MULTI-CHAIN CHIMERIC POLYPEPTIDES AND USE THEREOF IN THE TREATMENT OF LIVER DISEASES

1 . A method of treating a liver disease or a metabolic syndrome in a subject, wherein the method comprises administering to the subject a therapeutically effective amount of a multi-chain chimeric polypeptide comprising:

(a) a first chimeric polypeptide comprising:

(i) a first target-binding domain;

(ii) soluble tissue factor domain; and

(iii) a first domain of a pair of affinity domains;

(b) a second chimeric polypeptide comprising:

(i) a second domain of a pair of affinity domains; and

(ii) a second target-binding domain,

wherein:

the first chimeric polypeptide and the second chimeric polypeptide associate through the binding of the first domain and the second domain of the pair of affinity domains; and

the first target-binding domain binds specifically to a ligand of TGF-β receptor II (TGF-βRII) and the second target-binding domain binds specifically to a ligand of TGF-βRII.

2 . The method of claim 1 , wherein the liver disease is selected from the group consisting of: fatty liver disease, hepatic steatosis, acute hepatic porphyria, Alagille syndrome, alcohol-related liver disease, alpha-1 anti-trypsin deficiency, autoimmune hepatitis, benign liver tumors, cholangiocarcinoma, biliary atresia, Budd-Chiari syndrome, cirrhosis, Crigler-Najjar syndrome, galactosemia, Gilbert syndrome, hemochromatosis, hepatic encephalopathy, hepatitis A, hepatitis B, hepatitis C, hepatorenal syndrome, intrahepatic cholestasis of pregnancy (ICP), lysosomal acid lipase deficiency (LAL-D), liver cysts, liver cancer, newborn jaundice, non-alcoholic fatty liver disease, non-alcoholic steatohepatitis, primary biliary cholangitis (PBC), primary sclerosing cholangitis (PSC), progressive familial intrahepatic cholestasis (PFIC), Reye's syndrome, type 1 glycogen storage disease, and Wilson's disease.

3 . The method of claim 1 , wherein the metabolic syndrome is selected from the group consisting of: coronary heart disease, pulmonary disease, gall bladder disease, dyslipidemia, hypertension, type 2 diabetes, dementia, cancer, gynecological abnormalities including polycystic ovarian syndrome, osteoarthritis, pancreatitis, idiopathic intracranial hypertension, stroke, and cataracts.

4 . A method of reducing one or more of the rate of: progression from non-alcoholic fatty liver disease (NAFL) to non-alcoholic steatohepatitis (NASH), progression from NASH to cirrhosis, and progression from cirrhosis to hepatocellular carcinoma, comprising administering to a subject identified or diagnosed as having NAFL, NASH, or cirrhosis, a therapeutically effective amount of a multi-chain chimeric polypeptide comprising:

(a) a first chimeric polypeptide comprising:

(i) a first target-binding domain;

(ii) soluble tissue factor domain; and

(iii) a first domain of a pair of affinity domains;

(b) a second chimeric polypeptide comprising:

(i) a second domain of a pair of affinity domains; and

(ii) a second target-binding domain,

wherein:

the first chimeric polypeptide and the second chimeric polypeptide associate through the binding of the first domain and the second domain of the pair of affinity domains; and

the first target-binding domain binds specifically to a ligand of TGF-β receptor II (TGF-βRII) and the second target-binding domain binds specifically to a ligand of TGF-PRII.

5 .- 7 . (canceled)

8 . A method of reducing inflammation in a liver of a subject, wherein the method comprises administering to the subject a therapeutically effective amount of a multi-chain chimeric polypeptide comprising:

(a) a first chimeric polypeptide comprising:

(i) a first target-binding domain;

(ii) soluble tissue factor domain; and

(iii) a first domain of a pair of affinity domains;

(b) a second chimeric polypeptide comprising:

(i) a second domain of a pair of affinity domains; and

(ii) a second target-binding domain,

wherein:

the first chimeric polypeptide and the second chimeric polypeptide associate through the binding of the first domain and the second domain of the pair of affinity domains; and

the first target-binding domain binds specifically to a ligand of TGF-β receptor II (TGF-βRII) and the second target-binding domain binds specifically to a ligand of TGF-βRII.

9 . A method of decreasing gluconeogenesis in a liver of a subject, wherein the method comprises administering to the subject a therapeutically effective amount of a multi-chain chimeric polypeptide comprising:

(a) a first chimeric polypeptide comprising:

(i) a first target-binding domain;

(ii) soluble tissue factor domain; and

(iii) a first domain of a pair of affinity domains;

(b) a second chimeric polypeptide comprising:

(i) a second domain of a pair of affinity domains; and

(ii) a second target-binding domain,

wherein:

the first chimeric polypeptide and the second chimeric polypeptide associate through the binding of the first domain and the second domain of the pair of affinity domains; and

the first target-binding domain binds specifically to a ligand of TGF-β receptor II (TGF-βRII) and the second target-binding domain binds specifically to a ligand of TGF-βRII.

10 . A method of decreasing lipogenesis in a liver of a subject, wherein the method comprises administering to the subject a therapeutically effective amount of a multi-chain chimeric polypeptide comprising:

(a) a first chimeric polypeptide comprising:

(i) a first target-binding domain;

(ii) soluble tissue factor domain; and

(iii) a first domain of a pair of affinity domains;

(b) a second chimeric polypeptide comprising:

(i) a second domain of a pair of affinity domains; and

(ii) a second target-binding domain,

wherein:

the first chimeric polypeptide and the second chimeric polypeptide associate through the binding of the first domain and the second domain of the pair of affinity domains; and

the first target-binding domain binds specifically to a ligand of TGF-β receptor II (TGF-βRII) and the second target-binding domain binds specifically to a ligand of TGF-βRII.

11 . A method of decreasing hepatocytic senescence in a liver of a subject, wherein the method comprises administering to the subject a therapeutically effective amount of a multi-chain chimeric polypeptide comprising:

(a) a first chimeric polypeptide comprising:

(i) a first target-binding domain;

(ii) soluble tissue factor domain; and

(iii) a first domain of a pair of affinity domains;

(b) a second chimeric polypeptide comprising:

(i) a second domain of a pair of affinity domains; and

(ii) a second target-binding domain,

wherein:

the first chimeric polypeptide and the second chimeric polypeptide associate through the binding of the first domain and the second domain of the pair of affinity domains; and

the first target-binding domain binds specifically to a ligand of TGF-β receptor II (TGF-βRII) and the second target-binding domain binds specifically to a ligand of TGF-βRII.

12 . A method of rebalancing metabolic function in a liver of a subject, wherein the method comprises administering to the subject a therapeutically effective amount of a multi-chain chimeric polypeptide comprising:

(a) a first chimeric polypeptide comprising:

(i) a first target-binding domain;

(ii) soluble tissue factor domain; and

(iii) a first domain of a pair of affinity domains;

(b) a second chimeric polypeptide comprising:

(i) a second domain of a pair of affinity domains; and

(ii) a second target-binding domain,

wherein:

the first chimeric polypeptide and the second chimeric polypeptide associate through the binding of the first domain and the second domain of the pair of affinity domains; and

the first target-binding domain binds specifically to a ligand of TGF-β receptor II (TGF-βRII) and the second target-binding domain binds specifically to a ligand of TGF-βRII.

13 . A method of modulating expression of one or more genes in Tables 1-4 in a liver of a subject, wherein the method comprises administering to the subject a therapeutically effective amount of a multi-chain chimeric polypeptide comprising:

(a) a first chimeric polypeptide comprising:

(i) a first target-binding domain;

(ii) soluble tissue factor domain; and

(iii) a first domain of a pair of affinity domains;

(b) a second chimeric polypeptide comprising:

(i) a second domain of a pair of affinity domains; and

(ii) a second target-binding domain,

wherein:

the first chimeric polypeptide and the second chimeric polypeptide associate through the binding of the first domain and the second domain of the pair of affinity domains; and

the first target-binding domain binds specifically to a ligand of TGF-β receptor II (TGF-βRII) and the second target-binding domain binds specifically to a ligand of TGF-βRII.

14 .- 17 . (canceled)

18 . The method of claim 8 , wherein the subject has been previously identified or diagnosed as having a liver disease or a metabolic syndrome.

19 .- 22 . (canceled)

23 . The method of claim 1 , wherein the first target-binding domain and the soluble tissue factor domain directly abut each other in the first chimeric polypeptide.

24 . The method of claim 1 , wherein the first chimeric polypeptide further comprises a linker sequence between the first target-binding domain and the soluble tissue factor domain in the first chimeric polypeptide.

25 . The method of claim 1 , wherein the soluble tissue factor domain and the first domain of the pair of affinity domains directly abut each other in the first chimeric polypeptide.

26 . The method of claim 1 , wherein the first chimeric polypeptide further comprises a linker sequence between the soluble tissue factor domain and the first domain of the pair of affinity domains in the first chimeric polypeptide.

27 . The method of claim 1 , wherein the second domain of the pair of affinity domains and the second target-binding domain directly abut each other in the second chimeric polypeptide.

28 . The method of claim 1 , wherein second chimeric polypeptide further comprises a linker sequence between the second domain of the pair of affinity domains and the second target-binding domain in the second chimeric polypeptide.

29 . The method of claim 1 , wherein one or both of the first target-binding domain and the second target-binding domain is an antigen-binding domain.

30 . The method of claim 1 , wherein one or both of the first target-binding domain and the second target-binding domain is a soluble interleukin or cytokine receptor.

31 .- 32 . (canceled)

33 . The method of claim 1 , wherein the soluble tissue factor domain is a soluble human tissue factor domain.

34 . The method of claim 33 , wherein the soluble human tissue factor domain comprises a sequence that is at least 80% identical to SEQ ID NO: 1.

35 . The method of claim 1 , wherein the pair of affinity domains is a sushi domain from an alpha chain of human IL-15 receptor (IL-15Rα) and a soluble IL-15.

36 . The method of claim 1 , wherein the first target-binding domain comprises a soluble TGF-βRII.

37 . The method of claim 36 , wherein the first target-binding domain comprises a first sequence that is at least 80% identical to SEQ ID NO: 2 and a second sequence that is at least 80% identical to SEQ ID NO: 2, wherein the first and second sequence are separated by a linker.

38 .- 39 . (canceled)

40 . The method of claim 37 , wherein the linker comprises a sequence of SEQ ID NO: 3.

41 . The method of claim 36 , wherein the first target-binding domain comprises a sequence that is at least 80% identical to SEQ ID NO: 4.

42 .- 43 . (canceled)

44 . The method of claim 36 , wherein the first chimeric polypeptide comprises a sequence that is at least 80% identical to SEQ ID NO: 6.

45 .- 47 . (canceled)

48 . The method of claim 1 , wherein the second target-binding domain comprises a soluble TGF-βRII.

49 . The method of claim 48 , wherein the second target-binding domain comprises a first sequence that is at least 80% identical to SEQ ID NO: 2 and a second sequence that is at least 80% identical to SEQ ID NO: 2, wherein the first and second sequence are separated by a linker.

50 .- 51 . (canceled)

52 . The method of claim 49 , wherein the linker comprises a sequence of SEQ ID NO: 3.

53 . The method of claim 48 , wherein the second target-binding domain comprises a sequence that is at least 80% identical to SEQ ID NO: 4.

54 .- 55 . (canceled)

56 . The method of claim 48 , wherein the second chimeric polypeptide comprises a sequence that is at least 80% identical to SEQ ID NO: 5.

57 . The method of claim 56 , wherein the first chimeric polypeptide comprises a sequence that is at least 80% identical to SEQ ID NO: 6.

58 .- 61 . (canceled)