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US-20260125438-A1 - RECOMBINANT erIL-15 NK CELLS

US20260125438A1US 20260125438 A1US20260125438 A1US 20260125438A1US-20260125438-A1

Abstract

Systems and methods are presented that provide for improved NK cell function. In preferred aspects, NK-92 cells express recombinant er/LSP-IL-15 to so render the NK-92 cells independent of exogenous cytokines and to provide extracellular immune stimulation.

Inventors

  • Patrick Soon-Shiong
  • Shahrooz Rabizadeh
  • Kayvan Niazi
  • Hans G. Klingemann

Assignees

  • NANTCELL, INC.
  • NANTBIO, INC.
  • IMMUNITYBIO, INC.

Dates

Publication Date
20260507
Application Date
20260105

Claims (20)

  1. 1 . A method of stimulating a cell mediated immune response in a patient, the method comprising administering to the patient an effective amount of a composition comprising a genetically modified natural killer (NK) cell, wherein the genetically modified NK cell comprises a recombinant nucleic acid comprising a first segment encoding a long signal peptide LSP variant of Interleukin-15 (IL-15) as set forth in SEQ ID NO: 5, wherein the variant comprises an endoplasmic reticulum-retention signal (erLSP-IL-15).
  2. 2 . The method of claim 1 , wherein the composition further comprises at least one agent selected from the list consisting of: a drug that breaks into the tumor microenvironment (TME), a drug that reduces immune suppression, a drug that stimulates immune competent cells, a cancer vaccine, and a drug that promotes memory cell development.
  3. 3 . The method of claim 2 , wherein the drug that breaks into the TME comprises abraxane.
  4. 4 . The method of claim 2 , wherein the drug that reduces immune suppression is cyclophosphamide.
  5. 5 . The method of claim 2 , wherein the drug that stimulates immune competent cells is Alt-803.
  6. 6 . The method of claim 2 , wherein the cancer vaccine is an adenoviral V (Adv) based vaccine comprising a nucleic acid encoding at least one of a tumor associated antigen, a tumor specific antigen, or a neoantigen.
  7. 7 . The method of claim 2 , wherein the drug that promotes memory cell development is a tumor targeted IL-12 fusion construct.
  8. 8 . The method of claim 1 , wherein the NK cells are formulated for intravenous (IV) administration.
  9. 9 . The method of claim 1 , wherein the NK cells are formulated for administration at a dose between 5×10 7 cells/dose IV and 5×10 10 cells/dose IV.
  10. 10 . The method of claim 1 , wherein the nucleic acid comprises a second segment encoding at least one of a CD16, a chimeric antigen receptor (CAR), or an immune modulatory compound.
  11. 11 . The method of claim 10 , wherein the immune modulatory compound comprises a compound that interferes with checkpoint inhibition, is an immune stimulator, or inhibits a cytokine involved in immune suppression.
  12. 12 . The method of claim 10 , wherein the CAR comprises an ectodomain having binding specificity against a tumor associated antigen, a tumor specific antigen, or a neoepitope.
  13. 13 . The method of claim 1 , wherein the NK cell is an autologous NK cell.
  14. 14 . The method of claim 1 , wherein the NK cell is an NK-92 cell.
  15. 15 . A method of stimulating a cell mediated immune response in a patient, the method comprising administering to the patient an effective amount of a composition comprising a genetically modified natural killer NK cell, wherein the genetically modified NK cell comprises a recombinant nucleic acid comprising 1) a first segment encoding a long signal peptide LSP variant of Interleukin-15 (IL-15) as set forth in SEQ ID NO: 5, wherein the variant comprises an endoplasmic reticulum-retention signal (erLSP-IL-15); and 2) a second segment encoding a chimeric antigen receptor (CAR).
  16. 16 . The method of claim 15 , wherein the composition further comprises Alt-803.
  17. 17 . The method of claim 15 , wherein the NK cells are formulated for intravenous (IV) administration.
  18. 18 . The method of claim 15 , wherein the NK cells are formulated for administration at a dose between 5×10 7 cells/dose IV and 5×10 10 cells/dose IV.
  19. 19 . The method of claim 15 , wherein the CAR comprises an ectodomain with binding specificity against a tumor-associated antigen, a tumor-specific antigen, or a neoepitope.
  20. 20 . The method of claim 15 , wherein the CAR comprises binding specificity for PD-1, PD-L1, CTLA-4, CD19, CEA, MUC-1, CYPB1, PSA, Her-2, or Brachyury.

Description

This application is a continuation application of U.S. patent application Ser. No. 17/732,190 which was filed on Apr. 28, 2022, which is a continuation application of U.S. patent application Ser. No. 17/438,386, which was filed Sep. 10, 2021, and which is a 371 application of International application with the serial number PCT/US2020/19991, which was filed Feb. 27, 2020, and which claims the benefit of U.S. provisional patent application with the Ser. No. 62/819,256, filed on Mar. 15, 2019, all of which are incorporated herein by reference in their entirety. SEQUENCE LISTING The content of the XML file of the sequence listing named SEQ_102538.0075US3.XML, which is 12,111 bytes in size was created on Jan. 5, 2026 and electronically submitted via EFS-Web along with the present application is incorporated by reference in its entirety. FIELD OF THE INVENTION The present disclosure relates to genetically modified immune cells expressing IL-15, especially as they relate to NK cells that express and intracellularly retain a modified IL-15 and that further express at least one of a high affinity variant of CD16 and a CAR (chimeric antigen receptor). BACKGROUND OF THE INVENTION The background description includes information that may be useful in understanding the present disclosure. It is not an admission that any of the information provided herein is prior art or relevant to the presently claimed invention, or that any publication specifically or implicitly referenced is prior art. All publications and patent applications herein are incorporated by reference to the same extent as if each individual publication or patent application were specifically and individually indicated to be incorporated by reference. Where a definition or use of a term in an incorporated reference is inconsistent or contrary to the definition of that term provided herein, the definition of that term provided herein applies and the definition of that term in the reference does not apply. NK-92 cells are desirable in various aspects of cell-based therapies as they lack apparent toxicity against allogeneic cells in a recipient and have a relatively broad spectrum of cytotoxic activity towards a wide range of tumor cells. Moreover, NK-92 cells can be cultivated in a relatively simple manner and as such present an attractive option for adoptive cancer immunotherapy. Unfortunately, proliferation and function of NK-92 cells are highly dependent on IL-2, which increases cost where NK-92 cells are needed at large scale. To circumvent problems associated with such cytokine requirements, NK-92 cells have been transformed to express and intracellularly retain IL-2 (see e.g., Exp Hematol 33:159-164). While such modified cells indeed became independent of exogenous cytokines, various disadvantages remained. Among other things, IL-2 released from such modified cells may increase IL-2 mediated effects in vivo where such cells are used in a mammal, which is particularly undesirable where IL-2 stimulates immune suppression in a tumor microenvironment (typically via growth and expansion of myeloid derived suppressor cells (MDSCs) and T-regulatory cells (Tregs)). In another example, NK-92 cells were transfected to express IL-15 from a cDNA that was cloned into a pcDNA3 expression vector (see e.g., Haematologica, 2004; 89:338-347), and so transfected cells continuously produced high levels of IL-15 in the culture supernatant, which was thought to make the cells proliferate significantly more rapidly in response to stimulation with low doses of IL-2 or IL-15. Moreover, the cumulative number of cells in long-term culture was also significantly higher than with non-transfected cells. However, where such cells are used in vivo, the high levels of secreted IL-15 may become clinically problematic. Similarly, NK-92 cells were transformed using a viral transfection system to express a recombinant native form of IL-15 (see e.g., Cancer Immunol Immunother (2012) 61:1451-1461). While such recombinant cells were able to grow in the absence of exogenous cytokines and expressed a recombinant CAR, transfection efficacy was relatively low, produced relatively low quantities of IL-15 intracellularly, and secreted low quantities of IL-15 into the culture medium. Moreover, cytotoxicity of the recombinant cells was reduced as compared to the parental NK-92 cell line. Notably, where the same IL-15 was expressed from a plasmid, so generated NK-92 cells were not fully independent from exogenous growth factors, thus limiting in vivo use of such recombinant cells. Thus, even though various modified immune cells, and especially modified NK cells are known in the art, all or almost all of them suffer from various disadvantages. Consequently, there is a need to provide improved modified NK cells exhibit desirable growth characteristics while maintaining targeted cytotoxicity. SUMMARY OF THE INVENTION Various recombinant cells, compositions, and methods are disclosed herein where an NK c