

[0117] E139. The isolated antibody of E1, or an ALK2-binding fragment thereof, wherein the antibody has at least 90% (e.g., 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more) sequence identity to amino acids 1 to 332 of sequence SEQ ID NO:68, or at least 95% sequence identity to amino acids 1 to 332 of sequence SEQ ID NO:68, excluding light chain CDR1, CDR2, and CDR3 and heavy chain CDR1, CDR2, and CDR3.

[0118] E140. The isolated antibody of E1, or an ALK2-binding fragment thereof, wherein the antibody has at least 90% (e.g., 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more) sequence identity to amino acids 1 to 333 of sequence SEQ ID NO:69, or at least 95% sequence identity to amino acids 1 to 333 of sequence SEQ ID NO:69, excluding light chain CDR1, CDR2, and CDR3 and heavy chain CDR1, CDR2, and CDR3.

[0119] E141. The isolated antibody of E1, or an ALK2-binding fragment thereof, wherein the antibody has at least 90% (e.g., 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more) sequence identity to amino acids 1 to 332 of sequence SEQ ID NO: 70, or at least 95% sequence identity to amino acids 1 to 332 of sequence SEQ ID NO: 70, or at least 98% sequence identity to amino acids 1 to 332 of sequence SEQ ID NO: 70, excluding light chain CDR1, CDR2, and CDR3 and heavy chain CDR1, CDR2, and CDR3.

[0120] E142. The isolated antibody of E1, or an ALK2-binding fragment thereof, wherein the antibody has at least 90% (e.g., 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more) sequence identity to amino acids 1 to 337 of sequence SEQ ID NO: 71, or at least 95% sequence identity to amino acids 1 to 337 of sequence SEQ ID NO: 71, or at least 98% sequence identity to amino acids 1 to 337 of sequence SEQ ID NO: 71, excluding light chain CDR1, CDR2, and CDR3 and heavy chain CDR1, CDR2, and CDR3.

[0121] E143. The isolated antibody of E1, or an ALK2-binding fragment thereof, wherein said antibody comprises or consists of amino acids 1 to 433 of the sequence of SEQ ID NO: 67. E144. The isolated antibody of E1, or an ALK2-binding fragment thereof, wherein said antibody comprises or consists of amino acids 1 to 434 of the sequence of SEQ ID NO: 68.

[0122] E145. The isolated antibody of E1, or an ALK2-binding fragment thereof, wherein said antibody comprises or consists of amino acids 1 to 435 of the sequence of SEQ ID NO: 69. E146. The isolated antibody of E1, or an ALK2-binding fragment thereof, wherein said antibody comprises or consists of amino acids 1 to 434 of the sequence of SEQ ID NO: 70.

[0123] E147. The isolated antibody of E1, or an ALK2-binding fragment thereof, wherein said antibody comprises or consists of amino acids 1 to 439 of SEQ ID NO: 71. E148. An isolated antibody, or an ALK2-binding fragment thereof, comprising: (1) a light chain variable domain, the light chain complementarity determination comprising or consisting of an amino acid sequence selected from RASQGISGNWLT (SEQ ID NO: 40), SGDX1X2RX3X4X5X6H (SEQ ID NO: 64, wherein X1 is N or A, X2 is I or L, X3 is K or Y, X4 is K or Y, X5 is Y or I, and X6 is V or A), and SGSSSNIGQNYVS (SEQ ID NO: 58). Region (CDR) 1; a light chain CDR2 comprising or consisting of the amino acid sequence LX1IYX2X3X4X5X6X7S (SEQ ID NO:65, where X1 is V or L, X2 is D, R, or Y, X3 is A, D, or N, X4 is S or N, X5 is K or N, X6 is L or R, and X7 is Q or P); and a CDR2 selected from HQSYRGPM (SEQ ID NO:42), SSAGRDNY (SEQ ID NO:48), QSYGPGSV (SEQ ID NO:54), and SSWDLLSKSR (SEQ ID NO:60). and (2) a heavy chain CDR1 comprising or consisting of the amino acid sequence GX1TFX2X3X4X5X6X7 (SEQ ID NO:66, where X1 is F or G, X2 is G or S, X3 is R, S, D, or T, X4 is F, S, Y, or H, X5 is V or A, X6 is M or I, and X7 is H or S); a heavy chain CDR2 comprising or consisting of an amino acid sequence selected from: WVSX1IX2YX3X4SX5TYYADSVKG (SEQ ID NO:76, where X1 is V or S, X2 is G, H, or F, X3 is S or D, X4 is G or S, and X5 is S, E, or N); and WMGLIQPRFGTANYAQKFQR (SEQ ID NO:62);and a heavy chain variable domain comprising a heavy chain CDR3 comprising or consisting of an amino acid sequence selected from EPGYYYPSGYYRGPGYWMDV (SEQ ID NO: 45), DRYFFDV (SEQ ID NO: 51), PKSYASGPFAY (SEQ ID NO: 57), and DYYGGMAY (SEQ ID NO: 63);

[0124] E149. The isolated antibody of E148, or an ALK2-binding fragment thereof, wherein the light chain CDR1 comprises or consists of the sequence of SEQ ID NO: 64, and X1 is N. E150. The isolated antibody of E148, or an ALK2-binding fragment thereof, wherein the light chain CDR1 comprises or consists of the sequence of SEQ ID NO: 64, and X1 is A. E150.

[0125] E151. The isolated antibody of any one of E148 to E150, or an ALK2-binding fragment thereof, wherein the light chain CDR1 comprises or consists of the sequence of SEQ ID NO: 64, and X2 is I.

[0126] E152. The isolated antibody of any one of E148 to E150, or an ALK2-binding fragment thereof, wherein the light chain CDR1 comprises or consists of the sequence of SEQ ID NO: 64, and X2 is L.

[0127] E153. The isolated antibody of any one of E148 to E152, or an ALK2-binding fragment thereof, wherein the light chain CDR1 comprises or consists of the sequence of SEQ ID NO: 64 and X3 is K.

[0128] E154. The isolated antibody of any one of E148 to E152, or an ALK2-binding fragment thereof, wherein the light chain CDR1 comprises or consists of the sequence of SEQ ID NO: 64 and X3 is Y.

[0129] E155. The isolated antibody of any one of E148 to E154, or an ALK2-binding fragment thereof, wherein the light chain CDR1 comprises or consists of the sequence of SEQ ID NO: 64 and X4 is K.

[0130] E156. The isolated antibody of any one of E148 to E154, or an ALK2-binding fragment thereof, wherein the light chain CDR1 comprises or consists of the sequence of SEQ ID NO: 64, and X4 is Y.

[0131] E157. The isolated antibody of any one of E148 to E156, or an ALK2-binding fragment thereof, wherein the light chain CDR1 comprises or consists of the sequence of SEQ ID NO: 64, and X5 is Y.

[0132] E158. The isolated antibody of any one of E148 to E156, or an ALK2-binding fragment thereof, wherein the light chain CDR1 comprises or consists of the sequence of SEQ ID NO: 64, and X5 is I.

[0133] E159. The isolated antibody of any one of E148 to E158, or an ALK2-binding fragment thereof, wherein the light chain CDR1 comprises or consists of the sequence of SEQ ID NO: 64 and X6 is V.

[0134] E160. The isolated antibody of any one of E148 to E158, or an ALK2-binding fragment thereof, wherein the light chain CDR1 comprises or consists of the sequence of SEQ ID NO: 64 and X6 is A.

[0135] E161. The isolated antibody of any one of E148 to E160, or an ALK2-binding fragment thereof, wherein the light chain CDR2 comprises or consists of the sequence of SEQ ID NO: 65 and X1 is V.

[0136] E162. The isolated antibody of any one of E148 to E160, or an ALK2-binding fragment thereof, wherein the light chain CDR2 comprises or consists of the sequence of SEQ ID NO: 65, and X1 is L.

[0137] E163. The isolated antibody of any one of E148 to E162, or an ALK2-binding fragment thereof, wherein the light chain CDR2 comprises or consists of the sequence of SEQ ID NO: 65 and X2 is D.

[0138] E164. The isolated antibody of any one of E148 to E162, or an ALK2-binding fragment thereof, wherein the light chain CDR2 comprises or consists of the sequence of SEQ ID NO: 65, and X2 is R.

[0139] E165. The isolated antibody of any one of E148 to E162, or an ALK2-binding fragment thereof, wherein the light chain CDR2 comprises or consists of the sequence of SEQ ID NO: 65 and X2 is Y.

[0140] E166. The isolated antibody of any one of E148 to E165, or an ALK2-binding fragment thereof, wherein the light chain CDR2 comprises or consists of the sequence of SEQ ID NO: 65 and X3 is A.

[0141] E167. The isolated antibody of any one of E148 to E165, or an ALK2-binding fragment thereof, wherein the light chain CDR2 comprises or consists of the sequence of SEQ ID NO: 65, and X3 is D.

[0142] E168. An isolated antibody of any one of E148 to E165, or an ALK2-binding fragment thereof, wherein the light chain CDR2 comprises or consists of the sequence of SEQ ID NO: 65 and X3 is N.

[0143] E169. The isolated antibody of any one of E148 to E168, or an ALK2-binding fragment thereof, wherein the light chain CDR2 comprises or consists of the sequence of SEQ ID NO: 65 and X4 is S.

[0144] E170. The isolated antibody of any one of E148 to E168, or an ALK2-binding fragment thereof, wherein the light chain CDR2 comprises or consists of the sequence of SEQ ID NO: 65 and X4 is N.

[0145] E171. The isolated antibody of any one of E148 to E170, or an ALK2-binding fragment thereof, wherein the light chain CDR2 comprises or consists of the sequence of SEQ ID NO: 65, and X5 is K.

[0146] E172. The isolated antibody of any one of E148 to E170, or an ALK2-binding fragment thereof, wherein the light chain CDR2 comprises or consists of the sequence of SEQ ID NO: 65, and X5 is N.

[0147] E173. The isolated antibody of any one of E148 to E172, or an ALK2-binding fragment thereof, wherein the light chain CDR2 comprises or consists of the sequence of SEQ ID NO: 65, and X6 is L.

[0148] E174. The isolated antibody of any one of E148 to E172, or an ALK2-binding fragment thereof, wherein the light chain CDR2 comprises or consists of the sequence of SEQ ID NO: 65, and X6 is R.

[0149] E175. The isolated antibody of any one of E148 to E174, or an ALK2-binding fragment thereof, wherein the light chain CDR2 comprises or consists of the sequence of SEQ ID NO: 65, and X7 is Q.

[0150] E176. The isolated antibody of any one of E148 to E174, or an ALK2-binding fragment thereof, wherein the light chain CDR2 comprises or consists of the sequence of SEQ ID NO: 65, and X7 is P.

[0151] E177. The isolated antibody of any one of E148 to E176, or an ALK2-binding fragment thereof, wherein the heavy chain CDR1 comprises or consists of the sequence of SEQ ID NO: 66, and X1 is F.

[0152] E178. The isolated antibody of any one of E148 to E176, or an ALK2-binding fragment thereof, wherein the heavy chain CDR1 comprises or consists of the sequence of SEQ ID NO: 66, and X1 is G.

[0153] E179. The isolated antibody of any one of E148 to E178, or an ALK2-binding fragment thereof, wherein the heavy chain CDR1 comprises or consists of the sequence of SEQ ID NO: 66, and X2 is G.

[0154] E180. The isolated antibody of any one of E148 to E178, or an ALK2-binding fragment thereof, wherein the heavy chain CDR1 comprises or consists of the sequence of SEQ ID NO: 66, and X2 is S.

[0155] E181. The isolated antibody of any one of E148 to E180, or an ALK2-binding fragment thereof, wherein the heavy chain CDR1 comprises or consists of the sequence of SEQ ID NO: 66, and X3 is R.

[0156] E182. The isolated antibody of any one of E148 to E180, or an ALK2-binding fragment thereof, wherein the heavy chain CDR1 comprises or consists of the sequence of SEQ ID NO: 66, and X3 is S.

[0157] E183. The isolated antibody of any one of E148 to E180, or an ALK2-binding fragment thereof, wherein the heavy chain CDR1 comprises or consists of the sequence of SEQ ID NO: 66, and X3 is D.

[0158] E184. The isolated antibody of any one of E148 to E180, or an ALK2-binding fragment thereof, wherein the heavy chain CDR1 comprises or consists of the sequence of SEQ ID NO: 66, and X3 is T.

[0159] E185. The isolated antibody of any one of E148 to E184, or an ALK2-binding fragment thereof, wherein the heavy chain CDR1 comprises or consists of the sequence of SEQ ID NO: 66, and X4 is F.

[0160] E186. The isolated antibody of any one of E148 to E184, or an ALK2-binding fragment thereof, wherein the heavy chain CDR1 comprises or consists of the sequence of SEQ ID NO: 66 and X4 is S.

[0161] E187. The isolated antibody of any one of E148 to E184, or an ALK2-binding fragment thereof, wherein the heavy chain CDR1 comprises or consists of the sequence of SEQ ID NO: 66, and X4 is Y.

[0162] E188. The isolated antibody of any one of E148 to E184, or an ALK2-binding fragment thereof, wherein the heavy chain CDR1 comprises or consists of the sequence of SEQ ID NO: 66 and X4 is H.

[0163] E189. The isolated antibody of any one of E148 to E188, or an ALK2-binding fragment thereof, wherein the heavy chain CDR1 comprises or consists of the sequence of SEQ ID NO: 66, and X5 is V.

[0164] E190. An isolated antibody of any one of E148 to E188, or an ALK2-binding fragment thereof, wherein the heavy chain CDR1 comprises or consists of the sequence of SEQ ID NO: 66, and X5 is V or A.

[0165] E191. The isolated antibody of any one of E148 to E190, or an ALK2-binding fragment thereof, wherein the heavy chain CDR1 comprises or consists of the sequence of SEQ ID NO: 66, and X6 is M.

[0166] E192. The isolated antibody of any one of E148 to E190, or an ALK2-binding fragment thereof, wherein the heavy chain CDR1 comprises or consists of the sequence of SEQ ID NO: 66 and X6 is I.

[0167] E193. The isolated antibody of any one of E148 to E192, or an ALK2-binding fragment thereof, wherein the heavy chain CDR1 comprises or consists of the sequence of SEQ ID NO: 66, and X7 is H.

[0168] E194. An isolated antibody of any one of E148 to E192, or an ALK2-binding fragment thereof, wherein the heavy chain CDR1 comprises or consists of the sequence of SEQ ID NO: 66 and X7 is S.

[0169] E195. The isolated antibody of any one of E148 to E194, or an ALK2-binding fragment thereof, wherein the heavy chain CDR2 comprises or consists of the sequence of SEQ ID NO: 76, and X1 is V.

[0170] E196. The isolated antibody of any one of E148 to E194, or an ALK2-binding fragment thereof, wherein the heavy chain CDR2 comprises or consists of the sequence of SEQ ID NO: 76, and X1 is S.

[0171] E197. The isolated antibody of any one of E148 to E196, or an ALK2-binding fragment thereof, wherein the heavy chain CDR2 comprises or consists of the sequence of SEQ ID NO: 76, and X2 is G.

[0172] E198. The isolated antibody of any one of E148 to E196, or an ALK2-binding fragment thereof, wherein the heavy chain CDR2 comprises or consists of the sequence of SEQ ID NO: 76, and X2 is H.

[0173] E199. The isolated antibody of any one of E148 to E196, or an ALK2-binding fragment thereof, wherein the heavy chain CDR2 comprises or consists of the sequence of SEQ ID NO: 76, and X2 is F.

[0174] E200. The isolated antibody of any one of E148 to E199, or an ALK2-binding fragment thereof, wherein the heavy chain CDR2 comprises or consists of the sequence of SEQ ID NO: 76, and X3 is S.

[0175] E201. The isolated antibody of any one of E148 to E199, or an ALK2-binding fragment thereof, wherein the heavy chain CDR2 comprises or consists of the sequence of SEQ ID NO: 76 and X3 is D.

[0176] E202. An isolated antibody of any one of E148 to E201, or an ALK2-binding fragment thereof, wherein the heavy chain CDR2 comprises or consists of the sequence of SEQ ID NO: 76 and X4 is G.

[0177] E203. An isolated antibody of any one of E148 to E201, or an ALK2-binding fragment thereof, wherein the heavy chain CDR2 comprises or consists of the sequence of SEQ ID NO: 76 and X4 is S.

[0178] E204. An isolated antibody of any one of E148 to E203, or an ALK2-binding fragment thereof, wherein the heavy chain CDR2 comprises or consists of the sequence of SEQ ID NO: 76, and X5 is S.

[0179] E205. An isolated antibody of any one of E148 to E203, or an ALK2-binding fragment thereof, wherein the heavy chain CDR2 comprises or consists of the sequence of SEQ ID NO: 76, and X5 is E.

[0180] E206. An isolated antibody of any one of E148 to E203, or an ALK2-binding fragment thereof, wherein the heavy chain CDR2 comprises or consists of the sequence of SEQ ID NO: 76 and X5 is N.

[0181] E207. The isolated antibody of E148, or an ALK2-binding fragment thereof, wherein the light chain CDR1 comprises or consists of the sequence RASQGISGNWLT (SEQ ID NO: 40). E208. The isolated antibody of E148, or an ALK2-binding fragment thereof, wherein the light chain CDR1 comprises or consists of the sequence SGDNIRKKYVH (SEQ ID NO:46).

[0182] E209. The isolated antibody of E148, or an ALK2-binding fragment thereof, wherein the light chain CDR1 comprises or consists of the sequence SGDALRYYIAH (SEQ ID NO:52). E210. The isolated antibody of E148, or an ALK2-binding fragment thereof, wherein the light chain CDR1 comprises or consists of the sequence SGSSSNIGQNYVS (SEQ ID NO:58).

[0183] E211. An isolated antibody of any one of E148 and E207 to E210, or an ALK2-binding fragment thereof, wherein the light chain CDR2 comprises or consists of the sequence LLIYDASNLQS (SEQ ID NO: 41).

[0184] E212. An isolated antibody of any one of E148 and E207 to E210, or an ALK2-binding fragment thereof, wherein the light chain CDR2 comprises or consists of the sequence LVIYRDSNRPS (SEQ ID NO: 47).

[0185] E213. An isolated antibody of any one of E148 and E207 to E210, or an ALK2-binding fragment thereof, wherein the light chain CDR2 comprises or consists of the sequence LVIYYNNNRPS (SEQ ID NO:53).

[0186] E214. An isolated antibody of any one of E148 and E207 to E210, or an ALK2-binding fragment thereof, wherein the light chain CDR2 comprises or consists of the sequence LLIYDNSKRPS (SEQ ID NO:59).

[0187] E215. An isolated antibody of any one of E148 and E207 to E214, or an ALK2-binding fragment thereof, wherein the light chain CDR3 comprises or consists of the sequence HQSYRGPM (SEQ ID NO: 42).

[0188] E216. An isolated antibody of any one of E148 and E207 to E214, or an ALK2-binding fragment thereof, wherein the light chain CDR3 comprises or consists of the sequence SSAGRDNY (SEQ ID NO: 48).

[0189] E217. An isolated antibody of any one of E148 and E207 to E214, or an ALK2-binding fragment thereof, wherein the light chain CDR3 comprises or consists of the sequence QSYGPGSV (SEQ ID NO:54).

[0190] E218. An isolated antibody of any one of E148 and E207 to E214, or an ALK2-binding fragment thereof, wherein the light chain CDR3 comprises or consists of the sequence SSWDLLSKSR (SEQ ID NO: 60).

[0191] E219. An isolated antibody of any one of E148 and E207 to E218, or an ALK2-binding fragment thereof, wherein the heavy chain CDR1 comprises or consists of the sequence GFTFGRFVMH (SEQ ID NO: 43).

[0192] E220. An isolated antibody of any one of E148 and E207 to E218, or an ALK2-binding fragment thereof, wherein the heavy chain CDR1 comprises or consists of the sequence GFTFSSSAMH (SEQ ID NO: 49).

[0193] E221. An isolated antibody of any one of E148 and E207 to E218, or an ALK2-binding fragment thereof, wherein the heavy chain CDR1 comprises or consists of the sequence GFTFSDYAMH (SEQ ID NO:55).

[0194] E222. An isolated antibody of any one of E148 and E207 to E218, or an ALK2-binding fragment thereof, wherein the heavy chain CDR1 comprises or consists of the sequence GGTFSTHAIS (SEQ ID NO: 61).

[0195] E223. An isolated antibody of any one of E148 and E207 to E222, or an ALK2-binding fragment thereof, wherein the heavy chain CDR2 comprises or consists of the sequence WVSVIGYSGSSTYYADSVKG (SEQ ID NO: 44).

[0196] E224. An isolated antibody of any one of E148 and E207 to E222, or an ALK2-binding fragment thereof, wherein the heavy chain CDR2 comprises or consists of the sequence WVSVIHYDSSETYYADSVKG (SEQ ID NO:50).

[0197] E225. An isolated antibody of any one of E148 and E207 to E222, or an ALK2-binding fragment thereof, wherein the heavy chain CDR2 comprises or consists of the sequence WVSSIFYSGSNTYYADSVKG (SEQ ID NO:56).

[0198] E226. An isolated antibody of any one of E148 and E207 to E222, or an ALK2-binding fragment thereof, wherein the heavy chain CDR2 comprises or consists of the sequence WMGLIQPRFGTANYAQKFQR (SEQ ID NO: 62).

[0199] E227. An isolated antibody of any one of E148 and E207 to E226, or an ALK2-binding fragment thereof, wherein the heavy chain CDR3 comprises or consists of the sequence EPGYYYPSGYYRGPGYWMDV (SEQ ID NO: 45).

[0200] E228. An isolated antibody of any one of E148 and E207 to E226, or an ALK2-binding fragment thereof, wherein the heavy chain CDR3 comprises or consists of the sequence DRYFFDV (SEQ ID NO:51).

[0201] E229. An isolated antibody of any one of E148 and E207 to E226, or an ALK2-binding fragment thereof, wherein the heavy chain CDR3 comprises or consists of the sequence PKSYASGPFAY (SEQ ID NO:57).

[0202] E230. An isolated antibody of any one of E148 and E207 to E226, or an ALK2-binding fragment thereof, wherein the heavy chain CDR3 comprises or consists of the sequence DYYGGMAY (SEQ ID NO: 63).

[0203] E231. The isolated antibody of E148, or an ALK2-binding fragment thereof, wherein the light chain CDR1 comprises or consists of the amino acid sequence RASQGISGNWLT (SEQ ID NO: 40), the light chain CDR2 comprises or consists of the amino acid sequence LLIYDASNLQS (SEQ ID NO: 41), and the light chain CDR3 comprises or consists of the amino acid sequence HQSYRGPM (SEQ ID NO: 42).

[0204] E232. The isolated antibody of E148, or an ALK2-binding fragment thereof, wherein the heavy chain CDR1 comprises or consists of the amino acid sequence GFTFGRFVMH (SEQ ID NO: 43), the heavy chain CDR2 comprises or consists of the amino acid sequence WVSVIGYSGSSTYYADSVKG (SEQ ID NO: 44), and the heavy chain CDR3 comprises or consists of the amino acid sequence EPGYYYPSGYYRGPGYWMDV (SEQ ID NO: 45).

[0205] E233. The isolated antibody of E148, or an ALK2-binding fragment thereof, wherein the light chain CDR1 comprises or consists of the amino acid sequence RASQGISGNWLT (SEQ ID NO: 40), the light chain CDR2 comprises or consists of the amino acid sequence LLIYDASNLQS (SEQ ID NO: 41), the light chain CDR3 comprises or consists of the amino acid sequence HQSYRGPM (SEQ ID NO: 42), the heavy chain CDR1 comprises or consists of the amino acid sequence GFTFGRFVMH (SEQ ID NO: 43), the heavy chain CDR2 comprises or consists of the amino acid sequence WVSVIGYSGSSTYYADSVKG (SEQ ID NO: 44), and the heavy chain CDR3 comprises or consists of the amino acid sequence EPGYYYPSGYYRGPGYWMDV (SEQ ID NO: 45).

[0206] E234. The isolated antibody of E148, or an ALK2-binding fragment thereof, wherein the light chain CDR1 consists of the amino acid sequence RASQGISGNWLT (SEQ ID NO: 40), the light chain CDR2 consists of the amino acid sequence LLIYDASNLQS (SEQ ID NO: 41), the light chain CDR3 consists of the amino acid sequence HQSYRGPM (SEQ ID NO: 42), the heavy chain CDR1 consists of the amino acid sequence GFTFGRFVMH (SEQ ID NO: 43), the heavy chain CDR2 consists of the amino acid sequence WVSVIGYSGSSTYYADSVKG (SEQ ID NO: 44), and the heavy chain CDR3 consists of the amino acid sequence EPGYYYPSGYYRGPGYWMDV (SEQ ID NO: 45).

[0207] E235. The isolated antibody of E148, or an ALK2-binding fragment thereof, wherein the light chain CDR1 comprises or consists of the amino acid sequence SGDNIRKKYVH (SEQ ID NO: 46), the light chain CDR2 comprises or consists of the amino acid sequence LVIYRDSNRPS (SEQ ID NO: 47), and the light chain CDR3 comprises or consists of the amino acid sequence SSAGRDNY (SEQ ID NO: 48).

[0208] E236. The isolated antibody of E148, or an ALK2-binding fragment thereof, wherein the heavy chain CDR1 comprises or consists of the amino acid sequence GFTFSSSAMH (SEQ ID NO: 49), the heavy chain CDR2 comprises or consists of the amino acid sequence WVSVIHYDSSETYYADSVKG (SEQ ID NO: 50), and the heavy chain CDR3 comprises or consists of the amino acid sequence DRYFFDV (SEQ ID NO: 51).

[0209] E237. The isolated antibody of E148, or an ALK2-binding fragment thereof, wherein the light chain CDR1 comprises or consists of the amino acid sequence SGDNIRKKYVH (SEQ ID NO: 46), the light chain CDR2 comprises or consists of the amino acid sequence LVIYRDSNRPS (SEQ ID NO: 47), the light chain CDR3 comprises or consists of the amino acid sequence SSAGRDNY (SEQ ID NO: 48), the heavy chain CDR1 comprises or consists of the amino acid sequence GFTFSSSAMH (SEQ ID NO: 49), the heavy chain CDR2 comprises or consists of the amino acid sequence WVSVIHYDSSETYYADSVKG (SEQ ID NO: 50), and the heavy chain CDR3 comprises or consists of the amino acid sequence DRYFFDV (SEQ ID NO: 51).

[0210] E238. The isolated antibody of E148, or an ALK2-binding fragment thereof, wherein the light chain CDR1 consists of the amino acid sequence SGDNIRKKYVH (SEQ ID NO: 46), the light chain CDR2 consists of the amino acid sequence LVIYRDSNRPS (SEQ ID NO: 47), the light chain CDR3 consists of the amino acid sequence SSAGRDNY (SEQ ID NO: 48), the heavy chain CDR1 consists of the amino acid sequence GFTFSSSAMH (SEQ ID NO: 49), the heavy chain CDR2 consists of the amino acid sequence WVSVIHYDSSETYYADSVKG (SEQ ID NO: 50), and the heavy chain CDR3 consists of the amino acid sequence DRYFFDV (SEQ ID NO: 51).

[0211] E239. The isolated antibody of E148, or an ALK2-binding fragment thereof, wherein the light chain CDR1 comprises or consists of the amino acid sequence SGDALRYYIAH (SEQ ID NO: 52), the light chain CDR2 comprises or consists of the amino acid sequence LVIYYNNNRPS (SEQ ID NO: 53), and the light chain CDR3 comprises or consists of the amino acid sequence QSYGPGSV (SEQ ID NO: 54).

[0212] E240. The isolated antibody of E148, or an ALK2-binding fragment thereof, wherein the heavy chain CDR1 comprises or consists of the amino acid sequence GFTFSDYAMH (SEQ ID NO:55), the heavy chain CDR2 comprises or consists of the amino acid sequence WVSSIFYSGSNTYYADSVKG (SEQ ID NO:56), and the heavy chain CDR3 comprises or consists of the amino acid sequence PKSYASGPFAY (SEQ ID NO:57).

[0213] E241. The isolated antibody of E148, or an ALK2-binding fragment thereof, wherein the light chain CDR1 comprises or consists of the amino acid sequence SGDALRYYIAH (SEQ ID NO:52), the light chain CDR2 comprises or consists of the amino acid sequence LVIYYNNNRPS (SEQ ID NO:53), the light chain CDR3 comprises or consists of the amino acid sequence QSYGPGSV (SEQ ID NO:54), the heavy chain CDR1 comprises or consists of the amino acid sequence GFTFSDYAMH (SEQ ID NO:55), the heavy chain CDR2 comprises or consists of the amino acid sequence WVSSIFYSGSNTYYADSVKG (SEQ ID NO:56), and the heavy chain CDR3 comprises or consists of the amino acid sequence PKSYASGPFAY (SEQ ID NO:57).

[0214] E242. The isolated antibody of E148, or an ALK2-binding fragment thereof, wherein the light chain CDR1 consists of the amino acid sequence SGDALRYYIAH (SEQ ID NO: 52), the light chain CDR2 consists of the amino acid sequence LVIYYNNNRPS (SEQ ID NO: 53), the light chain CDR3 consists of the amino acid sequence QSYGPGSV (SEQ ID NO: 54), the heavy chain CDR1 consists of the amino acid sequence GFTFSDYAMH (SEQ ID NO: 55), the heavy chain CDR2 consists of the amino acid sequence WVSSIFYSGSNTYYADSVKG (SEQ ID NO: 56), and the heavy chain CDR3 consists of the amino acid sequence PKSYASGPFAY (SEQ ID NO: 57).

[0215] E243. The isolated antibody of E148, or an ALK2-binding fragment thereof, wherein the light chain CDR1 comprises or consists of the amino acid sequence SGSSSNIGQNYVS (SEQ ID NO:58), the light chain CDR2 comprises or consists of the amino acid sequence LLIYDNSKRPS (SEQ ID NO:59), and the light chain CDR3 comprises or consists of the amino acid sequence SSWDLLSKSR (SEQ ID NO:60).

[0216] E244. The isolated antibody of E148, or an ALK2-binding fragment thereof, wherein the heavy chain CDR1 comprises or consists of the amino acid sequence GGTFSTHAIS (SEQ ID NO: 61), the heavy chain CDR2 comprises or consists of the amino acid sequence WMGLIQPRFGTANYAQKFQR (SEQ ID NO: 62), and the heavy chain CDR3 comprises or consists of the amino acid sequence DYYGGMAY (SEQ ID NO: 63).

[0217] E245. The isolated antibody of E148, or an ALK2-binding fragment thereof, wherein the light chain CDR1 comprises or consists of the amino acid sequence SGSSSNIGQNYVS (SEQ ID NO:58), the light chain CDR2 comprises or consists of the amino acid sequence LLIYDNSKRPS (SEQ ID NO:59), the light chain CDR3 comprises or consists of the amino acid sequence SSWDLLSKSR (SEQ ID NO:60), the heavy chain CDR1 comprises or consists of the amino acid sequence GGTFSTHAIS (SEQ ID NO:61), the heavy chain CDR2 comprises or consists of the amino acid sequence WMGLIQPRFGTANYAQKFQR (SEQ ID NO:62), and the heavy chain CDR3 comprises or consists of the amino acid sequence DYYGGMAY (SEQ ID NO:63).

[0218] E246. The isolated antibody of E148, or an ALK2-binding fragment thereof, wherein the light chain CDR1 consists of the amino acid sequence SGSSSNIGQNYVS (SEQ ID NO:58), the light chain CDR2 consists of the amino acid sequence LLIYDNSKRPS (SEQ ID NO:59), the light chain CDR3 consists of the amino acid sequence SSWDLLSKSR (SEQ ID NO:60), the heavy chain CDR1 consists of the amino acid sequence GGTFSTHAIS (SEQ ID NO:61), the heavy chain CDR2 consists of the amino acid sequence WMGLIQPRFGTANYAQKFQR (SEQ ID NO:62), and the heavy chain CDR3 consists of the amino acid sequence DYYGGMAY (SEQ ID NO:63).

[0219] E247. The isolated antibody of E148, or an ALK2-binding fragment thereof, wherein the antibody has at least 90% (e.g., 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more) sequence identity with amino acids 1 to 344 of sequence SEQ ID NO: 72, or at least 95% sequence identity with amino acids 1 to 344 of sequence SEQ ID NO: 72, or at least 98% sequence identity with amino acids 1 to 344 of sequence SEQ ID NO: 72, excluding light chain CDR1, CDR2, and CDR3 and heavy chain CDR1, CDR2, and CDR3.

[0220] E248. The isolated antibody of E148, or an ALK2-binding fragment thereof, wherein the antibody has at least 90% (e.g., 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more) sequence identity with amino acids 1 to 327 of sequence SEQ ID NO: 73, or at least 95% sequence identity with amino acids 1 to 327 of sequence SEQ ID NO: 73, excluding light chain CDR1, CDR2, and CDR3 and heavy chain CDR1, CDR2, and CDR3.

[0221] E249. The isolated antibody of E148, or an ALK2-binding fragment thereof, wherein the antibody has at least 90% (e.g., 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more) sequence identity with amino acids 1 to 331 of sequence SEQ ID NO: 74, or at least 95% sequence identity with amino acids 1 to 331 of sequence SEQ ID NO: 74, excluding light chain CDR1, CDR2, and CDR3 and heavy chain CDR1, CDR2, and CDR3.

[0222] E250. The isolated antibody of E148, or an ALK2-binding fragment thereof, wherein the antibody has at least 90% (e.g., 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more) sequence identity with amino acids 1 to 332 of sequence SEQ ID NO: 75, or at least 95% sequence identity with amino acids 1 to 332 of sequence SEQ ID NO: 75, excluding light chain CDR1, CDR2, and CDR3 and heavy chain CDR1, CDR2, and CDR3.

[0223] E251. The isolated antibody of E148, or an ALK2-binding fragment thereof, wherein said antibody comprises or consists of amino acids 1 to 446 of the sequence of SEQ ID NO: 72. E252. The isolated antibody of E148, or an ALK2-binding fragment thereof, wherein the antibody comprises or consists of amino acids 1 to 429 of the sequence of SEQ ID NO: 73.

[0224] E253. The isolated antibody of E148, or an ALK2-binding fragment thereof, wherein said antibody comprises or consists of amino acids 1 to 433 of the sequence of SEQ ID NO: 74. E254. The isolated antibody of E148, or an ALK2-binding fragment thereof, wherein the antibody comprises or consists of amino acids 1 to 434 of the sequence of SEQ ID NO: 75.

[0225] E255. The isolated antibody of any one of E1-E254, or an ALK2-binding fragment thereof, wherein said antibody is a monoclonal antibody. E256. The isolated antibody of any one of E1-E255, or an ALK2-binding fragment thereof, wherein said antibody is a humanized antibody.

[0226] E257. The isolated antibody of any one of E1-E256, or an ALK2-binding fragment thereof, wherein the ALK2-binding fragment is a Fab. E258. The isolated antibody of any one of E1 to E256, or an ALK2-binding fragment thereof, wherein the ALK2-binding fragment is Fab'.

[0227] E259. The isolated antibody of any one of E1 to E256, or an ALK2-binding fragment thereof, wherein the ALK2-binding fragment is F(ab')2. E260. The isolated antibody of any one of E1 to E256, or an ALK2-binding fragment thereof, wherein the ALK2-binding fragment is an Fv.

[0228] E261. The antibody has a K of 14 nM or less. D An isolated antibody, any one of E1 to E260, or an ALK2-binding fragment thereof, which binds to human ALK2 at a value E262. The antibody has a K of 5 nM or less. D An isolated antibody, any one of E1 to E261, or an ALK2-binding fragment thereof, which binds to human ALK2 at a value

[0229] E263. The antibody has a K of 1 nM or less. D An isolated antibody, any one of E1 to E262, or an ALK2-binding fragment thereof, which binds to human ALK2 at a value E264. The antibody has a K of 0.5 nM or less. D An isolated antibody, any one of E1 to E263, or an ALK2-binding fragment thereof, which binds to human ALK2 at a value

[0230] E265. A nucleic acid molecule comprising a nucleotide sequence encoding any one of the antibodies E1 to E264 or an ALK2-binding fragment thereof. An expression vector comprising a nucleic acid molecule of E266.E265.

[0231] Cells containing an expression vector for E267.E266. A pharmaceutical composition comprising any one of the antibodies or ALK2-binding fragments thereof of E1 to E264, the nucleic acid molecule of E265, or the expression vector of E266, and one or more pharma- ceutically acceptable carriers or excipients.

[0232] E269. The pharmaceutical composition of E268, wherein said antibody or ALK2-binding fragment thereof is in a therapeutically effective amount. E270. A method for increasing bone mineral density in a subject in need thereof by administering to the subject a therapeutically effective amount of an antibody or ALK2-binding fragment thereof of any one of E1 to E264, a nucleic acid molecule of E265, an expression vector of E266, or a pharmaceutical composition of E268 or E269.

[0233] E271. A method for reducing bone resorption (e.g., bone loss) in a subject in need thereof by administering to the subject a therapeutically effective amount of an antibody or ALK2-binding fragment thereof of any one of E1 to E264, a nucleic acid molecule of E265, an expression vector of E266, or a pharmaceutical composition of E268 or E269.

[0234] E272. A method for increasing bone formation in a subject in need thereof by administering to the subject a therapeutically effective amount of an antibody or ALK2-binding fragment thereof of any one of E1 to E264, a nucleic acid molecule of E265, an expression vector of E266, or a pharmaceutical composition of E268 or E269.

[0235] E273. A method for increasing bone strength in a subject in need thereof by administering to the subject a therapeutically effective amount of any one of antibodies or ALK2-binding fragments thereof of E1 to E264, a nucleic acid molecule of E265, an expression vector of E266, or a pharmaceutical composition of E268 or E269.

[0236] E274. A method for reducing the risk of fracture in a subject in need thereof by administering to the subject a therapeutically effective amount of any one of antibodies E1 to E264 or ALK2-binding fragments thereof, a nucleic acid molecule of E265, an expression vector of E266, or a pharmaceutical composition of E268 or E269.

[0237] E275. Any one of the methods of E270-E274, wherein the subject has or is at risk of developing primary osteoporosis, secondary osteoporosis, osteopenia, osteopetrosis, fracture, bone cancer or cancer metastasis associated bone loss, Paget's disease, renal osteodystrophy, treatment-related bone loss, diet-related bone loss, bone loss associated with treatment for obesity, low gravity-related bone loss, or immobility-related bone loss.

[0238] E276. The method of E275, wherein the subject has or is at risk of developing primary osteoporosis. E277. The method of E275, wherein the subject has or is at risk of developing secondary osteoporosis.

[0239] E278. The method of E275, wherein the subject has or is at risk of developing osteopenia. E279. A method for treating a subject having or at risk of developing a bone disease by administering to the subject a therapeutically effective amount of an antibody or ALK2-binding fragment thereof of any one of E1 to E264, a nucleic acid molecule of E265, an expression vector of E266, or a pharmaceutical composition of E268 or E269.

[0240] E280. The method of E279, wherein the bone disease is primary osteoporosis, secondary osteoporosis, osteopenia, osteopetrosis, fracture, bone loss associated with bone cancer or cancer metastasis, Paget's disease, renal osteodystrophy, treatment-related bone loss, diet-related bone loss, bone loss associated with treatment for obesity, low gravity-related bone loss, or immobility-related bone loss.

[0241] E281. The method of E280, wherein the bone disease is primary osteoporosis. E282. The method of E280, wherein the bone disease is secondary osteoporosis. E283. The method of E280, wherein the bone disease is osteopenia.

[0242] E284. A method of treating a subject having or at risk of developing primary osteoporosis by administering to the subject an antibody or ALK2-binding fragment thereof of any one of E1-E264, a nucleic acid molecule of E265, an expression vector of E266, or a pharmaceutical composition of E268 or E269.

[0243] E285. A method of treating a subject having or at risk of developing secondary osteoporosis by administering to the subject an antibody or ALK2-binding fragment thereof of any one of E1 to E264, a nucleic acid molecule of E265, an expression vector of E266, or a pharmaceutical composition of E268 or E269.

[0244] E286. A method for treating a subject having or at risk of developing osteopenia by administering to the subject an antibody or ALK2-binding fragment thereof of any one of E1 to E264, a nucleic acid molecule of E265, an expression vector of E266, or a pharmaceutical composition of E268 or E269.

[0245] E287. A method for treating a subject having or at risk of developing a fracture by administering to the subject an antibody or ALK2-binding fragment thereof of any one of E1 to E264, a nucleic acid molecule of E265, an expression vector of E266, or a pharmaceutical composition of E268 or E269.

[0246] E288. A method for treating a subject having or at risk of developing bone loss associated with bone cancer or cancer metastasis by administering to the subject a therapeutically effective amount of any one of antibodies or ALK2-binding fragments thereof of E1-E264, a nucleic acid molecule of E265, an expression vector of E266, or a pharmaceutical composition of E268 or E269.

[0247] E289. A method for treating a subject having or at risk of developing Paget's disease by administering to the subject a therapeutically effective amount of an antibody or ALK2-binding fragment thereof of any one of E1 to E264, a nucleic acid molecule of E265, an expression vector of E266, or a pharmaceutical composition of E268 or E269.

[0248] E290. A method for treating a subject having or at risk of developing renal osteodystrophy by administering to the subject a therapeutically effective amount of any one of antibodies or ALK2-binding fragments thereof of E1 to E264, a nucleic acid molecule of E265, an expression vector of E266, or a pharmaceutical composition of E268 or E269.

[0249] E291. A method of treating a subject having or at risk of developing treatment-associated bone loss by administering to the subject a therapeutically effective amount of an antibody or ALK2-binding fragment thereof of any one of E1-E264, a nucleic acid molecule of E265, an expression vector of E266, or a pharmaceutical composition of E268 or E269.

[0250] E292. A method of treating a subject having or at risk of developing diet-related bone loss by administering to the subject a therapeutically effective amount of an antibody or ALK2-binding fragment thereof of any one of E1-E264, a nucleic acid molecule of E265, an expression vector of E266, or a pharmaceutical composition of E268 or E269.

[0251] E293. A method for treating a subject having or at risk of developing low gravity associated bone loss by administering to the subject a therapeutically effective amount of an antibody or ALK2-binding fragment thereof of any one of E1-E264, a nucleic acid molecule of E265, an expression vector of E266, or a pharmaceutical composition of E268 or E269.

[0252] E294. A method of treating a subject having or at risk of developing immobility-associated bone loss by administering to the subject a therapeutically effective amount of an antibody or ALK2-binding fragment thereof of any one of E1-E264, a nucleic acid molecule of E265, an expression vector of E266, or a pharmaceutical composition of E268 or E269.

[0253] E295. The method of any one of E275, E276, E280, E281, and E284, wherein the primary osteoporosis is age-related osteoporosis or hormone-related osteoporosis. E296. The method of any one of E275, E278, E280, E282, and E285, wherein the secondary osteoporosis is immobilization-induced osteoporosis or glucocorticoid-induced osteoporosis.

[0254] E297. The method of any one of E275, E280, and E288, wherein said cancer is multiple myeloma. E298. The method of any one of E275, E280, and E291, wherein said treatment is FGF-21 treatment, GLP-1 treatment, cancer treatment, or treatment of obesity or type 2 diabetes.

[0255] E299. The method of any one of E275, E280, and E292, wherein the diet-related bone loss is rickets. E300. Any one of the methods of E270 to E299, wherein said subject is at risk of fracture.

[0256] E301. The method of any one of E270 to E300, wherein said method increases bone formation in said subject. E302. The method of any one of E270-E301, wherein said method reduces bone resorption (eg, bone loss) in said subject.

[0257] E303. The method of any one of E270 to E302, wherein said method increases osteoblast activity or osteoblast formation. E304. The method of any one of E270 to E303, wherein said method reduces osteoclast activity or reduces osteoclast formation.

[0258] E305. Any one of the methods of E270-E304, wherein said method reduces the risk of fracture. E306. The method of any one of E270 to E305, wherein said method increases bone mineral density.

[0259] E307. Any one of the methods E270 to E306, wherein the bone is cortical bone. E308. Any one of E270 to E306, wherein the bone is trabecular bone. E309. Any one of the methods of E270 to E308, wherein the method further comprises assessing bone mineral density in the subject (e.g., assessing bone mineral density using a standard test such as dual energy x-ray absorptiometry) prior to administering the antibody or ALK2-binding fragment thereof, the nucleic acid molecule, the expression vector, or the pharmaceutical composition.

[0260] E310. Any one of the methods of E270 to E309, wherein the method further comprises assessing bone mineral density in the subject after administering the antibody or ALK2-binding fragment thereof, the nucleic acid molecule, the expression vector, or the pharmaceutical composition (e.g., assessing bone mineral density using a standard test such as dual-energy x-ray absorptiometry).

[0261] E311. A method for increasing red blood cell levels in a subject in need thereof by administering to the subject a therapeutically effective amount of an antibody or ALK2-binding fragment thereof of any one of E1 to E264, a nucleic acid molecule of E265, an expression vector of E266, or a pharmaceutical composition of E268 or E269.

[0262] E312. A method for increasing hemoglobin levels (e.g., serum hemoglobin levels) in a subject in need thereof by administering to the subject a therapeutically effective amount of an antibody or ALK2-binding fragment thereof of any one of E1 to E264, a nucleic acid molecule of E265, an expression vector of E266, or a pharmaceutical composition of E268 or E269.

[0263] E313. A method for increasing red blood cell count in a subject in need thereof by administering to the subject a therapeutically effective amount of any one of antibodies E1 to E264 or ALK2-binding fragments thereof, a nucleic acid molecule of E265, an expression vector of E266, or a pharmaceutical composition of E268 or E269.

[0264] E314. A method for promoting or increasing erythropoiesis in a subject in need thereof by administering to the subject a therapeutically effective amount of any one of antibodies or ALK2-binding fragments thereof of E1 to E264, a nucleic acid molecule of E265, an expression vector of E266, or a pharmaceutical composition of E268 or E269.

[0265] E315. A method for reducing hepcidin levels (e.g., serum hepcidin concentration) in a subject in need thereof by administering to the subject a therapeutically effective amount of an antibody or ALK2-binding fragment thereof of any one of E1 to E264, a nucleic acid molecule of E265, an expression vector of E266, or a pharmaceutical composition of E268 or E269.

[0266] E316. A method for increasing iron levels (e.g., serum iron levels) in a subject in need thereof by administering to the subject a therapeutically effective amount of an antibody or ALK2-binding fragment thereof of any one of E1 to E264, a nucleic acid molecule of E265, an expression vector of E266, or a pharmaceutical composition of E268 or E269.

[0267] E317. The method of any one of E311-E316, wherein said subject has or is at risk of developing anemia or blood loss. The method of E317, wherein said anemia or blood loss is associated with cancer, cancer treatment, renal disease or failure (e.g., chronic renal disease or acute renal disease or failure), myelodysplastic syndrome, thalassemia, nutritional deficiency, adverse reaction to medication, inflammatory or autoimmune disease, splenomegaly, porphyria, vasculitis, hemolysis, bone marrow deficiency, bone marrow transplant, diabetes, liver disease (e.g., acute liver disease or chronic liver disease), bleeding (e.g., acute or chronic bleeding), infection, hemoglobinopathies, drug use, alcohol abuse, advanced age, Churg-Strauss syndrome, Felty syndrome, graft versus host disease, hematopoietic stem cell transplant, myelofibrosis, pancytopenia, pure red blood cell aplasia, Henoch-Schönlein purpura, Schwachmann syndrome (e.g., Schwachmann-Diamond syndrome), contraindication to blood transfusion, surgery, trauma, wound, ulcer, urinary tract bleeding, gastrointestinal bleeding, frequent blood donation, or heavy menstrual bleeding.

[0268] E319. A method for treating a subject having or at risk of developing anemia by administering to the subject a therapeutically effective amount of an antibody or ALK2-binding fragment thereof of any one of E1 to E264, a nucleic acid molecule of E265, an expression vector of E266, or a pharmaceutical composition of E268 or E269.

[0269] E320. The method of E319, wherein said anemia is associated with cancer, cancer treatment, renal disease or failure (e.g., chronic renal disease or acute renal disease or failure), myelodysplastic syndrome, thalassemia, nutritional deficiency, adverse reaction to medication, inflammatory or autoimmune disease, splenomegaly, porphyria, vasculitis, hemolysis, bone marrow deficiency, bone marrow transplant, diabetes, liver disease (e.g., acute liver disease or chronic liver disease), bleeding (e.g., acute or chronic bleeding), infection, hemoglobinopathies, drug use, alcohol abuse, advanced age, Churg-Strauss syndrome, Felty syndrome, graft versus host disease, hematopoietic stem cell transplant, myelofibrosis, pancytopenia, pure red blood cell aplasia, Henoch-Schönlein purpura, Schwachmann syndrome (e.g., Schwachmann-Diamond syndrome), contraindication to blood transfusion, surgery, trauma, wound, ulcer, urinary tract bleeding, gastrointestinal bleeding, frequent blood donations, or heavy menstrual bleeding.

[0270] E321. The method of E317-320, wherein said anemia is aplastic anemia, iron deficiency anemia, vitamin deficiency anemia, inflammatory anemia (also called anemia of chronic disease, e.g., anemia caused by an inflammatory disease or condition such as an infection (e.g., chronic infection such as HIV / AIDS or tuberculosis), an autoimmune disease (e.g., rheumatoid arthritis or lupus), cancer (e.g., cancer or cancer treatment), inflammatory bowel disease (e.g., Crohn's disease or ulcerative colitis), and chronic kidney disease), anemia associated with myeloid disease, hemolytic anemia, sickle cell anemia, microcytic anemia, hypochromic anemia, sideroblastic anemia, Diamond-Blackfan anemia, Fanconi anemia, or refractory anemia with excess blasts.

[0271] E322. The method of E321, wherein said anemia is IRIDA. E323. The method of E322, wherein said anemia is inflammatory anemia. E324. The method of any one of E311-E323, wherein said anemia is associated with elevated hepcidin levels.

[0272] E325. The method of any one of E311-E324, wherein said subject does not respond adequately to treatment with erythropoietin (EPO) or is susceptible to the adverse effects of EPO. E326. The method of any one of E311 to E325, wherein said method increases red blood cell formation, red blood cell count, hematocrit, or hemoglobin level.

[0273] E327. The method of any one of E311 to E326, wherein said method prevents or reduces elevated hepcidin levels. E328. The method of any one of E311-E327, wherein said method increases iron levels.

[0274] E329. The method of any one of E311-E328, wherein said method reduces the subject's need for blood transfusions. E330. Any one of the methods of E311-E329, wherein the method further comprises assessing the subject's red blood cell levels, hemoglobin levels, red blood cell count, hepcidin levels, or iron levels (e.g., assessing blood cell levels, hemoglobin levels, red blood cell count, hepcidin levels, or iron levels using a standard test such as a blood test) prior to administering the antibody or ALK2-binding fragment thereof, the nucleic acid molecule, the expression vector, or the pharmaceutical composition.

[0275] E331. Any one of the methods of E311-E330, wherein the method further comprises evaluating the subject's red blood cell levels, hemoglobin levels, red blood cell count, hepcidin levels, or iron levels after administering the antibody or ALK2-binding fragment thereof, the nucleic acid molecule, the expression vector, or the pharmaceutical composition (e.g., evaluating the blood cell levels, hemoglobin levels, red blood cell count, hepcidin levels, or iron levels using a standard test such as a blood test).

[0276] E332. A method for preventing or reducing heterotopic ossification in a subject in need thereof by administering to the subject a therapeutically effective amount of any one of antibodies or ALK2-binding fragments thereof of E1 to E264, a nucleic acid molecule of E265, an expression vector of E266, or a pharmaceutical composition of E268 or E269.

[0277] E333. The method of E332, wherein said subject has fibrodysplasia ossificans progressiva (FOP). The method of E333, wherein FOP is hereditary FOP or sporadic FOP.

[0278] E335. A method for treating a subject having FOP by administering to the subject a therapeutically effective amount of an antibody or ALK2-binding fragment thereof of any one of E1 to E264, a nucleic acid molecule of E265, an expression vector of E266, or a pharmaceutical composition of E268 or E269.

[0279] The method of E335, wherein FOP is hereditary FOP or sporadic FOP. E337. The method of any one of E332 to E336, wherein said method slows or inhibits the progression of FOP.

[0280] E338. The method of any one of E332-E337, wherein said method delays the onset of FOP (eg, delays the onset of heterotopic ossification). E339. Any one of the methods of E332-E338, wherein the method prevents heterotopic ossification (e.g., prevents heterotopic ossification in a subject at risk of developing heterotopic ossification or prevents recurrence of heterotopic ossification following surgical resection of the heterotopic bone).

[0281] E340. Any one of the methods of E332-E339, wherein the method reduces heterotopic ossification (e.g., reduces the amount of heterotopic ossification in a subject, reduces the occurrence of heterotopic ossification in a subject, or reduces recurrence of heterotopic ossification following surgical resection of heterotopic bone).

[0282] E341. Any one of the methods of E332 to E340, wherein said method reduces heterotopic bone formation. E342. The method of any one of E332-E341, wherein said method reduces recurrence of heterotopic bone (eg, following surgical resection).

[0283] E343. Any one of the methods of E332-E342, wherein the method further comprises assessing the subject's ectopic ossification (e.g., amount or location of ectopic bone) (e.g., assessing ectopic ossification using standard tests such as radiography (e.g., x-ray), CT (computed tomography), and / or MRI (magnetic resonance imaging)) prior to administering the antibody or ALK2-binding fragment thereof, the nucleic acid molecule, the expression vector, or the pharmaceutical composition.

[0284] E344. Any one of the methods of E332-E343, wherein the method further comprises assessing the ectopic ossification (e.g., amount or location of ectopic bone) in the subject after administering the antibody or ALK2-binding fragment thereof, the nucleic acid molecule, the expression vector, or the pharmaceutical composition (e.g., assessing ectopic ossification using standard tests such as radiography (e.g., x-ray), CT (computed tomography), and / or MRI (magnetic resonance imaging)).

[0285] E345. Any one of the methods of E332 to E344, wherein the method further comprises determining whether the subject harbors an activating mutation in ALK2 (ACVR1) prior to administering the antibody or ALK2-binding fragment thereof, the nucleic acid molecule, the expression vector, or the pharmaceutical composition.

[0286] E346. A method for treating a subject having Sjogren's syndrome by administering to the subject a therapeutically effective amount of an antibody or ALK2-binding fragment thereof of any one of E1 to E264, a nucleic acid molecule of E265, an expression vector of E266, or a pharmaceutical composition of E268 or E269.

[0287] E347. The method of E346, wherein said method reduces or ameliorates a symptom of Sjogren's syndrome. E348. The method of E347, wherein said symptom is dry eyes, dry mouth, eye irritation, fibrosis of the lacrimal gland, inflammation of the lacrimal gland, blurred vision, systemic inflammation, joint pain, or fatigue.

[0288] E349. The method of E348, wherein said symptom is dry eye. E350. A method of treating or alleviating dry eye in a subject having Sjogren's syndrome (treating dry eye associated with Sjogren's syndrome) by administering to the subject a therapeutically effective amount of an antibody or ALK2-binding fragment thereof of any one of E1 to E264, a nucleic acid molecule of E265, an expression vector of E266, or a pharmaceutical composition of E268 or E269.

[0289] E351. The method of any one of E346 to E350, wherein said method increases tear production (eg, increases the amount of tear produced). E352. The method of any one of E346 to E351, wherein said method reduces eye irritation.

[0290] E353. Any one of the methods of E346 to E352, wherein the method improves vision (eg, improves eyesight). E354. Any one of the methods of E346 to E353, wherein said method reduces fibrosis of the lacrimal gland.

[0291] E355. Any one of the methods of E346 to E354, wherein the method reduces inflammation of the lacrimal gland. E356. The method of any one of E346 to E355, wherein said method increases saliva production or saliva flow.

[0292] E357. The method of any one of E346-E356, wherein said method reduces systemic inflammation. E358. The method of any one of E346 to E357, wherein said method reduces joint pain.

[0293] E359. The method of any one of E346 to E358, wherein said method reduces fatigue. E360. A method for treating a subject having multiple osteochondromas (MO) by administering to the subject a therapeutically effective amount of any one of antibodies or ALK2-binding fragments thereof of E1 to E264, a nucleic acid molecule of E265, an expression vector of E266, or a pharmaceutical composition of E268 or E269.

[0294] E361. A method for preventing or reducing the formation of osteochondroma in a subject with MO (e.g., preventing the formation of osteochondroma in a subject with MO who has not yet developed osteochondroma or reducing or preventing the formation of osteochondroma in a subject with MO who has already developed one or more osteochondromas), comprising administering to the subject a therapeutically effective amount of an antibody or ALK2-binding fragment thereof of any one of E1 to E264, a nucleic acid molecule of E265, an expression vector of E266, or a pharmaceutical composition of E268 or E269.

[0295] A method for reducing the size of an osteochondroma in a subject having E362.MO, comprising administering to the subject a therapeutically effective amount of any one of antibodies or ALK2-binding fragments thereof of E1 to E264, a nucleic acid molecule of E265, an expression vector of E266, or a pharmaceutical composition of E268 or E269.

[0296] A method for slowing or inhibiting the growth of osteochondroma in a subject with E363.MO, comprising administering to the subject a therapeutically effective amount of any one of antibodies or ALK2-binding fragments thereof of E1 to E264, a nucleic acid molecule of E265, an expression vector of E266, or a pharmaceutical composition of E268 or E269.

[0297] A method for reducing the number of osteochondromas in a subject with E364.MO, comprising administering to the subject a therapeutically effective amount of any one of antibodies E1 to E264 or ALK2-binding fragments thereof, a nucleic acid molecule of E265, an expression vector of E266, or a pharmaceutical composition of E268 or E269.

[0298] E365. Any one of the methods E360-E364, wherein the osteochondroma is formed on the long bone. E366. Any one of the methods E360-E364, wherein the osteochondroma is formed on the flat bone.

[0299] E367. Any one of E360-E366, in which the osteochondroma forms at the growing end (metaphysis) of the bone. E368. A method for treating a subject having diffuse intrinsic pontine glioma (DIPG), comprising administering to the subject a therapeutically effective amount of any one of antibodies or ALK2-binding fragments thereof of E1 to E264, a nucleic acid molecule of E265, an expression vector of E266, or a pharmaceutical composition of E268 or E269.

[0300] E369. Method E368, wherein the method extends survival time (eg, life span) of the subject. E370. The method of E368 or E369, wherein said method reduces tumor growth.

[0301] E371. The method of any one of E368 to E370, wherein said method reduces tumor volume. E372. The method of any one of E368-E371, wherein said method reduces tumor metastasis.

[0302] E373. Any one of the methods E368 to E372, wherein the method further comprises determining whether the subject has a mutation in ALK2 (ACVR1) prior to administering the antibody or ALK2-binding fragment thereof, the nucleic acid molecule, the expression vector, or the pharmaceutical composition.

[0303] E374. A method for reducing or preventing the onset of posterior capsule opacification (PCO) in a subject, comprising administering to a subject in need thereof a therapeutically effective amount of an antibody or ALK2-binding fragment thereof of any one of E1 to E264, a nucleic acid molecule of E265, an expression vector of E266, or a pharmaceutical composition of E268 or E269.

[0304] A method for treating a subject having or at risk of developing E375.PCO, comprising administering to the subject a therapeutically effective amount of any one of antibodies E1 to E264 or ALK2-binding fragments thereof, a nucleic acid molecule of E265, an expression vector of E266, or a pharmaceutical composition of E268 or E269.

[0305] E376. The method of E374 or E375, wherein said subject has had or is soon to have cataract surgery (e.g., the subject is scheduled to have cataract surgery in 6 months, 5 months, 4 months, 3 months, 2 months, 1 month, 2 weeks, 1 week, or less).

[0306] E377. The method of any one of E374 to E376, wherein said method improves vision. E378. The method of any one of E374 to E377, wherein the method reduces light sensitivity or glare.

[0307] E379. The method of any one of E374-E378, wherein the method reduces or inhibits fibrosis (eg, fibrosis on or near the posterior capsule, eg, fibrosis near an implanted lens).

[0308] E380. The method of any one of E374-E379, wherein said method reduces or inhibits ocular inflammation. E381. A method for treating a subject having or at risk of developing cardiac hypertrophy, comprising administering to the subject a therapeutically effective amount of an antibody or ALK2-binding fragment thereof of any one of E1 to E264, a nucleic acid molecule of E265, an expression vector of E266, or a pharmaceutical composition of E268 or E269.

[0309] E382. A method for delaying or preventing the onset of cardiac hypertrophy, comprising administering to a subject a therapeutically effective amount of any one of antibodies or ALK2-binding fragments thereof of E1 to E264, a nucleic acid molecule of E265, an expression vector of E266, or a pharmaceutical composition of E268 or E269.

[0310] E383. The method of E381 or E382, wherein said subject is identified as having hypertension or valvular disease. E384. A method for treating a subject having or at risk of developing cardiac fibrosis, comprising administering to the subject a therapeutically effective amount of an antibody or ALK2-binding fragment thereof of any one of E1-E264, a nucleic acid molecule of E265, an expression vector of E266, or a pharmaceutical composition of E268 or E269.

[0311] E385. A method for delaying or preventing the onset of cardiac fibrosis, comprising administering to a subject a therapeutically effective amount of any one of antibodies or ALK2-binding fragments thereof of E1 to E264, a nucleic acid molecule of E265, an expression vector of E266, or a pharmaceutical composition of E268 or E269.

[0312] E386. The method of E384 or E385, wherein said subject is identified as having hypertension, diabetic hypertrophic cardiomyopathy, or idiopathic dilated cardiomyopathy, or said subject has had a myocardial infarction.

[0313] E387. The method of any one of E381-E386, wherein said method reduces or reverses cardiac fibrosis. E388. The method of any one of E381-E387, wherein said method ameliorates one or more symptoms of cardiac hypertrophy or cardiac fibrosis.

[0314] E389. The method of E388, wherein the symptoms of cardiac hypertrophy or cardiac fibrosis are decreased exercise capacity, decreased blood ejection volume, increased left ventricular end diastolic pressure, increased pulmonary capillary wedge pressure, decreased cardiac output, decreased cardiac index, increased pulmonary artery pressure, increased left ventricular end systolic and diastolic dimensions, increased left and right ventricular wall stress, increased wall tension and / or wall thickness, decreased myocardial contractility, increased myocardial cell area, or increased extracellular matrix deposition in the myocardium.

[0315] E390. Any one of the methods of E270-E310, wherein the antibody or ALK2-binding fragment thereof, nucleic acid molecule, expression vector, or pharmaceutical composition is administered in an amount sufficient to increase bone mineral density, decrease bone resorption (e.g., decrease bone loss), reduce risk of fracture, increase bone strength, decrease the rate of bone resorption (e.g., decrease the rate of bone loss), increase bone formation, increase the rate of bone formation, decrease osteoclast activity, increase osteoblast activity, or treat a bone disease.

[0316] E391. Any one of the methods of E311-E331, wherein the antibody or ALK2-binding fragment thereof, nucleic acid molecule, expression vector, or pharmaceutical composition is administered in an amount sufficient to increase red blood cell levels, increase hemoglobin levels, increase red blood cell formation, increase red blood cell count, increase hematocrit, reduce the need for blood transfusions, increase iron levels, reduce iron deficiency, reduce increased hepcidin levels, or treat anemia.

[0317] E392. Any one of the methods of E332-E345, wherein said antibody or ALK2-binding fragment thereof, said nucleic acid molecule, said expression vector, or said pharmaceutical composition is administered in an amount sufficient to treat FOP, slow or stop the progression of FOP, delay the onset of FOP, prevent heterotopic ossification, reduce heterotopic ossification, prevent or reduce ectopic bone formation, or prevent or reduce the recurrence of ectopic bone.

[0318] E393. Any one of the methods of E346-E359, wherein the antibody or ALK2-binding fragment thereof, nucleic acid molecule, expression vector, or pharmaceutical composition is administered in an amount sufficient to treat Sjogren's syndrome, treat or alleviate dry eye associated with Sjogren's syndrome, reduce or ameliorate the symptoms of Sjogren's syndrome, increase tear production, reduce eye irritation, improve vision, reduce fibrosis of the lacrimal gland, reduce inflammation of the lacrimal gland, or increase saliva production or saliva flow.

[0319] E394. Any one of the methods of E360-E367, wherein the antibody or ALK2-binding fragment thereof, the nucleic acid molecule, the expression vector, or the pharmaceutical composition is administered in an amount sufficient to treat MO, prevent formation of osteochondroma, reduce formation of osteochondroma, reduce size of osteochondroma, reduce growth of osteochondroma, or reduce number of osteochondroma.

[0320] E395. Any one of the methods of E368-E373, wherein the antibody or ALK2-binding fragment thereof, the nucleic acid molecule, the expression vector, or the pharmaceutical composition is administered in an amount sufficient to treat DIPG, prolong survival (e.g., increase lifespan), reduce tumor growth, reduce tumor volume, or reduce tumor metastasis.

[0321] E396. Any one of the methods of E374-E380, wherein the antibody or ALK2-binding fragment thereof, the nucleic acid molecule, the expression vector, or the pharmaceutical composition is administered in an amount sufficient to treat PCO, prevent the onset of PCO, improve vision, reduce light sensitivity or glare, reduce or inhibit fibrosis, or reduce or inhibit ocular inflammation.

[0322] E397. Any one of the methods of E381-E3889, wherein the antibody or ALK2-binding fragment thereof, nucleic acid molecule, expression vector, or pharmaceutical composition is administered in an amount sufficient to treat cardiac hypertrophy, treat cardiac fibrosis, delay or prevent the onset of cardiac hypertrophy, delay or prevent the onset of cardiac fibrosis, or ameliorate a symptom of cardiac fibrosis.

[0323] definition As used herein, the term "about" refers to a value within a range of 10% above or below the stated value.

[0324] As used herein, "administration" refers to providing or giving a therapeutic agent (e.g., an ALK2 antibody or antigen-binding fragment thereof (e.g., an ALK2-binding fragment) described herein) to a subject by any effective route. Exemplary routes of administration are described herein below.

[0325] The term "antibody" is used in the broadest sense and specifically encompasses intact monoclonal antibodies, polyclonal antibodies, multispecific antibodies (e.g., bispecific antibodies) formed from at least two intact antibodies, and antibody fragments so long as they exhibit the desired biological activity.

[0326] An "antibody fragment" comprises a portion of an intact antibody, preferably the antigen-binding or variable region of the intact antibody. Examples of antibody fragments include Fab, Fab', F(ab')2, and Fv fragments; diabodies; linear antibodies (Zapata et al. Protein Eng. 8(10):1057-1062 (1995)); single-chain antibody molecules; and multispecific antibodies formed from antibody fragments.

[0327] The term "monoclonal antibody" as used herein refers to an antibody obtained from a population of substantially homogeneous antibodies, i.e., the individual antibodies within the population are identical except for possible minor mutations that may occur naturally. Monoclonal antibodies are highly specific, being directed against a single antigenic site.

[0328] The term "monoclonal antibody" as used herein specifically includes "chimeric" antibodies in which a portion of the heavy and / or light chains are identical or homologous to corresponding sequences in antibodies derived from a particular species or belonging to a particular antibody class or subclass, while the remainder of the chains are identical or homologous to corresponding sequences in antibodies derived from another species or belonging to another antibody class or subclass, as well as fragments of such antibodies, so long as they exhibit the desired biological activity (U.S. Pat. No. 4,816,567; Morrison et al., Proc. Natl. Acad. Sci. USA, 81:6851-6855 (1984)).

[0329] "Humanized" forms of non-human (e.g., murine) antibodies are chimeric antibodies, antibody chains, or fragments thereof (such as Fv, Fab, Fab', F(ab')2, or other antigen-binding subsequences of an antibody) that contain minimal sequence derived from a non-human antibody. In most cases, humanized antibodies are human antibodies (recipient antibody) in which residues from a complementarity-determining region (CDR) of the recipient are replaced by residues from a CDR of a non-human species (donor antibody) such as mouse, rat, or rabbit having the desired specificity, affinity, or capacity. In some instances, Fv framework region (FR) residues of the human antibody are replaced by corresponding non-human residues. Furthermore, humanized antibodies may comprise residues that are found neither in the recipient antibody nor in the imported CDR or framework sequences.

[0330] As used herein, the terms "bone mineral density (BMD)", "bone density" and "bone mass" refer to a measure of the amount of bone mineral (e.g., calcium) in bone tissue. BMD can be measured by well-established clinical techniques known to those skilled in the art (e.g., by single-energy or dual-energy photon or x-ray absorptiometry (DEXA)). The concept of BMD is related to the mass of mineral per unit volume of bone, but clinically it is measured by proxy according to the optical density per square centimeter of the bone surface during imaging. BMD measurements are used in clinical medicine as an indirect indicator of osteoporosis and fracture risk. In some embodiments, BMD test results are provided as a T-score, where the T-score represents the subject's BMD compared to the ideal or peak bone mineral density of a healthy 30-year-old adult. A score of 0 indicates that the BMD is equal to the normal reference value for healthy young adults. The difference between the subject's measured BMD and the BMD of the healthy young adult reference value is measured in standard deviation units (SD). Thus, a T-score of +1SD to -1SD may indicate normal BMD, a T-score of -1SD to -2.5SD may indicate low bone mass (e.g., osteopenia), and a T-score less than -2.5SD may indicate osteoporosis or severe osteoporosis. In some embodiments, an ALK2 antibody or antigen-binding fragment thereof (e.g., an ALK2 binding fragment) of the invention is administered to a subject in need thereof, where the patient has low bone mass (e.g., a T-score of -1SD to -2.5SD). In some embodiments, an ALK2 antibody or antigen-binding fragment thereof (e.g., an ALK2 binding fragment) of the invention is administered to a subject in need thereof, where the patient has osteoporosis (e.g., a T-score less than -2.5SD). In some embodiments, administration of an ALK2 antibody or antigen-binding fragment thereof (e.g., an ALK2 binding fragment) of the invention treats the subject by increasing the subject's BMD (e.g., increasing the subject's T-score).

[0331] As used herein, the term "bone strength" refers to bone measurements determined by bone mineral density as well as bone quality. Bone quality is influenced by bone geometry, microarchitecture, and tissue composition properties. Bone strength can be used to assess fracture risk.

[0332] As used herein, the term "bone disease" refers to a condition characterized by bone damage (e.g., decreased bone mineral density, decreased bone strength, and / or bone loss). Such diseases or conditions may be caused by an imbalance in osteoblast and / or osteoclast activity (e.g., increased bone resorption or decreased bone formation). Bone diseases include primary osteoporosis, secondary osteoporosis, osteopenia, osteopetrosis, bone fracture, bone loss associated with bone cancer or cancer metastasis (e.g., bone loss associated with multiple myeloma), Paget's disease, renal osteodystrophy, treatment-related bone loss, diet-related bone loss, bone loss associated with obesity treatment, low gravity-related bone loss, and immobility-related bone loss.

[0333] As used herein, the term "bone remodeling" or "bone metabolism" refers to the process for maintaining bone strength and ionic homeostasis by replacing discrete portions of old bone with packets of newly synthesized proteinaceous matrix. Bone is resorbed by osteoclasts and deposited by osteoblasts in a process called ossification. Bone cell activity plays an important role in this process. Conditions that result in bone loss can be caused by either increased resorption or decreased ossification. In healthy individuals, bone formation exceeds resorption during childhood. As the aging process begins, resorption exceeds formation. The rate of bone resorption is also usually much higher in older women after menopause due to estrogen deficiency associated with menopause.

[0334] As used herein, the term "bone resorption" or "bone catabolic activity" refers to the process by which osteoclasts break down bone tissue and release minerals, resulting in the movement of minerals (e.g., calcium) from bone tissue to the blood. Increased rates of bone resorption are associated with aging, including in postmenopausal women. High rates of bone resorption, or rates of bone resorption that exceed the rate of ossification, are associated with bone disorders, such as reduced bone mineral density, including osteopenia and osteoporosis, resulting in bone loss. In some embodiments, the ALK2 antibody or antigen-binding fragment thereof (e.g., ALK2-binding fragment) of the invention is administered to a subject in need thereof to reduce bone resorption (e.g., the rate of bone resorption in the subject).

[0335] As used herein, the terms "bone formation," "ossification," "osteogenesis," or "bone anabolic activity" refer to the process of forming new bone tissue by osteoblasts. In some embodiments, an ALK2 antibody or antigen-binding fragment thereof (e.g., an ALK2-binding fragment) of the invention is administered to a subject in need thereof to increase bone formation (e.g., to increase the rate of bone formation or bone generation in a subject).

[0336] As used herein, the term "cell type" refers to a group of cells that share a statistically separable phenotype based on gene expression data. For example, cells of a common cell type may share similar structural and / or functional characteristics, such as similar gene activation patterns and antigen presentation profiles. Cells of a common cell type may include cells isolated from a common tissue (e.g., epithelial, nervous, connective, bone, or muscle tissue) and / or cells isolated from a common organ, tissue system, blood vessel, or other structure and / or common region of a life cycle.

[0337] As used herein, the terms "conservative mutation," "conservative substitution," and "conservative amino acid substitution" refer to the replacement of one or more amino acids with one or more different amino acids that exhibit similar physicochemical properties, such as polarity, electrostatic charge, and steric bulk. These properties for each of the 20 naturally occurring amino acids are summarized in Table 1 below.

[0338] [Table 1]

[0339] From this table it can be seen that conservative amino acid families include: (i) G, A, V, L, and I; (ii) D and E; (iii) C, S, and T; (iv) H, K, and R; (v) N and Q; and (vi) F, Y, and W. Thus, a conservative mutation or substitution is one that replaces one amino acid with a member of the same amino acid family (e.g., Ser with Thr, Lys with Arg).

[0340] As used herein, the term "isolated antibody" refers to an antibody that is substantially free of other antibodies having different antigenic specificities (e.g., an isolated antibody that binds ALK2 is substantially free of contaminants, e.g., antibodies that do not bind ALK2). Moreover, an "isolated" antibody has been identified and separated and / or recovered from a component of its natural environment, which are materials that would interfere with diagnostic or therapeutic uses for the antibody, and may include enzymes, hormones, and other proteinaceous or non-proteinaceous solutes.

[0341] As used herein, the terms "increase" and "decrease" refer to modulation resulting in a greater or lesser amount of the function, expression, or activity of the subject, respectively, compared to a reference. For example, after administration of an ALK2 antibody or antigen-binding fragment thereof (e.g., an ALK2-binding fragment) of the present invention in a manner described herein, the amount of a marker measured herein (e.g., red blood cell level, bone mineral density, tear production, survival time) may increase in the subject relative to the amount of the marker before administration or relative to an untreated subject. Alternatively, the amount of a marker measured herein (e.g., heterotopic ossification, or osteochondroma size, number, or formation) may decrease in the subject relative to the amount of the marker before administration or relative to an untreated subject.

[0342] Typically, the measurements are taken after administration at a time when the administration has had the described effect, for example, at least one week, one month, three months, or six months after the treatment regimen has been initiated. As used herein, the terms "increasing red blood cell levels" and "promoting red blood cell formation" refer to clinically observable measures such as increased hematocrit, red blood cell count, and hemoglobin measurements, and are intended to be neutral as to the mechanism by which such changes occur. As used herein, the term "low red blood cell levels" refers to red blood cell count, hematocrit, and / or hemoglobin measurements that are below the range of values ​​considered normal for a subject's age and sex. In some embodiments, an ALK2 antibody or antigen-binding fragment thereof (e.g., an ALK2-binding fragment) of the invention is administered to a subject (e.g., a subject with anemia) to increase red blood cell levels (e.g., increase hemoglobin levels or red blood cell count).

[0343] As used herein, the terms "erythropoiesis" and "erythropoiesis" refer to the production of red blood cells, such as the process of erythropoiesis whereby red blood cells are produced in the bone marrow. As used herein, the term "anemia" refers to a hemoglobin or red blood cell abnormality that results in a reduced level of oxygen in the blood. Anemia may be related to abnormal production, processing, or function of red blood cells and / or hemoglobin. The term anemia refers to a reduced number of red blood cells and / or a reduced level of hemoglobin in the blood compared to normal blood levels.

[0344] As used herein, the term "inflammatory anemia" refers to a type of anemia caused by inflammatory cytokines and characterized by hypoferremia, mainly due to elevated hepcidin, despite sufficient iron stores. Bone morphogenetic proteins (BMPs) upregulate hepcidin by activating the SMAD signaling pathway through ALK2. Inflammatory anemia is also called anemia of chronic disease. Chronic conditions that cause inflammatory anemia include infectious diseases (e.g., chronic infectious diseases such as HIV / AIDS or tuberculosis), autoimmune diseases (e.g., rheumatoid arthritis or lupus), cancer (e.g., cancer or cancer treatment), inflammatory bowel disease (e.g., Crohn's disease or ulcerative colitis), and chronic kidney disease.

[0345] As used herein, the terms "iron-refractory iron deficiency anemia" and "IRIDA" refer to inherited forms of iron deficiency anemia. The main features of IRIDA include lifelong anemia (hemoglobin 6-9 g / dL); very small red blood cell size (microcytic) with a mean corpuscular volume (MCV) of 45-65 fL; very low levels of iron in the blood (transferrin saturation <5%); abnormal oral iron absorption - no response to oral iron supplements or failure of an "oral iron challenge"; abnormal iron utilization - slow, incomplete, and temporary response to parenteral iron (iron injected intravenously); and the presence of other affected family members with an autosomal recessive inheritance pattern. The diagnosis of IRIDA can be confirmed by measuring the levels of hepcidin in the blood. Mutations in TMPRSS6 have been found to be associated with IRIDA.

[0346] As used herein, the terms "heterotopic ossification" and "ectopic bone formation" refer to the abnormal growth of bone in non-skeletal tissues, such as muscles, tendons, and other soft tissues. "Ectopic bone" refers to bone formed in non-skeletal tissues. Heterotopic ossification can develop in subjects with FOP.

[0347] As used herein, the terms "fibrodysplasia ossificans progressiva" and "FOP" refer to a disorder in which skeletal muscles and connective tissues such as tendons and ligaments are gradually replaced by bone (ossification). The condition leads to bone formation outside of the skeleton (extraskeletal or heterotopic bone), restricting movement. The process is typically evident during early childhood, beginning in the neck and shoulders and working its way down the torso and into the limbs. FOP is easy to diagnose, as people with FOP are born with an abnormal big toe (hallux valgus). Trauma, such as a fall or invasive medical procedure, or a viral illness, can cause symptoms of muscle swelling and inflammation (myositis). These flare-ups can last from days to months and often result in permanent bone growth in the damaged area. FOP is caused by a mutation in the ACVR1 gene and is inherited in an autosomal dominant manner.

[0348] As used herein, the term "fibrosis" refers to a pathological process of excessive formation of fibrous connective tissue. Fibrosis is characterized by fibroblast accumulation and collagen deposition beyond normal deposition in a particular tissue. In response to inflammation or tissue injury, nearby fibroblasts can migrate to the wound, proliferate, and generate large amounts of collagenous extracellular matrix. When fibrosis occurs in response to injury, the term "scar" can be used synonymously. Fibrosis can develop in many tissues of the body, including, for example, lung, skin, liver, kidney, heart, eye, lacrimal gland, tendon, cartilage, pancreatic tissue, uterine tissue, nervous tissue, testis, ovary, adrenal gland, artery, vein, colon, small and large intestine, biliary tract, and intestine.

[0349] As used herein, the term "cardiac hypertrophy" refers to the abnormal enlargement or thickening of the myocardium resulting from the process by which adult cardiomyocytes respond to stress through hypertrophic growth. Such growth is characterized by an increase in cell size without cell division, the assembly of additional sarcomeres within the cell to maximize force production, and the activation of fetal cardiac gene programs. Cardiac hypertrophy is often associated with increased risk of morbidity and mortality, and has been found to be associated with fibrotic tissue deposition in the cardiac interstitium with alterations in the extracellular matrix scaffold (e.g., cardiac fibrosis).

[0350] As used herein, the term "cardiac fibrosis", also known as myocardial fibrosis, refers to fibrosis of the myocardium, characterized by wall stiffening, reduced contractility, and overall deterioration of cardiac function. Cardiac myofibroblasts mediate fibrosis by excessive deposition of connective tissue (e.g., extracellular matrix) in the interstitial space. Cardiac fibrosis is often found in association with cardiac hypertrophy.

[0351] As used herein, the term "dry eye" refers to a chronic condition that occurs when the eye does not produce enough tears (e.g., reduced tear production or tear volume) or when tears evaporate rapidly. Dry eye can be accompanied by ocular discomfort and abnormal visual function.

[0352] As used herein, the term "Sjogren's syndrome" refers to a systemic inflammatory disorder characterized by dry mouth, decreased tearing, and dry mucous membranes. Dry eyes and mouth are the most common symptoms of this syndrome.

[0353] As used herein, "dry eye associated with Sjogren's syndrome" refers to dry eye associated with primary Sjogren's syndrome and dry eye associated with secondary Sjogren's syndrome, which is generally classified as dry eye with reduced tear production.

[0354] As used herein, the term "multiple osteochondromas" or "MO" refers to a condition or disease associated with the formation of osteochondromas in bones, such as the ends of long bones or flat bones. Subjects with MO often carry loss-of-function mutations in exostosin genes, such as EXT1 or EXT2. MO is also known as multiple congenital exostoses, Bessel-Hagen disease, multiple osteochondral exostoses, multiple chondromatous exostoses, multiple congenital exostoses, and multiple congenital osteochondromas.

[0355] As used herein, the term "osteochondroma" refers to a benign (non-cancerous) tumor that develops during childhood or adolescence. It is an abnormal overgrowth of cartilage and bone, usually forming on the surface of bones near the growth plates. Osteochondromas most often form on the long bones of the legs and arms, and on flat bones such as the pelvis and shoulder blade.

[0356] As used herein, the term "diffuse intrinsic pontine glioma" refers to a highly aggressive and difficult to treat brain tumor that arises in the pons and accounts for approximately 20% of all pediatric brain tumors. The median overall survival for children with diffuse intrinsic pontine glioma (DIPG) is 9-11 months, with 1-year and 2-year survival rates of approximately 30% and less than 10%, respectively. DIPGs have been found to be molecularly distinct from adult gliomas, and frequently harbor mutations in genes that code for histone variants. Mutations in ALK2 (ACVR1) have also been observed in DIPGs.

[0357] As used herein, the term "posterior capsule opacification" refers to the most common postoperative complication of cataract surgery, occurring in 20%-40% of patients. In posterior capsule opacification (PCO), the posterior capsule undergoes secondary opacification due to migration, proliferation, and differentiation of lens epithelial cells. PCO can cause significant visual symptoms, especially when the central visual axis is involved, and can be identified by the formation of fibrosis (e.g., scar tissue) in the posterior capsule (e.g., posterior to the lens implant). Symptoms of PCO include a gradual decrease in visual acuity, blurred vision, sensitivity to sunlight, and the appearance of glare or halos around lights.

[0358] "Percent (%) amino acid sequence identity" to a reference polypeptide sequence is defined as the percentage of amino acid residues in a candidate sequence that are identical to the amino acid residues in the reference polypeptide sequence after aligning the sequences and introducing gaps as necessary to achieve the maximum percent sequence identity over the entire length of comparison. Conservative substitutions are not considered as part of the sequence identity. Alignment for purposes of determining percent amino acid sequence identity can be accomplished in a variety of ways within the skill of the art, for example, using publicly available computer software such as BLAST, BLAST-2, ALIGN, and Megalign (DNASTAR) software. In certain embodiments, the amino acid sequence identity is at least 80%, 90%, 95%, 98%, or 99%.

[0359] As used herein, the term "affinity" or "binding affinity" refers to the strength of the binding interaction between two molecules. Primarily, binding affinity refers to the strength of the sum of non-covalent interactions between a molecule and its binding partner, e.g., between an ALK2 antibody or antigen-binding fragment thereof (e.g., an ALK2-binding fragment) and an ALK2 protein. Unless otherwise specified, binding affinity refers to the intrinsic binding affinity that reflects a 1:1 interaction between members of a binding pair. The binding affinity between two molecules is generally measured by the dissociation constant (K D ) or affinity constant (K A ) Two molecules that have low binding affinity for each other generally bind slowly and tend to dissociate easily, with a large K D Two molecules that have a high affinity for each other generally bind easily, tend to bind for a long time, and have a small K D The K of two interacting molecules is D can be determined using methods and techniques well known in the art, for example, surface plasmon resonance. D is k off / k on It is calculated as the ratio of

[0360] As used herein, the term "specific binding" of an antibody or ALK2-binding fragment thereof (e.g., Fab, Fab', F(ab')2, or Fv fragment) is binding to a target molecule that is measurably different from binding to a molecule that is not the target molecule. As used herein, specific binding refers to greater than 95% preference for binding of a specific antigen over background ("non-specific") binding. "Substantially specific" binding refers to greater than about 80% preference for binding of a specific antigen over background. Binding can be measured using a variety of methods, including, but not limited to, Western blot, immunoblot, enzyme-linked immunosorbent assay ("ELISA"), radioimmunoassay ("RIA"), immunoprecipitation, surface plasmon resonance, biolayer interferometry, chemiluminescence, fluorescence polarization, phosphorescence, immunohistochemistry, matrix-assisted laser desorption / ionization time-of-flight ("MALDI-TOF") mass spectrometry, microcytometry, microarrays, microscopy, fluorescence-activated cell sorting ("FACS"), and flow cytometry. In certain embodiments, an antibody that specifically binds to a target (e.g., ALK2) has a dissociation constant (K) of up to 0.1 nM, 0.15 nM, 0.2 nM, 0.3 nM, 0.5 nM, 0.7 nM, 1.0 nM, 2.0 nM, 4.0 nM, 5.0 nM, 7.0 nM, 10.0 nM, 14.0 nM, or 15.0 nM. D ).

[0361] As used herein, the term "polypeptide" refers to a single polymer in which the monomers are amino acid residues covalently linked to one another through amide bonds. Polypeptide is intended to encompass any amino acid sequence, whether naturally occurring, recombinant, or synthetically produced.

[0362] As used herein, the terms "effective amount," "therapeutically effective amount," and "sufficient amount" of a composition described herein or an ALK2 antibody or antigen-binding fragment thereof (e.g., an ALK2-binding fragment) refer to an amount sufficient to effect beneficial or desired results, including clinical results, when administered to a subject. Thus, the "effective amount" or its synonyms will depend on the context in which it is being applied. For example, when treating a patient having or at risk of developing a disease such as a bone disease (e.g., osteoporosis, or a condition involving bone damage, e.g., primary osteoporosis, secondary osteoporosis, osteopenia, osteopetrosis, bone fracture, bone loss associated with bone cancer or cancer metastasis, Paget's disease, renal osteodystrophy, treatment-related bone loss, diet-related bone loss, bone loss associated with the treatment of obesity, low gravity-related bone loss, or immobility-related bone loss), a disease or condition involving low red blood cell levels (e.g., anemia or blood loss), heterotopic ossification (e.g., heterotopic ossification due to FOP), Sjogren's syndrome (e.g., dry eye associated with Sjogren's syndrome), DIPG, MO, PCO, or cardiac hypertrophy and / or cardiac fibrosis, it is an amount of the composition or ALK2 antibody or antigen-binding fragment thereof (e.g., ALK2-binding fragment) sufficient to achieve a therapeutic response compared to the response obtained without administration of the composition or ALK2 antibody or antigen-binding fragment thereof (e.g., ALK2-binding fragment). The amount of a given composition described herein that corresponds to such an amount will vary depending on a variety of factors, such as the given agent, formulation, route of administration, type of disease or disorder, subject identity (e.g., age, sex, weight) or host being treated, but can nevertheless be determined by one of skill in the art by routine methods known in the art. Dosage regimens can be adjusted to provide the optimal therapeutic response.

[0363] As used herein, "locally" or "local administration" refers to administration to a particular site in the body for a local rather than a systemic effect. Examples of local administration are transdermal, inhalation, intra-articular, intrathecal, intravaginal, intravitreal, intrauterine, intralesional administration, lymph node administration, intratumoral administration, and administration to a mucous membrane of a subject, where administration is for a local rather than a systemic effect.

[0364] As used herein, the term "pharmaceutical composition" refers to a mixture containing a therapeutic agent, optionally in combination with one or more pharma- ceutical acceptable excipients, diluents, and / or carriers, that is administered to a subject to prevent, treat, or control a particular disease or condition that affects or may affect the subject. The pharmaceutical composition may be in tablet or capsule form for oral administration, or in aqueous form for intravenous or subcutaneous administration.

[0365] As used herein, the term "pharmaceutical acceptable carrier or excipient" refers to an excipient or diluent in a pharmaceutical composition. A pharmaceutical acceptable carrier should be compatible with other ingredients of the formulation, free from undue toxicity, irritation, allergic reactions, and other problematic complications commensurate with a reasonable benefit / risk ratio, and suitable for contact with the tissues of a subject. In the present invention, a pharmaceutical acceptable carrier or excipient should provide sufficient pharmaceutical stability for the ALK2 antibody or antigen-binding fragment thereof (e.g., ALK2-binding fragment) of the present invention. The nature of the carrier or excipient varies depending on the method of administration. For example, aqueous carriers are mainly used for intravenous administration, while solid carriers are preferred for oral administration.

[0366] As used herein, the term "sample" refers to a specimen isolated from a subject, such as blood, blood components (e.g., serum or plasma), urine, saliva, amniotic fluid, cerebrospinal fluid, tissue (e.g., neural tissue, placental tissue, or skin tissue), pancreatic juice, chorionic villus specimens, and cells (e.g., blood cells or bone cells).

[0367] As used herein, "treatment" with respect to a disease or condition refers to an approach to obtain beneficial or desired results, e.g., clinical results. Beneficial or desired results include, but are not limited to, alleviation or amelioration of one or more symptoms or conditions; reduction in the extent of the disease or condition; a stable (i.e., not worsening) state of the disease, disorder, or condition; inhibition of the spread of the disease or condition; delay or slowing of the progression of the disease or condition; amelioration or palliation of the disease or condition; and remission (partial or complete), whether detectable or undetectable. "Ameliorating" or "alleviating" a disease or condition means that the extent and / or undesirable clinical symptoms of the disease, disorder, or condition are lessened compared to the extent in the absence of treatment, and / or the time course of progression is slowed or prolonged compared to the time course in the absence of treatment. "Treatment" can also mean extending survival compared to the expected survival in the absence of treatment. Those in need of treatment include those already with the disease or disorder, as well as those susceptible to the disease or disorder or those in need of prevention of the disease or disorder.

[0368] As used herein, the terms "subject" and "patient" refer to a mammal, e.g., a human. Mammals include, but are not limited to, humans and domestic animals and livestock, such as monkeys (e.g., cynomolgus monkeys), mice, dogs, cats, horses, and cattle. The subject treated according to the methods described herein may be a subject diagnosed with a bone disease (e.g., a disease or condition involving bone damage, such as osteoporosis, osteopenia, osteopetrosis, bone fracture, bone loss associated with bone cancer or cancer metastasis, Paget's disease, renal osteodystrophy, treatment-related bone loss, diet-related bone loss, bone loss associated with the treatment of obesity, low gravity-related bone loss, or immobility-related bone loss), a disease involving low blood cell levels (e.g., anemia or blood loss), heterotopic ossification (e.g., heterotopic ossification due to FOP), Sjogren's syndrome (e.g., dry eye associated with Sjogren's syndrome), DIPG, MO, PCO, or cardiac hypertrophy and / or cardiac fibrosis, or a subject diagnosed as at risk of developing these conditions. Diagnosis can be performed by any method or technique known in the art, and one of skill in the art will appreciate that the subject to be treated in accordance with the present disclosure may have undergone standard testing, or may be identified as being at risk without testing due to the presence of one or more risk factors associated with the disease or condition. [Brief description of the drawings]

[0369] [Figure 1A] Figures 1A-1B are a series of graphs showing BMP6 inhibition dose-response curves of anti-ALK2 fABs in C2C12 cells using the SMAD1 luciferase reporter. The 50% inhibition values ​​(IC50) of the 3 and 4 parameter curve fits in Graphpad Prism are shown next to each graph. Figure 1A is a graph of the dose-response curves of the first four fABs generated. [Figure 1B]Figures 1A-1B are a series of graphs showing BMP6 inhibition dose-response curves of anti-ALK2 fABs in C2C12 cells using the SMAD1 luciferase reporter. The 50% inhibition values ​​(IC50) of the 3 and 4 parameter curve fits in Graphpad Prism are shown next to each graph. Figure 1B is a graph of the dose-response curves of five second round fABs generated from Rapid Pool Maturation. [Diagram 2] 1 is a graph showing the effect of treatment with an ALK2 antibody on hemoglobin levels in a mouse model of IRIDA. Treatment with an ALK2 antibody prevents the decrease in hemoglobin associated with TMPRSS6 deficiency (*=p≦0.05; **=p≦0.01). [Figure 3A] 3A-3C are a series of graphs showing the effect of ALK2 antibody treatment on hemoglobin levels, hepcidin concentrations, and iron levels in a mouse model of IRIDA. Treatment with ALK2 antibody reverses the reduction in hemoglobin associated with TMPRSS6 deficiency (*=p≦0.05; **=p≦0.01). [Figure 3B] 3A-3C are a series of graphs showing the effect of ALK2 antibody treatment on hemoglobin levels, hepcidin concentrations, and iron levels in a mouse model of IRIDA. Treatment with ALK2 antibody reverses the increase in serum hepcidin associated with TMPRSS6 deficiency (**=p≦0.01). [Figure 3C] 3A-3C are a series of graphs showing the effect of ALK2 antibody treatment on hemoglobin levels, hepcidin concentrations, and iron levels in a mouse model of IRIDA. Treatment with ALK2 antibody reverses the reduction in serum iron associated with TMPRSS6 deficiency (*=p≦0.05; **=p≦0.01). [Figure 4] 1 is a graph showing the effect of ALK2 antibody treatment on bone mineral density in a mouse model of osteoporosis. Treatment with ALK2 antibody increases bone mineral density (reduces bone loss) associated with orchiectomy (*=p≦0.05; ****=p≦0.0001). [Diagram 5]6 shows the amino acid sequence of Antibody 1 (SEQ ID NO:67), with the CDR1, CDR2, and CDR3 domains of the light and heavy chains highlighted and the amino acid positions provided. Also provided are the variable light chain, constant light chain, variable heavy chain, CH1 domain, EcoRI linker, Flag tag, tag linker, and the start and end amino acids of the 6 amino acid histidine tag. [Figure 6] 6 shows the amino acid sequence of Antibody 2 (SEQ ID NO:68), with the CDR1, CDR2, and CDR3 domains of the light and heavy chains highlighted and the amino acid positions provided. Also provided are the variable light chain, constant light chain, variable heavy chain, CH1 domain, EcoRI linker, Flag tag, tag linker, and the start and end amino acids of the 6 amino acid histidine tag. [Figure 7] 6 shows the amino acid sequence of Antibody 3 (SEQ ID NO:69), with the CDR1, CDR2, and CDR3 domains of the light and heavy chains highlighted and the amino acid positions provided. Also provided are the variable light chain, constant light chain, variable heavy chain, CH1 domain, EcoRI linker, Flag tag, tag linker, and the start and end amino acids of the 6 amino acid histidine tag. [Figure 8] 7 shows the amino acid sequence of Antibody 4 (SEQ ID NO:70), with the CDR1, CDR2, and CDR3 domains of the light and heavy chains highlighted and the amino acid positions provided. Also provided are the variable light chain, constant light chain, variable heavy chain, CH1 domain, EcoRI linker, Flag tag, tag linker, and the start and end amino acids of the 6 amino acid histidine tag. [Figure 9] 7 shows the amino acid sequence of Antibody 5 (SEQ ID NO:71), with the light and heavy chain CDR1, CDR2, and CDR3 domains highlighted and the amino acid positions provided. Also provided are the variable light chain, constant light chain, variable heavy chain, CH1 domain, EcoRI linker, Flag tag, tag linker, and the start and end amino acids of the 6 amino acid histidine tag. [Figure 10]7 shows the amino acid sequence of Antibody 6 (SEQ ID NO:72), with the light and heavy chain CDR1, CDR2, and CDR3 domains highlighted and the amino acid positions provided. Also provided are the variable light chain, constant light chain, variable heavy chain, CH1 domain, EcoRI linker, Flag tag, tag linker, and the start and end amino acids of the 6 amino acid histidine tag. [Figure 11] 7 shows the amino acid sequence of Antibody 7 (SEQ ID NO:73), with the light and heavy chain CDR1, CDR2, and CDR3 domains highlighted and the amino acid positions provided. Also provided are the variable light chain, constant light chain, variable heavy chain, CH1 domain, EcoRI linker, Flag tag, tag linker, and the start and end amino acids of the 6 amino acid histidine tag. [Figure 12] 7 shows the amino acid sequence of Antibody 8 (SEQ ID NO:74), with the light and heavy chain CDR1, CDR2, and CDR3 domains highlighted and the amino acid positions provided. Also provided are the variable light chain, constant light chain, variable heavy chain, CH1 domain, EcoRI linker, Flag tag, tag linker, and the start and end amino acids of the 6 amino acid histidine tag. [Figure 13] 7 shows the amino acid sequence of Antibody 9 (SEQ ID NO:75), with the light and heavy chain CDR1, CDR2, and CDR3 domains highlighted and the amino acid positions provided. Also provided are the variable light chain, constant light chain, variable heavy chain, CH1 domain, EcoRI linker, Flag tag, tag linker, and the start and end amino acids of the 6 amino acid histidine tag. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0370] Detailed Description of the Invention Described herein are compositions and methods for treating bone disease, anemia, heterotopic ossification (e.g., heterotopic ossification due to FOP), Sjogren's syndrome (e.g., dry eye associated with Sjogren's syndrome), DIPG, MO, PCO, or cardiac hypertrophy and / or cardiac fibrosis. The invention features ALK2 antibodies (e.g., neutralizing antibodies) and antigen-binding fragments thereof (e.g., ALK2-binding fragments), as well as nucleic acid molecules comprising a nucleotide sequence encoding the antibody or the ALK2-binding fragment thereof, and expression vectors comprising a nucleic acid molecule comprising a nucleotide sequence encoding the antibody or the ALK2-binding fragment thereof. The invention also features methods of treating or preventing a bone disease (e.g., a disease or condition involving bone damage such as primary osteoporosis, secondary osteoporosis, osteopenia, osteopetrosis, bone fracture, bone loss associated with bone cancer or cancer metastasis, Paget's disease, renal osteodystrophy, treatment-related bone loss, diet-related bone loss, bone loss associated with the treatment of obesity, low gravity-related bone loss, or immobility-related bone loss), low red blood cell levels (e.g., low hemoglobin levels or low red blood cell count, e.g., anemia), ectopic ossification (e.g., ectopic ossification due to FOP), Sjogren's syndrome (e.g., dry eye associated with Sjogren's syndrome), DIPG, MO, PCO, or cardiac hypertrophy and / or cardiac fibrosis in a subject, such as a human subject, using the ALK2 antibodies or antigen-binding fragments thereof, nucleic acid molecules, expression vectors, and compositions described herein.

[0371] ALK2 antibodies ALK2 (also known as ACVR1) is a bone morphogenetic protein (BMP) receptor of the transforming growth factor-β (TGF-β) superfamily. ALK2 is widely expressed in many tissues during embryogenesis and highly expressed in bone during postnatal development, where it is thought to play essential roles in both osteogenesis and chondrogenesis. Gain-of-function mutations in ALK2, including c.617G>A (p.R206H), c.619C>G (p.Q207E), c.1067G>A (p.G356D), c.982G>T (p.G328W), c.983G>A (p.G328E), c.982G>A (p.G328R), c.774G>C / c.774G>T (P.R258S), c.1124G>C (p.R375P), c.587T>C (p.L196P), c.590-592delCTT (p.P197_F198delinsL), and c.605G>T (p.R202l), have been found in subjects with FOP and are associated with Alk2 R206HStudies using mutant mice suggest that ALK2 regulates osteogenic differentiation of mesenchymal stem cells. Activating mutations in ALK2 have also been observed in approximately 25% of DIPG patients, and small molecule ALK2 inhibitors have been found to prolong survival and reduce cellularity in an orthotopic DIPG xenograft model. ALK2 may also be involved in anemia, as BMP activation of the SMAD signaling pathway via ALK2 induces upregulation of hepcidin, a master regulator of iron homeostasis involved in anemia of inflammation (also known as anemia of chronic disease) and iron-refractory iron deficiency anemia (IRIDA). ALK2 also promotes the proliferation of developing lens epithelial cells. Thus, inhibition of ALK2 may prevent or reduce the abnormal proliferation of lens epithelial cells that leads to PCO. Additionally, small molecule ALK2 inhibitors have been found to reduce inflammation and treat symptoms of Sjögren's syndrome in established mouse models, and to reduce osteochondroma formation, growth, and size in multiple studies using mouse models of MO. Furthermore, both treatment with small molecule ALK2 inhibitors and cardiomyocyte-specific deletion of ALK2 reduced cardiac hypertrophy and left ventricular fibrosis in mice. Furthermore, the fibrosis-promoting effects of ALK2 ligands suggest that inhibition of ALK2 could be used to treat diseases or conditions in which fibrosis contributes to the pathology (e.g., posterior capsule fibrosis in PCO, lacrimal gland fibrosis in Sjögren's syndrome, and fibrosis in cardiac fibrosis and / or cardiac hypertrophy).

[0372] The present invention is based in part on the discovery that the ALK2 antibodies described herein restore hemoglobin levels, hepcidin concentrations, and iron levels in a mouse model of IRIDA and increase bone density (e.g., reduce bone loss) in a mouse model of osteoporosis. Without wishing to be bound by theory, because TMPRSS6 acts to suppress hepcidin secretion and anemia, and inflammatory anemia is often characterized by elevated hepcidin levels, the discovery that the ALK2 antibodies described herein restore hemoglobin levels in the TMPRSS6 knockdown model of IRIDA suggests that the ALK2 antibodies described herein can be used to treat inflammatory anemia. Given the gain-of-function mutations in ALK2 identified in patients with FOP and DIPG, the ALK2 antibodies or antigen-binding fragments thereof described herein can also be used to treat FOP and / or to prevent or treat heterotopic ossification, and to treat DIPG or prolong the survival of subjects with DIPG. Previous studies have also shown that small molecule ALK2 inhibitors can be used to treat symptoms of Sjogren's syndrome, MO (e.g., osteochondroma formation, growth, or size reduction), and cardiac hypertrophy and cardiac fibrosis. Thus, the ALK2 antibodies or antigen-binding fragments thereof described herein can also be used to treat these conditions. Finally, the requirement of ALK2 to promote the proliferation of lens epithelial cells suggests that ALK2 inhibition may be effective in treating PCO. Thus, the ALK2 antibodies or antigen-binding fragments thereof, nucleic acid molecules, expression vectors, and compositions described herein can be administered to a subject, such as a human subject, to treat or prevent a bone disease (e.g., a disease or condition involving bone damage such as primary osteoporosis, secondary osteoporosis, osteopenia, osteopetrosis, bone fracture, bone loss associated with bone cancer or cancer metastasis, Paget's disease, renal osteodystrophy, treatment-related bone loss, diet-related bone loss, bone loss associated with the treatment of obesity, low gravity-related bone loss, or immobility-related bone loss), anemia (e.g., IRIDA, anemia of inflammation), heterotopic ossification (e.g., heterotopic ossification due to FOP), Sjogren's syndrome (e.g., dry eye associated with Sjogren's syndrome), DIPG, MO, PCO, or cardiac hypertrophy and / or cardiac fibrosis.

[0373] Vectors, host cells, and protein production The ALK2 antibodies and antigen-binding fragments thereof (e.g., ALK2-binding fragments) described herein can be produced from host cells. A host cell refers to a vehicle that contains the necessary cellular components, e.g., organelles, required to express the antibodies and antigen-binding fragments thereof described herein from the corresponding nucleic acid. The nucleic acid may be contained in a nucleic acid vector, which can be introduced into the host cell by conventional techniques known in the art (e.g., transformation, transfection, electroporation, calcium phosphate precipitation, direct microinjection, infection, etc.). The choice of nucleic acid vector depends in part on the host cell to be used. In general, preferred host cells are of either eukaryotic (e.g., mammalian) or prokaryotic (e.g., bacterial) origin.

[0374] Construction of Nucleic Acid Vectors and Host Cells Nucleic acid sequences encoding the amino acid sequence of the ALK2 antibodies or antigen-binding fragments thereof (e.g., ALK2-binding fragments) described herein can be prepared by various methods known in the art. These methods include, but are not limited to, oligonucleotide-mediated (or site-directed) mutagenesis and PCR mutagenesis. Nucleic acid molecules encoding the ALK2 antibodies or antigen-binding fragments thereof described herein can be obtained using standard techniques, e.g., gene synthesis. Alternatively, nucleic acid molecules encoding the ALK2 antibodies or antigen-binding fragments thereof described herein can be mutated to contain specific amino acid substitutions using standard techniques in the art, e.g., QuikChange™ mutagenesis. Nucleic acid molecules can be synthesized using nucleotide synthesizers or PCR techniques.

[0375] A nucleic acid sequence encoding an ALK2 antibody or antigen-binding fragment thereof (e.g., an ALK2-binding fragment) described herein can be inserted into a vector capable of replicating and expressing the nucleic acid molecule in a prokaryotic or eukaryotic host cell. Many vectors are available in the art and can be used for the purposes of the present invention. Each vector can contain various components that can be tailored and optimized for compatibility with a particular host cell. For example, vector components can include, but are not limited to, an origin of replication, a selectable marker gene, a promoter, a ribosome binding site, a signal sequence, a nucleic acid sequence encoding a protein of interest, and a transcription termination sequence.

[0376] In some embodiments, mammalian cells can be used as host cells of the present invention. Examples of mammalian cell types include, but are not limited to, human embryonic kidney (HEK) (e.g., HEK293, HEK293F), Chinese hamster ovary (CHO), HeLa, COS, PC3, Vero, MC3T3, NS0, Sp2 / 0, VERY, BHK, MDCK, W138, BT483, Hs578T, HTB2, BT20, T47D, NS0 (a mouse myeloma cell line that does not endogenously produce immunoglobulin chains), CRL7O3O, and HsS78Bst cells. In some embodiments, E. coli cells can also be used as host cells of the present invention. Examples of E. coli strains include, but are not limited to, E. coli 294 (ATCC® 31,446), E. coli λ1776 (ATCC® 31,537), E. coli BL21(DE3) (ATCC® BAA-1025), and E. coli RV308 (ATCC® 31,608). Different host cells have characteristic and specific mechanisms for post-translational processing and modification (e.g., glycosylation) of protein products. Appropriate cell lines or host systems can be selected to ensure correct modification and processing of the expressed polypeptide. The above expression vectors can be introduced into suitable host cells using conventional techniques in the art, such as transformation, transfection, electroporation, calcium phosphate precipitation, and direct microinjection. Once the vector has been introduced into the host cells for protein production, the host cells are cultured in conventional nutrient media modified as appropriate for inducing promoters, selecting transformants, or amplifying the genes encoding the desired sequences.Methods for expressing therapeutic proteins are known in the art and are described, for example, in Paulina Baibas, Argelia Lorence (eds.) Recombinant Gene Expression: Reviews and Protocols (Methods in Molecular Biology), Humana Press; 2nd Edition 2004 and Vladimir Voynov and Justin A. Caravella (eds.) Therapeutic Proteins: Methods and Protocols (Methods in Molecular Biology). (Biology) Humana Press; 2nd Edition 2012.

[0377] Protein Production, Recovery, and Purification The host cells used to produce the ALK2 antibodies or antigen-binding fragments thereof (e.g., ALK2-binding fragments) described herein can be grown in a medium known in the art that is suitable for culturing the selected host cells. Examples of suitable media for mammalian host cells include Minimum Essential Medium (MEM), Dulbecco's Modified Eagle Medium (DMEM), Expi293™ Expression Medium, DMEM supplemented with fetal bovine serum (FBS), and RPMI-1640. Examples of suitable media for bacterial host cells include Luria Broth (LB) with necessary additives, such as a selection agent, e.g., ampicillin. The host cells are cultured at an appropriate temperature, such as about 20° C. to about 39° C., e.g., 25° C. to about 37° C., preferably 37° C., and at a CO2 level, such as 5-10%. The pH of the medium is often about 6.8 to 7.4, e.g., 7.0, depending mainly on the host organism. When an inducible promoter is used in the expression vector of the present invention, protein expression is induced under conditions suitable for activation of the promoter.

[0378] In some embodiments, depending on the expression vector and host cell used, the expressed protein may be secreted from the host cell (e.g., mammalian host cell) into the cell culture medium. Protein recovery may include filtering the cell culture medium to remove cellular debris. The protein may be further purified. The ALK2 antibodies or antigen-binding fragments thereof (e.g., ALK2-binding fragments) described herein can be purified by any method of protein purification known in the art, for example, by chromatography (e.g., ion exchange, affinity, and size exclusion column chromatography), centrifugation, differential solubility, or other standard techniques for protein purification. For example, the protein can be isolated and purified by appropriately selecting an affinity column, such as a Protein A column (e.g., POROS Protein A chromatography), in combination with a chromatography column (e.g., POROS HS-50 cation exchange chromatography), filtration, ultrafiltration, salting out, and dialysis procedures.

[0379] In other embodiments, the host cells may be disrupted, for example, by osmotic shock, sonication, or lysis, and the expressed protein may be recovered. Once the cells are disrupted, cell debris may be removed by centrifugation or filtration. In some cases, the polypeptide may be linked to a marker sequence, such as a peptide, to facilitate purification. One example of a marker amino acid sequence is a hexahistidine peptide (His-tag), which binds to nickel-functionalized agarose affinity columns with micromolar affinity. Other peptide tags useful for purification include, but are not limited to, the hemagglutinin "HA" tag, which corresponds to an epitope derived from the influenza hemagglutinin protein (Wilson et al., Cell 37:767, 1984).

[0380] Alternatively, the ALK2 antibodies or antigen-binding fragments thereof (e.g., ALK2-binding fragments) described herein can be produced by cells of a subject (e.g., a human) by administering a vector (e.g., a viral vector (e.g., a retroviral vector, an adenoviral vector, a poxvirus vector (e.g., a vaccinia virus vector such as Modified Vaccinia Ankara (MVA)), an adeno-associated virus vector, and an alphavirus vector) containing a nucleic acid molecule encoding an antibody or antigen-binding fragment thereof described herein, e.g., in a gene therapy context. Once inside the subject's cells (e.g., by transformation, transfection, electroporation, calcium phosphate precipitation, direct microinjection, infection, etc.), the vector promotes expression of the antibody or antigen-binding fragment thereof, which is then secreted from the cells. If treatment of a disease or disorder is the desired outcome, no further measures may be necessary. If collection of protein is desired, blood can be collected from the subject and the protein purified from the blood by methods known in the art.

[0381] Pharmaceutical Compositions and Preparations The ALK2 antibodies or antigen-binding fragments thereof (e.g., ALK2-binding fragments) described herein can be incorporated into a vehicle for administration to a patient, such as a human patient suffering from bone disease, low red blood cell levels (e.g., low hemoglobin levels or low red blood cell count, e.g., anemia), heterotopic ossification (e.g., heterotopic ossification due to FOP), Sjogren's syndrome (e.g., dry eye associated with Sjogren's syndrome), MO, DIPG, PCO, or cardiac hypertrophy and / or cardiac fibrosis. In some embodiments, pharmaceutical compositions comprising the ALK2 antibodies or antigen-binding fragments thereof (e.g., ALK2-binding fragments) described herein can be used in combination with other agents (e.g., biological therapeutics and / or small molecules) or compositions in therapy. Pharmaceutical compositions comprising the ALK2 antibodies or antigen-binding fragments thereof can be prepared using methods known in the art. For example, such compositions can be prepared in a desired form, e.g., a lyophilized formulation or an aqueous solution, using, for example, a physiologically acceptable carrier, excipient, or stabilizer (Remington: The Science and Practice of Pharmacology, 22nd Edition, Allen, L. Ed., 2013, incorporated herein by reference). In some embodiments, a pharmaceutical composition of the invention comprises a nucleic acid molecule (DNA or RNA, e.g., mRNA) encoding an antibody or ALK2-binding fragment thereof described herein, or a vector comprising such a nucleic acid molecule.

[0382] Acceptable carriers and excipients in pharmaceutical compositions are non-toxic to recipients at the dosages and concentrations used. Acceptable carriers and excipients may include buffers such as phosphate, citrate, HEPES, and TAE, antioxidants such as ascorbic acid and methionine, preservatives such as hexamethonium chloride, octadecyldimethylbenzyl ammonium chloride, resorcinol, and benzalkonium chloride, proteins such as human serum albumin, gelatin, dextran, and immunoglobulins, hydrophilic polymers such as polyvinylpyrrolidone, amino acids such as glycine, glutamine, histidine, lysine, and carbohydrates such as glucose, mannose, sucrose, and sorbitol. The pharmaceutical composition of the present invention can be administered parenterally in the form of an injectable formulation. Pharmaceutical compositions for injection can be formulated using a sterile solution or any pharma- ceutical acceptable liquid as a vehicle. Pharmaceutically acceptable media include, but are not limited to, sterile water, saline, and cell culture media (e.g., Dulbecco's Modified Eagle Medium (DMEM), alpha-modified Eagle Medium (a-MEMI).

[0383] Mixtures of ALK2 antibodies or antigen-binding fragments thereof (e.g., ALK2-binding fragments) can be prepared in water, suitably mixed with one or more excipients, carriers, or diluents. Dispersions can also be prepared in glycerol, liquid polyethylene glycols, and mixtures thereof, as well as in oils. Under ordinary conditions of storage and use, these preparations may contain a preservative to prevent the growth of microorganisms. Suitable dosage forms for injectable use include sterile aqueous solutions or dispersions and sterile powders for the extemporaneous preparation of sterile injectable solutions or dispersions (as described in U.S. Pat. No. 5,466,468, the disclosure of which is incorporated herein by reference). In any event, the preparations can be sterile and fluid to the extent that easy syringability exists. The preparations can be stable under the conditions of manufacture and storage and can be preserved against the contaminating action of microorganisms, such as bacteria and fungi. The carrier can be, for example, a solvent or dispersion medium containing water, ethanol, a polyol (e.g., glycerol, propylene glycol, and liquid polyethylene glycol, etc.), suitable mixtures thereof, and / or vegetable oils. Proper fluidity can be maintained, for example, by the use of a coating such as lecithin, by the maintenance of the required particle size in the case of dispersion, and by the use of surfactants. Prevention of the action of microorganisms can be brought about by various antibacterial and antifungal agents, for example, parabens, chlorobutanol, phenol, sorbic acid, thimerosal, and the like. In many cases, it will be preferable to include isotonic agents, for example, sugars or sodium chloride. Prolonged absorption of injectable compositions can be brought about by the use in the composition of agents delaying absorption, for example, aluminum monostearate and gelatin.

[0384] For example, the solutions containing the pharmaceutical compositions described herein may be suitably buffered if necessary, and the liquid diluent is first made isotonic with sufficient saline or glucose. These particular aqueous solutions are particularly suitable for intravenous, intramuscular, subcutaneous, and intraperitoneal administration. In this regard, the sterile aqueous media that can be used will be known to those skilled in the art in light of this disclosure. For example, one dose may be dissolved in 1 ml of isotonic NaCl solution and added to 1000 ml of subcutaneous injection fluid, or may be injected at the proposed site of injection. Some variation in dosage will necessarily occur depending on the condition of the subject being treated.

[0385] The pharmaceutical composition of the present invention can be prepared in microcapsules, such as hydroxymethylcellulose or gelatin-microcapsules and poly(methyl methacrylate) microcapsules. The pharmaceutical composition of the present invention can also be prepared in other drug delivery systems, such as liposomes, albumin microspheres, microemulsions, nanoparticles, and nanocapsules. Such techniques are described in Remington: The Science and Practice of Pharmacology, 22nd Edition, Edited by Allen, L. (2013). Pharmaceutical compositions used for in vivo administration must be sterile. This is easily achieved by filtration through sterile filtration membranes.

[0386] The pharmaceutical composition of the present invention may also be prepared as a sustained release formulation. Suitable examples of sustained release preparations include semipermeable matrices of solid hydrophobic polymers containing the polypeptide of the present invention. Examples of sustained release matrices include polyesters, hydrogels, polylactides, copolymers of L-glutamic acid and gamma-ethyl-L-glutamic acid, non-degradable ethylene-vinyl acetate, degradable lactic acid-glycolic acid copolymers such as LUPRON DEPOT™, and poly-D-(-)-3-hydroxybutyric acid. Some sustained release formulations allow the release of molecules over several months, e.g., 1-6 months, while other formulations release the pharmaceutical composition of the present invention over shorter periods, e.g., days to weeks.

[0387] The pharmaceutical compositions can be formulated into unit dosage forms as desired. The amount of active ingredient described herein, e.g., an ALK2 antibody or antigen-binding fragment thereof (e.g., an ALK2-binding fragment) contained in the pharmaceutical preparation is such that a suitable dosage within the specified range is provided (e.g., a dosage within the range of 0.01 to 100 mg / kg body weight).

[0388] The pharmaceutical composition for gene therapy may be in an acceptable diluent or may comprise a slow release matrix in which the gene delivery vehicle is embedded. When hydrodynamic injection is used as a delivery method, the pharmaceutical composition comprising the nucleic acid molecule encoding the antibody or its ALK2-binding fragment described herein, or the vector (e.g., viral vector) comprising the nucleic acid molecule, is rapidly delivered intravenously in a large volume of fluid. Vectors that can be used as in vivo gene delivery vehicles include, but are not limited to, retroviral vectors, adenoviral vectors, poxvirus vectors (e.g., vaccinia virus vectors such as modified vaccinia Ankara), adeno-associated virus vectors, and alphavirus vectors.

[0389] Route, Dosage, and Administration Pharmaceutical compositions comprising the ALK2 antibody or antigen-binding fragment thereof (e.g., ALK2-binding fragment) of the present invention as a therapeutic agent can be administered by various routes, such as intravenous, parenteral, intradermal, transdermal, intramuscular, intranasal, subcutaneous, percutaneous, topical, intratracheal, intraperitoneal, intraarterial, intravascular, intrathecal, intraventricular, inhalation, perfusion, lavage, and oral. Pharmaceutical compositions can also be formulated for or administered via oral, ocular, nasal, spray, aerosol, rectal, or vaginal administration. For injectable formulations, various effective pharmaceutical carriers are known in the art. See, for example, ASHP Handbook on Injectable Drugs, Toissel, 18th Edition (2014). For ocular administration, the formulation can be delivered systemically by injection (e.g., intraocular injection) or locally (e.g., by instillation into the eye (e.g., eye drops) as a solution, suspension, or ointment, etc.).

[0390] In some embodiments, pharmaceutical compositions comprising nucleic acid molecules encoding the antibodies or ALK2-binding fragments thereof described herein or vectors comprising such nucleic acid molecules can be administered by gene delivery. Methods of gene delivery are well known to those skilled in the art. Vectors that can be used for in vivo gene delivery and expression include, but are not limited to, retroviral vectors, adenoviral vectors, poxvirus vectors (e.g., vaccinia virus vectors such as modified vaccinia Ankara (MVA)), adeno-associated virus vectors, and alphavirus vectors. In some embodiments, mRNA molecules encoding the polypeptides of the present invention can be administered directly to the subject.

[0391] In some embodiments of the present invention, nucleic acid molecules encoding the polypeptides described herein or vectors comprising such nucleic acid molecules may be administered using a hydrodynamic injection platform. In hydrodynamic injection, a nucleic acid molecule encoding an antibody or ALK2-binding fragment thereof described herein is placed under the control of a strong promoter in an engineered plasmid (e.g., a viral plasmid). The plasmid is often rapidly delivered intravenously in a large volume of fluid. Hydrodynamic injection uses controlled hydrodynamic pressure in the vein to increase cell permeability, such that the high pressure from the rapid injection of a large volume of fluid results in extravasation of the fluid and plasmid from the vein. Expression of the nucleic acid molecule is primarily driven by the liver. In mice, hydrodynamic injection is often performed by injecting the plasmid into the tail vein. In certain embodiments, an mRNA molecule encoding an antibody or ALK2-binding fragment thereof described herein may be administered using hydrodynamic injection.

[0392] The most appropriate route and dose for administration in a given case will depend on the particular composition administered, the patient, the formulation method, the method of administration (e.g., time and route of administration), the patient's age, weight, sex, severity of the disease being treated, the patient's diet, and the patient's excretion rate. The pharmaceutical composition of the present invention may include a dose of the ALK2 antibody or antigen-binding fragment thereof (e.g., ALK2-binding fragment) of the present invention in the range of 0.01 to 500 mg / kg (e.g., 0.01, 0.1, 0.2, 0.3, 0.4, 0.5, 1, 2, 3, 4, 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 100, 150, 200, 250, 300, 350, 400, 450, or 500 mg / kg). Also, in more specific embodiments, the dose is about 0.1 to about 30 mg / kg, and in more specific embodiments, about 0.3 to about 30 mg / kg. The dosage can be adapted by the physician according to conventional factors such as the degree of the disease and the different parameters of the subject.

[0393] Pharmaceutical compositions are administered in a manner compatible with the dosage form and in such amount as is therapeutically effective to result in improvement or alleviation of symptoms. Pharmaceutical compositions are administered in a variety of dosage forms, for example, intravenous, subcutaneous, and oral dosage forms (e.g., ingestible solutions, drug release capsules). Generally, therapeutic antibodies are administered at 0.1-100 mg / kg, for example, 1-50 mg / kg. Pharmaceutical compositions comprising the ALK2 antibodies or antigen-binding fragments thereof (e.g., ALK2-binding fragments) of the invention can be administered to a subject in need thereof, for example, once or more times (e.g., 1-10 times or more) daily, weekly, biweekly, monthly, bimonthly, quarterly, biennially, yearly, or as medically necessary. In some embodiments, pharmaceutical compositions comprising the ALK2 antibodies or antigen-binding fragments thereof (e.g., ALK2-binding fragments) of the invention can be administered to a subject in need thereof weekly, biweekly, monthly, bimonthly, or quarterly. Doses can be provided in either single or multiple dosing regimens. The timing of administration may be decreased as the condition improves or increased as the patient's health declines.

[0394] treatment method The compositions and methods described herein can be used to treat and / or prevent (e.g., prevent the onset of or treat a subject diagnosed with) a condition, such as bone disease, anemia, or low red blood cell levels (e.g., low hemoglobin levels or low red blood cell count), heterotopic ossification (e.g., heterotopic ossification due to FOP), Sjogren's syndrome (e.g., dry eye associated with Sjogren's syndrome), DIPG, MO, PCO, or cardiac hypertrophy and / or cardiac fibrosis. In some embodiments, an ALK2 antibody (e.g., a neutralizing antibody) or antigen-binding fragment thereof (e.g., an ALK2-binding fragment) described herein is administered to increase bone mineral density, increase bone formation, increase bone strength, decrease bone resorption (e.g., bone loss), or reduce the risk of fracture in a subject in need thereof. The ALK2 antibodies or antigen-binding fragments thereof (e.g., ALK2-binding fragments) described herein may increase bone mineral density, increase bone formation, increase bone strength, decrease bone resorption (e.g., bone loss), or decrease the risk of fracture compared to measurements obtained before treatment or compared to bone mineral density, bone strength, bone formation, bone resorption, or risk of fracture normally observed in untreated subjects. In some embodiments, the subject may have or be at risk of developing a disease that results in bone damage (e.g., osteoporosis or osteopenia). In some embodiments of the methods described herein, the subject has or is at risk of developing a disease or condition involving bone damage (e.g., primary osteoporosis, secondary osteoporosis, osteopenia, osteopetrosis, fracture, bone loss associated with bone cancer or cancer metastasis, Paget's disease, renal osteodystrophy, treatment-related bone loss, diet-related bone loss, bone loss associated with treatment of obesity, low gravity-related bone loss, or immobility-related bone loss).The invention also includes methods of treating a subject having or at risk of developing primary osteoporosis, secondary osteoporosis, osteopenia, osteopetrosis, bone fracture, bone cancer or cancer metastasis associated bone loss, Paget's disease, renal osteodystrophy, treatment-related bone loss, diet-related bone loss, bone loss associated with the treatment of obesity, low gravity-related bone loss, or immobility-related bone loss, by administering to the subject an effective amount of an ALK2 antibody or antigen-binding fragment thereof described herein (e.g., an ALK2-binding fragment).

[0395] In any of the methods described herein, the subject having or at risk of developing a bone disease (e.g., bone injury) has or is at risk of developing a disease or condition including primary osteoporosis, secondary osteoporosis, osteopenia, osteopetrosis, bone fracture, bone loss associated with bone cancer or cancer metastasis, Paget's disease, renal osteodystrophy, treatment-related bone loss, diet-related bone loss, bone loss associated with the treatment of obesity, low gravity-related bone loss, or immobility-related bone loss. In some embodiments, the primary osteoporosis is age-related osteoporosis or hormone-related osteoporosis (e.g., associated with estrogen decline). In some embodiments, the secondary osteoporosis is immobilization-induced osteoporosis or glucocorticoid-induced osteoporosis. In some embodiments, the bone cancer is multiple myeloma, or the cancer metastasis-related bone loss is caused by multiple myeloma. In some embodiments, the treatment-related bone loss results from treatment with FGF-21 or GLP-1, treatment with FGF-21 or GLP-1 including a therapeutic agent, treatment of type 2 diabetes and / or obesity, or cancer treatment (e.g., chemotherapy or radiation therapy). In some embodiments, the diet-related bone loss is rickets (e.g., vitamin D deficiency). In some embodiments, the low gravity-related bone loss is the absence of load-related bone loss. In some embodiments, the methods described herein increase bone mineral density (e.g., increase bone mass). In some embodiments, the methods described herein decrease bone resorption (e.g., decrease bone catabolic activity or decrease bone loss), e.g., decrease bone resorption compared to measurements obtained before treatment or compared to bone resorption normally observed in untreated subjects. In some embodiments, the methods described herein increase bone formation (e.g., increase bone anabolic activity or increase bone generation), e.g., increase bone formation compared to measurements obtained before treatment or compared to bone formation normally observed in untreated subjects. In some embodiments, the methods described herein increase osteoblast activity or osteoblast formation, e.g., increase osteoblast activity or osteoblast formation compared to measurements obtained before treatment or compared to osteoblast activity or osteoblast formation normally observed in untreated subjects.In some embodiments, the methods described herein decrease osteoclast activity or osteoclast formation, e.g., decrease osteoclast activity or osteoclast formation, compared to measurements obtained before treatment, or compared to osteoclast activity or osteoclast formation normally observed in untreated subjects, hi some embodiments, the bone is cortical or trabecular bone.

[0396] In some embodiments, an ALK2 antibody or antigen-binding fragment thereof (e.g., an ALK2-binding fragment) described herein is administered to increase red blood cell levels (e.g., increase hemoglobin levels, increase red blood cell count, or increase red blood cell formation or production) in a subject in need thereof. An ALK2 antibody or antigen-binding fragment thereof (e.g., an ALK2-binding fragment) described herein may increase red blood cell levels (e.g., increase hemoglobin levels, red blood cell count, or red blood cell formation) compared to measurements obtained before treatment or compared to red blood cell levels normally observed in an untreated subject. In some embodiments, the subject may have a disease or condition associated with low red blood cell levels (e.g., anemia or blood loss). In some embodiments of the methods described herein, the subject has or is at risk of developing a disease or condition associated with low red blood cell levels (e.g., anemia or blood loss, e.g., aplastic anemia, iron deficiency anemia, iron refractory iron deficiency anemia (IRIDA), or inflammatory anemia (also called anemia of chronic disease, e.g., anemia caused by infectious disease (e.g., chronic infectious disease such as HIV / AIDS or tuberculosis), autoimmune disease (e.g., rheumatoid arthritis or lupus), cancer (e.g., cancer or cancer treatment), inflammatory bowel disease (e.g., Crohn's disease or ulcerative colitis), and chronic kidney disease)). The invention also includes methods of treating a subject having or at risk of developing low red blood cell levels (e.g., low hemoglobin levels or low red blood cell count, e.g., anemia) by administering to the subject an effective amount of an ALK2 antibody or antigen-binding fragment thereof (e.g., an ALK2-binding fragment) described herein. The methods described herein may include screening the subject for one or more mutations in genes known to be associated with anemia (e.g., a mutation in TMPRSS6 associated with IRIDA) prior to treatment with or administration of a composition described herein. Subjects can be screened for gene mutations using standard methods known to those of skill in the art (e.g., genetic testing).

[0397] The invention also includes methods of treating a subject having or at risk of developing anemia or blood loss by administering to the subject an effective amount of an ALK2 antibody or antigen-binding fragment thereof (e.g., an ALK2-binding fragment) described herein. In any of the methods described herein, the subject having or at risk of developing low red blood cell levels (e.g., low hemoglobin level or low red blood cell count) has or is at risk of developing anemia or blood loss. In some embodiments, the anemia is due to a nutritional deficiency (e.g., vitamin deficiency), bone marrow deficiency (e.g., paroxysmal nocturnal hemoglobinuria), an adverse reaction to medication (e.g., anti-retroviral HIV drugs), myelodysplastic syndrome, bone marrow transplant, cancer (e.g., solid tumors such as breast cancer, lung cancer, colon cancer, etc.; tumors of the lymphatic system such as chronic lymphocytic leukemia, non-Hodgkin's lymphoma, Hodgkin's lymphoma; or tumors of the hematopoietic system such as leukemia or multiple myeloma), cancer treatment (e.g., radiation therapy or chemotherapy, e.g., chemotherapy with platinum-containing agents), inflammatory or autoimmune diseases (e.g., rheumatoid arthritis, other inflammatory arthritis, systemic lupus erythematosus (SLE), acute or chronic skin diseases (e.g., psoriasis), or inflammatory bowel disease (e.g., Crohn's disease or ulcerative colitis), cystitis, gastritis), idiopathic or congenital conditions, acute or chronic renal disease or disorder (e.g., chronic kidney disease), acute or chronic liver disease, acute or chronic bleeding, infection (e.g., malarial osteomyelitis), splenomegaly, porphyria, vasculitis, hemolysis, urinary tract infection, hemoglobinopathies (e.g., sickle cell disease), thalassemia, Churg-Strauss syndrome, Felty syndrome, graft-versus-host disease, hematopoietic stem cell transplantation, myelofibrosis, pancytopenia, pure red blood cell aplasia, Henoch-Schönlein purpura, Schwachmann syndrome (e.g., Schwachmann-Diamond syndrome), drug use or abuse (e.g., alcohol abuse), or associated with contraindications to transfusion (e.g., elderly patients, patients with allo- or auto-antibodies, pediatric patients, patients with cardiopulmonary disease, patients who oppose transfusions for religious reasons (e.g., some Jehovah's Witnesses)).In some embodiments, the anemia is aplastic anemia, iron deficiency anemia, iron refractory iron deficiency anemia (IRIDA), vitamin deficiency anemia, inflammatory anemia (also called anemia of chronic disease, e.g., anemia caused by an inflammatory disease or condition, such as an infection (e.g., chronic infection such as HIV / AIDS or tuberculosis), an autoimmune disease (e.g., rheumatoid arthritis or lupus), cancer, inflammatory bowel disease (e.g., Crohn's disease or ulcerative colitis), and chronic kidney disease), anemia associated with myeloid disease, hemolytic anemia, sickle cell anemia, microcytic anemia, hypochromic anemia, sideroblastic anemia, Diamond-Blackfan anemia, Fanconi anemia, or refractory anemia with excess blasts. In some embodiments, the anemia is associated with elevated hepcidin levels (e.g., elevated compared to hepcidin levels in a subject without anemia). The compositions and methods described herein can also be used to treat subjects who do not respond adequately to erythropoietin (EPO) or who are susceptible to adverse effects of EPO (e.g., hypertension, headaches, vascular thrombosis, flu-like syndrome, shunt occlusion, and myocardial infarction). In some embodiments, the blood loss is due to surgery, trauma, wounds, ulcers, urinary tract bleeding, gastrointestinal bleeding, frequent blood donations, or heavy menstrual bleeding (e.g., menorrhagia). In some embodiments, the methods described herein increase red blood cell levels (e.g., hemoglobin levels or red blood cell counts) compared to measurements obtained prior to treatment or compared to red blood cell levels normally observed in untreated subjects. In some embodiments, the methods described herein increase or induce red blood cell formation compared to measurements obtained prior to treatment or compared to red blood cell formation normally observed in untreated subjects. In some embodiments, the compositions and methods described herein reduce the subject's need for blood transfusions (e.g., the subject no longer requires blood transfusions or the subject requires less frequent transfusions than prior to treatment with the compositions and methods described herein). Subjects with normal red blood cell levels may also be treated using the methods and compositions described herein to increase red blood cell levels so that blood can be collected and stored for later use in transfusions.

[0398] In some embodiments, the ALK2 antibodies or antigen-binding fragments thereof (e.g., ALK2-binding fragments) described herein may be administered to a subject to treat FOP, to slow or stop the progression of FOP, to delay the onset of FOP (e.g., to delay the onset of heterotopic ossification), or to prevent or reduce heterotopic ossification, such as ectopic ossification, in a subject with FOP (e.g., to prevent or reduce bone formation in muscles, tendons, ligaments, or other connective tissues, e.g., to prevent or reduce heterotopic ossification in a subject with FOP who has already experienced heterotopic ossification, or to prevent the onset of heterotopic ossification in a subject who has not yet developed heterotopic ossification). FOP may be hereditary FOP (e.g., FOP associated with germline transmission of a mutation, such as an autosomal dominant mutation) or sporadic FOP (e.g., FOP associated with a spontaneous, non-hereditary mutation). The ALK2 antibodies or antigen-binding fragments thereof (e.g., ALK2-binding fragments) described herein may reduce the amount of ectopic bone, reduce the formation of ectopic bone, or reduce the recurrence of ectopic bone (e.g., recurrence following surgical resection) compared to the amount of ectopic bone, ectopic bone formation, or ectopic bone recurrence observed in subjects who have received an untreated or control treatment. The ALK2 antibodies or antigen-binding fragments thereof (e.g., ALK2-binding fragments) described herein may prevent the formation of ectopic bone (e.g., in subjects at risk of developing ectopic ossification, such as subjects with FOP, or may prevent the recurrence of ectopic bone, such as recurrence after surgical resection. In some embodiments, the subject is at risk for developing ectopic ossification (e.g., the subject has a genetic mutation associated with FOP but has not yet exhibited symptoms of FOP, e.g., has not yet exhibited ectopic ossification). The methods described herein may include screening the subject for one or more mutations in genes known to be associated with FOP (e.g., mutations in ALK2, also known as activin A type I receptor (ACVR1)) prior to treatment with or administration of a composition described herein. Subjects can be screened for genetic mutations using standard methods known to those of skill in the art (e.g., genetic testing).

[0399] In some embodiments, an ALK2 antibody or antigen-binding fragment thereof (e.g., an ALK2-binding fragment) described herein may be administered to a subject to treat Sjogren's syndrome (e.g., to treat dry eye associated with Sjogren's syndrome) or to alleviate one or more symptoms of Sjogren's syndrome (e.g., to alleviate dry eye or dry mouth associated with Sjogren's syndrome). The ALK2 antibodies or antigen-binding fragments thereof described herein (e.g., ALK2-binding fragments) may reduce ocular irritation, reduce fibrosis of the lacrimal gland, reduce inflammation of the lacrimal gland, improve vision, increase saliva production, increase tear production (e.g., increase tear volume), reduce systemic inflammation, reduce joint pain, reduce fatigue in a subject with Sjogren's syndrome, or reduce the severity or occurrence of one or more symptoms of Sjogren's syndrome (e.g., reduce dry eye, dry mouth, or joint pain associated with Sjogren's syndrome) compared to the severity or occurrence of ocular irritation, vision, inflammation or fibrosis of the lacrimal gland, saliva production, tear production, systemic inflammation, joint pain, fatigue, or a symptom of Sjogren's syndrome in a pre-treated subject or an untreated subject with Sjogren's syndrome. In some embodiments, an ALK2 antibody or antigen-binding fragment thereof (e.g., an ALK2-binding fragment) described herein may be administered to a subject to treat DIPG, prolong survival (e.g., survival time, e.g., life span) of a subject with DIPG, prolong progression-free survival of a subject with DIPG, reduce DIPG tumor growth, reduce size or volume of a DIPG tumor, or prevent or reduce metastasis of a DIPG tumor. An ALK2 antibody or antigen-binding fragment thereof (e.g., an ALK2-binding fragment) described herein prolong survival (e.g., survival time, e.g., life span) of a subject with DIPG, prolong progression-free survival, reduce DIPG tumor growth, reduce size or volume of a DIPG tumor, or prevent or reduce metastasis of a DIPG tumor compared to the survival, progression-free survival, tumor growth, tumor size or volume, or tumor metastasis of an untreated or control treated subject, or compared to the tumor growth, tumor size or volume, or tumor metastasis of the subject prior to treatment.In some embodiments, the subject has an activating mutation in ALK2 (ACVR1). The methods described herein can include screening the subject for mutations in ALK2 (ACVR1) prior to treatment with or administration of a composition described herein. Subjects can be screened for genetic mutations using standard methods known to those of skill in the art (e.g., genetic testing).

[0400] In some embodiments, an ALK2 antibody or antigen-binding fragment thereof (e.g., an ALK2-binding fragment) described herein can be administered to a subject to treat MO, reduce or prevent the formation of osteochondroma in a subject with MO, reduce the number of osteochondromas in a subject with MO, reduce the size of osteochondromas in a subject with MO, or slow the growth of osteochondromas in a subject with MO. Osteochondromas can form in the growing ends of bones (metaphysis), long bones (e.g., long bones of the legs, arms, or fingers), and / or flat bones (e.g., hip bone (pelvic bone), ribs, or scapula). The ALK2 antibodies or antigen-binding fragments thereof (e.g., ALK2-binding fragments) described herein may reduce or prevent the formation of osteochondroma (e.g., prevent the formation of osteochondroma in a subject with MO who has not yet developed an osteochondroma, or reduce or prevent the formation of osteochondroma, reduce the number of osteochondroma, reduce the size of osteochondroma, or slow the growth of osteochondroma, compared to the formation of osteochondroma, number of osteochondroma, size of osteochondroma, or growth of osteochondroma in a subject prior to treatment or an untreated subject with MO. The methods described herein may include screening the subject for mutations in EXT1 or EXT2 prior to treatment with or administration of a composition described herein. Subjects can be screened for genetic mutations using standard methods known to those of skill in the art (e.g., genetic testing).

[0401] In some embodiments, the ALK2 antibodies or antigen-binding fragments thereof described herein (e.g., ALK2-binding fragments) are used to treat PCO, reduce or prevent the onset of PCO in subjects who have PCO or are at risk of developing PCO (e.g., subjects who have had or will soon have cataract surgery, e.g., subjects who will have cataract surgery in 6 months, 5 months, 4 months, 3 months, 2 months, 1 month, 2 weeks, 1 week, or more). It may be administered to a subject who is to undergo cataract surgery, e.g., a subject at risk of developing PCO, to prevent the onset of PCO, improve vision in a subject with PCO (e.g., reduce blurry or cloudy vision), reduce light sensitivity or glare in a subject with PCO, reduce or prevent fibrosis (e.g., fibrosis near the implanted lens, e.g., fibrosis on or near the posterior capsule), lens fiber differentiation, lens epithelial cell proliferation, or reduce or prevent ocular inflammation compared to the onset of PCO in an untreated subject, fibrosis (e.g., fibrosis near the implanted lens, e.g., fibrosis on or near the posterior capsule), lens fiber differentiation, lens epithelial cell proliferation, or ocular inflammation. The ALK2 antibodies or antigen-binding fragments thereof (e.g., ALK2-binding fragments) described herein may reduce or prevent the onset of PCO in a subject at risk of developing PCO, reduce or prevent fibrosis (e.g., fibrosis near an implanted lens, e.g., fibrosis on or near the posterior capsule), reduce or prevent lens fiber differentiation, reduce or inhibit lens epithelial cell proliferation, or reduce or prevent ocular inflammation, or may improve visual acuity (e.g., reduce blurry or cloudy vision), or reduce light sensitivity or glare, reduce fibrosis, reduce ocular inflammation, reduce lens epithelial cell proliferation, or reduce lens fiber differentiation, compared to the visual acuity, light sensitivity or glare, fibrosis, ocular inflammation, lens epithelial cell proliferation, or lens fiber differentiation in the subject prior to treatment.

[0402] In some embodiments, the ALK2 antibodies or antigen-binding fragments thereof (e.g., ALK2-binding fragments) described herein treat cardiac hypertrophy, treat cardiac fibrosis, delay or prevent the onset of cardiac hypertrophy (e.g., in a subject at risk of developing cardiac hypertrophy, e.g., a subject with hypertension or valvular disease), delay or prevent the onset of cardiac fibrosis (e.g., in a subject at risk of developing cardiac fibrosis, e.g., a subject with hypertension, a subject who has suffered a myocardial infarction, a subject with diabetic hypertrophic cardiomyopathy, or a subject with idiopathic dilated cardiomyopathy), reverse cardiac fibrosis, They may be administered to a subject to reduce or inhibit cardiac scar formation, increase or induce cardiac regeneration, or ameliorate one or more symptoms of cardiac hypertrophy or cardiac fibrosis (e.g., improving exercise capacity, increasing blood ejection volume, decreasing left ventricular end diastolic pressure, decreasing pulmonary capillary wedge pressure, increasing cardiac output, increasing cardiac index, decreasing pulmonary artery pressure, decreasing left ventricular end systolic and diastolic dimensions, decreasing left and right ventricular wall stress, decreasing wall tension and / or wall thickness, increasing myocardial contractility, decreasing myocardial cell area, decreasing cardiomyocyte extracellular matrix deposition, improving quality of life, and / or reducing disease-related morbidity and mortality). The ALK2 antibodies or antigen-binding fragments thereof (e.g., ALK2-binding fragments) described herein delay or prevent the onset of cardiac hypertrophy, delay or prevent the onset of cardiac fibrosis, reverse cardiac fibrosis, reduce or inhibit cardiac scar formation, increase or induce cardiac regeneration, or ameliorate one or more symptoms of cardiac hypertrophy or cardiac fibrosis in a subject having or at risk of developing cardiac hypertrophy or cardiac fibrosis, compared to the onset of cardiac hypertrophy, the onset of cardiac fibrosis, the extent of cardiac fibrosis, cardiac scar formation, cardiac regeneration, or one or more symptoms of cardiac hypertrophy or cardiac fibrosis in subjects prior to treatment or in untreated or control-treated subjects.

[0403] The compositions described herein may increase bone mineral density, increase bone strength, density, decrease bone resorption (e.g., bone loss), decrease the rate of bone resorption (e.g., bone loss), increase bone formation, increase the rate of bone formation, decrease osteoclast activity, increase osteoblast activity, decrease the risk of fracture, increase red blood cell levels, increase hemoglobin levels, decrease the need for blood transfusions, increase red blood cell formation or increase red blood cell count, treat anemia, increase iron levels, decrease iron deficiency, decrease elevated hepcidin levels, treat FOP, and increase ectopic bone the formation of heterotopic bone in a subject (e.g., preventing the initial formation of heterotopic bone in a subject or preventing the recurrence of heterotopic bone following surgical resection), slowing or halting the progression of FOP, delaying the onset of FOP (e.g., delaying the onset of heterotopic ossification), reducing heterotopic ossification (e.g., reducing the amount of heterotopic bone in a subject or reducing the formation of heterotopic bone, such as the initial formation of heterotopic bone, or the recurrence of heterotopic bone following surgical resection), treating Sjogren's syndrome, relieving dry eye associated with Sjogren's syndrome, relieving dry mouth associated with Sjogren's syndrome, reducing eye inflammation, and increasing saliva production. and / or increasing tear production, reducing systemic inflammation, reducing joint pain, reducing fatigue, reducing the severity or occurrence of symptoms of Sjogren's syndrome (e.g., reducing dry eye, dry mouth, or joint pain associated with Sjogren's syndrome), treating DIPGs, prolonging survival (e.g., survival time, e.g., life span) in a subject having DIPGs, prolonging progression-free survival in a subject having DIPGs, reducing the growth of DIPG tumors, reducing the size or volume of DIPG tumors, preventing or reducing metastasis of DIPG tumors, treating MO, and / or reducing the progression of osteochondroma in a subject having MO. reducing or preventing the formation of osteochondromas in a subject with MO, reducing the number of osteochondromas in a subject with MO, reducing the size of osteochondromas in a subject with MO, slowing the growth of osteochondromas in a subject with MO, reducing or preventing the onset of PCO, improving vision in a subject with PCO (e.g., reducing blurred or cloudy vision), reducing light sensitivity or glare in a subject with PCO, reducing or preventing fibrosis (e.g., fibrosis near an implanted lens, e.g., fibrosis on or near the posterior capsule), reducing or inhibiting the proliferation of lens epithelial cells, and reducing or preventing differentiation of lens fibers;or PCO, reduce or prevent ocular inflammation in a subject with PCO, treat cardiac hypertrophy, treat cardiac fibrosis, delay or prevent the onset of cardiac hypertrophy, delay or prevent the onset of cardiac fibrosis, reverse cardiac fibrosis, reduce or inhibit cardiac scar formation, increase or induce cardiac regeneration, or ameliorate one or more symptoms of cardiac hypertrophy or cardiac fibrosis (e.g., improve exercise capacity, increase blood ejection volume, decrease left ventricular end diastolic pressure, decrease pulmonary capillary wedge pressure, increase cardiac output, increase cardiac index, decrease pulmonary artery pressure, decrease left ventricular end systolic and diastolic dimensions, decrease left and right ventricular wall stress, decrease wall tension and / or wall thickness, increase myocardial contractility, decrease cardiomyocyte extracellular matrix deposition, improve quality of life, and / or decrease disease-related morbidity and mortality). Bone mineral density can be assessed using well-established clinical techniques known to those skilled in the art (e.g., by dual energy x-ray absorptiometry). Red blood cell levels can be assessed using standard blood tests that measure red blood cell counts and hemoglobin levels. FOP symptoms (e.g., heterotopic ossification) and MO (e.g., osteochondroma) can be assessed using standard imaging methods such as radiography (e.g., x-ray), CT (computed tomography), and / or MRI (magnetic resonance imaging). DIPG tumor size, growth, volume, or metastasis can be assessed using imaging methods such as MRI. Cardiac hypertrophy and / or cardiac fibrosis symptoms can be assessed using clinical approaches to evaluate cardiac function and morphology (e.g., stress tests, echocardiography, CT, MRI, cardiac magnetic resonance imaging, single photon emission computed tomography, positron emission tomography, balloon catheter-based approaches, or histological analysis of biopsies). Sjogren's syndrome and PCO symptoms can be assessed by evaluating visual function and imaging the eye to evaluate inflammation and / or fibrosis (e.g., using slit lamp images to evaluate fibrosis formed in the posterior capsule). The methods described herein can also include assessing bone mineral density, red blood cell levels, or symptoms of FOP, DIPG, MO, PCO, Sjogren's syndrome, or cardiac hypertrophy and / or cardiac fibrosis in a subject prior to treatment or administration of a composition described herein, or after administration or treatment with a composition described herein.Depending on the dose and route of administration used for treatment, the evaluation can be performed 1 month, 2 months, 3 months, 4 months, 5 months, 6 months or more after administration of the ALK2 antibody or antigen-binding fragment thereof (e.g., ALK2-binding fragment) or pharmaceutical composition. Depending on the results of the evaluation, the subject may receive additional treatment.

[0404] Nucleic acids encoding the ALK2 antibodies or antigen-binding fragments thereof (e.g., ALK2-binding fragments) described herein, or expression vectors containing said nucleic acids, can also be administered according to any of the methods described herein. In any of the methods described herein, the polypeptides, nucleic acids, or vectors can be administered as part of a pharmaceutical composition.

[0405] kit The compositions described herein can be provided as a kit for use in treating bone disease, anemia, FOP, Sjogren's syndrome (e.g., dry eye associated with Sjogren's syndrome), MO, DIPG, PCO, or cardiac hypertrophy and / or cardiac fibrosis. The compositions can include an ALK2 antibody or antigen-binding fragment thereof (e.g., ALK2-binding fragment) described herein, and can be provided in a unit dosage form, optionally in a pharma- ceutically acceptable excipient (e.g., saline), in an amount sufficient to treat bone disease, anemia, FOP, Sjogren's syndrome (e.g., dry eye associated with Sjogren's syndrome), MO, DIPG, PCO, or cardiac hypertrophy and / or cardiac fibrosis. The kit can further include a package insert instructing a user of the kit, such as a physician, to practice the methods described herein. The kit can optionally include a syringe or other device for administering the composition.

[0406] Working Example The following examples are provided to further illustrate certain embodiments of the present invention, but are not intended to limit the scope of the invention, and by their illustrative nature, it will be understood that other procedures, methodologies, or techniques known to those skilled in the art may alternatively be used.

[0407] Example 1 - Generation of ALK2 Antibodies Antibodies that recognize ALK2 were generated by selecting antibodies from a human combinatorial phage library, followed by two rounds of enrichment and counterselection against closely related antigens to deplete nonspecific antibodies. After the second round of enrichment, Rapid Pool Maturation "RAPMAT™" (Prassler et al., Immunotherapy, 1(4), 571-583, 2009) was performed (RAPMAT is a registered trademark of MorphoSys AG). A highly diverse set of CDR variability was cloned into the selected antibody pool, further expanding the diversity of the preselected antibody pool. The resulting expanded antibody library was selected with ALK2, where it was enriched two more times using high stringency washing conditions, reduced antigen amounts during the selection stage, and counterselected against closely related antigens to deplete nonspecific antibodies. After selection, the enriched antibody gene pool was subcloned from the phage display vector into a bacterial expression vector, and the final Fab antibody format was determined as shown in Table 2 below.

[0408] [Table 2]

[0409] Example 2 - Assessment of ALK2 antibody binding affinity by surface plasmon resonance (SPR) A GE Biacore 3000 was used to measure the kinetics of the interaction between anti-ALK2 antibody (ligand) and ALK2-His / ALK2-mFC (analyte). Flow cells 1-4 were immobilized with GE's anti-human capture antibody using an amine coupling kit. Anti-ALK2 protein was then captured on the chip in flow cells 2-4, and flow cell 1 was left empty as a reference cell to measure and subtract nonspecific binding. When performing kinetic assays on the Biacore, it is best to allow a maximum analyte binding response (RMax) of 100 resonance units (RU). This is achieved by the formula RMax100=(kDa analyte / kDa ligand)×immobilized ligand RU. Too high a density of ligand in the flow cell will result in an artificially low concentration of unbound analyte near the chip surface, and the resulting sensorgram will not represent the true binding between the ligand and analyte. This is called the mass transfer effect and was prevented by calculating the appropriate level of ligand binding to the chip surface and by using a high flow rate of 40 μL / min. HBS-EP+ was used for running buffer and protein dilution. Each ligand was run in duplicate concentration series. At the end of each cycle, all bound anti-ALK2 and analyte were removed with 3M MgCl2 to regenerate the chip. The kinetics of the interaction between monovalent fAB antibodies in the fAB-FH format and ALK2-FC was also measured using a ForteBio Octet RED384.

[0410] Table 3 below shows the K of fAB of Antibody 1, Antibody 2, and Antibody 3 measured against ALK2-FC by ForteBio Octet. D values, and K of intact antibodies measured against ALK2-His by GE Biacore D Table 4 shows the K values ​​of Antibody 2 measured against ALK2-mFC by GE Biacore. D Table 5 shows the k values ​​for the interaction between monovalent fAB antibodies in the fAB-FH format and ALK2-FC, measured using a ForteBio Octet RED384. a , k d , and K DIndicates the value.

[0411] [Table 3]

[0412] [Table 4]

[0413] [Table 5]

[0414] Example 3 - Evaluation of ALK2 antibodies using a genetic luciferase reporter assay C2C12-BRE-luciferase (SMAD1 reporter) cells were plated in 96-well plates in DMEM supplemented with 2% FBS and placed in an incubator for >3 hours to acclimate to the plate surface. For each fAb variant, a dilution series was made in 2% DMEM and dosed to the cells. The remaining wells were used for positive control and background replicates. The cells were then placed in a 37°C incubator for 45 minutes. Wells were treated with either vehicle (to determine background values), BMP6 (to determine ALK2 stimulation values), or fAb followed by 40ng / mL BMP6 (to determine IC50 of fAb). The degree of BRE-stimulated luciferase was measured as a proxy for SMAD1 phosphorylation. Plates were incubated overnight and then read using a Promega Steady Glo and a Molecular Devices Spectramax M5e. Inhibition was calculated as the percentage of signal loss compared to positive control wells. Figures 1A-1B show BMP6 inhibition dose response curves of anti-ALK2 fABs performed in C2C12 cells using the SMAD1 luciferase reporter. The median inhibition values ​​(IC50) for both the 3- and 4-parameter curve fits in Graphpad Prism are displayed next to the curves. Figure 1A shows results from the first four fABs generated, and Figure 1B shows results from five second round fABs generated using Rapid Pool Maturation.

[0415] Example 4 - Effect of ALK2 antibodies in a mouse model of IRIDA Eight-week-old male C57BL / 6 mice were treated with vehicle or ALK2 antibody (5 mg / kg antibody 2, twice weekly, intraperitoneal injection). Six days after the start of treatment, vehicle-treated mice were intravenously administered lipid-encapsulated siRNA targeting either luciferase (control) or TMPRSS6 (0.3 mg / kg). TMPRSS6 encodes a transmembrane type II serine protease that inhibits hepcidin secretion by cleaving hemojuvelin, and mutations in TMPRSS6 have been observed in human subjects with IRIDA. ALK2 antibody-treated mice were administered TMPRSS6 siRNA (0.3 mg / kg). Mice were euthanized 48 hours after siRNA administration. Whole blood was collected and assayed for hematological parameters. As shown in Figure 2, treatment with ALK2 antibody prevents the reduction in hemoglobin associated with TMPRSS6 deficiency ( * = p ≤ 0.05; ** =p≦0.01).

[0416] In the second experiment, 8-week-old female albino C57BL / 6 mice were intravenously administered lipid-encapsulated siRNA targeting either luciferase (control) or TMPRSS6 (0.75 mg / kg) every 3 days. Mice received siRNA treatment until the time when hepcidin increased and serum hemoglobin and serum iron decreased in the cohort that received TMPRSS6 siRNA. At this point, mice were further randomized to receive either vehicle or anti-ALK2 antibody (Antibody 2, 5 mg / kg, administered intraperitoneally). Mice were euthanized 12 days after the first siRNA administration. Whole blood was collected and assayed for hematological parameters. Additionally, serum was collected and analyzed for hepcidin concentration by ELISA and total iron content by colorimetry. As shown in Figure 3A, treatment with ALK2 antibody reversed the hemoglobin reduction associated with TMPRSS6 deficiency ( * = p ≤ 0.05; ** =p≦0.01). ALK2 antibody treatment also demonstrated increased serum hepcidin levels associated with TMPRSS6 deficiency ( ** = p ≤ 0.01, Figure 3B ) and the reduction in serum iron associated with TMPRSS6 deficiency ( * = p ≤ 0.05;** = p ≦ 0.01, Figure 3C).

[0417] Example 5 - Effect of ALK2 antibodies in a mouse model of osteoporosis C57BL / 6 mice were orchiectomized (ORX) or sham-operated at 9 weeks of age. During the subsequent 6-week recovery period, ORX mice developed an osteoporotic phenotype, and ORX mice were injected intraperitoneally twice weekly with either vehicle or ALK2 antibody (5 mg / kg antibody 2). Whole-body dual-energy X-ray absorptiometry (Lunar PIXImus, GE Lunar Corp.) was performed 16 weeks after the start of treatment. High-resolution digital images were captured and BMD (software version 2.10) was determined using a hydroxyapatite phantom as a reference. As shown in Figure 4, ORX mice treated with ALK2 antibody had greater BMD than mice treated with vehicle, indicating that treatment with ALK2 antibody improves BMD or reduces bone loss.

[0418] Example 6 - Effect of ALK2 antibodies in a mouse model of MO The effect of ALK2 antibody on osteochondroma formation is determined using a mouse model of MO. The mice have an inducible, chondrocyte-specific, homozygous disruption of the Ext1 gene, and when lactating dams are administered a single dose of doxycycline at P8, osteochondroma formation is observed in 100% of mutants. Mice are divided into two groups that receive twice-weekly intraperitoneal injections of either ALK2 antibody (5 mg / kg) or vehicle starting at P8. Micro-CT imaging is performed weekly to monitor osteochondroma in the knee joint and thorax. At the end of 4 weeks of treatment, mice are sacrificed. From a portion of each group, whole-body skeletons are prepared and stained with alizarin red and alcian blue for imaging purposes. In the remaining mice, samples of the chondro-osseous junctions of the hind limbs and thorax are taken. Tissues are fixed in 4% paraformaldehyde, decalcified in EDTA, embedded in paraffin, and cut at a thickness of 5 μm. Safranin O / Fast Green staining is then used to measure changes in cartilage morphology and the development of osteochondroma.

[0419] Example 7 - Effect of ALK2 antibody on posterior capsule opacification The effect of ALK2 antibody on the formation of posterior capsule opacification (PCO) after cataract surgery is determined using a mouse model of injury-induced PCO. Briefly, 10-16 week old mice are divided into two groups that receive either ALK2 antibody (5 mg / kg) or vehicle twice weekly for 2 weeks prior to surgery. Cataract surgery is performed under anesthesia and one of the pupils is dilated with 2.5% phenylephrine hydrochloride and 1% tropicamide. An ophthalmic scalpel is used to incise the central cornea extending into the lens capsule. Hydrodissection with balanced salt solution is used to separate the lens fibrous mass from the lens capsule, the lens fibrous mass is removed, the incision is closed, and the anterior chamber is refilled with balanced salt solution. In each group, some mice are euthanized immediately after surgery and at 1, 5, and 14 days after surgery. Mice continue to receive either ALK2 antibody or vehicle twice weekly until euthanasia. Immediately prior to dissection, animals are anesthetized and assessed for corneal opacity, presence or absence of neovascularization, and degree of pupil dilation. At dissection, both the operated and non-operated (control) eyes are harvested, embedded, snap frozen, and stored at -80°. The degree of presence of inflammatory cells, fibrotic markers (αSMA, fibronectin), and lens fiber differentiation markers (Prox1, cMaf) are then assessed using immunohistochemistry.

[0420] Example 8 - Effect of ALK2 antibody in a mouse orthotopic xenograft model of DIPG The effect of ALK2 antibodies on inhibiting glioma growth in diffuse intrinsic pontine gliomas (DIPGs) is evaluated using a mouse orthotopic xenograft model of DIPGs. This model incorporates the implantation of a patient-derived cell line (HSJD-DIPG-007) containing mutations in the HK27M and ACRV1 genes into the fourth ventricle of immunodeficient NOD-SCID mice. The primary outcome of this study is the sustained survival of transplanted pups. Briefly, 3-week-old female NOD-SCID mice are implanted with HSJD-DIPG-007 cells via stereotactic surgery. 14–21 days after inoculation, mice are randomized into two groups and administered vehicle or ALK2 antibodies (5 mg / kg) twice weekly. Mice are expected to develop symptoms within 21 days after inoculation, with a mean lifespan of approximately 60 days without intervention. Mice are closely monitored for weight, health, and condition throughout the study. Mice were euthanized and removed from the study if they lost more than 20% of their body weight or showed signs of deterioration or distress. After completion, mouse brains were carefully dissected and fixed in formalin for later histological evaluation and to measure in vivo proliferation of HSJD-DIPG-007 cells.

[0421] Example 9 - Effect of ALK2 antibodies in a mouse model of Sjogren's syndrome The effect of ALK2 antibodies on improving symptoms of Sjögren's syndrome is determined in a non-obese diabetic (NOD) autoimmune mouse model. In addition to insulitis, NOD mice develop immune infiltration in exocrine and salivary glands, ultimately leading to secretory dysfunction. The primary phenotypic outcome measured in these mice is the amount of tear produced. This is tested weekly to give a longitudinal analysis of disease progression and efficacy of pharmacological intervention. The antibody is administered at 5 mg / kg twice weekly by intraperitoneal injection. Briefly, NOD mice (12 weeks old) are assessed for tear production at baseline and randomized into two groups to receive either vehicle or ALK2 antibody for 8 weeks. During this period, mice are tested weekly for tear production. At the end of the study, lacrimal glands are harvested to assess mRNA expression of fibrosis markers (MMP9, MMP2, COL1A1, TIMPs) and inflammatory markers (MCP1, IL6, TNF-α). The contralateral lacrimal gland is dissected and fixed in formalin for histological analysis of inflammation and fibrosis.

[0422] Example 10 - Effect of ALK2 antibodies in a mouse model of cardiac hypertrophy The effect of ALK2 antibodies on the development of cardiac hypertrophy and fibrosis is determined using an angiotensin II (A2)-induced cardiac hypertrophy mouse model. Mice are divided into groups that receive either ALK2 antibodies (5 mg / kg) or vehicle twice weekly for 2 weeks prior to surgery or immediately after surgery. Briefly, anesthetized mice are implanted with osmotic mini-pumps filled with A2, which release a constant amount of A2 for 2 or 4 weeks to induce cardiac hypertrophy. At the end of the 2 or 4 weeks, mice undergo echocardiography under sedation. The weight of the whole heart and the left ventricle (LV) are measured, and the LV is divided in half and stored in liquid nitrogen or formalin. The tibial length (TL) is also measured. Changes in heart weight, heart weight to TL ratio, LV weight, and LV to TL ratio are assessed. In addition, the thickness of the interventricular septum and posterior wall are measured, as well as the area and fibrosis of cardiomyocytes.

[0423] Example 11 - Treatment of anemia by administration of ALK2 antibodies According to the methods disclosed herein, a physician of the art can treat a subject, such as a human patient, with anemia (e.g., IRIDA or anemia of inflammation) to increase hemoglobin levels, increase red blood cell count, or reduce iron deficiency. The treatment method can include diagnosing or identifying the subject as a candidate for treatment based on a blood test that measures a hematological parameter. To treat the subject, a physician of the art can administer a composition comprising an ALK2 antibody or an antigen-binding fragment thereof (e.g., an ALK2-binding fragment) to the subject. The composition comprising an ALK2 antibody or an antigen-binding fragment thereof (e.g., an ALK2-binding fragment) can be administered to the subject, for example, by parenteral injection (e.g., intravenous injection) to treat anemia. The ALK2 antibody or antigen-binding fragment thereof (e.g., an ALK2-binding fragment) is administered in a therapeutically effective amount, e.g., 0.01-500 mg / kg (e.g., 0.01, 0.1, 0.2, 0.3, 0.4, 0.5, 1, 2, 3, 4, 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 100, 150, 200, 250, 300, 350, 400, 450, or 500 mg / kg). In some embodiments, the ALK2 antibody or antigen-binding fragment thereof (e.g., an ALK2-binding fragment) is administered every other month, once a month, once every two weeks, or at least once a week or more (e.g., 1, 2, 3, 4, 5, 6, or 7 times a week or more). The ALK2 antibody or antigen-binding fragment thereof (eg, an ALK2-binding fragment) is administered in an amount sufficient to increase hemoglobin levels, increase red blood cell count, or reduce iron deficiency.

[0424] After administering the composition to the patient, practitioners in the art can monitor the patient's improvement in response to treatment in various ways.For example, physicians can monitor the patient's hemoglobin level, red blood cell count, or iron deficiency by performing blood tests.The finding that the patient shows improved hemoglobin level, red blood cell count, or iron deficiency after administration of the composition compared to the test results before administration of the composition indicates that the patient is responding favorably to treatment.Subsequent dosages can be determined and administered as necessary.

[0425] Example 12 - Treatment of bone disease by administration of ALK2 antibodies According to the methods disclosed herein, a physician of the art can treat a subject, such as a human patient, with a bone disease (e.g., osteoporosis or osteopenia) to increase bone mineral density, increase bone formation, decrease bone resorption (e.g., bone loss), or reduce the risk of fracture. The treatment method can include diagnosing or identifying the subject as a candidate for treatment based on a standard clinical test of bone mineral density (e.g., dual-energy x-ray absorptiometry). To treat the subject, a physician of the art can administer a composition comprising an ALK2 antibody or an antigen-binding fragment thereof (e.g., an ALK2-binding fragment) to the subject. The composition comprising an ALK2 antibody or an antigen-binding fragment thereof (e.g., an ALK2-binding fragment) can be administered to the subject, for example, by parenteral injection (e.g., intravenous injection) to treat the bone disease. The ALK2 antibody or antigen-binding fragment thereof (e.g., an ALK2-binding fragment) is administered in a therapeutically effective amount, e.g., 0.01-500 mg / kg (e.g., 0.01, 0.1, 0.2, 0.3, 0.4, 0.5, 1, 2, 3, 4, 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 100, 150, 200, 250, 300, 350, 400, 450, or 500 mg / kg). In some embodiments, the ALK2 antibody or antigen-binding fragment thereof (e.g., an ALK2-binding fragment) is administered every other month, once a month, once every two weeks, or at least once a week or more (e.g., 1, 2, 3, 4, 5, 6, or 7 times a week or more). The ALK2 antibody or antigen-binding fragment thereof (e.g., an ALK2-binding fragment) is administered in an amount sufficient to increase bone mineral density, increase bone formation, decrease bone resorption (e.g., bone loss), or decrease the risk of fracture.

[0426] After administering the composition to the patient, practitioners in the art can monitor the patient's improvement in response to treatment in a variety of ways. For example, physicians can monitor the patient's bone mineral density by performing dual-energy X-ray absorptiometry. Finding that the patient shows increased bone mineral density, increased bone formation, decreased bone resorption (e.g., bone loss), or decreased risk of fracture after administration of the composition compared to test results before administration of the composition indicates that the patient is responding favorably to treatment. Subsequent dosages can be determined and administered as necessary.

[0427] Example 13 - Treatment of FOP by administration of ALK2 antibodies According to the methods disclosed herein, a practitioner of the art can treat a subject, such as a human patient with FOP (e.g., a patient with a gain-of-function mutation in ALK2), to prevent or reduce heterotopic ossification, slow the progression of FOP, or slow the onset of FOP. The method of treatment can include diagnosing or identifying the subject as a candidate for treatment based on genetic testing for ALK2 mutations or performing radiological imaging, CT, or MRI. To treat the subject, a practitioner of the art can administer a composition comprising an ALK2 antibody or an antigen-binding fragment thereof (e.g., an ALK2-binding fragment) to the subject. A composition comprising an ALK2 antibody or an antigen-binding fragment thereof (e.g., an ALK2-binding fragment) can be administered to the subject, for example, by parenteral injection (e.g., intravenous injection) to treat FOP. The ALK2 antibody or antigen-binding fragment thereof (e.g., an ALK2-binding fragment) is administered in a therapeutically effective amount, e.g., 0.01-500 mg / kg (e.g., 0.01, 0.1, 0.2, 0.3, 0.4, 0.5, 1, 2, 3, 4, 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 100, 150, 200, 250, 300, 350, 400, 450, or 500 mg / kg). In some embodiments, the ALK2 antibody or antigen-binding fragment thereof (e.g., an ALK2-binding fragment) is administered every other month, once a month, once every two weeks, or at least once a week or more (e.g., 1, 2, 3, 4, 5, 6, or 7 times a week or more). The ALK2 antibody or antigen-binding fragment thereof (eg, an ALK2-binding fragment) is administered in an amount sufficient to reduce heterotopic ossification, slow the progression of FOP, or delay the onset of FOP.

[0428] After administering the composition to the patient, practitioners in the art can monitor the patient's improvement in response to treatment in a variety of ways. For example, physicians can use radiography, CT, and / or MRI to monitor the patient's heterotopic ossification or disease progression. Findings that the patient shows a reduction in heterotopic ossification (e.g., a reduction in the amount of heterotopic ossification or the formation of additional heterotopic bone), no development of heterotopic ossification, or a delay in the onset or slowing of progression of FOP after administration of the composition, compared to test results before administration of the composition, indicate that the patient is responding favorably to treatment. Subsequent dosages can be determined and administered as necessary.

[0429] Example 14 - Treatment of Sjogren's syndrome by administration of ALK2 antibodies According to the methods disclosed herein, a practitioner of the art can treat a subject, such as a human patient, with Sjogren's syndrome to improve or reduce one or more symptoms of Sjogren's syndrome (e.g., dry eye, dry mouth, eye irritation, blurred vision, lacrimal gland fibrosis, lacrimal gland inflammation, systemic inflammation, joint pain, or fatigue). To treat the subject, a practitioner of the art can administer a composition comprising an ALK2 antibody or an antigen-binding fragment thereof (e.g., an ALK2-binding fragment) to the subject. A composition comprising an ALK2 antibody or an antigen-binding fragment thereof (e.g., an ALK2-binding fragment) can be administered to the subject, for example, by parenteral injection (e.g., intravenous injection) to treat Sjogren's syndrome. The ALK2 antibody or antigen-binding fragment thereof (e.g., an ALK2-binding fragment) is administered in a therapeutically effective amount, e.g., 0.01-500 mg / kg (e.g., 0.01, 0.1, 0.2, 0.3, 0.4, 0.5, 1, 2, 3, 4, 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 100, 150, 200, 250, 300, 350, 400, 450, or 500 mg / kg). In some embodiments, the ALK2 antibody or antigen-binding fragment thereof (e.g., an ALK2-binding fragment) is administered every other month, once a month, once every two weeks, or at least once a week or more (e.g., 1, 2, 3, 4, 5, 6, or 7 times a week or more). The ALK2 antibody or antigen-binding fragment thereof (e.g., an ALK2-binding fragment) is administered in an amount sufficient to ameliorate or reduce one or more symptoms of Sjogren's syndrome (e.g., by increasing tear or saliva production, e.g., dry eyes, dry mouth, eye irritation, blurred vision, lacrimal gland fibrosis, lacrimal gland inflammation, systemic inflammation, joint pain, or fatigue).

[0430] After administering the composition to the patient, practitioners in the art can monitor the patient's improvement in response to the treatment in a variety of ways. For example, a physician can monitor the patient's symptoms of Sjogren's syndrome by evaluating the patient's reports of tear production (e.g., tear production rate), saliva production, vision, eye irritation, inflammation or fibrosis of the lacrimal gland, or joint pain or fatigue. Finding that the patient shows a reduction in one or more symptoms of Sjogren's syndrome (e.g., dry eye, dry mouth, eye irritation, blurred vision, fibrosis of the lacrimal gland, inflammation of the lacrimal gland, systemic inflammation, joint pain, or fatigue) after administration of the composition compared to symptoms before administration of the composition indicates that the patient is responding favorably to the treatment. Subsequent dosages can be determined and administered as necessary.

[0431] Example 15 - Treatment of MO by administration of ALK2 antibodies According to the methods disclosed herein, a practitioner of the art can treat a subject, such as a human patient, with MO to prevent or reduce the formation of osteochondroma, reduce the size of osteochondroma, reduce the number of osteochondroma, or slow the growth of osteochondroma. The treatment method can include diagnosing or identifying the subject as a candidate for treatment based on genetic testing for mutations in EXT1 or EXT2, or based on performing radiological imaging, CT, or MRI. To treat the subject, a practitioner of the art can administer a composition comprising an ALK2 antibody or an antigen-binding fragment thereof (e.g., an ALK2-binding fragment) to the subject. A composition comprising an ALK2 antibody or an antigen-binding fragment thereof (e.g., an ALK2-binding fragment) can be administered to the subject, for example, by parenteral injection (e.g., intravenous injection) to treat MO. The ALK2 antibody or antigen-binding fragment thereof (e.g., an ALK2-binding fragment) is administered in a therapeutically effective amount, e.g., 0.01-500 mg / kg (e.g., 0.01, 0.1, 0.2, 0.3, 0.4, 0.5, 1, 2, 3, 4, 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 100, 150, 200, 250, 300, 350, 400, 450, or 500 mg / kg). In some embodiments, the ALK2 antibody or antigen-binding fragment thereof (e.g., an ALK2-binding fragment) is administered every other month, once a month, once every two weeks, or at least once a week or more (e.g., 1, 2, 3, 4, 5, 6, or 7 times a week or more). The ALK2 antibody or antigen-binding fragment thereof (e.g., an ALK2-binding fragment) is administered in an amount sufficient to reduce the formation of osteochondroma, reduce the size of osteochondroma, reduce the number of osteochondroma, or slow the growth of osteochondroma.

[0432] After administering the composition to the patient, practitioners in the art can monitor the patient's improvement in response to treatment in a variety of ways. For example, the physician can use radiography, CT, and / or MRI to monitor the growth, number, or size of the patient's osteochondroma. Finding that the patient shows reduced osteochondroma formation (e.g., no new osteochondroma formation or less osteochondroma formation compared to untreated subjects or osteochondroma formation before treatment), reduced osteochondroma size, reduced osteochondroma number, or delayed osteochondroma growth after administration of the composition, compared to test results before administration of the composition or compared to untreated subjects, indicates that the patient is responding favorably to the treatment. Subsequent dosages can be determined and administered as necessary.

[0433] Example 16 - Treatment of DIPG by administration of ALK2 antibody According to the methods disclosed herein, a physician of the art can treat a subject, such as a human patient, with DIPG to improve survival time, reduce tumor growth, reduce tumor size or volume, or prevent or reduce tumor metastasis. The treatment method can include diagnosing or identifying a subject as a candidate for treatment based on genetic testing for activating mutations in ALK2. To treat a subject, a physician of the art can administer a composition comprising an ALK2 antibody or an antigen-binding fragment thereof (e.g., an ALK2-binding fragment) to the subject. The composition comprising an ALK2 antibody or an antigen-binding fragment thereof (e.g., an ALK2-binding fragment) can be administered to the subject, for example, by parenteral injection (e.g., intravenous injection) or by local administration to the central nervous system (e.g., intraventricular, intrathecal, or intracisternal injection). The ALK2 antibody or antigen-binding fragment thereof (e.g., an ALK2-binding fragment) is administered in a therapeutically effective amount, e.g., 0.01-500 mg / kg (e.g., 0.01, 0.1, 0.2, 0.3, 0.4, 0.5, 1, 2, 3, 4, 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 100, 150, 200, 250, 300, 350, 400, 450, or 500 mg / kg). In some embodiments, the ALK2 antibody or antigen-binding fragment thereof (e.g., an ALK2-binding fragment) is administered every other month, once a month, once every two weeks, or at least once a week or more (e.g., 1, 2, 3, 4, 5, 6, or 7 times a week or more). The ALK2 antibody or antigen-binding fragment thereof (e.g., an ALK2-binding fragment) is administered in an amount sufficient to improve survival time, reduce tumor growth, reduce tumor size or volume, or prevent or reduce tumor metastasis.

[0434] After administering the composition to the patient, practitioners of the art can monitor the patient's improvement in response to the treatment in a variety of ways. For example, a physician can use imaging (e.g., MRI) to monitor the patient's survival time and tumor size, growth, or metastasis. The finding that the patient exhibits reduced tumor growth, reduced tumor size or volume, reduced tumor metastasis, or increased survival time compared to untreated subjects or compared to tumor growth, size, or metastasis in the subject prior to administration indicates that the patient is responding favorably to the treatment. Subsequent dosages can be determined and administered as necessary.

[0435] Example 17 - Treatment or prevention of PCO by administration of ALK2 antibodies According to the methods disclosed herein, a practitioner of the art can treat a subject, such as a human patient, who has or is at risk of developing PCO (e.g., a patient with PCO or a patient who has undergone or will soon undergo cataract surgery, e.g., a patient who is scheduled to undergo cataract surgery in the next 6 months, 5 months, 4 months, 3 months, 2 months, 1 month, 2 weeks, 1 week, or sooner) to prevent the onset of PCO, reduce PCO, improve vision (e.g., reduce blurry or cloudy vision), reduce light sensitivity or glare, reduce or prevent fibrosis (e.g., fibrosis on or near the posterior capsule, e.g., near an implanted lens), reduce or prevent differentiation of lens fibers, reduce or inhibit proliferation of lens epithelial cells, or reduce or prevent ocular inflammation. To treat a subject, a practitioner of the art can administer a composition comprising an ALK2 antibody or an antigen-binding fragment thereof (e.g., an ALK2-binding fragment) to the subject. A composition comprising an ALK2 antibody or an antigen-binding fragment thereof (e.g., an ALK2-binding fragment) can be administered to a subject, for example, by parenteral injection (e.g., intravenous injection) or ocular administration (e.g., intraocular injection or topical administration) to treat PCO. The ALK2 antibody or antigen-binding fragment thereof (e.g., an ALK2-binding fragment) is administered in a therapeutically effective amount, for example, 0.01 to 500 mg / kg (e.g., 0.01, 0.1, 0.2, 0.3, 0.4, 0.5, 1, 2, 3, 4, 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 100, 150, 200, 250, 300, 350, 400, 450, or 500 mg / kg). In some embodiments, the ALK2 antibody or antigen-binding fragment thereof (e.g., an ALK2-binding fragment) is administered every other month, once a month, once every two weeks, or at least once a week (e.g., 1, 2, 3, 4, 5, 6, or 7 or more times per week).The ALK2 antibody or antigen-binding fragment thereof (e.g., an ALK2-binding fragment) is administered in an amount sufficient to prevent the onset of PCO, reduce PCO, improve vision (e.g., reduce blurred or cloudy vision), reduce light sensitivity or glare, reduce or prevent fibrosis (e.g., fibrosis on or near the posterior capsule, e.g., near an implanted lens), reduce or prevent lens fiber differentiation, reduce or inhibit lens epithelial cell proliferation, or reduce or prevent ocular inflammation.

[0436] After administering the composition to the patient, practitioners of the art can monitor the patient's improvement in response to the treatment in a variety of ways. For example, the physician can monitor the patient's vision, light sensitivity or glare, posterior capsule opacity, or ocular inflammation. A finding that the patient does not develop PCO or exhibits improved vision, reduced light sensitivity or glare, reduced fibrosis, or reduced ocular inflammation after administration of the composition, compared to untreated subjects or compared to the patient's measurements before treatment, indicates that the patient is responding favorably to the treatment. Subsequent dosages can be determined and administered as necessary.

[0437] Example 18 - Treatment of cardiac hypertrophy and / or cardiac fibrosis by administration of ALK2 antibodies According to the methods disclosed herein, a practitioner of the art can treat a subject, such as a human patient, with cardiac hypertrophy and / or cardiac fibrosis to delay or prevent the onset of cardiac hypertrophy or cardiac fibrosis, reverse cardiac fibrosis, reduce or inhibit cardiac scar formation, increase or induce cardiac regeneration, or ameliorate one or more symptoms of cardiac hypertrophy or cardiac fibrosis (e.g., improve exercise capacity, increase blood ejection volume, decrease left ventricular end diastolic pressure, decrease pulmonary capillary wedge pressure, increase cardiac output, increase cardiac index, decrease pulmonary artery pressure, decrease left ventricular end systolic and diastolic dimensions, decrease left and right ventricular wall stress, decrease wall tension and / or wall thickness, increase myocardial contractility, decrease cardiomyocyte extracellular matrix deposition, or decrease fibrosis). To treat a subject, a practitioner of the art can administer a composition comprising an ALK2 antibody or an antigen-binding fragment thereof (e.g., an ALK2-binding fragment) to the subject. A composition comprising an ALK2 antibody or an antigen-binding fragment thereof (e.g., an ALK2-binding fragment) can be administered to a subject, for example, by parenteral injection (e.g., intravenous injection) to treat cardiac hypertrophy and / or cardiac fibrosis. The ALK2 antibody or antigen-binding fragment thereof (e.g., an ALK2-binding fragment) is administered in a therapeutically effective amount, for example, 0.01 to 500 mg / kg (e.g., 0.01, 0.1, 0.2, 0.3, 0.4, 0.5, 1, 2, 3, 4, 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 100, 150, 200, 250, 300, 350, 400, 450, or 500 mg / kg). In some embodiments, the ALK2 antibody or antigen-binding fragment thereof (e.g., an ALK2-binding fragment) is administered every other month, once a month, once every two weeks, or at least once a week (e.g., 1, 2, 3, 4, 5, 6, or 7 or more times per week).The ALK2 antibody or antigen-binding fragment thereof (e.g., an ALK2-binding fragment) is administered in an amount sufficient to delay or prevent the onset of cardiac hypertrophy or cardiac fibrosis, reverse cardiac fibrosis, reduce or inhibit cardiac scar formation, increase or induce cardiac regeneration, or ameliorate one or more symptoms of cardiac hypertrophy or cardiac fibrosis (e.g., improved exercise capacity, increased blood ejection volume, decreased left ventricular end diastolic pressure, decreased pulmonary capillary wedge pressure, increased cardiac output, increased cardiac index, decreased pulmonary artery pressure, decreased left ventricular end systolic and diastolic dimensions, decreased left and right ventricular wall stress, decreased wall tension and / or wall thickness, increased myocardial contractility, decreased cardiomyocyte extracellular matrix deposition, or decreased fibrosis).

[0438] After administering the composition to the patient, practitioners in the art can monitor the patient's improvement in response to treatment in a variety of ways. For example, physicians can monitor the patient's condition of cardiac hypertrophy or cardiac fibrosis (e.g., exercise capacity, blood ejection volume, left ventricular end-diastolic pressure, pulmonary capillary wedge pressure, cardiac output, cardiac index, pulmonary artery pressure, left ventricular end-systolic and diastolic dimensions, left and right ventricular wall stress, wall tension and / or wall thickness, or myocardial contractility) using stress tests, echocardiography, MRI, or other approaches. A finding that the patient exhibits reduced cardiac fibrosis, increased cardiac regeneration, or improvement in one or more symptoms of cardiac hypertrophy or cardiac fibrosis (e.g., improved exercise capacity, increased blood ejection volume, decreased left ventricular end diastolic pressure, decreased pulmonary capillary wedge pressure, increased cardiac output, increased cardiac index, decreased pulmonary artery pressure, decreased left ventricular end systolic and diastolic dimensions, decreased left and right ventricular wall stress, decreased wall tension and / or wall thickness, increased myocardial contractility, decreased cardiomyocyte extracellular matrix deposition, or decreased fibrosis) after administration of the composition compared to test results prior to administration of the composition or compared to untreated subjects indicates that the patient is responding favorably to the treatment. Subsequent doses can be determined and administered as necessary.

[0439] Other embodiments While the invention has been described with respect to particular embodiments thereof, it is capable of further modifications, it being understood that this application is intended to cover any variations, uses, or adaptations of the invention generally in accordance with the principles of the invention, including such departures from the disclosure as are within known or customary practice within the art to which the invention pertains and which may be applicable to the essential features described above.

[0440] All publications, patents, and patent applications are herein incorporated by reference in their entirety to the same extent as if each individual publication, patent, or patent application was specifically and individually indicated to be incorporated by reference in its entirety.

[0441] Other embodiments are within the scope of the following claims.

Claims

1. 1. An isolated antibody, or an ALK2-binding fragment thereof, comprising: (1) a light chain variable domain comprising: SGSSSNIGSNYVS (SEQ ID NO: 1) and SGDX １ X ２ X ３ X ４ X ５ X ６ X ７ X ８ (SEQ ID NO:2, where X １ is S or N, and X ２ is I or L, and X ３ is P, G, or R; X ４ is S, T, or K; X ５ is F, K, or Y; X ６ is F, Y, or S; X is ７ is A or V, and X ８ a light chain complementarity determining region (CDR) 1 comprising an amino acid sequence selected from the group consisting of: １ X ２ IYX ３ X ４ X ５ X ６ RPS (SEQ ID NO: 3, where X １ is V or L; X ２ is V or L; X ３ is K, R, G or Y; X ４ is N or D, and X ５ is N or S, and X ６ (1) a light chain variable domain comprising a light chain CDR2 comprising the amino acid sequence GXDHSDRFYV (SEQ ID NO: 4), YVTAPWKSIW (SEQ ID NO: 5), YSADAQQMKA (SEQ ID NO: 6), QVYASVHRM (SEQ ID NO: 7), and QTYDWSHFGW (SEQ ID NO: 8); and (2) a light chain CDR3 comprising the amino acid sequence selected from ASWDHSDRFYV (SEQ ID NO: 4), YVTAPWKSIW (SEQ ID NO: 5), YSADAQQMKA (SEQ ID NO: 6), QVYASVHRM (SEQ ID NO: 7), and QTYDWSHFGW (SEQ ID NO: 8). １ TFX ２ S.X. ３ X ４ X ５ X ６ (SEQ ID NO: 9, where X １ is G or F, and X ２ is S or N, and X ３ is Y, H, S, or A; X ４ is G or A, and X ５ is V, M, or I; X ６ is S or H); １ IIPX ２ FGX ３ ANYAQKFQG (SEQ ID NO: 10, where: １ is G or R, and X ２ is H or D, and X ３ is I or T), WVGRIKS KX １ D.X. ２ X ３ TTDYAAPVKG (SEQ ID NO: 11, where: １ is A or R, and X ２ is S or G, and X ３ is G or Y), and a heavy chain CDR2 comprising an amino acid sequence selected from the group consisting of WVSVISSDGGSTYYADSVKG (SEQ ID NO: 12); and a heavy chain CDR3 comprising an amino acid sequence selected from the group consisting of EIGSLDI (SEQ ID NO: 13), DYGVAFAY (SEQ ID NO: 14), DYGGLKFDY (SEQ ID NO: 15), GPTQAIHYFAY (SEQ ID NO: 16), and AGFILGSLGVAWMDV (SEQ ID NO: 17).

2. 2. The isolated antibody or ALK2-binding fragment thereof of claim 1, wherein the light chain CDR1 comprises the sequence SGSSSNIGSNYVS (SEQ ID NO:1).

3. 2. The isolated antibody or ALK2-binding fragment thereof of claim 1, wherein the light chain CDR1 comprises the sequence SGDSIPSFFAS (SEQ ID NO: 18).

4. 2. The isolated antibody or ALK2-binding fragment thereof of claim 1, wherein the light chain CDR1 comprises the sequence SGDNIGTKYAY (SEQ ID NO: 19).

5. 2. The isolated antibody or ALK2-binding fragment thereof of claim 1, wherein the light chain CDR1 comprises the sequence SGDNLRKYSAH (SEQ ID NO: 20).

6. 2. The isolated antibody or ALK2-binding fragment thereof of claim 1, wherein the light chain CDR1 comprises the sequence SGDSLGSKSVH (SEQ ID NO:21).

7. 7. The isolated antibody or ALK2-binding fragment thereof according to any one of claims 1 to 6, wherein the light chain CDR2 comprises the sequence VLIYKNNHRPS (SEQ ID NO:24).

8. 7. The isolated antibody or ALK2-binding fragment thereof according to any one of claims 1 to 6, wherein the light chain CDR2 comprises the sequence LVIYRDSNRPS (SEQ ID NO:25).

9. 7. The isolated antibody or ALK2-binding fragment thereof according to any one of claims 1 to 6, wherein the light chain CDR2 comprises the sequence LVIYGDSDRPS (SEQ ID NO:26).

10. 7. The isolated antibody or ALK2-binding fragment thereof according to any one of claims 1 to 6, wherein the light chain CDR2 comprises the sequence LVIYYDNKRPS (SEQ ID NO:27).

11. 7. The isolated antibody or ALK2-binding fragment thereof according to any one of claims 1 to 6, wherein the light chain CDR2 comprises the sequence LVIYRDSKRPS (SEQ ID NO:28).

12. 12. The isolated antibody or ALK2-binding fragment thereof according to any one of claims 1 to 11, wherein the light chain CDR3 comprises the sequence ASWDHSDRFYV (SEQ ID NO: 4).

13. 12. The isolated antibody or ALK2-binding fragment thereof according to any one of claims 1 to 11, wherein the light chain CDR3 comprises the sequence YVTAPWKSIW (SEQ ID NO:5).

14. 12. The isolated antibody or ALK2-binding fragment thereof according to any one of claims 1 to 11, wherein the light chain CDR3 comprises the sequence YSADAQQMKA (SEQ ID NO: 6).

15. 12. The isolated antibody or ALK2-binding fragment thereof according to any one of claims 1 to 11, wherein the light chain CDR3 comprises the sequence QVYASVHRM (SEQ ID NO:7).

16. 12. The isolated antibody or ALK2-binding fragment thereof of any one of claims 1 to 11, wherein the light chain CDR3 comprises QTYDWSHFGW (SEQ ID NO: 8).

17. 17. The isolated antibody or ALK2-binding fragment thereof according to any one of claims 1 to 16, wherein the heavy chain CDR1 comprises the sequence GGTFSSYGVS (SEQ ID NO:31).

18. 17. The isolated antibody or ALK2-binding fragment thereof according to any one of claims 1 to 16, wherein the heavy chain CDR1 comprises the sequence GFTFSSHAMS (SEQ ID NO:32).

19. 17. The isolated antibody or ALK2-binding fragment thereof according to any one of claims 1 to 16, wherein the heavy chain CDR1 comprises the sequence GFTFNSSAMS (SEQ ID NO:33).

20. 17. The isolated antibody or ALK2-binding fragment thereof according to any one of claims 1 to 16, wherein the heavy chain CDR1 comprises the sequence GGTFSSYAIH (SEQ ID NO: 34).

21. 17. The isolated antibody or ALK2-binding fragment thereof according to any one of claims 1 to 16, wherein the heavy chain CDR1 comprises the sequence GFTFSSAAMH (SEQ ID NO:35).

22. 22. The isolated antibody or ALK2-binding fragment thereof according to any one of claims 1 to 21, wherein the heavy chain CDR2 comprises the sequence WMGGIIPHFGIANYAQKFQG (SEQ ID NO: 36).

23. 22. The isolated antibody or ALK2-binding fragment thereof of any one of claims 1 to 21, wherein the heavy chain CDR2 comprises the sequence WVGRIKSKADSGTTDYAAPVKG (SEQ ID NO: 37).

24. 22. The isolated antibody or ALK2-binding fragment thereof according to any one of claims 1 to 21, wherein the heavy chain CDR2 comprises the sequence WVGRIKSKRDGYTTDYAAPVKG (SEQ ID NO: 38).

25. 22. The isolated antibody or ALK2-binding fragment thereof according to any one of claims 1 to 21, wherein the heavy chain CDR2 comprises the sequence WMGRIIPDFGTANYAQKFQG (SEQ ID NO: 39).

26. 22. The isolated antibody or ALK2-binding fragment thereof according to any one of claims 1 to 21, wherein the heavy chain CDR2 comprises the sequence WVSVISSDGGSTYYADSVKG (SEQ ID NO: 12).

27. 27. The isolated antibody or ALK2-binding fragment thereof according to any one of claims 1 to 26, wherein the heavy chain CDR3 comprises the sequence EIGSLDI (SEQ ID NO: 13).

28. 27. The isolated antibody or ALK2-binding fragment thereof according to any one of claims 1 to 26, wherein the heavy chain CDR3 comprises the sequence DYGVAFAY (SEQ ID NO: 14).

29. 27. The isolated antibody or ALK2-binding fragment thereof according to any one of claims 1 to 26, wherein the heavy chain CDR3 comprises the sequence DYGGLKFDY (SEQ ID NO: 15).

30. 27. The isolated antibody or ALK2-binding fragment thereof according to any one of claims 1 to 26, wherein the heavy chain CDR3 comprises the sequence GPTQAIHYFAY (SEQ ID NO: 16).

31. 27. The isolated antibody or ALK2-binding fragment thereof according to any one of claims 1 to 26, wherein the heavy chain CDR3 comprises the sequence AGFILGSLGVAWMDV (SEQ ID NO: 17).

32. Light chain CDR2 has the sequence LVIYX １ D.X. ２ X ３ RPS (SEQ ID NO: 22, where X １ is R, G, or Y; X ２ is S or N, and X ３ is N, D, or K).

33. The light chain CDR2 has the sequence LVIYRDSX １ RPS (SEQ ID NO:23, where X １ The isolated antibody or ALK2-binding fragment thereof of claim 1 , comprising:

34. Heavy chain CDR1 has the sequence GFTFSSX １ AMX ２ (SEQ ID NO:29, where X １ is H or A, and X ２ The isolated antibody or ALK2-binding fragment thereof of claim 1 , comprising:

35. The heavy chain CDR1 has the sequence GFTFX １ S.X. ２ AMS (SEQ ID NO: 30, where X １ is S or N, and X ２ The isolated antibody or ALK2-binding fragment thereof of claim 1 , comprising:

36. 2. The isolated antibody or ALK2-binding fragment thereof of claim 1, wherein the light chain CDR1 comprises the amino acid sequence SGSSSNIGSNYVS (SEQ ID NO: 1), the light chain CDR2 comprises the amino acid sequence VLIYKNNHRPS (SEQ ID NO: 24), and the light chain CDR3 comprises the amino acid sequence ASWDHSDRFYV (SEQ ID NO: 4).

37. 37. The isolated antibody or ALK2-binding fragment thereof of claim 1 or 36, wherein the heavy chain CDR1 comprises the amino acid sequence GGTFSSYGVS (SEQ ID NO: 31), the heavy chain CDR2 comprises the amino acid sequence WMGGIIPHFGIANYAQKFQG (SEQ ID NO: 36), and the heavy chain CDR3 comprises the amino acid sequence EIGSLDI (SEQ ID NO: 13).

38. 2. The isolated antibody or ALK2-binding fragment thereof of claim 1, wherein the light chain CDR1 comprises the amino acid sequence SGDSIPSFFAS (sequence number 18), the light chain CDR2 comprises the amino acid sequence LVIYRDSNRPS (sequence number 25), and the light chain CDR3 comprises the amino acid sequence YVTAPWKSIW (sequence number 5).

39. 40. The isolated antibody or ALK2-binding fragment thereof of claim 1 or 38, wherein the heavy chain CDR1 comprises the amino acid sequence GFTFSSHAMS (SEQ ID NO: 32), the heavy chain CDR2 comprises the amino acid sequence WVGRIKSKADSGTTDYAAPVKG (SEQ ID NO: 37), and the heavy chain CDR3 comprises the amino acid sequence DYGVAFAY (SEQ ID NO: 14).

40. 2. The isolated antibody or ALK2-binding fragment thereof of claim 1, wherein the light chain CDR1 comprises the amino acid sequence SGDNIGTKYAY (sequence number 19), the light chain CDR2 comprises the amino acid sequence LVIYGDSDRPS (sequence number 26), and the light chain CDR3 comprises the amino acid sequence YSADAQQMKA (sequence number 6).

41. 41. The isolated antibody of claim 1 or 40, or an ALK2-binding fragment thereof, wherein the heavy chain CDR1 comprises the amino acid sequence GFTFNSSAMS (SEQ ID NO: 33), the heavy chain CDR2 comprises the amino acid sequence WVGRIKSKRDGYTTDYAAPVKG (SEQ ID NO: 38), and the heavy chain CDR3 comprises the amino acid sequence DYGGLKFDY (SEQ ID NO: 15).

42. 2. The isolated antibody or ALK2-binding fragment thereof of claim 1, wherein the light chain CDR1 comprises the amino acid sequence SGDNLRKYSAH (SEQ ID NO: 20), the light chain CDR2 comprises the amino acid sequence LVIYYDNKRPS (SEQ ID NO: 27), and the light chain CDR3 comprises the amino acid sequence QVYASVHRM (SEQ ID NO: 7).

43. 43. The isolated antibody of claim 1 or 42, or an ALK2-binding fragment thereof, wherein the heavy chain CDR1 comprises the amino acid sequence GGTFSSYAIH (SEQ ID NO: 34), the heavy chain CDR2 comprises the amino acid sequence WMGRIIPDFGTANYAQKFQG (SEQ ID NO: 39), and the heavy chain CDR3 comprises the amino acid sequence GPTQAIHYFAY (SEQ ID NO: 16).

44. 2. The isolated antibody or ALK2-binding fragment thereof of claim 1, wherein the light chain CDR1 comprises the amino acid sequence SGDSLGSKSVH (sequence number 21), the light chain CDR2 comprises the amino acid sequence LVIYRDSKRPS (sequence number 28), and the light chain CDR3 comprises the amino acid sequence QTYDWSHFGW (sequence number 8).

45. 45. The isolated antibody of claim 1 or 44, or an ALK2-binding fragment thereof, wherein the heavy chain CDR1 comprises the amino acid sequence GFTFSSAAMH (SEQ ID NO: 35), the heavy chain CDR2 comprises the amino acid sequence WVSVISSDGGSTYYADSVKG (SEQ ID NO: 12), and the heavy chain CDR3 comprises the amino acid sequence AGFILGSLGVAWMDV (SEQ ID NO: 17).

46. 2. The isolated antibody of claim 1, wherein the antibody has at least 90% sequence identity with amino acids 1 to 331 of sequence SEQ ID NO:67, 1 to 332 of sequence SEQ ID NO:68, 1 to 333 of sequence SEQ ID NO:69, 1 to 332 of sequence SEQ ID NO:70, or 1 to 337 of sequence SEQ ID NO:71, excluding light chain CDR1, CDR2, and CDR3 and heavy chain CDR1, CDR2, and CDR3.

47. 2. The isolated antibody of claim 1, wherein the antibody comprises amino acids 1 to 433 of sequence SEQ ID NO: 67, amino acids 1 to 434 of sequence SEQ ID NO: 68, amino acids 1 to 435 of sequence SEQ ID NO: 69, amino acids 1 to 434 of sequence SEQ ID NO: 70, or amino acids 1 to 439 of sequence SEQ ID NO:

71.

48. An isolated antibody, or an ALK2-binding fragment thereof, comprising: (1) a light chain variable domain, comprising RASQGISGNWLT (SEQ ID NO: 40), SGDX １ X ２ R.X. ３ X ４ X ５ X ６ H (SEQ ID NO:64, where X １ is N or A, and X ２ is I or L, and X ３ is K or Y, and X ４ is K or Y, and X ５ is Y or I, and X ６ a light chain complementarity determining region (CDR) 1 comprising an amino acid sequence selected from the group consisting of SGSSSNIGQNYVS (SEQ ID NO:58); １ IYX ２ X ３ X ４ X ５ X ６ X ７ S (SEQ ID NO: 65, where X １ is V or L; X ２ is D, R, or Y; X ３ is A, D, or N, and X ４ is S or N, and X ５ is K or N, and X ６ is L or R, and X ７ (1) a light chain variable domain comprising a light chain CDR2 comprising or consisting of the amino acid sequence HQSYRGPM (SEQ ID NO:42), SSAGRDNY (SEQ ID NO:48), QSYGPGSV (SEQ ID NO:54), and SSWDLLSKSR (SEQ ID NO:60); and (2) a light chain CDR3 comprising the amino acid sequence GX １ TFX ２ X ３ X ４ X ５ X ６ X ７ (SEQ ID NO:66, where X １ is F or G, and X ２ is G or S, and X ３ is R, S, D, or T; X ４ is F, S, Y, or H; X ５ is V or A, and X ６ is M or I, and X ７ is H or S); １ IX ２ YX ３ X ４ S.X. ５ TYYADSVKG (SEQ ID NO: 76, where: １ is V or S, and X ２ is G, H, or F; ３ is S or D, and X ４ is G or S, and X ５ is S, E, or N), and WVS-X １ IX ２ YX ３ X ４ S.X. ５ TYYADSVKG (SEQ ID NO: 76, where: １ is V or S, and X ２ is G, H, or F; ３ is S or D, and X ４ is G or S, and X ５ is S, E, or N), and WMGLIQPRFGTANYAQKFQR (SEQ ID NO:62); and a heavy chain CDR3 comprising an amino acid sequence selected from the group consisting of EPGYYYPSGYYRGPGYWMDV (SEQ ID NO:45), DRYFFDV (SEQ ID NO:51), PKSYASGPFAY (SEQ ID NO:57), and DYYGGMAY (SEQ ID NO:63), or an ALK2-binding fragment thereof.

49. 49. The isolated antibody or ALK2-binding fragment thereof of claim 48, wherein the light chain CDR1 comprises the sequence RASQGISGNWLT (SEQ ID NO: 40).

50. 49. The isolated antibody or ALK2-binding fragment thereof of claim 48, wherein the light chain CDR1 comprises the sequence SGDNIRKKYVH (SEQ ID NO:46).

51. 49. The isolated antibody or ALK2-binding fragment thereof of claim 48, wherein the light chain CDR1 comprises the sequence SGDALRYYIAH (SEQ ID NO:52).

52. 49. The isolated antibody or ALK2-binding fragment thereof of claim 48, wherein the light chain CDR1 comprises the sequence SGSSSNIGQNYVS (SEQ ID NO:58).

53. 53. The isolated antibody or ALK2-binding fragment thereof according to any one of claims 48 to 52, wherein the light chain CDR2 comprises the sequence LLIYDASNLQS (SEQ ID NO:41).

54. 53. The isolated antibody or ALK2-binding fragment thereof according to any one of claims 48 to 52, wherein the light chain CDR2 comprises the sequence LVIYRDSNRPS (SEQ ID NO:47).

55. 53. The isolated antibody or ALK2-binding fragment thereof according to any one of claims 48 to 52, wherein the light chain CDR2 comprises the sequence LVIYYNNNRPS (SEQ ID NO:53).

56. 53. The isolated antibody or ALK2-binding fragment thereof according to any one of claims 48 to 52, wherein the light chain CDR2 comprises the sequence LLIYDNSKRPS (SEQ ID NO:59).

57. 57. The isolated antibody or ALK2-binding fragment thereof according to any one of claims 48 to 56, wherein the light chain CDR3 comprises the sequence HQSYRGPM (SEQ ID NO:42).

58. 57. The isolated antibody or ALK2-binding fragment thereof according to any one of claims 48 to 56, wherein the light chain CDR3 comprises the sequence SSAGRDNY (SEQ ID NO:48).

59. 57. The isolated antibody or ALK2-binding fragment thereof according to any one of claims 48 to 56, wherein the light chain CDR3 comprises the sequence QSYGPGSV (SEQ ID NO:54).

60. 57. The isolated antibody or ALK2-binding fragment thereof of any one of claims 48 to 56, wherein the light chain CDR3 comprises the sequence SSWDLLSKSR (SEQ ID NO: 60).

61. 61. The isolated antibody or ALK2-binding fragment thereof according to any one of claims 48 to 60, wherein the heavy chain CDR1 comprises the sequence GFTFGRFVMH (SEQ ID NO:43).

62. 61. The isolated antibody or ALK2-binding fragment thereof of any one of claims 48 to 60, wherein the heavy chain CDR1 comprises the sequence GFTFSSSSAMH (SEQ ID NO:49).

63. 61. The isolated antibody or ALK2-binding fragment thereof of any one of claims 48 to 60, wherein the heavy chain CDR1 comprises the sequence GFTFSDYAMH (SEQ ID NO:55).

64. 61. The isolated antibody or ALK2-binding fragment thereof according to any one of claims 48 to 60, wherein the heavy chain CDR1 comprises the sequence GGTFSTHAIS (SEQ ID NO:61).

65. 65. The isolated antibody or ALK2-binding fragment thereof of any one of claims 48 to 64, wherein the heavy chain CDR2 comprises the sequence WVSVIGYSGSSTYYADSVKG (SEQ ID NO:44).

66. 65. The isolated antibody or ALK2-binding fragment thereof of any one of claims 48 to 64, wherein the heavy chain CDR2 comprises the sequence WVSVIHYDSSETYYADSVKG (SEQ ID NO:50).

67. 65. The isolated antibody or ALK2-binding fragment thereof of any one of claims 48 to 64, wherein the heavy chain CDR2 comprises the sequence WVSSIFYSGSNTYYADSVKG (SEQ ID NO:56).

68. 65. The isolated antibody or ALK2-binding fragment thereof of any one of claims 48 to 64, wherein the heavy chain CDR2 comprises the sequence WMGLIQPRFGTANYAQKFQR (SEQ ID NO: 62).

69. 69. The isolated antibody or ALK2-binding fragment thereof of any one of claims 48 to 68, wherein the heavy chain CDR3 comprises the sequence EPGYYYPSGYYRGPGYWMDV (SEQ ID NO: 45).

70. 69. The isolated antibody or ALK2-binding fragment thereof according to any one of claims 48 to 68, wherein the heavy chain CDR3 comprises the sequence DRYFFDV (SEQ ID NO:51).

71. 69. The isolated antibody or ALK2-binding fragment thereof according to any one of claims 48 to 68, wherein the heavy chain CDR3 comprises the sequence PKSYASGPFAY (SEQ ID NO:57).

72. 69. The isolated antibody or ALK2-binding fragment thereof according to any one of claims 48 to 68, wherein the heavy chain CDR3 comprises the sequence DYYGGMAY (SEQ ID NO: 63).

73. 49. The isolated antibody or ALK2-binding fragment thereof of claim 48, wherein the light chain CDR1 comprises the amino acid sequence RASQGISGNWLT (sequence number 40), the light chain CDR2 comprises the amino acid sequence LLIYDASNLQS (sequence number 41), and the light chain CDR3 comprises the amino acid sequence HQSYRGPM (sequence number 42).

74. 74. The isolated antibody or ALK2-binding fragment thereof of claim 48 or 73, wherein the heavy chain CDR1 comprises the amino acid sequence GFTFGRFVMH (SEQ ID NO: 43), the heavy chain CDR2 comprises the amino acid sequence WVSVIGYSGSSTYYADSVKG (SEQ ID NO: 44), and the heavy chain CDR3 comprises the amino acid sequence EPGYYYPSGYYRGPGYWMDV (SEQ ID NO: 45).

75. 49. The isolated antibody or ALK2-binding fragment thereof of claim 48, wherein the light chain CDR1 comprises the amino acid sequence SGDNIRKKYVH (sequence number 46), the light chain CDR2 comprises the amino acid sequence LVIYRDSNRPS (sequence number 47), and the light chain CDR3 comprises the amino acid sequence SSAGRDNY (sequence number 48).

76. 76. The isolated antibody or ALK2-binding fragment thereof of claim 48 or 75, wherein the heavy chain CDR1 comprises the amino acid sequence GFTFSSSSAMH (SEQ ID NO: 49), the heavy chain CDR2 comprises the amino acid sequence WVSVIHYDSSETYYADSVKG (SEQ ID NO: 50), and the heavy chain CDR3 comprises the amino acid sequence DRYFFDV (SEQ ID NO: 51).

77. 49. The isolated antibody or ALK2-binding fragment thereof of claim 48, wherein the light chain CDR1 comprises the amino acid sequence SGDALRYYIAH (SEQ ID NO: 52), the light chain CDR2 comprises the amino acid sequence LVIYYNNNRPS (SEQ ID NO: 53), and the light chain CDR3 comprises the amino acid sequence QSYGPGSV (SEQ ID NO: 54).

78. 78. The isolated antibody or ALK2-binding fragment thereof of claim 48 or 77, wherein the heavy chain CDR1 comprises the amino acid sequence GFTFSDYAMH (SEQ ID NO: 55), the heavy chain CDR2 comprises the amino acid sequence WVSSIFYSGSNTYYADSVKG (SEQ ID NO: 56), and the heavy chain CDR3 comprises the amino acid sequence PKSYASGPFAY (SEQ ID NO: 57).

79. 49. The isolated antibody or ALK2-binding fragment thereof of claim 48, wherein the light chain CDR1 comprises the amino acid sequence SGSSSNIGQNYVS (SEQ ID NO:58), the light chain CDR2 comprises the amino acid sequence LLIYDNSKRPS (SEQ ID NO:59), and the light chain CDR3 comprises the amino acid sequence SSWDLLSKSR (SEQ ID NO:60).

80. 80. The isolated antibody or ALK2-binding fragment thereof of claim 48 or 79, wherein the heavy chain CDR1 comprises the amino acid sequence GGTFSTHAIS (SEQ ID NO: 61), the heavy chain CDR2 comprises the amino acid sequence WMGLIQPRFGTANYAQKFQR (SEQ ID NO: 62), and the heavy chain CDR3 comprises the amino acid sequence DYYGGMAY (SEQ ID NO: 63).

81. 49. The isolated antibody of claim 48, wherein the antibody has at least 90% sequence identity with amino acids 1 to 344 of sequence SEQ ID NO: 72, amino acids 1 to 327 of sequence SEQ ID NO: 73, amino acids 1 to 331 of sequence SEQ ID NO: 74, or amino acids 1 to 332 of sequence SEQ ID NO: 75, excluding light chain CDR1, CDR2, and CDR3 and heavy chain CDR1, CDR2, and CDR3.

82. 49. The isolated antibody of claim 48, wherein the antibody comprises amino acids 1 to 446 of sequence SEQ ID NO: 72, amino acids 1 to 429 of sequence SEQ ID NO: 73, amino acids 1 to 433 of sequence SEQ ID NO: 74, or amino acids 1 to 434 of sequence SEQ ID NO:

75.

83. A nucleic acid molecule comprising a nucleotide sequence encoding the antibody or ALK2-binding fragment thereof according to any one of claims 1 to 82.

84. 84. An expression vector comprising the nucleic acid molecule of claim 83.

85. A cell comprising the expression vector of claim 84.

86. A pharmaceutical composition comprising an antibody or an ALK2-binding fragment thereof according to any one of claims 1 to 82, a nucleic acid molecule according to claim 83, or an expression vector according to claim 84, together with one or more pharma- ceutically acceptable carriers or excipients.

87. 87. The pharmaceutical composition of claim 86, wherein the antibody or ALK2 binding fragment thereof is in a therapeutically effective amount.

88. A method for increasing bone mineral density in a subject in need thereof comprising administering to the subject a therapeutically effective amount of an antibody or ALK2-binding fragment thereof described in any one of claims 1 to 82, a nucleic acid molecule described in claim 83, an expression vector described in claim 84, or a pharmaceutical composition described in claim 86 or 87.

89. A method for reducing bone resorption, increasing bone formation, increasing bone strength or reducing risk of fracture in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of an antibody or ALK2-binding fragment thereof described in any one of claims 1 to 82, a nucleic acid molecule described in claim 83, an expression vector described in claim 84, or a pharmaceutical composition described in claim 86 or 87.

90. A method for treating a subject at risk of developing a bone disease, comprising administering to the subject a therapeutically effective amount of an antibody or ALK2-binding fragment thereof described in any one of claims 1 to 82, a nucleic acid molecule described in claim 83, an expression vector described in claim 84, or a pharmaceutical composition described in claim 86 or 87.

91. 91. The method of claim 90, wherein the bone disease is primary osteoporosis, secondary osteoporosis, osteopenia, osteopetrosis, bone fracture, bone loss associated with bone cancer or cancer metastasis, Paget's disease, renal osteodystrophy, treatment-related bone loss, diet-related bone loss, bone loss associated with the treatment of obesity, low gravity-related bone loss, or immobility-related bone loss.

92. A method of increasing red blood cell levels, increasing hemoglobin levels, increasing red blood cell count, promoting or increasing red blood cell formation, and decreasing hepcidin levels in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of an antibody or ALK2-binding fragment thereof described in any one of claims 1 to 82, a nucleic acid molecule described in claim 83, an expression vector described in claim 84, or a pharmaceutical composition described in claim 86 or 87.

93. A method of treating a subject having or at risk of developing anemia, comprising administering to the subject a therapeutically effective amount of an antibody or ALK2-binding fragment thereof described in any one of claims 1 to 82, a nucleic acid molecule described in claim 83, an expression vector described in claim 84, or a pharmaceutical composition described in claim 86 or 87.

94. 94. The method of claim 93, wherein the anemia is associated with cancer, cancer treatment, chronic kidney disease, acute kidney disease or failure, chronic renal disease or failure, myelodysplastic syndrome, thalassemia, nutritional deficiency, adverse reaction to medication, inflammatory or autoimmune disease, splenomegaly, porphyria, vasculitis, hemolysis, bone marrow deficiency, bone marrow transplantation, acute liver disease, chronic liver disease, acute bleeding, chronic bleeding, infection, hemoglobinopathies, drug use, alcohol abuse, Churg-Strauss syndrome, Felty syndrome, graft versus host disease, hematopoietic stem cell transplantation, myelofibrosis, pancytopenia, pure red blood cell aplasia, Henoch-Schonlein purpura, Schwachmann syndrome, advanced age, contraindications to blood transfusion, surgery, trauma, wound, ulcer, urinary tract bleeding, gastrointestinal bleeding, frequent blood donations, or heavy menstrual bleeding.

95. 95. The method of claim 93 or 94, wherein the anemia is aplastic anemia, iron deficiency anemia, vitamin deficiency anemia, inflammatory anemia, IRIDA, anemia associated with myeloid disease, hemolytic anemia, sickle cell anemia, microcytic anemia, hypochromic anemia, sideroblastic anemia, Diamond-Blackfan anemia, Fanconi anemia, or refractory anemia with excess blasts.

96. 96. The method of claim 95, wherein the anemia is IRIDA.

97. 96. The method of claim 95, wherein the anemia is anemia of inflammation.

98. 98. The method of any one of claims 93 to 97, wherein the anemia is associated with elevated hepcidin levels.

99. A method for preventing or reducing heterotopic ossification in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of an antibody or ALK2-binding fragment thereof described in any one of claims 1 to 82, a nucleic acid molecule described in claim 83, an expression vector described in claim 84, or a pharmaceutical composition described in claim 86 or 87.

100. 100. The method of claim 99, wherein the subject has Fibrodysplasia Ossificans Progressiva (FOP).

101. 82. A method of treating a subject having FOP, comprising administering to the subject a therapeutically effective amount of an antibody or ALK2-binding fragment thereof according to any one of claims 1 to 82, a nucleic acid molecule according to claim 83, an expression vector according to claim 84, or a pharmaceutical composition according to claim 86 or 87.

102. 102. The method of claim 100 or 101, wherein the FOP is hereditary FOP or sporadic FOP.

103. A method of treating a subject having Sjogren's syndrome, multiple osteochondromas, diffuse intrinsic pontine glioma, PCO, cardiac hypertrophy, or cardiac fibrosis, comprising administering to the subject a therapeutically effective amount of an antibody or ALK2-binding fragment thereof according to any one of claims 1 to 82, a nucleic acid molecule according to claim 83, an expression vector according to claim 84, or a pharmaceutical composition according to claim 86 or claim 87.

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