- Document Number:
20250041344
- Appl. No:
18/793364
- Application Filed:
August 02, 2024
- Abstract:
The present disclosure provides gene editing methods for modulating the expression of an inhibitor of DNA-binding E-protein transcription factors, namely Id3, and thereby affecting T cell function. First, it provides experimental evidence that Id3 is critical to the persistence and function of tissue-infiltrating GVHD T cells in a mouse model. Id3 reduces chromatin accessibility (ChrAcc) of transcription factors (TFs) that drive T cell PD-1 transcription, differentiation and dysfunction. Id3 loss increases PD-1 expression and impairs tissue-infiltrating Th1 cells. Second, it provides proof-of-concept that targeting ID3 in human T cells using a CRIPSR/Cas9 knockout (KO) prevents xeno-GVHD but preserves the anti-leukemic activity of chimeric antigen receptor (CAR)-T cells. Third, it provides experimental evidence that ectopic expression of Id3 in engineered human CAR-T cells enhances the ability of these cells to eliminate tumors.
- Assignees:
Hackensack Meridian Health, Inc. (Edison, NJ, US)
- Claim:
1. An immunotherapy method for treating a recipient subject with a hematologic cancer comprising administering to the recipient subject an activated and expanded purified population of genetically engineered CD3+ T cells derived from a healthy donor, a. wherein the genetic engineering of the donor CD3+ T cells in vitro reduces expression of Id3, an inhibitor of DNA-binding E-protein transcription factors, by at least 25%, compared to a control; and b. wherein the method poses a decreased risk of graft versus host reaction while preserving graft versus tumor immunity in the recipient subject.
- Claim:
2. The immunotherapy method according to claim 1, wherein the administering is by infusion.
- Claim:
3. The immunotherapy method according to claim 1, wherein a. The subject is a mammal; or b. The subject is a human.
- Claim:
4. The immunotherapy method according to claim 1, wherein the hematologic cancer is a leukemia, a myelodysplastic neoplasm, a myeloma, or a lymphoma.
- Claim:
5. The immunotherapy method according to claim 1 wherein the donor T cells are allogeneic to the recipient subject.
- Claim:
6. The immunotherapy method according to claim 1, wherein a. the CD3+ T cells are purified from mononuclear cells collected from umbilical cord blood or adult peripheral blood; and b. the CD3+ T cells comprise CD4+ T cells, CD8+ T cells or both; and c. the CD3+ T cells comprising the edited Id3 gene are expanded and activated in vitro in presence of a cytokine.
- Claim:
7. The immunotherapy method according to claim 6, wherein the cytokine is selected from the group consisting of IL-2, IL-7, IL-15, IL-18, IL-21, or a combination thereof.
- Claim:
8. The method according to claim 1, wherein the reducing of the expression of the Id3 gene of the CD3+ T cells is accomplished by CRISPR/Cas9.
- Claim:
9. The method according to claim 6, wherein the activated and expanded purified population of genetically engineered CD3+ T cells comprising CD4+ T cells, CD8+ T cells or both comprising the edited Id3 gene is characterized by an improved ability to secrete effector cytokines, an improved cytotoxicity, or both against tumor cells compared to a control population of mononuclear cells.
- Claim:
10. An immunotherapy method for treating a recipient subject with a hematologic cancer comprising a. genetically engineering a population of CD3+ T cells derived from a healthy donor: i. to express a chimeric antigen receptor (CAR) that specifically binds a tumor antigen; and ii. to ectopically express Id3 (Id30E); b. activating and expanding the purified population of genetically engineered CD3+CAR−, IdOE T-cells of (a); and c. administering the activated and expanded purified population of genetically engineered population of CD3+CAR, IdOE T-cells of (b) wherein the cell population is characterized by: a lower frequency of terminally exhausted Tcells of phenotype PD-1+TIM3+; or a higher frequency of progenitor exhausted cells (TPEX) of phenotype PD1+TIM3−; or an enhanced persistence and enhanced ability to expand in vitro; or a higher frequency of cells of a central memory cell phenotype (CD62+CD45RA−) upon ex vivo culture in IL-2, IL-7 and IL-15; or an enhanced ability to produce IL-2 and to proliferate upon antigen challenge in ex vivo culture; or augmented memory protection against tumor challenge in a mouse leukemia model; or improved overall survival; or a combination thereof, compared to control CAR-T cells.
- Claim:
11. The immunotherapy method according to claim 10, wherein a. the genetic engineering to express a CAR comprises transducing the CD3+ T cells with a retroviral vector comprising a nucleic acid encoding a synthetic CAR to stably express the CAR; and b. the CAR comprises an extracellular antigen recognition domain, a spacer/hinge region and transmembrane domain, and an intracellular signal transduction domain; and c. the therapeutic dose of the CAR-T cells is about 1×10E6 to 20×10E6 CAR-T cells/m2 body surface area.
- Claim:
12. The immunotherapy method according to claim 11, wherein a. the extracellular antigen recognition domain of the CAR comprising an scFv fragment derived from a monoclonal antibody binds specifically to CD19, CD20, CD22, CD33, or CD30; and b. the intracellular signal transduction domain of the CAR comprises a CD3ζ activation chain and one or more costimulatory molecules.
- Claim:
13. The immunotherapy method according to claim 12, wherein the costimulatory molecule comprises 4-1BB.
- Claim:
14. The immunotherapy method according to claim 10, further comprising administering a short course of chemotherapy to reduce the T cell population of the subject prior to the administering of the population of CAR-T ID3OE cells.
- Claim:
15. The method according to claim 10, wherein the human CAR-T cells engineered to ectopically express Id3 have an enhanced ability to eliminate tumors compared to a CAR-T cell control that does not ectopically express Id3.
- Claim:
16. The immunotherapy method according to claim 1, wherein the administering is by infusion.
- Claim:
17. The immunotherapy method according to claim 1, wherein a. The subject is a mammal; or b. The subject is a human.
- Claim:
18. The immunotherapy method according to claim 10, wherein the hematologic cancer is a leukemia, a myelodysplastic neoplasm, a myeloma, or a lymphoma.
- Claim:
19. The method according to claim 10 wherein the donor T cells are allogeneic to the recipient subject.
- Claim:
20. The method according to claim 10, wherein a. the CD3+ T cells are purified from mononuclear cells collected from umbilical cord blood or adult peripheral blood; b. the CD3+ T cells comprise CD4+ T cells, CD8+ T cells or both and c. the CD3+ T cells are expanded and activated in vitro in presence of a cytokine selected from IL-2, IL-7, IL-15, IL-18, IL-21, or a combination thereof.
- Claim:
21. The method according claim 10, further comprising administering an additional agent.
- Claim:
22. The method according to claim 21, wherein the additional agent comprises: an approved immune checkpoint inhibitor at a dose standard for the cancer indication, or rituxuximab (anti-CD20); or alemtuzumab (antiCD52); or epratuzumab (anti-CD-22); or a clinical grade alpha-1-antitrypsin.
- Claim:
23. The method according to claim 22, wherein the immune checkpoint inhibitor is an anti-PD-1 inhibitor; an anti-PD-L1 inhibitor, or an anti-CTLA-4 inhibitor.
- Claim:
24. The method according to claim 23, wherein the anti-PD-1 inhibitor is lambrolizumab/pembrolizumab or nivolumab.
- Claim:
25. The method according to claim 23, wherein the anti-PDL-1 inhibitor is atezolizumab.
- Claim:
26. The method according to claim 23, wherein the anti-CTLA-4 inhibitor is ipilimumab.
- Current International Class:
61; 61; 61; 61; 07; 12; 12; 12; 12; 12
- Accession Number:
edspap.20250041344
Processing Request
No Comments.