Multi-chain chimeric polypeptides and uses thereof
Provided herein are multi-chain chimeric polypeptides that include: (a) a first chimeric polypeptide including a first target-binding domain, a soluble tissue factor domain, and a first domain of a pair of affinity domains; and (b) a second chimeric polypeptide including a second domain of a pair of affinity domains and a second target-binding domain, where 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. Also provided here are methods of using these multi-chain chimeric polypeptides and nucleic acids encoding these multi-chain chimeric polypeptides.
1 . A method of stimulating a natural killer (NK) cell, the method comprising contacting the NK cell with an effective amount of a multi-chain chimeric polypeptide comprising:
(a) a first chimeric polypeptide comprising:
(i) a first target-binding domain comprising SEQ ID NO: 16;
(ii) a soluble tissue factor domain comprising SEQ ID NO: 1; and
(iii) a first domain of a pair of affinity domains comprising SEQ ID NO: 14; and
(b) a second chimeric polypeptide comprising:
(i) a second domain of a pair of affinity domains comprising SEQ ID NO: 28; and
(ii) a second target-binding domain comprising a first sequence comprising SEQ ID NO: 66 and a second sequence comprising SEQ ID NO: 68, wherein the first and second sequences are separated by a linker,
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.
2 . 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.
3 . 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.
4 . 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.
5 . 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.
6 . 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.
7 . 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.
8 . The method of claim 1 , wherein:
the first chimeric polypeptide comprises SEQ ID NO: 70; and
the second chimeric polypeptide comprises SEQ ID NO: 74.
9 . A method of activating an NK cell in a subject in need thereof, the method comprising 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 comprising SEQ ID NO: 16;
(ii) a soluble tissue factor domain comprising SEQ ID NO: 1; and
(iii) a first domain of a pair of affinity domains comprising SEQ ID NO: 14; and
(b) a second chimeric polypeptide comprising:
(i) a second domain of a pair of affinity domains comprising SEQ ID NO: 28; and
(ii) a second target-binding domain comprising a first sequence of SEQ ID NO: 66 and a second sequence of SEQ ID NO: 68, wherein the first and second sequences are separated by a linker,
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.
10 . The method of claim 9 , wherein the subject has been identified or diagnosed as having a cancer, an aging-related disease or condition, or an infectious disease.
11 . The method of claim 10 , wherein the infectious disease is an infection with human immunodeficiency virus, cytomegalovirus, adenovirus, coronavirus, rhinovirus, rotavirus, smallpox, herpes simplex virus, hepatitis B virus, hepatitis A virus, hepatitis C virus, papillomavirus, and influenza virus.
12 . The method of claim 9 , wherein the first target-binding domain and the soluble tissue factor domain directly abut each other in the first chimeric polypeptide.
13 . The method of claim 9 , 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.
14 . The method of claim 9 , 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.
15 . The method of claim 9 , 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.
16 . The method of claim 9 , 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.
17 . The method of claim 9 , wherein the 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.
18 . The method of claim 9 , wherein:
the first chimeric polypeptide comprises SEQ ID NO: 70; and
the second chimeric polypeptide comprises SEQ ID NO: 74.