Modified natural killer cells that express IL15 and uses thereof
The present invention provides, in certain aspects, a natural killer (NK) cell that expresses all or a functional portion of interleukin-15 (IL-15), and methods for producing such cells. The invention further provides methods of using a natural killer (NK) cell that expresses all or a functional portion of interleukin-15 (IL-15) to treat cancer in a subject or to enhance expansion and/or survival of NK cells.
1. A natural killer (NK) cell that expresses all or a functional portion of interleukin-15 (IL-15), wherein the all or a functional portion of the IL-15 is fused to all or a portion of a transmembrane protein that anchors the expressed IL-15 as a cell membrane-bound polypeptide (mbIL15) of the NK cell, and wherein the all or a functional portion of IL-15 promotes one or more of:
(i) NK cell survival,
(ii) regulation of NK cell and T cell activation and proliferation, and
(iii) support of NK cell development from hematopoietic stem cells.
2. The NK cell of claim 1 wherein the NK cells exhibits enhanced survival and expansion in the absence of IL-2, as compared to an NK cell not expressing mbIL-15.
3. The natural killer (NK) cell of claim 1 , wherein the NK cell is produced by a method comprising:
introducing a nucleic acid encoding all or a functional portion of IL-15 fused to all or a portion of a transmembrane protein into the NK cell; and
maintaining the NK cell under conditions in which the all or a functional portion of the IL-15 is expressed, thereby producing a NK cell that expresses the all or a functional portion of IL-15 fused to all or a portion of a transmembrane protein.
4. A composition comprising a Natural Killer (NK) cell and a physiologically acceptable carrier or excipient,
wherein the NK cell expresses all or a functional portion of interleukin-15 (IL-15),
wherein the all or a functional portion of the IL-15 is fused to all or a portion of a transmembrane protein that anchors the expressed IL-15 as a cell membrane-bound polypeptide (mbIL15) of the NK cell, and
wherein the all or a functional portion of IL-15 promotes one or more of:
(i) NK cell survival,
(ii) regulation of NK cell and T cell activation and proliferation, and
(iii) support of NK cell development from hematopoietic stem cells.
5. A pharmaceutical composition comprising a Natural Killer (NK) cell and a pharmaceutically acceptable carrier,
wherein the NK cell expresses all or a functional portion of interleukin-15 (IL-15),
wherein the all or a functional portion of the IL-15 is fused to all or a portion of a transmembrane protein that anchors the expressed IL-15 as a cell membrane-bound polypeptide (mbIL15) of the NK cell, and
wherein the all or a functional portion of IL-15 promotes one or more of:
(i) NK cell survival,
(ii) regulation of NK cell and T cell activation and proliferation, and
(iii) support of NK cell development from hematopoietic stem cells.
6. A pharmaceutical composition comprising a Natural Killer (NK) cell, a pharmaceutically acceptable carrier, and all or a functional portion of IL-2,
wherein the NK cell expresses all or a functional portion of interleukin-15 (IL-15),
wherein the all or a functional portion of the IL-15 is fused to all or a portion of a transmembrane protein that anchors the expressed IL-15 as a cell membrane-bound polypeptide (mbIL15) of the NK cell,
wherein the all or a functional portion of IL-15 promotes one or more of:
(i) NK cell survival,
(ii) regulation of NK cell and T cell activation and proliferation, and
(iii) support of NK cell development from hematopoietic stem cells, and
wherein the IL-2, or functional portion of IL-2, is present in an amount effective to increase expansion of the NK cell in vitro or in vivo.
7. A natural killer (NK) cell that expresses all or a functional portion of interleukin-15 (IL-15) as a membrane-bound polypeptide,
wherein the all or a functional portion of the IL-15 is fused to all or a portion of a transmembrane protein that anchors the expressed IL-15 as a cell membrane-bound polypeptide (mb15) of the NK cell,
wherein the transmembrane protein is selected from the group consisting of CD8α, CD4, CD3ε, CD3γ, CD3δ, CD3ζ, CD28, CD137, glycophorin A, glycophorin D, nicotinic acetylcholine receptor, a GABA receptor, FcεRIγ, and a T-cell receptor, and
wherein the NK cell exhibits enhanced survival and expansion in the absence of IL-2, as compared to NK cells not expressing membrane-bound IL15.
8. The NK cell of claim 7 , wherein the membrane-bound IL15 activates IL-15 signaling and/or anti-apoptotic signaling in an autocrine manner.
9. The NK cell of claim 7 , wherein the NK cell exhibits enhanced cytotoxicity against leukemia cells, lymphoma cells, and solid tumor cells as compared to a NK cell not expressing membrane-bound IL15.
10. The NK cell of claim 7 , wherein the enhanced survival and expansion is in vitro, and wherein the NK cell can be maintained in vitro for 2 months in the absence of exogenous IL-2.
11. The NK cell of claim 7 , wherein the enhanced survival and expansion is in vivo.
12. The NK cell of claim 7 , wherein the NK cell infiltrates one or more tissues to a greater extent as compared to a NK cell not expressing membrane-bound IL15, wherein the one or more tissues is selected from the group consisting of bone marrow, liver, spleen, kidney and lung, and wherein the NK cell expressing all or a functional portion of IL-15 as a membrane-bound polypeptide exhibits enhanced expression of one or more activation receptors as compared to a NK cell not expressing membrane-bound IL15, wherein the one or more activation receptors are selected from the group consisting of NKG2D, NKp44 (CD336) and NKp30 (CD337), CD16 and CD56.
13. The NK cell of claim 7 , wherein the NK cell exhibits enhanced phosphorylation of one or more of STAT1, STAT3 and STAT5, Src, Erk1/2 or Mek as compared to a NK cell not expressing membrane-bound IL15, and wherein the NK cell expressing all or a functional portion of IL-15 as a membrane-bound polypeptide exhibits enhanced anti-apoptotic signaling as compared to a NK cell not expressing membrane-bound IL15.
14. A Natural Killer (NK) cell that expresses all or a functional portion of interleukin-15 (IL-15), wherein the all or a functional portion of the IL-15 is fused to all or a portion of a transmembrane protein that anchors the expressed IL-15 as a cell membrane-bound polypeptide (mbIL15) of the NK cell, and wherein the all or a functional portion of IL-15 promotes one or more of:
(i) NK cell survival,
(ii) regulation of NK cell and T cell activation and proliferation, and
(iii) support of NK cell development from hematopoietic stem cells, and
wherein the all or a functional portion of the IL-15 comprises the amino acid sequence of SEQ ID NO: 4.
15. A Natural Killer (NK) cell that expresses all or a functional portion of interleukin-15 (IL-15), wherein the all or a functional portion of the IL-15 is fused to all or a portion of a transmembrane protein that anchors the expressed IL-15 as a cell membrane-bound polypeptide (mbIL15) of the NK cell, and wherein the all or a functional portion of IL-15 promotes one or more of:
(i) NK cell survival,
(ii) regulation of NK cell and T cell activation and proliferation, and
(iii) support of NK cell development from hematopoietic stem cells,
wherein the NK cell is produced by a method comprising:
introducing a nucleic acid encoding all or a functional portion of IL-15 fused to all or a portion of a transmembrane protein into the NK cell; and
maintaining the NK cell under conditions in which the all or a functional portion of the IL-15 is expressed, thereby producing a NK cell that expresses the all or a functional portion of IL-15 fused to all or a portion of a transmembrane protein, and wherein the nucleic acid comprises SEQ ID NO: 1.