Dual CAR-T cells
Inventors
Choulika, André • Poirot, Laurent • ARANDA ORGILLES, Beatriz • Duchateau, Philippe
Assignees
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Publication Number
US-12618048-B2
Publication Date
2026-05-05
Expiration Date
Abstract
The present invention concerns new engineered immune cells expressing two CARs directed against two different targets, polynucleotides for preparing said immune cells, pharmaceutical compositions comprising said immune cells, and the use of said immune cells in the treatment of cancers.
Core Innovation
The invention relates to a genetically engineered immune cell that expresses a chimeric antigen receptor (CAR) specific for CD22 and a chimeric antigen receptor (CAR) specific for CD20 at its cell surface. The CAR22 and CAR20 are encoded by an exogenous nucleic acid incorporated in the genome of the immune cell, and the same promoter controls expression of CAR20 and CAR22. The exogenous nucleic acid includes, from 5′ to 3′, a promoter, a nucleic acid encoding CAR20, a nucleic acid encoding a self-cleaving peptide, and a nucleic acid encoding CAR22, and comprises SEQ ID NO: 32.
The disclosed immune cell further includes embodiments in which CAR20 and CAR22 lack a leader sequence when present at the cell surface. The CAR20 and CAR22 may be defined by specific amino acid sequences, including SEQ ID NO: 18 for CAR20 and SEQ ID NO: 14 for CAR22, optionally without the leader sequence of SEQ ID NO: 1. In some embodiments, the engineered immune cell comprises at least one allele encoding TCR alpha, TCR beta, or CD3 that has been inactivated, and has at least one inactivated CD52 allele.
The document also discloses an isolated polynucleotide in which CAR20 is under a promoter and CAR22 is on a single nucleic acid molecule with a self-cleaving peptide located between the CAR20- and CAR22-encoding regions. The polynucleotide comprises SEQ ID NO: 32, and a vector comprising the isolated polynucleotide is included. Ex vivo introduction into a T cell is described for preparing the genetically engineered immune cell, and the document further describes dual CAR activation behavior, increased IFN-γ release, and tumor control in mouse lymphoma models.
Claims Coverage
The partial content provides three independent claims, covering: a genetically engineered immune cell with genome-incorporated dual CAR22/CAR20 expression controlled by a shared promoter and including a self-cleaving peptide and SEQ ID NO: 32; an isolated polynucleotide with promoter-controlled CAR20 followed by a self-cleaving peptide between CAR20 and CAR22 on a single nucleic acid molecule, comprising SEQ ID NO: 32; and a genetically engineered immune cell with specific CAR amino acid sequences plus inactivated TCR alpha/beta/CD3 alleles and an inactivated CD52 allele. Across these independent claims, the inventive features center on dual CAR expression from a single genome-incorporated construct and specified immune cell engineering constraints.
Dual CAR22/CAR20 cell surface expression encoded by a single genome-incorporated exogenous nucleic acid
A genetically engineered immune cell expressing CAR22 and CAR20 at its cell surface, wherein the CAR22 and CAR20 are encoded by an exogenous nucleic acid incorporated in the genome of the immune cell, and wherein the exogenous nucleic acid comprises, from 5′ to 3′, a promoter, a nucleic acid encoding CAR20, a nucleic acid encoding a self-cleaving peptide, and a nucleic acid encoding CAR22, with the same promoter controlling expression of CAR20 and CAR22 and the exogenous nucleic acid comprising SEQ ID NO: 32.
Single-molecule CAR20-self-cleaving peptide-CAR22 polynucleotide with shared promoter and SEQ ID NO: 32
An isolated polynucleotide comprising, from 5′ to 3′, a promoter that controls expression of CAR20, followed by a nucleic acid encoding CAR20, and a nucleic acid encoding CAR22 on a single nucleic acid molecule, with a nucleic acid sequence encoding a self-cleaving peptide located between the CAR20- and CAR22-encoding regions, and wherein the single nucleic acid molecule comprises SEQ ID NO: 32.
Dual CAR22/CAR20 immune cell with specified CAR amino acid sequences and inactivated TCR/CD52 alleles
A genetically engineered immune cell expressing CAR20 and CAR22 at its cell surface, wherein CAR20 comprises SEQ ID NO: 18, optionally without the leader sequence of SEQ ID NO: 1; CAR22 comprises SEQ ID NO: 14, optionally without the leader sequence of SEQ ID NO: 1; the immune cell comprises at least one allele encoding TCR alpha, TCR beta, or CD3 that has been inactivated; the immune cell has at least one inactivated CD52 allele; and CAR20 and CAR22 are encoded by an exogenous nucleic acid incorporated in the genome of the immune cell, wherein the exogenous nucleic acid comprises SEQ ID NO: 32.
Across the independent claims, the document claims dual CAR22/CAR20 expression from a single genome-incorporated exogenous nucleic acid using one promoter and a self-cleaving peptide, the corresponding isolated polynucleotide on one molecule, and a dual CAR immune cell further defined by CAR amino acid sequence identities and inactivated TCR alpha/beta/CD3 alleles and CD52 allele.
Stated Advantages
Increased IFN-γ release, consistent with dual CAR activity.
In vivo tumor burden control and survival in mouse lymphoma models.
Documented Applications
Treating a patient having a cancer associated with CD20 expression, CD22 expression, or CD20 expression and CD22 expression by administering an effective amount of the genetically engineered immune cell.
Treating cancers selected from lymphoma, Hodgkin lymphoma, non-Hodgkin lymphoma, leukemia, multiple myeloma, B-chronic lymphocytic leukemia, hairy cell leukemia, acute lymphocytic leukemia, acute lymphocytic cancer, or acute myeloid leukemia.
In vivo evaluation in mouse lymphoma models for tumor burden control and survival.
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