Cytotoxic molecules responsive to intracellular ligands for selective T cell mediated killing

Patent metadata

Field	Value
Publication number	US-12570968-B2
Application number	US-202117357704-A
Country	US
Kind code	B2
Filing date	Jun 24, 2021
Priority date	Dec 15, 2014
Publication date	Mar 10, 2026
Grant date	Mar 10, 2026

Abstract

Compositions and methods are provided for the cell-mediated targeted killing of diseased cells based on the presence of an intracellular antigen, rather than a surface-bound marker. The targeting cells are modified to express a cytotoxic protein that is delivered into a targeted cell, and after delivery is selectively activated by the presence of a cytoplasmic protein of interest. In one embodiment of the invention, the cytotoxic molecule is a Granzyme B (GrB) polypeptide. In the compositions of the invention, GrB is modified to render its cytotoxic enzymatic functions inactive, until the presence of an intracellular antigen unlocks the GrB molecule to enable enzymatic activities.

First claim

What is claimed is:

1 . A method of treating an individual for cancer, the method comprising administering an effective dose of a cell comprising (i) Granzyme B (GrB) fused to a truncated oncoprotein and an oncoprotein specific binding partner; and/or (ii) a nucleic acid encoding for Granzyme B (GrB) fused to a truncated oncoprotein and an oncoprotein specific binding partner;
wherein the oncoprotein is human telomerase reverse transcriptase (hTERT), and the oncoprotein specific binding partner is an antibody, scFv or nanobody.

2 . The method of claim 1 , wherein the oncoprotein specific binding partner is fused through a linker internal to or at the amino termini of GrB and the truncated oncoprotein is fused through a linker to the carboxyl termini of GrB.

3 . The method of claim 2 , wherein the oncoprotein specific binding partner is an scFv or a nanobody.

4 . The method of claim 1 , wherein a secretion signal peptide and a 6×-histidine tag are attached to the N-terminus of the engineered GrB for purification.

5 . The method of claim 1 , wherein GrB signal peptide is attached to the N-terminus of the engineered GrB.

6 . The method of claim 1 , wherein a secretion peptide is attached to the N-terminus of the engineered GrB.

7 . The method of claim 1 , wherein the oncoprotein comprises hTERT that lacks a catalytic domain.

8 . The method of claim 1 , wherein the oncoprotein comprises hTERT long N-terminal extension (NTE) and/or short C-terminal extension (CTE) in the absence of the catalytic domain.

Assignees

• Univ California

Inventors

• Chen Yvonne Y
• Ho Patrick

Classifications

• C12N2510/00

  Genetically modified cells · CPC title

• C12N5/0636

  T lymphocytes · CPC title

• C07K2319/95

  containing a motif/fusion for degradation (ubiquitin fusions, PEST sequence) · CPC title

• C07K2317/622

  Single chain antibody (scFv) · CPC title

• C07K16/40

  against enzymes · CPC title