JP-2026076273-A - Anti-dinitrophenol chimeric antigen receptor

Abstract

[Problem] To provide a method and composition containing an anti-dinitrophenol chimeric antigen receptor (CAR). [Solution] The present invention includes nucleic acids encoding anti-dinitrophenol chimeric antigen receptors (CARs), polypeptides encoded by this nucleic acid, cells containing this nucleic acid or polypeptide, and methods utilizing these cells. Some embodiments further include the use of dinitrophenol (DNP) and its derivatives. [Selection Diagram] None

Inventors

• ジェンセン，マイケル シー．
• マッセイ，ジェイムズ エフ．
• チェン，ジョセフ ケー．

Dates

Publication Date

20260511

Application Date

20260126

Priority Date

20200204

Claims (1)

1. A nucleic acid encoding a chimeric antigen receptor (CAR), wherein the CAR is A ligand-binding domain that specifically binds to the dinitrophenol (DNP) moiety; Spacer; Nucleic acids, including a transmembrane domain and an intracellular signaling domain.

Description

This application, which is cross-referenced to a related application , claims priority to U.S. Provisional Patent Application No. 62/969,931, “Anti-Dinitrophenol Chimeric Antigen Receptor,” filed on 4 February 2020, which is incorporated herein by reference in its entirety. This application, along with a reference to a sequence listing, was filed with an electronic sequence listing. This sequence listing was provided as a file of approximately 43kb created on February 2, 2021, with the filename SCRI269WOSEQLIST. The information contained in this electronic sequence listing is incorporated herein by reference in its entirety. The embodiments provided herein include methods and compositions comprising an anti-dinitrophenol chimeric antigen receptor (CAR). Some embodiments include nucleic acids encoding this CAR, polypeptides encoded by this nucleic acid, cells containing this polypeptide, and methods utilizing these cells. Some embodiments further include the use of dinitrophenol (DNP) and its derivatives. Immunotherapy utilizing adoptive cell transfer of T cells possessing chimeric antigen receptors (CARs) is an effective treatment method for cancer. The structure of a CAR includes an antigen-binding domain, a spacer domain, a transmembrane domain, and one or more costimulatory activation domains. CAR T cells may be generated from T cells obtained from patients or donors. In some cases, CARs exert their function by binding to specific antigens on the cell surface, causing lysis of cells presenting that antigen. While various studies have focused on designing the ligand-binding domain of CARs to target desired cell surface antigens and reduce toxicity, further therapeutic approaches using CAR T cells are still needed. An embodiment of a DNP-ether phospholipid (DNP-PLE) is shown, comprising (i) a DNP portion; (ii) a polyethylene glycol (PEG) portion; (iii) a polar head; and (iv) a hydrophobic tail. Flow cytometry data for MDA-MB-231 control cells and MDA-MB-231 control cells incubated with anti-DNP-Alexa Fluor 488 antibody are shown. Flow cytometry data for MDA-MB-231 control cells, MDA-MB-231 control cells incubated with anti-DNP-Alexa Fluor 488 antibody, and MDA-MB-231 cells labeled with 5 μM DNP-PLE and stained with anti-DNP-Alexa Fluor 488 antibody are shown. Flow cytometry data for MDA-MB-231 control cells, MDA-MB-231 control cells incubated with anti-DNP-Alexa Fluor 488 antibody, and MDA-MB-231 cells labeled with 500 nM DNP-PLE and stained with anti-DNP-Alexa Fluor 488 antibody are shown. Flow cytometry data are shown for MDA-MB-231 control cells, MDA-MB-231 control cells incubated with anti-DNP-Alexa Fluor 488 antibody, and MDA-MB-231 cells labeled with 50 nM DNP-PLE and stained with anti-DNP-Alexa Fluor 488 antibody. Figures 2A to 2D show histogram plots of the flow cytometry data. Confocal images of MDA-MB-231 control cells incubated with anti-DNP-Alexa Fluor 488 antibody are shown. This image shows a confocal image of MDA-MB-231 cells incubated with 5 μM DNP-PLE. The image shows confocal images of MDA-MB-231 cells incubated with 5 μM DNP-PLE and stained with anti-DNP-Alexa Fluor 488 antibody. The image shows confocal images of MDA-MB-231 cells incubated with 1 μM DNP-PLE and stained with anti-DNP-Alexa Fluor 488 antibody. A schematic diagram of a second-generation CAR cassette with a long spacer for expressing anti-DNP CARs is shown. Confocal images of MDA-MB-231 control cells co-cultured with anti-DNP CAR H9 cells are shown. The image shows a confocal image of MDA-MB-231 cells stained with 5 μM DNP-PLE and co-cultured with anti-DNP CAR H9 cells. The embodiments provided herein include methods and compositions comprising an anti-dinitrophenol chimeric antigen receptor (CAR). Some embodiments include nucleic acids encoding this CAR, polypeptides encoded by this nucleic acid, cells containing this polypeptide, and methods utilizing these cells. Some embodiments further include the use of dinitrophenol (DNP) and its derivatives. Adoptive transfer of T cells modified by transgenes has been successful under selected conditions (e.g., malignancies of the CD19 B-cell lineage), but generalizing T-cell adoptive transfer therapy to other types of cancer is difficult because there is no single target antigen that is found in all forms of cancer but not in normal healthy cells. CAR T-cell therapy can be used to treat various types of cancer that afflict many people, but its development is hindered by the daunting challenge of identifying and meticulously investigating tens of thousands of antigens (e.g., antigens targeted by CARs) presented on cancer cells. In some embodiments described herein, this challenge is solved by using DNP-ether phospholipid (DNP-PLE). DNP-PLE associates with tumor cells and interacts with T cells possessing DNP-specific CARs by presenting a specific target molecule to those CARs, thereby inducing the lysis of DNP-presenting tumor cells. This approach eliminates the need to identify and