Lentiviral vectors for regulated expression of a chimeric antigen receptor molecule

Patent metadata

Field	Value
Publication number	US-2017166623-A1
Application number	US-201515327517-A
Country	US
Kind code	A1
Filing date	Jul 24, 2015
Priority date	Jul 25, 2014
Publication date	Jun 15, 2017
Grant date	—

Abstract

The invention relates to the regulated expression of a chimeric antigen receptor (CAR) within a lentiviral vector. The CAR comprises a hook-binding domain that interacts with a hook, preferably encoded by the same lentiviral vector, which prevents proper processing and release of the CAR to the cell membrane. The invention encompasses vectors, methods of making the vectors, and methods of using them, including medicinal uses. The vectors can be used for administration to humans to induce immune responses and to treat cancers and tumors.

First claim

1 . A chimeric antigen receptor comprising:
a binding domain; a transmembrane domain; a hook-binding domain comprising a streptavidin-binding peptide; and an activation domain comprising a T cell activating fragment of at least 100 amino acids of SEQ ID NO:3; SEQ ID NO:4; SEQ ID NO:5; SEQ ID NO:6, SEQ ID NO:7, or SEQ ID NO:8, SEQ ID NO:33, or SEQ ID NO:34.

2 . A lentiviral vector expressing a chimeric antigen receptor comprising:
a binding domain; a transmembrane domain; a hook-binding domain comprising a streptavidin-binding peptide; and an activation domain comprising a T cell activating fragment of at least 100 amino acids of SEQ ID NO:3; SEQ ID NO:4; SEQ ID NO:5; SEQ ID NO:6, SEQ ID NO:7, or SEQ ID NO:8, SEQ ID NO:33, or SEQ ID NO:34.

3 . A nucleic acid vector encoding a chimeric antigen receptor comprising:
a binding domain; a transmembrane domain; a hook-binding domain comprising a streptavidin-binding peptide; and an activation domain comprising a T cell activating fragment of at least 100 amino acids of SEQ ID NO:3; SEQ ID NO:4; SEQ ID NO:5; SEQ ID NO:6, SEQ ID NO:7, or SEQ ID NO:8, SEQ ID NO:33, or SEQ ID NO:34.

4 . The chimeric antigen receptor of claim 1 , wherein the binding domain comprises a single-chain Fv antibody or a nanobody.

5 . The lentiviral vector of claim 2 , wherein the vector encodes a hook comprising the amino acid sequence of SEQ ID NO:31 or SEQ ID NO:32.

6 . The lentiviral vector of any of claim 2 , wherein the vector comprises a β2-microglobulin, ubiquitin, MHCI, or MHCII promoter.

7 . The lentiviral vector of claim 2 , wherein the vector is a DNA.

8 . A lentiviral vector particle encoding a chimeric antigen receptor comprising:
a binding domain; a transmembrane domain; a hook-binding domain comprising a streptavidin-binding peptide; and an activation domain comprising a T cell activating fragment of at least 100 amino acids of SEQ ID NO:3; SEQ ID NO:4; SEQ ID NO:5; SEQ ID NO:6, SEQ ID NO:7, or SEQ ID NO:8, SEQ ID NO:33, or SEQ ID NO:34.

9 . The lentiviral particle of claim 8 , wherein the binding domain comprises a single-chain Fv antibody or a nanobody.

10 . The lentiviral vector particle of claim 8 , wherein the lentiviral vector particle encodes a hook comprising the amino acid sequence of SEQ ID NO:31 or SEQ ID NO:32.

11 . The lentiviral vector particle of claim 8 , wherein the lentiviral vector particle comprises a vesicular stomatitis virus glycoprotein.

12 . The lentiviral vector particle of claim 8 , wherein the lentiviral vector particle comprises HIV-1 subtype D Gag and Pol proteins.

13 . An isolated cell comprising the vector of claim 2 .

14 . (canceled)

15 . A method for inducing an immune response in a human comprising intramuscularly administering the lentiviral vector particle or cells transduced by the lentiviral vector particle of claim 8 to a human and subsequently administering biotin to the human.

16 . The lentiviral vector of claim 2 , comprising the nucleotide sequence of any of SEQ ID NO:45, SEQ ID NO:47, or SEQ ID NO:49.

17 . The chimeric antigen receptor of claim 1 , wherein the chimeric antigen receptor comprises the amino acid sequence of any of SEQ ID NO:46, SEQ ID NO:48, or SEQ ID NO:50.

18 . The lentiviral vector or the nucleic acid vector of claim 16 , further encoding the amino acid sequence of SEQ ID NO:42 or comprising the nucleic acid sequence of SEQ ID NO:43 and/or SEQ ID NO:44.

19 . The lentiviral vector of claim 2 , wherein the binding domain comprises a single-chain Fv antibody or a nanobody.

20 . The nucleic acid vector of claim 3 , wherein the binding domain comprises a single-chain Fv antibody or a nanobody.

21 . The nucleic acid vector of claim 3 , wherein the vector encodes a hook comprising the amino acid sequence of SEQ ID NO:31 or SEQ ID NO:32.

22 . The nucleic acid vector of claim 3 , comprising the nucleotide sequence of any of SEQ ID NO:45, SEQ ID NO:47, or SEQ ID NO:49.

Assignees

• Theravectys
• Inst Curie
• Centre Nat Rech Scient
• Univ Paris Curie
• Inst Nat De La Sante Et De La Res Medical (Inserm)

Inventors

• Agaugue Sophie
• Tibaldi Lorenzo
• Even-Desrumaux Klervi
• Trubetskoy Dmitry
• Perez Franck
• Amigorena Sebastian
• Bauche Cécile

Classifications

• A61P37/04

  Immunostimulants · CPC title

• C07K14/70503 Primary

  Immunoglobulin superfamily · CPC title

• C07K2319/03

  containing a transmembrane segment · CPC title

• C07K2317/622

  Single chain antibody (scFv) · CPC title

• C12N2799/027

  where the vector is derived from a retrovirus · CPC title