Targeted gene insertion for improved immune cells therapy
Inventors
BUSSER, Brian • Duchateau, Philippe • Juillerat, Alexandre • Poirot, Laurent • Valton, Julien
Assignees
Interested in licensing this patent?
MTEC can help explore whether this patent might be available for licensing for your application.
Publication Number
US-12391933-B2
Publication Date
2025-08-19
Expiration Date
Abstract
The invention pertains to the field of adaptive cell immunotherapy. It provides with the genetic insertion of exogenous coding sequence(s) that help the immune cells to direct their immune response against infected or malignant cells. These exogenous coding sequences are more particularly inserted under the transcriptional control of endogenous gene promoters that are sensitive to immune cells activation. Such method allows the production of safer immune primary cells of higher therapeutic potential.
Core Innovation
The invention relates to preparing a population of engineered primary human NK or T cells for immunotherapy by introducing an exogenous coding sequence encoding an interleukin selected from IL-15, IL-12, or IL-2 into the cells using a sequence-specific endonuclease that targets an endogenous gene. The endogenous gene encodes PD1, and the endogenous gene is cleaved and disrupted while the interleukin coding sequence is inserted into the endogenous gene.
The interleukin is under transcriptional control of the promoter of the endogenous gene, while the coding sequence of the endogenous gene is disrupted. In this PD1-targeted configuration, insertion places the interleukin coding sequence under transcriptional control of the PD1 endogenous promoter while disrupting PD1 coding sequence.
The preparation further includes introducing an exogenous coding sequence encoding a chimeric antigen receptor (CAR) or a recombinant TCR into the primary human NK or T cells. The engineered primary human NK or T cells are configured to secrete a level of the interleukin sufficient to enhance the antitumor activity of the cells.
Claims Coverage
The independent claim coverage identifies one immunotherapy manufacturing method for engineered primary human NK or T cells with three inventive features: PD1-targeted interleukin insertion under endogenous promoter control, interleukin secretion sufficient to enhance antitumor activity, and introduction of a CAR or recombinant TCR.
Endonuclease-mediated interleukin insertion into an endogenous PD1 gene under endogenous promoter control
Introducing an exogenous coding sequence encoding an interleukin selected from IL-15, IL-12, or IL-2 with a sequence-specific endonuclease targeting an endogenous gene into the primary human NK or T cells; cleaving the endogenous gene and inserting the exogenous coding sequence into the endogenous gene such that the interleukin is under transcriptional control of the promoter of the endogenous gene, while disrupting the coding sequence of the endogenous gene, wherein the endogenous gene encodes PD1.
Interleukin secretion sufficient to enhance antitumor activity
The engineered primary human NK or T cells secrete a level of the interleukin sufficient to enhance the antitumor activity of the cells.
CAR or recombinant TCR introduction
Introducing an exogenous coding sequence encoding a chimeric antigen receptor (CAR) or a recombinant TCR into the primary human NK or T cells.
Anti-CD22 CAR variant
The method is carried out using an anti-CD22 CAR.
Interleukin variant selection
The interleukin used is IL-2; and/or the method is carried out using interleukin IL-15.
Engineered cell composition thresholds
More than 50% of said engineered primary human NK or T cells are TCR negative T-cells and/or more than 50% of said engineered primary human NK or T cells are CAR positive cells.
The claims focus on PD1-locus engineering that places IL-15, IL-12, or IL-2 under transcriptional control of the endogenous PD1 promoter with PD1 coding disruption, together with CAR or recombinant TCR introduction, to produce engineered primary human NK or T cells with interleukin secretion sufficient to enhance antitumor activity.
Stated Advantages
Enhance the antitumor activity of the cells by secretion of a level of the interleukin sufficient to enhance antitumor activity.
Documented Applications
Immunotherapy using engineered primary human NK or T cells prepared by introducing an exogenous interleukin into an endogenous PD1 gene locus and introducing a CAR or recombinant TCR.
Tumor engagement evaluation of engineered T-cells expressing CAR and interleukin matrices, including serial killing against tumor cells.
Interested in licensing this patent?