Monoclonal antibodies that specifically bind to matrilin 3 and their use

Abstract

Monoclonal antibodies and antibody fragments that specifically bind to matrilin-3, conjugates including these molecules, and nucleic acid molecules encoding the antibodies, antigen binding fragments and conjugates, are disclosed. Also disclosed are compositions including the disclosed antibodies, antigen binding fragments, conjugates, and nucleic acid molecules. Methods of treating or inhibiting a cartilage disorder in a subject, as well as methods of increasing chondrogenesis in cartilage tissue are further provided. The methods can be used, for example, for treating or inhibiting a growth plate disorder in a subject, such as a skeletal dysplasia or short stature.

We claim: 
     
         1 . A method of increasing chondrogenesis in cartilage tissue, comprising:
 contacting cartilage tissue with a therapeutically effective amount of a conjugate comprising a monoclonal antibody or antigen binding molecule and an effector molecule, under conditions sufficient to form an immune complex, thereby increasing chondrogenesis in the cartilage tissue, and wherein:   (a) the monoclonal antibody and antigen binding fragment comprise: a heavy chain variable region comprising a heavy chain complementarity determining region (HCDR) 1 , a HCDR2, and a HCDR3, and a light chain variable region comprising a light chain complementarity determining region (LCDR) 1 , a LCDR2, and a L-CDR3, of the amino acid sequences set forth as one of:
 (i) SEQ ID NO: 1 and SEQ ID NO: 2, respectively (clone 13); 
 (ii) SEQ ID NO: 3 and SEQ ID NO: 4, respectively (clone 22); or 
 (iii) SEQ ID NO: 5 and SEQ ID NO: 6, respectively (clone 26); 
   
       and wherein the monoclonal antibody and antigen binding molecule specifically binds to matrilin-3; and
 (b) the effector molecule is a growth hormone, an insulin-like growth factor (IGF)-1, an Indian hedgehog polypeptide, a bone morphogenetic protein, a C-type natriuretic protein (CNP), a Wnt protein, or a biologically active fragment of the Wnt protein that induces chondrogenesis, or a steroid. 
 
     
     
         2 . The method of  claim 1 , wherein:
 (i) the HCDR1, HCDR2, and HCDR3 comprise the amino acid sequences set forth as residues 26-33, 51-58, and 97-106 of SEQ ID NO: 1, respectively, and the LCDR1, LCDR2, and LCDR3 comprise the amino acid sequence set forth as residues 27-32, 50-52, and 89-97 of SEQ ID NO: 2, respectively;   (ii) the HCDR1, HCDR2, and HCDR3 comprise the amino acid sequences set forth as residues 26-33, 53-61, and 100-109 of SEQ ID NO: 3, respectively, and the LCDR1, LCDR2, and LCDR3 comprise the amino acid sequence set forth as residues 26-31, 49-51, and 88-97 of SEQ ID NO: 4, respectively; or   (iii) the HCDR1, HCDR2, and HCDR3 comprise the amino acid sequences set forth as residues 26-33, 51-58, 97-108 of SEQ ID NO: 5, respectively, and the LCDR1, LCDR2, and LCDR3 comprise the amino acid sequence set forth as residues 26-33, 51-53, and 90-100 of SEQ ID NO: 6, respectively.   
     
     
         3 . The method of  claim 1 , wherein the heavy and light chain variable regions comprise the amino acid sequences set forth as one of:
 (i) SEQ ID NO: 1 and SEQ ID NO: 2, respectively;   (ii) SEQ ID NO: 3 and SEQ ID NO: 4, respectively; or   (iii) SEQ ID NO: 5 and SEQ ID NO: 5, respectively.   
     
     
         4 . The method of  claim 1 , wherein the antigen binding molecule is a Fv, a Fab, a F(ab′) 2 , an scFv, or an scFV 2 . 
     
     
         5 . The method of  claim 1 , wherein the effector molecule is the growth hormone, the IGF-1, or the CNP. 
     
     
         6 . The method of  claim 5 , wherein the effector molecule is the growth hormone. 
     
     
         7 . The method of  claim 5 , wherein the effector molecule is the IGF-1. 
     
     
         8 . The method of  claim 5 , wherein the effector molecule is the CNP. 
     
     
         9 . The method of  claim 1 , wherein the subject has a cartilage disorder, short stature or skeletal dysplasia. 
     
     
         10 . The method of  claim 9 , wherein the subject has the short stature. 
     
     
         11 . The method of  claim 9 , wherein:
 (a) the subject has the short stature and wherein the short stature is idiopathic short stature, short stature due to systemic disease, short stature associated with a dysmorphic syndrome, short stature due to chromosomal abnormalities, iatrogenic short stature, short stature due to radiation or medications, short stature in children born small for gestational age, or short stature due to undernutrition; or   (b) the subject has the skeletal dysplasia and wherein the skeletal dysplasia is achondroplasia, hypochondroplasia, or short stature homeobox gene (SHOX) deficiency.   
     
     
         12 . A method of targeting an effector molecule to cartilage tissue in a subject, comprising:
 administering to the subject a therapeutically effective amount of a conjugate comprising a monoclonal antibody or antigen binding molecule, and an effector molecule, under conditions sufficient to form an immune complex, wherein formation of the immune complex targets the heterologous effector molecule to the cartilage tissue in the subject, and wherein:   (a) the monoclonal antibody and antigen binding fragment comprise: a heavy chain variable region comprising a heavy chain complementarity determining region (HCDR) 1 , a HCDR2, and a HCDR3, and a light chain variable region comprising a light chain complementarity determining region (LCDR) 1 , a LCDR2, and a L-CDR3, of the amino acid sequences set forth as one of:
 (i) SEQ ID NO: 1 and SEQ ID NO: 2, respectively (clone 13); 
 (ii) SEQ ID NO: 3 and SEQ ID NO: 4, respectively (clone 22); or 
 (iii) SEQ ID NO: 5 and SEQ ID NO: 6, respectively (clone 26); 
   
       and wherein the monoclonal antibody and antigen binding molecule specifically binds to matrilin-3; and
 (b) the effector molecule is a growth hormone, an insulin-like growth factor (IGF)-1, an Indian hedgehog polypeptide, a bone morphogenetic protein, a C-type natriuretic protein (CNP), a Wnt protein, or a biologically active fragment of the Wnt protein that induces chondrogenesis, or a steroid. 
 
     
     
         13 . The method of  claim 12 , wherein:
 (i) the HCDR1, HCDR2, and HCDR3 comprise the amino acid sequences set forth as residues 26-33, 51-58, and 97-106 of SEQ ID NO: 1, respectively, and the LCDR1, LCDR2, and LCDR3 comprise the amino acid sequence set forth as residues 27-32, 50-52, and 89-97 of SEQ ID NO: 2, respectively;   (ii) the HCDR1, HCDR2, and HCDR3 comprise the amino acid sequences set forth as residues 26-33, 53-61, and 100-109 of SEQ ID NO: 3, respectively, and the LCDR1, LCDR2, and LCDR3 comprise the amino acid sequence set forth as residues 26-31, 49-51, and 88-97 of SEQ ID NO: 4, respectively; or   (iii) the HCDR1, HCDR2, and HCDR3 comprise the amino acid sequences set forth as residues 26-33, 51-58, 97-108 of SEQ ID NO: 5, respectively, and the LCDR1, LCDR2, and LCDR3 comprise the amino acid sequence set forth as residues 26-33, 51-53, and 90-100 of SEQ ID NO: 6, respectively.   
     
     
         14 . The method of  claim 12 , wherein the effector molecule is the growth hormone, the IGF-1, or the CNP. 
     
     
         15 . The method of  claim 14 , wherein the effector molecule is the growth hormone. 
     
     
         16 . The method of  claim 15 , wherein the effector molecule is the IGF-1. 
     
     
         17 . The method of  claim 15 , wherein the effector molecule is the CNP. 
     
     
         18 . The method of  claim 12 , wherein the subject has a cartilage disorder, short stature or skeletal dysplasia. 
     
     
         19 . The method of  claim 18 , wherein the subject has the short stature. 
     
     
         20 . The method of  claim 18 , wherein:
 (a) the subject has the short stature and wherein the short stature is idiopathic short stature, short stature due to systemic disease, short stature associated with a dysmorphic syndrome, short stature due to chromosomal abnormalities, iatrogenic short stature, short stature due to radiation or medications, short stature in children born small for gestational age, or short stature due to undernutrition; or   (b) the subject has the skeletal dysplasia and wherein the skeletal dysplasia is achondroplasia, hypochondroplasia, or short stature homeobox gene (SHOX) deficiency.