# US 2021/0275592 A1 · Immune Cells with DNMT3A Gene Modifications and Methods Related Thereto > Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt - Number: US 2021/0275592 A1 - Document ID: US20210275592A1 - Status: Published application - Kind: A1 - Application number: 17/330,378 - Filed: 2021-05-25 - Published: 2021-09-09 - Earliest claimed: 2015-11-04 - Assignees: Emory University; St. Jude Children's Hospital - Applicants: Emory University; St. Jude Children's Hospital, Inc. - Inventors: Rafi Ahmed; Benjamin Youngblood - Web page: https://patentgrep.com/patent/US20210275592A1 ## CPC - C12N 9/1007: {Methyltransferases (general) (2.1.1.)} - A61K 35/17: Lymphocytes; B-cells; T-cells; Natural killer cells; Interferon-activated or cytokine-activated lymphocytes (when activated by a specific antigen A61K39/00) - A61K 40/11: T-cells, e.g. tumour infiltrating lymphocytes [TIL] or regulatory T [Treg] cells; Lymphokine-activated killer [LAK] cells - A61K 40/31: Chimeric antigen receptors [CAR] - A61K 40/4211: {CD19 or B4} - A61P 31/14: for RNA viruses - A61P 35/00: Antineoplastic agents - C07K 14/005: from viruses - C07K 14/70596: {Molecules with a "CD"-designation not provided for elsewhere} - C12N 5/0636: {T lymphocytes} - C12N 5/0638: {Cytotoxic T lymphocytes [CTL] or lymphokine activated killer cells [LAK]} - C12Y 201/01037: DNA (cytosine-5-)-methyltransferase (2.1.1.37) - C12N 2510/00: Genetically modified cells - C12N 2740/16033: Use of viral protein as therapeutic agent other than vaccine, e.g. apoptosis inducing or anti-inflammatory - C12N 2740/16111: concerning HIV env - C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00) - A61K: PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES (devices or methods specially adapted for bringing pharmaceutical products into particular physical or administering forms A61J3/00; chemical aspects of, or use of materials for deodorisation of air, for disinfection or sterilisation, or for bandages, dressings, absorbent pads or surgical articles A61L; soap compositions C11D) - A61P: SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS - C07K: PEPTIDES (peptides in foodstuffs A23; obtaining protein compositions for foodstuffs, working-up proteins for foodstuffs A23J; preparations for medicinal purposes A61K; peptides containing beta-lactam rings C07D; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, C07D; ergot alkaloids of the cyclic peptide type C07D519/02; macromolecular compounds having statistically distributed amino acid units in their molecules, i.e. when the preparation does not provide for a specific; but for a random sequence of the amino acid units, homopolyamides and block copolyamides derived from amino acids C08G69/00; macromolecular products derived from proteins C08H1/00; preparation of glue or gelatine C09H; single cell proteins, enzymes C12N; genetic engineering processes for obtaining peptides C12N15/00; compositions for measuring or testing processes involving enzymes C12Q; investigation or analysis of biological material G01N33/00) - C12Y: ENZYMES ## Abstract This disclosure relates to the genetic modification of DNMT3A gene in immune cells. In certain embodiments, the modified immune cells may be used in adoptive T cells therapies to enhance immune responses against cancer or chronic infections. In certain embodiments, the disclosure relates to deleting, changing, or inserting nucleotides within the DNMT3A gene in immune cells, e.g., human CD8 T cells, such that the DNMT3A gene product does not function for methylation. In certain embodiments, modification of the DNMT3A gene provides an improvement in antigen-specific T cells functions and/or an enhancement of the longevity of the cells. ## Claims ### Claim 1 A method of treating cancer comprising a) collecting CD8 T cells from a subject diagnosed with cancer providing isolated CD8 T cells; b) modifying a DNMT3a gene in the isolated CD8 T cells such that the DNMT3a gene does not expresses a protein or a non-functional protein is expressed providing CD8 T cells with a non-functioning DNMT3a gene; and c) administering or implanting an effective amount of the CD8 T cells with a non-functioning DNMT3a gene into the subject diagnosed with cancer. ### Claim 2 (depends on 1) The method of claim 1, wherein the CD8 T cells with an non-functioning DNMT3a gene is further mixed with a recombinant virus that encodes a chimeric tumor associated antigen receptor under conditions such that the chimeric tumor associated antigen receptor is capable of expression on the surface of the CD8 T cells and capable of binding the tumor associated antigen. ### Claim 3 (depends on 1) The method of claim 1, wherein the isolated CD8 T cells is further mixed with a recombinant virus that encodes a tumor antigen receptor under conditions such that the tumor antigen receptor is capable of expression on the surface of the CD8 T cells and binding tumor cells. ### Claim 4 (depends on 1) The method of claim 1, wherein modifying a DNMT3A gene in the isolated CD8 T cells is done by making a double stranded cut of the DNMT3A gene under conditions such that repair of the double stranded results in a non-functioning mutation in the DNMT3A gene. ### Claim 5 (depends on 4) The method of claim 4, wherein the mutation results in an insertion, replacement or deletion of at least on nucleotide. ### Claim 6 (depends on 4) The method of claim 4, wherein the double stranded cut is created by a zinc finger nuclease. ### Claim 7 (depends on 4) The method of claim 4, wherein the double stranded cut is created by a Cas protein conjugated to a guide RNA. ### Claim 8 (depends on 1) The method of claim 1, wherein the subject is diagnosed with a hematological cancer. ### Claim 9 (depends on 8) The method of claim 8, wherein the hematological cancer is leukemia, lymphoma, or multiple myeloma. ### Claim 10 (depends on 1) The method of claim 1 further comprising administering to the subject an anti-CTLA-4 antibody, anti-PD-1 antibody, anti-PD-L1 antibody, or combinations thereof.