US 11,851,672 B2
CAR T-cells comprising transcriptional circuits
Martin Pulé, London (GB); James Sillibourne, London (GB); and Shaun Cordoba, London (GB)
Assigned to AUTOLUS LIMITED, London (GB)
Appl. No. 16/759,222
Filed by AUTOLUS LIMITED, London (GB)
PCT Filed Oct. 24, 2018, PCT No. PCT/GB2018/053088
§ 371(c)(1), (2) Date Apr. 24, 2020,
PCT Pub. No. WO2019/081935, PCT Pub. Date May 2, 2019.
Claims priority of application No. 1717524 (GB), filed on Oct. 25, 2017.
Prior Publication US 2020/0308600 A1, Oct. 1, 2020
Int. Cl. C12N 15/86 (2006.01); A61K 35/17 (2015.01); A61K 48/00 (2006.01)
CPC C12N 15/86 (2013.01) [A61K 35/17 (2013.01); A61K 48/00 (2013.01); C12N 2740/10043 (2013.01); C12N 2800/40 (2013.01)] 2 Claims
 
1. A method for treating a disease by killing target cells in a subject, which comprises the following steps:
(a) isolation of a T cell-containing sample from a subject,
(b) transduction or transfection of T cells of the sample with a kit of vectors comprising:
(i) a first vector which comprises a first transgene which encodes a chimeric antigen receptor (CAR) which binds an antigen expressed on the target cells and a nucleotide sequence encoding a transcription factor, and
(ii) a second vector which comprises a second transgene which encodes an entity of interest (EOI) and a marker gene
wherein expression of the second transgene within the T cell is dependent upon expression of the transcription factor,
or
(i) a first vector which comprises a first transgene which encodes a chimeric antigen receptor (CAR) which binds an antigen expressed on the target cells and a nucleotide sequence encoding a first transcription factor,
(ii) a second vector which comprises a second transgene which encodes an entity of interest (EOI) and a nucleotide sequence encoding a second transcription factor, and
(iii) a third vector which comprises a third transgene which encodes an entity of interest (EOI) and a marker gene
wherein expression of the second transgene within the T cell is dependent upon expression of the first transcription factor, and expression of the third transgene within the T cell is dependent upon expression of the second transcription factor,
(c) selecting T cells which express the marker gene, and
(d) administering the T cells selected from step (c) to the subject to kill the target cells.