Compositions and methods for targeted delivery of tgfbeta
A polypeptide complex with a target-binding polypeptide and dimer latency-associated polypeptide enhances targeted delivery of TGFβ, addressing inefficiencies and off-target issues in existing methods, resulting in improved therapeutic efficacy.
Patent Information
- Application Number
- HK62026122802
- Authority / Receiving Office
- HK · HK
- Patent Type
- Applications
- Current Assignee / Owner
- Priority Date
- 2023-07-11
- Filing Date
- 2026-04-30
- Publication Date
- 2026-07-31
AI Technical Summary
Current methods for targeted delivery of transforming growth factor β (TGFβ) are inefficient and lack specificity, leading to off-target effects and reduced efficacy.
A polypeptide complex comprising a target-binding polypeptide and a small latency complex, specifically a dimer latency-associated polypeptide (LAP) and a dimer mature TGFβ polypeptide, is developed to enhance targeted delivery to target cells or the extracellular matrix (ECM), with associated polynucleotides, carriers, and pharmaceutical compositions for improved therapeutic applications.
The polypeptide complex achieves precise targeting and enhanced therapeutic efficacy by binding specifically to target cells or ECM, reducing off-target effects and improving treatment outcomes.
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Abstract
Description
HKS260156-EP Compositions and Methods for Targeted Delivery of TGFβ This disclosure provides a polypeptide complex comprising a target-binding polypeptide that binds to molecules on target cells or in the extracellular matrix (ECM); and a small latency complex, particularly comprising a dimer latency-associated polypeptide (LAP) or a fragment or derivative thereof, and a dimer mature transforming growth factor β (TGFP) family polypeptide or a fragment or derivative thereof. Polypeptides (e.g., fusion polypeptides) and associated polynucleotides, carriers, cells, and pharmaceutical compositions are also provided. Methods for treating a subject using, for example, these polypeptide complexes and / or these fusion polypeptides or pharmaceutical compositions thereof are also provided. Abstract
Claims
Attorney Docket No: 250298.000604 Claims 1. A polypeptide complex comprising: a. a target-binding polypeptide that binds a molecule on a target cell or a molecule in an extracellular matrix (ECM); and b. a small latent complex (SLC) comprising: i. a dimeric latency associated polypeptide (LAP), or a fragment or derivative thereof; and ii. a dimeric mature Transforming Growth Factor β (TGFβ) family polypeptide, or a fragment or derivative thereof, wherein the mature TGFβ family polypeptide, or the fragment or derivative thereof, is inactive as a result of an interaction with the dimeric LAP, or the fragment or derivative thereof.
2. The polypeptide complex of claim 1, wherein the LAP, or the fragment or derivative thereof, is covalently attached to the target-binding polypeptide.
3. The polypeptide complex of claim 2, wherein the LAP, or the fragment or derivative thereof, is covalently attached to the target-binding polypeptide via a linker.
4. The polypeptide complex of claim 3, wherein the linker comprises the sequence (GGGGS)n (SEQ ID NO: 46), (GGGS)n (SEQ ID NO: 51), or GSGESGGGSG (SEQ ID NO: 96).
5. The polypeptide complex of claim 4, wherein the linker consists of the sequence (GGGGS)n (SEQ ID NO: 46), (GGGS)n (SEQ ID NO: 51), or GSGESGGGSG (SEQ ID NO: 96).
6. The polypeptide complex of claim 3, wherein the linker comprises the sequence GGGSGGGSGGGS (SEQ ID NO: 19).
7. The polypeptide complex of claim 6, wherein the linker consists of the sequence GGGSGGGSGGGS (SEQ ID NO: 19).
8. The polypeptide complex of claim 3, wherein the linker comprises the sequence GSGESGGGSG (SEQ ID NO: 96).Attorney Docket No: 250298.000604 9. The polypeptide complex of claim 8, wherein the linker consists of the sequence GSGESGGGSG (SEQ ID NO: 96).
10. The polypeptide complex of claim 7 or claim 9, wherein the target-binding polypeptide binds both the LAP, or the fragment or derivative thereof, and the molecule on the target cell or the molecule in the ECM.
11. The polypeptide complex of claim 10, wherein the target-binding polypeptide is an antibody or a fragment or derivative thereof.
12. The polypeptide complex of any one of claims 1-11, wherein the mature TGFβ family polypeptide, or the fragment or derivative thereof, and the LAP, or the fragment or derivative thereof, are associated via a noncovalent interaction.
13. The polypeptide complex of any one of claims 1-11, wherein the mature TGFβ family polypeptide, or the fragment or derivative thereof, and the LAP, or the fragment or derivative thereof, are separated by a protease cleavage site.
14. The polypeptide complex of claim 13, wherein the protease cleavage site is a furin cleavage site.
15. The polypeptide complex of claim 14, wherein the furin cleavage site comprises the sequence RXXR (SEQ ID NO: 35).
16. The polypeptide complex of claim 15, wherein the furin cleavage site consists of the sequence RXXR (SEQ ID NO: 35).
17. The polypeptide complex of claim 16, wherein the RXXR (SEQ ID NO: 35) is RHRR (SEQ ID NO: 85), RRKR (SEQ ID NO: 86), or RKKR (SEQ ID NO: 87).
18. The polypeptide complex of any one of claims 1-17, wherein the mature TGFβ family polypeptide, or the fragment or derivative thereof, binds a Transforming Growth Factor β Receptor (TGFβR) upon release of the mature TGFβ family polypeptide, or the fragment or derivative thereof, from the SLC.
19. The polypeptide complex of any one of claims 1-18, wherein the mature TGFβ family polypeptide, or the fragment or derivative thereof, induces Smad2 / 3 signaling in the target cell or a cell adjacent to the target cell upon release from the SLC.Attorney Docket No: 250298.000604 20. The polypeptide complex of any one of claims 1-19, wherein the LAP, or the fragment or derivative thereof, comprises an integrin binding motif.
21. The polypeptide complex of claim 20, wherein the integrin binding motif comprises the sequence RGD.
22. The polypeptide complex of claim 20, wherein the integrin is ⍺vβ6 integrin or ⍺vβ8 integrin.
23. The polypeptide complex of any one of claims 1-19, wherein the LAP, or the fragment or derivative thereof, does not comprise an integrin binding motif.
24. The polypeptide complex of any one of claims 1-23, wherein the target-binding polypeptide is an antigen-binding polypeptide or antigen-binding fragment thereof.
25. The polypeptide complex of claim 24, wherein the antigen-binding polypeptide is an antibody or antigen-binding fragment thereof.
26. The polypeptide complex of claim 25, wherein the antibody or antigen-binding fragment thereof comprises a heavy chain variable region.
27. The polypeptide complex of claim 25 or claim 26, wherein the antibody or antigen- binding fragment thereof comprises a light chain variable region.
28. The polypeptide complex of any one of claims 25-27, wherein the antibody or antigen-binding fragment thereof comprises an immunoglobulin heavy chain constant domain.
29. The polypeptide complex of claim 28, wherein the immunoglobulin heavy chain constant domain is an IgG1 domain.
30. The polypeptide complex of claim 28, wherein the immunoglobulin heavy chain constant domain is an IgG4 domain.
31. The polypeptide complex of any one of claims 25-30, wherein the antigen-binding polypeptide or the antigen-binding fragment thereof binds to CD63.Attorney Docket No: 250298.000604 32. The polypeptide complex of any one of claims 25-30, wherein the antigen-binding polypeptide or the antigen-binding fragment thereof binds to extra domain B of fibronectin (EDB-FN).
33. The polypeptide complex of any one of claims 25-30, wherein the antigen-binding polypeptide or the antigen-binding fragment thereof binds to epithelial cell adhesion molecule (Epcam).
34. The polypeptide complex of any one of claims 25-30, wherein the antigen-binding polypeptide or the antigen-binding fragment thereof binds to C-type lectin domain family 9 member A (Clec9a).
35. The polypeptide complex of any one of claims 1-34, wherein the target-binding polypeptide is not internalizing.
36. The polypeptide complex of any one of claims 1-35, wherein the mature TGFβ family polypeptide is a mature TGFβ polypeptide.
37. The polypeptide complex of claim 36, wherein the mature TGFβ polypeptide is a mature TGFβ1 polypeptide.
38. The polypeptide complex of claim 37, wherein the mature TGFβ1 polypeptide comprises the sequence of SEQ ID NO:
23.
39. The polypeptide complex of claim 38, wherein the mature TGFβ1 polypeptide consists of the sequence of SEQ ID NO:
23.
40. The polypeptide complex of claim 37, wherein the mature TGFβ1 polypeptide comprises the sequence of SEQ ID NO:
90.
41. The polypeptide complex of claim 40, wherein the mature TGFβ1 polypeptide consists of the sequence of SEQ ID NO:
90.
42. The polypeptide complex of claim 36, wherein the mature TGFβ polypeptide is a mature TGFβ2 polypeptide.
43. The polypeptide complex of claim 42, wherein the mature TGFβ2 polypeptide comprises the sequence of SEQ ID NO: 27.Attorney Docket No: 250298.000604 44. The polypeptide complex of claim 43, wherein the mature TGFβ2 polypeptide consists of the sequence of SEQ ID NO:
27.
45. The polypeptide complex of any one of claims 1-35, wherein the mature TGFβ family polypeptide is a mature Growth Differentiation Factor 8 (GDF8), a mature Growth Differentiation Factor 11 (GDF11) polypeptide, or a mature Bone Morphogenetic Protein 4 (BMP4).
46. The polypeptide complex of any one of claims 1-44, wherein the LAP comprises the sequence of positions 30-274 of the sequence of SEQ ID NO:
82.
47. The polypeptide complex of claim 46, wherein the LAP consists of the sequence of positions 30-274 of the sequence of SEQ ID NO:
82.
48. The polypeptide complex of any one of claims 1-44, wherein the LAP comprises the sequence of positions 21-298 of the sequence of SEQ ID NO:
116.
49. The polypeptide complex of claim 48, wherein the LAP consists of the sequence of positions 21-298 of the sequence of SEQ ID NO:
116.
50. The polypeptide complex of any one of claims 1-49, wherein the LAP, or the fragment or derivative thereof, is heterologous to the mature TGFβ family polypeptide, or the fragment or derivative thereof.
51. The polypeptide complex of any one of claims 1-50, wherein the LAP, or the fragment or derivative thereof, comprises one or more mutations, wherein the one or more mutations allow for proteolytic activation of the mature TGFβ family polypeptide, or the fragment or derivative thereof.
52. The polypeptide complex of any one of claims 1-51, wherein the LAP, or the fragment or derivative thereof, comprises one or more mutations, wherein the one or more mutations allow for mechanical activation of the mature TGFβ family polypeptide, or the fragment or derivative thereof.
53. The polypeptide complex of any one of claims 1-52, wherein the LAP, or the fragment or derivative thereof, comprises one or more mutations, wherein the one or moreAttorney Docket No: 250298.000604 mutations eliminate one or more protease cleavage sites within the LAP, or the fragment or derivative thereof.
54. The polypeptide complex of any one of claims 1-53, wherein the LAP, or the fragment or derivative thereof, comprises one or more mutations, wherein the one or more mutations decrease binding of the LAP, or the fragment or derivative thereof, to a latency associated binding protein (LTBP).
55. The polypeptide complex of claim 54, wherein the one or more mutations comprise a C33S mutation, wherein position 33 is in relation to the sequence of SEQ ID NO:
82.
56. The polypeptide complex of claim 54, wherein the one or more mutations comprise a C24S mutation, wherein position 24 is in relation to SEQ ID NO:
116.
57. The polypeptide complex of any one of claims 1-44 and 54-55, wherein the LAP comprises the sequence of SEQ ID NO:
31.
58. The polypeptide complex of claim 57, wherein the LAP consists of the sequence of SEQ ID NO:
31.
59. The polypeptide complex of any one of claims 1-44, 54, and 56, wherein the LAP comprises the sequence of SEQ ID NO:
94.
60. The polypeptide complex of claim 59, wherein the LAP consists of the sequence of SEQ ID NO:
94.
61. The polypeptide complex of any one of claims 1-60, wherein the mature TGFβ family polypeptide, or the fragment or derivative thereof, is chemically dissociated from the LAP, or the fragment or derivative thereof, and released in an active form from the SLC.
62. The polypeptide complex of claim 61, wherein the chemical dissociation comprises a protease treatment, a temperature treatment, an acid treatment, or any combination thereof.
63. The polypeptide complex of any one of claims 1-60, wherein the mature TGFβ family polypeptide, or the fragment or derivative thereof, is mechanically dissociated from the LAP, or the fragment or derivative thereof, and released in an active form from the SLC.Attorney Docket No: 250298.000604 64. The polypeptide complex of claim 63, wherein the mechanical dissociation occurs a result of an interaction between the LAP, or the fragment or derivative thereof, and an integrin polypeptide.
65. A pharmaceutical composition comprising the polypeptide complex of any one of claims 1-64.
66. The pharmaceutical composition of claim 65, further comprising a pharmaceutically acceptable carrier or diluent.
67. A fusion polypeptide comprising: a. a target-binding polypeptide that binds a molecule on a target cell or a molecule in an extracellular matrix (ECM); b. a latency associated polypeptide (LAP), or a fragment or derivative thereof; and c. a mature Transforming Growth Factor β (TGFβ) family polypeptide, or a fragment or derivative thereof.
68. The fusion polypeptide of claim 67, wherein the mature TGFβ family polypeptide, or the fragment or derivative thereof, is inactive as a result of an interaction with the LAP, or the fragment or derivative thereof.
69. The fusion polypeptide of claim 67 or claim 68, wherein the mature TGFβ family polypeptide, or the fragment or derivative thereof, binds a Transforming Growth Factor β Receptor (TGFβR) upon release of the mature TGFβ family polypeptide, or the fragment or derivative thereof, from the LAP, or the fragment or derivative thereof.
70. The fusion polypeptide of any one of claims 67-69, wherein the mature TGFβ family polypeptide, or the fragment or derivative thereof, induces Smad2 / 3 signaling in the target cell or a cell adjacent to the target cell upon release of the mature TGFβ family polypeptide, or the fragment or derivative thereof, from the LAP, or the fragment or derivative thereof.
71. The fusion polypeptide of any one of claims 67-70, wherein the fusion polypeptide comprises a linker.
72. The fusion polypeptide of claim 71, wherein the linker is located between the target- binding polypeptide and the LAP, or the fragment or derivative thereof.Attorney Docket No: 250298.000604 73. The fusion polypeptide of claim 72, wherein the fusion polypeptide comprises, from N-terminus to C-terminus, (i) the target-binding polypeptide, (ii) the linker, (iii) the LAP, or the fragment or derivative thereof, and (iv) the mature TGFβ family polypeptide, or the fragment or derivative thereof.
74. The fusion polypeptide of claim 72, wherein the fusion polypeptide comprises, from N-terminus to C-terminus, (i) the mature TGFβ family polypeptide, or the fragment or derivative thereof, (ii) the LAP, or the fragment or derivative thereof, (iii) the linker, and (iv) the target-binding polypeptide.
75. The fusion polypeptide of any one of claims 71-74, wherein the linker comprises the sequence (GGGGS)n (SEQ ID NO: 46), (GGGS)n (SEQ ID NO: 51), or GSGESGGGSG (SEQ ID NO: 96).
76. The fusion polypeptide of claim 75, wherein the linker consists of the sequence (GGGGS)n (SEQ ID NO: 46), (GGGS)n (SEQ ID NO: 51), or GSGESGGGSG (SEQ ID NO: 96).
77. The fusion polypeptide of claim 75, wherein the linker comprises the sequence GGGSGGGSGGGS (SEQ ID NO: 19).
78. The fusion polypeptide of claim 77, wherein the linker consists of the sequence GGGSGGGSGGGS (SEQ ID NO: 19).
79. The fusion polypeptide complex of claim 75, wherein the linker comprises the sequence GSGESGGGSG (SEQ ID NO: 96).
80. The fusion polypeptide complex of claim 79, wherein the linker consists of the sequence GSGESGGGSG (SEQ ID NO: 96).
81. The fusion polypeptide of any one of claims 67-80, wherein the mature TGFβ family polypeptide, or the fragment or derivative thereof, and the LAP, or the fragment or derivative thereof, are separated by a protease cleavage site.
82. The fusion polypeptide of claim 81, wherein the protease cleavage site is a furin cleavage site.Attorney Docket No: 250298.000604 83. The fusion polypeptide of claim 82, wherein the furin cleavage site comprises the sequence RXXR (SEQ ID NO: 35).
84. The fusion polypeptide of claim 83, wherein the furin cleavage site consists of the sequence RXXR (SEQ ID NO: 35).
85. The fusion polypeptide of claim 84, wherein the RXXR (SEQ ID NO: 35) is RHRR (SEQ ID NO: 85), RRKR (SEQ ID NO: 86), or RKKR (SEQ ID NO: 87).
86. The fusion polypeptide of any one of claims 67-84, further comprising a signal peptide.
87. The fusion polypeptide of claim 86, wherein the signal peptide is mROR signal peptide.
88. The fusion polypeptide of claim 87, wherein the mROR signal peptide comprises the sequence MHRPRRRGTRPPPLALLAALLLAARGADA (SEQ ID NO: 1).
89. The fusion polypeptide of claim 88, wherein the mROR signal peptide consists of the sequence MHRPRRRGTRPPPLALLAALLLAARGADA (SEQ ID NO: 1).
90. The fusion polypeptide of any one of claims 67-89, wherein the LAP, or the fragment or derivative thereof, comprises an integrin binding motif.
91. The fusion polypeptide of claim 90, wherein the integrin binding motif comprises the sequence RGD.
92. The fusion polypeptide of claim 90, wherein the integrin is ⍺vβ6 integrin or ⍺vβ8 integrin.
93. The fusion polypeptide of any one of claims 67-89, wherein the LAP, or the fragment or derivative thereof, does not comprise an integrin binding motif.
94. The fusion polypeptide of any one of claims 67-93, wherein the target-binding polypeptide is an antigen-binding polypeptide or antigen-binding fragment thereof.
95. The fusion polypeptide of claim 94, wherein the antigen-binding polypeptide is an antibody or antigen-binding fragment thereof.Attorney Docket No: 250298.000604 96. The fusion polypeptide of claim 95, wherein the antibody or antigen-binding fragment thereof comprises a heavy chain variable region.
97. The fusion polypeptide of claim 95 or claim 96, wherein the antibody or antigen- binding fragment thereof comprises a light chain variable region.
98. The fusion polypeptide of any one of claims 95-97, wherein the antibody or antigen- binding fragment thereof comprises an immunoglobulin heavy chain constant domain.
99. The fusion polypeptide of claim 98, wherein the immunoglobulin heavy chain constant domain is an IgG1 domain.
100. The fusion polypeptide of claim 98, wherein the immunoglobulin heavy chain constant domain is an IgG4 domain.
101. The fusion polypeptide of any one of claims 94-100, wherein the antigen-binding polypeptide or the antigen-binding fragment thereof binds to CD63.
102. The fusion polypeptide of any one of claims 94-100, wherein the antigen binding polypeptide of the antigen-binding fragment thereof binds to extra domain B of fibronectin (EDB-FN).
103. The fusion polypeptide of any one of claims 94-100, wherein the antigen-binding polypeptide or the antigen-binding fragment thereof binds to epithelial cell adhesion molecule (Epcam).
104. The fusion polypeptide of any one of claims 94-100, wherein the antigen-binding polypeptide or the antigen-binding fragment thereof binds to C-type lectin domain family 9 member A (Clec9a).
105. The fusion polypeptide of any one of claims 67-104, wherein the target-binding polypeptide is not internalizing.
106. The fusion polypeptide of any one of claims 67-105, wherein the mature TGFβ family polypeptide is a mature TGFβ polypeptide.
107. The fusion polypeptide of claim 106, wherein the mature TGFβ polypeptide is a mature TGFβ1 polypeptide.Attorney Docket No: 250298.000604 108. The fusion polypeptide of claim 107, wherein the mature TGFβ1 polypeptide comprises the sequence of SEQ ID NO:
23.
109. The fusion polypeptide of claim 108, wherein the mature TGFβ1 polypeptide consists of the sequence of SEQ ID NO:
23.
110. The fusion polypeptide of claim 107, wherein the mature TGFβ1 polypeptide comprises the sequence of SEQ ID NO:
90.
111. The fusion polypeptide of claim 110, wherein the mature TGFβ1 polypeptide comprises the sequence of SEQ ID NO:
90.
112. The fusion polypeptide of claim 106, wherein the mature TGFβ polypeptide is a mature TGFβ2 polypeptide.
113. The fusion polypeptide of claim 112, wherein the mature TGFβ2 polypeptide comprises the sequence of SEQ ID NO:
27.
114. The fusion polypeptide of claim 113, wherein the mature TGFβ2 polypeptide consists of the sequence of SEQ ID NO:
27.
115. The fusion polypeptide of any one of claims 67-105, wherein the mature TGFβ family polypeptide is a mature Growth Differentiation Factor 8 (GDF8), a mature Growth Differentiation Factor 11 (GDF11) polypeptide, or a mature Bone Morphogenetic Protein 4 (BMP4).
116. The fusion polypeptide of any one of claims 67-114, wherein the LAP comprises the sequence of positions 30-274 of the sequence of SEQ ID NO:
82.
117. The fusion polypeptide of claim 116, wherein the LAP consists of the sequence of positions 30-274 of the sequence of SEQ ID NO:
82.
118. The fusion polypeptide of any one of claims 67-114, wherein the LAP comprises the sequence of positions 21-298 of the sequence of SEQ ID NO:
116.
119. The fusion polypeptide of claim 118, wherein the LAP consists of the sequence of positions 21-298 of the sequence of SEQ ID NO: 116.Attorney Docket No: 250298.000604 120. The fusion polypeptide of any one of claims 67-119, wherein the LAP, or the fragment or derivative thereof, is heterologous to the mature TGFβ family polypeptide, or the fragment or derivative thereof.
121. The fusion polypeptide of any one of claims 67-120, wherein the LAP, or the fragment or derivative thereof, comprises one or more mutations, wherein the one or more mutations allow for proteolytic activation of the mature TGFβ family polypeptide, or the fragment or derivative thereof.
122. The fusion polypeptide of any one of claims 67-121, wherein the LAP, or the fragment or derivative thereof, comprises one or more mutations, wherein the one or more mutations allow for mechanical activation of the mature TGFβ family polypeptide, or the fragment or derivative thereof.
123. The fusion polypeptide of any one of claims 67-122, wherein the LAP, or the fragment or derivative thereof, comprises one or more mutations, wherein the one or more mutations eliminate one or more protease cleavage sites within the LAP, or the fragment or derivative thereof.
124. The fusion polypeptide of any one of claims 67-123, wherein the LAP, or the fragment or derivative thereof, comprises one or more mutations, wherein the one or more mutations decrease binding of the LAP, or the fragment or derivative thereof, to a latency associated binding protein (LTBP).
125. The fusion polypeptide of claim 124, wherein the one or more mutations comprise a C33S mutation, wherein position 33 is in relation to the sequence of SEQ ID NO:
82.
126. The fusion polypeptide of claim 124, wherein the one or more mutations comprise a C24S mutation, wherein position 24 is in relation to SEQ ID NO: 116 127. The fusion polypeptide of any one of claims 67-114 and 124-125, wherein the LAP comprises the sequence of SEQ ID NO:
31.
128. The fusion polypeptide of claim 127, wherein the LAP consists of the sequence of SEQ ID NO: 31.Attorney Docket No: 250298.000604 129. The fusion polypeptide of any one of claims 67-114, 124, and 126 wherein the LAP comprises the sequence of SEQ ID NO:
94.
130. The fusion polypeptide of claim 129, wherein the LAP consists of the sequence of SEQ ID NO:
94.
131. A polynucleotide encoding the fusion polypeptide of any one of claims 67-128.
132. A vector comprising the polynucleotide of claim 131.
133. The vector of claim 132, wherein the sequence encoding the fusion polypeptide is operably linked to a promoter, wherein the promoter mediates expression of the fusion polypeptide.
134. The vector of claim 132 or claim 133, wherein the vector is a viral vector.
135. The vector of claim 134, wherein the viral vector is an adeno-associated virus (AAV) vector.
136. A cell comprising the polypeptide complex of any one of claims 1-64, the fusion polypeptide of any one of claims 67-130, the polynucleotide of claim 131, or the vector of any one of claims 132-135.
137. A method of making the polypeptide complex of any one of claims 1-64, comprising incubating the cell comprising the polynucleotide of claim 131, or the vector of any one of claims 132-135, under conditions allowing for production of the polypeptide complex.
138. The method of claim 137, further comprising collecting the cell culture medium and isolating the produced polypeptide complex by a process comprising affinity chromatography.
139. The method of claim 138, wherein the affinity chromatography comprises a Protein A or a Protein G column or beads.
140. A method for treating a TGFβ dysregulation disorder in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of the polypeptide complex of any one of claims 1-64, the pharmaceutical composition of claim 65 or claim 66, the polynucleotide of claim 131, or the vector of any one of claims 132-135.Attorney Docket No: 250298.000604 141. The method of claim 140, wherein the polypeptide complex, pharmaceutical composition, polynucleotide, or vector is administered via injection.
142. The method of claim 141, wherein the injection is intravenous, intramuscular, subcutaneous, or intraperitoneal.
143. The method of any one of claims 140-142, wherein the polypeptide complex, pharmaceutical composition, polynucleotide, or vector is administered via hydrodynamic delivery (HDD).
144. The method of any one of claims 140-143, wherein the polypeptide complex, pharmaceutical composition, polynucleotide, or vector is administered to the liver of the subject.
145. The method of any one of claims 140-144, wherein the polypeptide complex, pharmaceutical composition, polynucleotide, or vector is administered in combination with an additional therapeutic agent.
146. The method of any one of claims 140-145, wherein the TGFβ dysregulation disorder is an inflammatory bowel disease (IBD).
147. The method of any one of claims 140-145, wherein the TGFβ dysregulation disorder is Marfan syndrome.
148. The method of any one of claims 140-145, wherein the TGFβ dysregulation disorder is an autoimmune disorder.
149. The method of any one of claims 140-145, wherein the TGFβ dysregulation disorder is a wound healing disorder.
150. A method for promoting wound healing in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of the polypeptide complex of any one of claims 1-64, the pharmaceutical composition of claim 65 or claim 66, the polynucleotide of claim 131, or the vector of any one of claims 132-135.
151. The method of claim 150, wherein the polypeptide complex, pharmaceutical composition, polynucleotide, or vector is administered to the wound of the subject.Attorney Docket No: 250298.000604 152. The method of any one of claims 140-151, wherein the subject is human.