	US 12,215,137 B2
	Compositions and methods for T cell delivery of therapeutic molecules
Yangbing Zhao, Lumberton, NJ (US); Carl H. June, Merion Station, PA (US); and Xiaojun Liu, Wallingford, PA (US)
Assigned to The Trustees of the University of Pennsylvania, Philadelphia, PA (US)
Filed by The Trustees of the University of Pennsylania, Philadelphia, PA (US)
Filed on Sep. 30, 2020, as Appl. No. 17/038,676.
Application 17/038,676 is a division of application No. 15/544,816, granted, now 10,828,353, previously published as PCT/US2015/058192, filed on Oct. 30, 2015.
Claims priority of provisional application 62/110,489, filed on Jan. 31, 2015.
Prior Publication US 2021/0008188 A1, Jan. 14, 2021
Int. Cl. C07K 14/705 (2006.01); A61K 39/00 (2006.01); A61K 39/395 (2006.01); C07K 14/71 (2006.01); C07K 14/715 (2006.01); C07K 16/28 (2006.01); C12N 5/0783 (2010.01); C12N 15/00 (2006.01)

CPC C07K 14/70578 (2013.01) [A61K 39/39558 (2013.01); A61K 39/4611 (2023.05); A61K 39/4631 (2023.05); A61K 39/464406 (2023.05); A61K 39/464412 (2023.05); C07K 14/70521 (2013.01); C07K 14/71 (2013.01); C07K 14/7155 (2013.01); C07K 16/2818 (2013.01); C07K 16/2827 (2013.01); C07K 16/2863 (2013.01); C12N 5/0636 (2013.01); C12N 15/00 (2013.01); C07K 2317/31 (2013.01); C07K 2317/622 (2013.01); C07K 2319/02 (2013.01); C07K 2319/10 (2013.01); C07K 2319/74 (2013.01); C12N 2501/515 (2013.01); C12N 2501/599 (2013.01); C12N 2510/00 (2013.01)]

21 Claims

1. A method for enhancing T cell antitumor activity in a subject in need thereof, the method comprising parenterally administering to the subject a composition comprising modified CAR T cells comprising a nucleic acid sequence encoding a switch molecule and a nucleic acid sequence encoding a CAR comprising an antigen binding domain that targets a tumor antigen, a transmembrane domain, a costimulatory domain, and an intracellular signaling domain;

wherein the switch molecule comprises:

(a) an extracellular domain of a transforming growth factor-beta receptor (TGF-beta-R), an intracellular domain of a signaling receptor selected from the group consisting of interleukin-12 receptor (IL-12R), CD3, CD28, CD137, CD27, ICOS, and OX40, and a transmembrane domain derived from the same molecule as the extracellular domain or the same molecule as the intracellular domain;

(b) an extracellular domain of a programmed cell death 1 (PD1) receptor, an intracellular domain of a signaling receptor selected from the group consisting of interleukin-12 receptor (IL-12R), CD3, CD28, CD137, CD27, ICOS, and OX40, and a transmembrane domain derived from the same molecule as the extracellular domain or the same molecule as the intracellular domain;

(c) an extracellular domain of a programmed cell death ligand 1 (PDL1) receptor, an intracellular domain of a signaling receptor selected from the group consisting of interleukin-12 receptor (IL-12R), CD3, CD28, CD137, CD27, ICOS, and OX40, and a transmembrane domain derived from the same molecule as the extracellular domain or the same molecule as the intracellular domain; or

(d) an extracellular domain of an interferon-gamma receptor (IFN-gamma) receptor, an intracellular domain of a signaling receptor selected from the group consisting of interleukin-12 receptor (IL-12R), CD3, CD28, CD137, CD27, ICOS, and OX40, and a transmembrane domain derived from the same molecule as the extracellular domain or the same molecule as the intracellular domain;

wherein the modified CAR T cells home to a target tumor site; and

wherein when the modified CAR T cells express the switch molecule, interaction of the extracellular domain of the switch molecule with TGFβ, PD1, PDL1, or IFN-γ in the tumor microenvironment converts an inhibitory signal of the TGFβ receptor, the PD1 receptor, the PDL1 receptor, or the IFN-γ receptor into an IL-12R, CD3, CD28, CD137, CD27, ICOS, or OX40 stimulatory signal thereby enhancing the antitumor activities of the modified CAR T cells.

21. A method for enhancing T cell antitumor activity in a subject in need thereof, the method comprising parenterally administering to the subject a composition comprising modified T cells comprising a nucleic acid sequence encoding a switch molecule and a nucleic acid sequence encoding a chimeric antigen receptor (CAR) that targets a tumor antigen,

wherein the tumor antigen is selected from the group consisting of p53, Ras, beta-Catenin, CDK4, alpha-Actinin-4, Tyrosinase, TRP1/gp75, TRP2, gp100, Melan-A/MART1, Gangliosides, PSMA, HER2, WT1, EphA3, EGFR, CD19, CD20, MAGE, BAGE, GAGE, NY-ESO-1, telomerase, and survivin;

wherein the switch molecule comprises:

(c) an extracellular domain of a programmed cell death ligand 1 (PDL1) receptor, an intracellular domain of a signaling receptor selected from the group consisting of interleukin-12 receptor (IL-12R), CD3, CD28, CD137, CD27, ICOS, and OX40, and a transmembrane domain derived from the same molecule as the extracellular domain, or the same molecule as the intracellular domain; or

wherein the modified T cells home to a target tumor site; and

wherein when the modified T cells express the switch molecule, interaction of the extracellular domain of the switch molecule with TGFβ, PD1, PDL1, or IFN-γ in the tumor site converts the inhibitory signal of the TGFβ receptor, the PD1 receptor, the PDL1 receptor, or the IFN-γ receptor into the IL-12R, the CD3, the CD28, the CD137, the CD27, the ICOS, or the OX40 stimulatory signal, thereby enhancing antitumor activities of the modified T cells.