Multipartite signaling proteins and uses thereof

• US 10,428,142 B2
• Filed: 01/28/2015
• Issued: 10/01/2019
• Est. Priority Date: 07/29/2013
• Status: Active Grant

• Current Assignee: Regeneron Pharmaceuticals Incorporated
• Original Assignee: bluebird bio, Inc.
• Inventors: Jarjour, Jordan, Astrakhan, Alexander, Certo, Michael
• Primary Examiner(s): Leavitt, Maria G
• Application Number:
  US14/608,098
• Publication Number:
  US 20150266973A1
• Time in Patent Office: 1,707 Days
• Field of Search: None
• CPC Class Codes: A61K 35/17 Lymphocytes; B-cells; T-cells; Natural killer cells; Interferon-activated or cytokine-activated lymphocytes when activated by a specific antigen A61K39/00
• CPC Class Codes: A61K 39/0005 Vertebrate antigens from snakes A61K39/38
• CPC Class Codes: A61K 39/4611 T-cells, e.g. tumor infiltrating lymphocytes [TIL], lymphokine-activated killer cells [LAK] or regulatory T cells [Treg]
• CPC Class Codes: A61K 39/4631 Chimeric Antigen Receptors [CAR]
• CPC Class Codes: A61K 39/464412 CD19 or B4
• CPC Class Codes: A61K 39/464419 Receptors for interleukins [IL]
• CPC Class Codes: A61K 39/464424 CD20
• CPC Class Codes: A61K 45/06 Mixtures of active ingredients without chemical characterisation, e.g. antiphlogistics and cardiaca
• CPC Class Codes: A61P 29/00 Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
• CPC Class Codes: A61P 35/00 Antineoplastic agents
• CPC Class Codes: A61P 37/02 Immunomodulators
• CPC Class Codes: A61P 37/06 Immunosuppressants, e.g. drugs for graft rejection
• CPC Class Codes: A61P 43/00 Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
• CPC Class Codes: C07H 21/04 with deoxyribosyl as saccharide radical
• CPC Class Codes: C07K 14/70503 Immunoglobulin superfamily
• CPC Class Codes: C07K 14/7051 T-cell receptor (TcR)-CD3 complex
• CPC Class Codes: C07K 14/70514 CD4
• CPC Class Codes: C07K 14/70517 CD8
• CPC Class Codes: C07K 14/70521 CD28, CD152
• CPC Class Codes: C07K 14/70535 Fc-receptors, e.g. CD16, CD32, CD64 (CD2314/705F)
• CPC Class Codes: C07K 14/7056 : Lectin superfamily, e.g. CD23, CD72
• CPC Class Codes: C07K 14/70578 : NGF-receptor/TNF-receptor superfamily, e.g. CD27, CD30, CD40, CD95 NGF-receptor C07K14/71, TNF-receptor C07K14/7151
• CPC Class Codes: C07K 14/70596 : Molecules with a "CD"-designation not provided for elsewhere
• CPC Class Codes: C07K 16/2803 : against the immunoglobulin superfamily
• CPC Class Codes: C07K 16/2896 : against molecules with a "CD"-designation, not provided for elsewhere
• CPC Class Codes: C07K 16/40 : against enzymes
• CPC Class Codes: C07K 2317/622 : Single chain antibody (scFv)
• CPC Class Codes: C07K 2319/00 : Fusion polypeptide
• CPC Class Codes: C07K 2319/03 : containing a transmembrane segment
• CPC Class Codes: C07K 2319/70 : containing domain for protein-protein interaction
• CPC Class Codes: C12N 15/85 : for animal cells
• CPC Class Codes: C12N 2510/00 : Genetically modified cells
• CPC Class Codes: C12N 9/003 : Dihydrofolate reductase [DHFR] (1.5.1.3)
• CPC Class Codes: C12N 9/12 : transferring phosphorus containing groups, e.g. kinases (2.7)
• CPC Class Codes: C12N 9/16 : acting on ester bonds (3.1)
• CPC Class Codes: C12N 9/90 : Isomerases (5.)
• CPC Class Codes: C12Y 207/11001 : Non-specific serine/threonine protein kinase (2.7.11.1), i.e. casein kinase or checkpoint kinase
• CPC Class Codes: C12Y 502/01008 : Peptidylprolyl isomerase (5.2.1.8), i.e. cyclophilin

Abstract

The present disclosure relates to compositions and methods for using cells having chemically-induced fusion protein complexes to spatially and temporally control immune cell signal initiation and downstream responses for treating disease.

Claims

1. An isolated non-natural cell comprising:

(a) a first nucleic acid molecule encoding a first fusion protein comprising a first multimerization domain comprising an FRB T2098L polypeptide, a CD8a transmembrane domain, a costimulatory domain of 4-1BB, and an actuator domain of CD3, wherein the first multimerization domain localizes extracellularly when the first fusion protein is expressed; and

(b) a second nucleic acid molecule encoding a second fusion protein comprising a binding domain comprising a single chain antibody variable region (scFv) specific for CD19 or B cell maturation antigen (BCMA), a second multimerization domain comprising an FKBP12 polypeptide, and a CD4, a CD154 or a CD71 transmembrane domain, wherein the second multimerization domain localizes extracellularly when the first fusion protein is expressed;

wherein the first fusion protein and the second fusion protein are each expressed as separate fusion proteins and form a polypeptide complex on the non-natural cell surface in the presence of a bridging factor, rapalog AP21967; and

wherein the bridging factor is associated with and disposed between the first and second multimerization domains.

2. An isolated non-natural cell comprising:

(a) a first nucleic acid molecule encoding a first fusion protein comprising a first multimerization domain comprising an FRB polypeptide, a CD8a transmembrane domain, a costimulatory domain of 4-1BB, and an actuator domain of CD3, wherein the first multimerization domain localizes extracellularly when the first fusion protein is expressed; and

(b) a second nucleic acid molecule encoding a second fusion protein comprising a binding domain comprising an scFv specific for CD19 or BCMA, a second multimerization domain comprising an FKBP12 polypeptide, and a CD4, a CD154 or a CD71 transmembrane domain as a second transmembrane domain, wherein the second multimerization domain localizes extracellularly when the first fusion protein is expressed;

wherein the first fusion protein and the second fusion protein are each expressed as separate fusion proteins and form a polypeptide complex on the non-natural cell surface in the presence of a bridging factor selected from the group consisting of;

Rapamycin, temsirolimus or everolimus; and

wherein the bridging factor is associated with and disposed between the first and second multimerization domains.

3. An isolated non-natural cell comprising:

(a) a first nucleic acid molecule encoding a first fusion protein comprising a first multimerization domain comprising an FRB T2098L polypeptide, a AMN transmembrane domain, a costimulatory domain of 4-1BB, and an actuator domain of CD3, wherein the first multimerization domain localizes extracellularly when the first fusion protein is expressed; and

(b) a second nucleic acid molecule encoding a second fusion protein comprising a binding domain comprising an scFv specific for CD19 or BCMA, a second multimerization domain comprising an FKBP12 polypeptide, and a CD154 transmembrane domain as a second transmembrane domain, wherein the second multimerization domain localizes extracellularly when the first fusion protein is expressed;

wherein the first fusion protein and the second fusion protein are each expressed as separate fusion proteins and form a polypeptide complex on the non-natural cell surface in the presence of a bridging factor, rapalog AP21967; and

wherein the bridging factor is associated with and disposed between the first and second multimerization domains.

4. An isolated non-natural cell comprising:

(a) a first nucleic acid molecule encoding a first fusion protein comprising a first multimerization domain comprising an FRB polypeptide, a AMN transmembrane domain, a costimulatory domain of 4-1BB, and an actuator domain of CD3, wherein the first multimerization domain localizes extracellularly when the first fusion protein is expressed; and

(b) a second nucleic acid molecule encoding a second fusion protein comprising a binding domain comprising an scFv specific for CD19 or BCMA, a second multimerization domain comprising an FKBP12 polypeptide, and a CD154 transmembrane domain as a second transmembrane domain, wherein the second multimerization domain localizes extracellularly when the first fusion protein is expressed;

wherein the first fusion protein and the second fusion protein are each expressed as separate fusion proteins and form a polypeptide complex on the non-natural cell surface in the presence of a bridging factor selected from the group consisting of;

Rapamycin, temsirolimus or everolimus; and

wherein the bridging factor is associated with and disposed between the first and second multimerization domains.