TARGETING CYTOTOXIC CELLS WITH CHIMERIC RECEPTORS FOR ADOPTIVE IMMUNOTHERAPY

• US 20140322183A1
• Filed: 03/15/2014
• Published: 10/30/2014
• Est. Priority Date: 03/15/2013
• Status: Active Grant

• Current Assignee: Novartis Ag, Trustees Of The University Of Pennsylvania (University of Pennsylvania)
• Original Assignee: Trustees Of The University Of Pennsylvania (University of Pennsylvania)
• Inventors: Milone, Michael C., Wang, Enxiu
• Granted Patent: US 9,745,368 B2
• Time in Patent Office: Days
• US Class Current: 424/93.21
• CPC Class Codes: C07K 14/70503 Immunoglobulin superfamily
• CPC Class Codes: C07K 14/70535 Fc-receptors, e.g. CD16, CD32, CD64 (CD2314/705F)
• CPC Class Codes: C07K 16/18 against material from animals or humans
• CPC Class Codes: C07K 2319/03 containing a transmembrane segment
• CPC Class Codes: C07K 2319/33 fusions for targeting to specific cell types, e.g. tissue specific targeting, targeting of a bacterial subspecies
• CPC Class Codes: C07K 2319/70 containing domain for protein-protein interaction

Abstract

The present invention provides compositions and methods for regulating the specificity and activity of T cells. In one embodiment, the invention provides a type of chimeric antigen receptor (CAR) wherein the CAR is termed a “KIR-CAR” which is a CAR design comprising a component of a receptor naturally found on natural killer (NK) cells. In one embodiment, the NK receptor includes but is not limited to a naturally occurring activating and inhibitory receptor of NK cells known as a killer cell immunoglobulin-like receptor (KIR).

Claims

1. A purified, or non-naturally occurring, NK cell receptor chimeric antigen receptor (NKR-CAR) comprising one, two or all of:

an extra-cellular antigen binding domain; and

a transmembrane domain; and

a cytoplasmic domain.

5. A nucleic acid comprising a sequence that encodes a NKR-CAR of claim 1.

6. The nucleic acid of claim 5, further comprising a sequence that encodes an adaptor molecule or intracellular signaling domain that interacts with said NKR-CAR.

7. A cytotoxic cell comprising a NKR-CAR of claim 1.

9. A method of making a cell of claim 7, comprising, introducing into a cytotoxic cell a nucleic acid of claim 5, or forming in a cytotoxic cell a NKR-CAR of claim 1.

11. A method of treating a mammal comprising administering to the mammal an effective amount of a cell of claim 7 thereby providing an anti-tumor immunity in the mammal.

2-4. -4. (canceled)

8. (canceled)

10. (canceled)

12-13. -13. (canceled)

14. A purified, or non-naturally occurring, killer cell immunoglobulin-like receptor chimeric antigen receptor (KIR-CAR) comprising:

an extra-cellular antigen binding domain; and

a transmembrane domain or cytoplasmic domain.

115. A cytotoxic cell comprising:

(a) a first KIR-CAR of claim 14.

149. A method of making a cell, comprising, forming in a cytotoxic cell, a KIR-CAR of claim 14.

15-66. -66. (canceled)

67. A nucleic acid comprising:

(a) sequence that encodes a first killer cell immunoglobulin-like receptor chimeric antigen receptor (KIR-CAR), wherein the first KIR-CAR comprises an extra-cellular antigen binding domain; and

a transmembrane domain, or cytoplasmic domain.

68. The nucleic acid of claim 67, wherein said first KIR-CAR is an actKIR-CAR.

69. The nucleic acid of claim 67, wherein said KIR-CAR is an inhKIR-CAR.

70. The nucleic acid of claim 67, wherein said nucleic acid comprises a DNA sequence or an RNA sequence.

72. The nucleic acid of claim 67, further comprising:

(b) sequence that encodes a second KIR-CAR that is different from said first KIR-CAR.

81. The nucleic acid of claim 67, further comprising:

sequence that encodes an intracellular signaling domain.

82. The nucleic acid of claim 81, wherein said intracellular signaling domain comprises an ITAM motif.

83. The nucleic acid of claim 81, wherein the sequence that encodes an intracellular signaling domain comprises a DAP 12 polypeptide comprising a DAP 12 intracellular signaling domain.

84. The nucleic acid of claim 83, wherein said DAP 12 polypeptide further comprises one or both of a transmembrane domain and an extracellular domain.

86. The nucleic acid of any of claim 81, wherein each of (a), (b), and (c) are present on the same nucleic acid molecule.

87. The nucleic acid of claim 81, wherein:

one of (a), (b), and (c) is encoded on a first nucleic acid molecule; and

and a second and third of (a), (b), and (c) is encoded on a second nucleic acid molecule.

91. The nucleic acid of claim 81, wherein each of (a), (b), and (c) are present on different nucleic acid molecules.

92. The nucleic acid of claim 72, wherein:

(i) the antigen binding domain of one of said first KIR-CAR and said second KIR-CAR does not comprise a light chain variable domain and a heavy chain variable domain;

(ii) the antigen binding domain of one of said first KIR-CAR and said second KIR-CAR is an scFv, and the other is other is other than an scFv;

(iii) when present on the surface of a cell, the antigen binding domains of said first KIR-CAR and said second KIR-CAR, associate with one another less than if both were scFv antigen binding domains;

(iv) wherein, when present on the surface of a cell, binding of the antigen binding domain of said first KIR-CAR to its cognate antigen is not substantially reduced by the presence of said second KIR-CAR;

(v) the antigen binding domain of one of said first KIR-CAR and said second KIR-CAR, comprises a single VH domain;

(vi) the antigen binding domain of one of said first KIR-CAR and said second KIR-CAR, is an scFv, and the other comprises a single VH domain;

(vii) the antigen binding domain of one of said first KIR-CAR and said second KIR-CAR, is an scFv, and the other comprises a nanobody;

or

(viii) the antigen binding domain of one of said first KIR-CAR and said second KIR-CAR, is an scFv, and the other comprises a camelid VHH domain.

101. The nucleic acid of claim 67, further comprising a sequence that encodes a T cell chimeric antigen receptor (TCAR).

104. The nucleic acid of claim 101, wherein said TCAR comprises one or both of an antigen binding domain and an activating cytoplasmic domain from the T cell receptor complex with CD3, and a costimulatory domain from costimulatory receptor.

106. The nucleic acid of claim 101, wherein:

(i) the antigen binding domain of one of said KIR-CAR and said TCAR does not comprise a light chain variable domain and a heavy chain variable domain;

(ii) the antigen binding domain of one of said KIR-CAR and said TCAR is an scFv, and the other is other than an scFv;

(iii) when present on the surface of a cell, the antigen binding domains of said KIR-CAR and said TCAR, associate with one another less than if both were scFv antigen binding domains;

(iv) when present on the surface of a cell, binding of the antigen binding domain of said KIR-CAR to its cognate antigen is not substantially reduced by the presence of said second TCAR;

(v) the antigen binding domain of one of said KIR-CAR and said TCAR, comprises a single VH domain;

(vi) the antigen binding domain of one of said KIR-CAR and said TCAR, is an scFv, and the other comprises a single VH domain;

(vii) the antigen binding domain of one of said KIR-CAR and said TCAR, is an scFv, and the other comprises a nanobody;

or

(viii) wherein, the antigen binding domain of one of said KIR-CAR and said TCAR, is an scFv, and the other comprises a camelid VHH domain.

144. A cytotoxic cell comprising:

a nucleic acid of claim 67;

or

a KIR-CAR encoded by a nucleic acid of claim 67.

145. The cytotoxic cell of claim 144, wherein said cytotoxic cell is a T cell or an NK cell.

148. A method of making a cell, comprising, introducing into a cytotoxic cell, a nucleic acid of claim 67.

150. A method of treating a mammal comprising administering to the mammal an effective amount of a cell of claim 144 thereby providing an anti-tumor immunity in the mammal.

71. (canceled)

73-80. -80. (canceled)

85. (canceled)

88-90. -90. (canceled)

93-100. -100. (canceled)

102-103. -103. (canceled)

105. (canceled)

107-114. -114. (canceled)

116-143. -143. (canceled)

146-147. -147. (canceled)

151-167. -167. (canceled)

168. A purified, or non-naturally occurring, natural cytotoxicity receptor chimeric antigen receptor (NCR-CAR) comprising:

an extra-cellular antigen binding domain;

a transmembrane domain; and

a cytoplasmic domain.

169-187. -187. (canceled)

188. A purified, or non-naturally occurring, signaling lymphocyte activation molecule family chimeric antigen receptor (SLAMF-CAR) comprising:

an extra-cellular antigen binding domain;

a transmembrane domain; and

a SLAMF cytoplasmic domain.

189-197. -197. (canceled)

198. A purified, or non-naturally occurring, Fc receptor chimeric antigen receptor (FcR-CAR) comprising:

an extra-cellular antigen binding domain;

a transmembrane domain a transmembrane domain comprising a positively charged moiety; and

a CD16 or CD64 cytoplasmic domain.

199-212. -212. (canceled)

213. A purified, or non-naturally occurring, Ly49 receptor chimeric antigen receptor (Ly49-CAR) comprising:

an extra-cellular antigen binding domain; and

a transmembrane domain.

214-232. -232. (canceled)

233. A cytotoxic cell comprising:

a first non-naturally occurring chimeric membrane embedded receptor comprising an antigen binding domain; and

a second non-naturally occurring chimeric membrane embedded receptor comprising an antigen binding domain,

wherein;

(i) the antigen binding domain of one of said first and said second non-naturally occurring chimeric membrane embedded receptor does not comprise a light chain variable domain and a heavy chain variable domain;

(ii) the antigen binding domain of one of said first and said second non-naturally occurring chimeric membrane embedded receptor comprises an scFv, and the other is other than an scFv;

(iii) when present on the surface of a cell, the antigen binding domains of said first and said second non-naturally occurring chimeric membrane embedded receptor, associate with one another less than if both were scFv antigen binding domains;

(iv) when present on the surface of a cell, binding of the antigen binding domain of said first non-naturally occurring chimeric membrane embedded receptor to its cognate antigen is not substantially reduced by the presence of said second non-naturally occurring chimeric membrane embedded receptor;

(v) the antigen binding domain of one of said first and said second non-naturally occurring chimeric membrane embedded receptor, comprises a single VH domain;

(vi) the antigen binding domain of one of said first and said second non-naturally occurring chimeric membrane embedded receptor, comprises an scFv, and the other comprises a single VH domain;

(vii) the antigen binding domain of one of said first and said second non-naturally occurring chimeric membrane embedded receptor, comprises an scFv, and the other comprises a nanobody;

or

(viii) the antigen binding domain of one of said first and said second non-naturally occurring chimeric membrane embedded receptor, comprises an scFv, and the other comprises a camelid VHH domain.

237. The cell of claim 233, wherein one or both of said first and said second non-naturally occurring chimeric membrane embedded receptors is a TCAR.

239. The cell of claim 233, wherein one or both of said first and said second non-naturally occurring chimeric membrane embedded receptors is a NKR-CAR.

249. A method of making a cell of claim 233, comprising, introducing into a cytotoxic cell, a nucleic acid of claim 248.

251. A method of treating a mammal comprising administering to the mammal an effective amount of a cell of claim 233 thereby providing an anti-tumor immunity in the mammal.

234-236. -236. (canceled)

238. (canceled)

240-247. -247. (canceled)

248. A nucleic acid comprising a sequence that encodes a first and second non-naturally occurring chimeric membrane embedded receptor comprising an antigen binding domain,

wherein:

(i) the antigen binding domain of one of said first and said second non-naturally occurring chimeric membrane embedded receptor does not comprise a light chain variable domain and a heavy chain variable domain;

(ii) the antigen binding domain of one of said first and said second non-naturally occurring chimeric membrane embedded receptor comprises an scFv, and the other is other than an scFv;

(iii) when present on the surface of a cell, the antigen binding domains of said first and said second non-naturally occurring chimeric membrane embedded receptor, associate with one another less than if both were scFv antigen binding domains;

(iv) when present on the surface of a cell, binding of the antigen binding domain of said first non-naturally occurring chimeric membrane embedded receptor to its cognate antigen is not substantially reduced by the presence of said second non-naturally occurring chimeric membrane embedded receptor;

(v) the antigen binding domain of one of said first and said second non-naturally occurring chimeric membrane embedded receptor, comprises a single VH domain;

(vi) the antigen binding domain of one of said first and said second non-naturally occurring chimeric membrane embedded receptor, comprises an scFv, and the other comprises a single VH domain;

(vii) the antigen binding domain of one of said first and said second non-naturally occurring chimeric membrane embedded receptor, comprises an scFv, and the other comprises a nanobody;

or

(viii) the antigen binding domain of one of said first and said second non-naturally occurring chimeric membrane embedded receptor, comprises an scFv, and the other comprises a camelid VHH domain.

250. (canceled)

252-253. -253. (canceled)