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CN-122003245-A - Genetically engineered mucosa-associated constant T (MAIT) cells for adoptive transfer cell therapy

CN122003245ACN 122003245 ACN122003245 ACN 122003245ACN-122003245-A

Abstract

The present disclosure relates generally to the field of genetic engineering of immune cells, and in particular to mucosal-related constant T (MAIT) cells genetically engineered to express exogenous T Cell Receptors (TCRs) and uses thereof. More specifically, the invention relates in its embodiments to cell compositions adapted for adoptive transfer cell therapy (ACT) providing improved therapeutic modalities.

Inventors

  • A. McClenkin
  • Mikal Lautem
  • H. L. Shakeed
  • Hillary Klein Ben Silbermann

Assignees

  • 普拉里生物科技有限公司
  • 哈达西特医疗研究服务和开发有限公司

Dates

Publication Date
20260508
Application Date
20240709
Priority Date
20230710

Claims (20)

  1. 1. A cell composition comprising a population of engineered mucosa-associated constant T (MAIT) cells that express an exogenous T Cell Receptor (TCR), wherein the MAIT cells are derived from the placenta, the cell composition optionally further comprising a pharmaceutically acceptable carrier.
  2. 2. The cell composition of claim 1, wherein the MAIT cells are derived from placental inter-villus blood (IVB).
  3. 3. The cell composition according to claim 1 or 2, which is adapted for adoptive transfer cell therapy (ACT).
  4. 4. The cell composition of any one of the preceding claims, comprising 10 9 -10 11 living cells of the engineered MAIT cell population.
  5. 5. The cell composition of any one of the preceding claims, comprising at least 90% tcrvα7.2 + CD161 High height cells.
  6. 6. The cell composition of any one of the preceding claims, wherein the TCR recognizes a tumor antigen.
  7. 7. The cell composition of claim 6, wherein the tumor antigen is selected from the group consisting of ：NY-ESO-1、KRAS、p53、PIK3CA、PTEN、ERBB2 (HER2)、AFP、KK-LC-1、RAC1-P29S、LAGE-1A、COL6A3、HA-2、HERV-E、BRAF、gp100、alpha- fetal protein, desmosomal binding protein/AHNAKS 2580F, cancer/testis antigen 1, ERBB2H473Y, ERBB IPE805G, minor H antigen (HA-1), PRAME, PSMA, TPBG, 5T4, MAGEA1, MAGE-A3/A6, MAGEA4/8, melan-a/MART-1, NRAS, and Wilms tumor 1 (WT-1).
  8. 8. The cellular composition of claim 7, wherein the tumor antigen is selected from the group consisting of NY-ESO-1 and age-1A.
  9. 9. The cell composition of any one of the preceding claims, wherein the TCR is capable of specifically binding to an epitope presented by HLA-A 2.
  10. 10. The cell composition of claim 9, wherein the TCR comprises a TCR a chain comprising CDR1 having the amino acid sequence of SEQ ID No. 1, CDR2 having the amino acid sequence of SEQ ID No. 2, and CDR3 having the amino acid sequence of SEQ ID No. 3, and a TCR β chain comprising CDR1 having the amino acid sequence of SEQ ID No. 4, CDR2 having the amino acid sequence of SEQ ID No. 5, and CDR3 having the amino acid sequence of SEQ ID No. 6.
  11. 11. The cell composition of claim 11, wherein the TCR comprises a TCR a chain having an amino acid sequence as set forth in SEQ ID No. 7, optionally excluding the signal peptide of SEQ ID No. 7 at positions 1-20, and a TCR β chain having an amino acid sequence as set forth in SEQ ID No. 8, optionally excluding the signal peptide of SEQ ID No. 8 at positions 1-21.
  12. 12. The cell composition according to any one of the preceding claims for use in therapy.
  13. 13. The cell composition for use according to claim 12, wherein the use is the treatment of a subject with a tumor or malignancy.
  14. 14. The cell composition for use according to claim 13, wherein the subject has a tumor selected from the group consisting of melanoma, breast cancer, colon cancer, kidney cancer, liver cancer, lung cancer, ovarian cancer, pancreatic cancer, prostate cancer, uterine cancer, cervical cancer, bladder cancer, gastric cancer, head and neck cancer, brain cancer, skin cancer and sarcoma.
  15. 15. The cell composition for use according to claim 13 or 14, wherein the subject has a treatment resistant tumor or is otherwise unsuitable for treatment with an immunotherapy comprising Chimeric Antigen Receptor (CAR) T cells and/or therapeutic antibodies.
  16. 16. The cell composition for use according to claim 15, wherein the use is in the treatment of a subject suffering from a condition associated with the expression of HLA class I restricted antigens.
  17. 17. The cell composition for use according to any one of claims 12 to 16, wherein the antigen is a surface expressed low density antigen characterized by less than 50 antigen molecules per cell.
  18. 18. A cellular composition adapted for adoptive transfer cell therapy (ACT), the composition comprising a substantially purified population of mucosa-associated constant T (MAIT) cells engineered to express an exogenous T Cell Receptor (TCR), wherein the TCR recognizes a tumor antigen selected from the group consisting of NY-ESO-1 and rage-1A.
  19. 19. The cell composition of claim 18, wherein the TCR comprises a TCR a chain comprising CDR1 having the amino acid sequence of SEQ ID No.1, CDR2 having the amino acid sequence of SEQ ID No. 2, and CDR3 having the amino acid sequence of SEQ ID No. 3, and a TCR β chain comprising CDR1 having the amino acid sequence of SEQ ID No. 4, CDR2 having the amino acid sequence of SEQ ID No.5, and CDR3 having the amino acid sequence of SEQ ID No. 6.
  20. 20. The cell composition of claim 19, wherein the TCR comprises a TCR a chain having an amino acid sequence as set forth in SEQ ID No. 7, optionally excluding the signal peptide of SEQ ID No. 7 at positions 1-20, and a TCR β chain having an amino acid sequence as set forth in SEQ ID No. 8, optionally excluding the signal peptide of SEQ ID No. 8 at positions 1-21.

Description

Genetically engineered mucosa-associated constant T (MAIT) cells for adoptive transfer cell therapy Technical Field The present disclosure relates generally to the field of genetic engineering of immune cells. More specifically, the present disclosure provides mucosal-related constant T (MAIT) cells genetically engineered to express exogenous T Cell Receptors (TCRs) and uses thereof. Background Mucosal associated constant T (MAIT) cells were first identified as populations of αβ T cells in humans, mice and cattle, enriched in the double negative (CD 4 -CD8-) subset, expressing constant vα7.2-jα33T cell receptor (TCR) in humans. The term MAIT is established due to the relative enrichment of these T cells within mucosal tissue. The limiting factor of MAIT cells is the MHC-1B molecule MR1 (MHC associated protein 1), which presents intermediates of the vitamin B (both riboflavin (vitamin B2) and folic acid (vitamin B9) synthesis pathways to MAIT cells. Strong stimulatory ligands in the riboflavin synthesis pathway include 5- (2-oxopropyleneamino) -6-D-ribosyl amino uracil (5-OP-RU) and 5- (2-oxoethyleneamino) -6-D-ribosyl amino uracil (5-OE-RU) produced by a wide variety of bacteria, mycobacteria, and yeasts during riboflavin synthesis. This pathway is not present in mammals and thus its immunological detection allows for efficient host-pathogen discrimination. It has been found that MAIT cells can recognize cells infected with bacteria and produce IFN-gamma in response. MAIT cells have been shown to protect mice from bacterial infection. Early phenotypic work established that human MAIT cells are CD8 + or double negative, have a major CCR7 - effector memory phenotype, and express high levels of CD161. The detailed phenotype of MAIT cells also demonstrates that they share several features with constant natural killer T (iNKT) cells, including the expression of PLZF, a transcription factor that controls the innate-like functionality of iNKT cells. Expression of PLZF also confers naive functionality to the MAIT cells, as demonstrated by the ability of cytokines to induce IFN- γ production in the absence of TCR stimulation. Subsequent studies found that human MAIT cells do not express a single constant TCR, but instead express a restricted TCR including V.alpha.7.2-J.alpha.33, V.alpha.7.2-J.alpha.12 or V.alpha.7.2-J.alpha.20, which is primarily associated with a limited pool (limited repertoire) of human beta chains such as V.beta.2/V.beta.13. Thus, the current understanding regards MAIT cells as T cells that (a) express a semi-constant V.alpha.7.2-J.alpha.33/12/20 TCR, (B) are activated by microbial vitamin B metabolite antigens presented by MR1 to perform both type 1 and type 17 effector functions, and (c) exhibit congenital-like characteristics, controlled by the expression of PLZFs, including the ability to be activated by cytokines that do not rely on their TCRs. MAIT cells are in contrast to conventional T cells, which possess highly variable TCRs capable of targeting a large number of peptide epitopes produced by viruses, bacteria and malignant cells. Thus, conventional T cells have precise specificity for a single peptide, and a single clone can undergo large-scale expansion to provide T cell memory. However, the frequency of any single peptide-specific T cell will be very low when it first encounters a pathogen. In contrast, the MAIT cell TCR provides an innate ability to respond to a particular ligand set without the need for amplification. Several properties of MAIT cells suggest that they play a fundamental role in mammalian immunity. First, MAIT cells have an intrinsic effector-memory phenotype, typically CD45RA -CD45RO+CD95HiCD62LLoCD44Hi, with the ability to rapidly secrete several pro-inflammatory cytokines. Second, MAIT cells are very abundant in human tissue, typically comprising 1-4% of all T cells in peripheral blood and up to 10% of airway T cells and 20-40% of hepatic T cells. Furthermore, since each TCR recognizes the same ligand, the number of MAIT cells will be significantly greater than the number of conventional antigen-specific T cells in response to the cognate antigen in the early stages of the immune response. Recent studies using single cell RNA sequencing technology and immunological technology revealed that MAIT cells exhibit significant heterogeneity, which summarizes conventional T cell biology. This significant heterogeneity has been shown to include the different CD4 + and CD8 + lineages, as well as "killing", "helper", and "modulating" cell phenotypes-an indication that a MAIT cell is performing a complex function. The conservation and abundance of MAIT cells may be explained by their broad functionality due to different activation patterns, each triggering a different transcriptome program. Because of their ability to respond to diverse functions in diverse immunological contexts, these interesting cells now appear to be multifunctional effectors, at a core location at the i