US 10,870,864 B2 · Methods for engineering T cells for immunotherapy by using RNA-guided Cas nuclease system

Abstract

The present invention relates to methods of developing genetically engineered, preferably non-alloreactive T-cells for immunotherapy. This method involves the use of RNA-guided endonucleases, in particular Cas9/CRISPR system, to specifically target a selection of key genes in T-cells. The engineered T-cells are also intended to express chimeric antigen receptors (CAR) to redirect their immune activity towards malignant or infected cells. The invention opens the way to standard and affordable adoptive immunotherapy strategies using T-Cells for treating cancer and viral infections.

Claims18

Dependent claims

A method of preparing and administering T-cells for cancer immunotherapy comprising the steps of:

(a) providing primary human T-cells from a donor,
(b) genetically modifying the primary human T-cells to eliminate expression of a gene encoding an immune checkpoint protein, comprising expressing in the cells
(i) a Cas9 endonuclease fused to a nuclear localization signal (NLS), and
(ii) a guide RNA that directs said endonuclease to at least one targeted locus in the gene encoding the immune checkpoint protein in the T-cell genome,
(c) expanding the genetically modified T-cells, and
(d) administering at least 104 of the expanded genetically modified T-cells to a cancer patient.

3 more elements

Dependencies are inferred from the claim text.

Facts

Application

16/793,896Filed 18 Feb 2020 · continuation

Priority

DK 20137029729 May 2013

Published

US 2020/0277625 A13 Sep 2020

Granted

22 Dec 2020

Term ends

Not estimatedThe source records don’t support an estimate.

Classification26 codes

C12N 15/85
Vectors or expression systems specially adapted for eukaryotic hosts: for animal cells
A61K 40/11
Characterised by the cell type used: t-cells, e.g. tumour infiltrating lymphocytes [TIL] or regulatory T [Treg] cells; Lymphokine-activated killer…
A61K 40/22
Characterised by the effect or the function of the cells: immunosuppressive or immunotolerising
A61K 40/31
Characterised by the recombinant expression of specific molecules in the cells of the immune system: chimeric antigen receptors [CAR]

Family

Application61/888,259Us provisional application · filed 8 Oct 2013

Application61/907,858Us provisional application · filed 22 Nov 2013

Application61/907,874Us provisional application · filed 22 Nov 2013

PCT/EP2014/056534Parent of this continuation · filed 1 Apr 2014

Application15/891,496Parent of this continuation · filed 8 Feb 2018

US 9,890,393 B2Parent of this continuation · filed 13 Feb 2018

US 2020/0277625 A1Pre-grant publication · published 3 Sep 2020

US 10,584,352 B2Parent of this continuation

Application14/892,934Parent of this continuation

US 2016/0272999 A1Published application

US 2018/0237798 A1Published application

US 10,870,864 B2This patent

From the US filing history. Foreign family members aren’t listed.

Methods for engineering T cells for immunotherapy by using RNA-guided Cas nuclease system

Life of this patent

Methods for engineering T cells for immunotherapy by using RNA-guided Cas nuclease system

Life of this patent

Abstract

Claims18