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EP-4741415-A2 - CHIMERIC MYD88 RECEPTORS FOR REDIRECTING IMMUNOSUPPRESSIVE SIGNALING AND RELATED COMPOSITIONS AND METHODS

EP4741415A2EP 4741415 A2EP4741415 A2EP 4741415A2EP-4741415-A2

Abstract

The present disclosure relates in some aspects to chimeric signaling receptors containing an extracellular domain capable of binding a molecule, such as an immunosuppressive cytokine, and a MyD88-containing intracellular domain capable of engaging a signaling pathway to activate an immune cell. In some aspects, the disclosure further relates to engineered cells, such as T cells, and compositions comprising the chimeric signaling receptors or engineered cells, and methods and uses thereof. In some embodiments, the cells may further express a genetically engineered recombinant antigen receptor directed against an antigen, such as a chimeric antigen receptor (CAR) or recombinant T cell receptor (TCR) or, in some cases, secrete a recombinant molecule, for example, a bispecific antibody.

Inventors

  • ASGHARZADEH, Shahab
  • MOGHIMI, Babak
  • HADJIDANIEL, Michael
  • SHIRINBAK, Soheila

Assignees

  • Simurx, Inc.
  • Children's Hospital Los Angeles

Dates

Publication Date
20260513
Application Date
20210716

Claims (15)

  1. A chimeric signaling receptor comprising, from N to C- terminus: (a) an extracellular domain that binds to an immunosuppressive cytokine selected from IL10, TGFβ, IL4, and IL1Ra; (b) a transmembrane domain; (c) a linker sequence; and (d) an intracellular signaling domain comprising two MyD88 polypeptides linked in tandem, wherein the two MyD88 polypeptides lack the full-length TIR domain.
  2. The chimeric signaling receptor of claim 1, wherein the extracellular domain is or contains an extracellular domain of a naturally occurring cytokine receptor or cytokine binding fragment thereof that is able to specifically bind the immunosuppressive cytokine.
  3. The chimeric signaling receptor of claim 1, wherein the extracellular domain is an antibody or an antigen binding fragment that is able to specifically bind the immunosuppressive cytokine.
  4. The chimeric signaling receptor of any of claims 1-3, wherein extracellular domain is: (i) an extracellular domain of TGFβ receptor (TGFβR) or a portion thereof that binds TGFβ; (ii) an extracellular domain of an IL10 receptor (IL10R) or a portion thereof that binds IL10; (iii) an extracellular domain of an IL4 receptor (IL4R) or a portion thereof that binds IL4; or (iv) an extracellular domain that is an antibody or antigen-binding fragment that binds to IL1Ra, a scFv directed against IL1Ra, or an sdAb directed against IL1Ra.
  5. The chimeric signaling receptor of any of claims 1, 2 or 4, wherein the extracellular domain: (i) is or comprises a portion of an amino acid sequence that exhibits at least or about 85%, 90%, 92%, 95%, or 97% sequence identity to SEQ ID NO: 20 and that binds to TGFβ; (ii) is or comprises a portion of an amino acid sequence that exhibits at least or about 85%, 90%, 92%, 95%, or 97% sequence identity to SEQ ID NO: 4 and that binds to TGFβ; (iii) is or comprises a portion of an amino acid sequence that exhibits at least or about 85%, 90%, 92%, 95%, or 97% sequence identity to SEQ ID NO: 119 and that binds to IL10; (iv) is or comprises a portion of an amino acid sequence that exhibits at least or about 85%, 90%, 92%, 95%, or 97% sequence identity to SEQ ID NO: 122 and that binds to IL10; or (v) is or comprises a portion of an amino acid sequence that exhibits at least or about 85%, 90%, 92%, 95%, or 97% sequence identity to SEQ ID NO: 125 and that binds to IL4.
  6. The chimeric signaling receptor of any of claims 1-5, wherein the transmembrane domain: (i) comprises the native transmembrane domain of the TGFβR; (ii) comprises the sequence of amino acids set forth in SEQ ID NO: 22 or a sequence of amino acids that exhibits at least or about 85%, at least or about 90%, at least or about 92%, at least or about 95%, at least or about 97% sequence identity to the sequence set forth in SEQ ID NO: 22; and/or (iii) is a heterologous transmembrane domain from a transmembrane protein other than the TGFβR.
  7. The chimeric signaling receptor of any of claims 1-6, wherein: (a) the two MyD88 polypeptides each comprise the death domain (DD), the intermediate domain (ID) and a portion of a TIR domain; and/or (b) (i) the two MyD88 polypeptides each independently is a sequence of amino acids selected from the group consisting of amino acids 2-155, 2-156, 2-157, 2-158, 2-159, 2-160, 2-161, 2-162, 2-163, 2-164, 2-165, 2-166, 2-167, 2-168, 2-169, 2-170, 2-171, 2-172, 2-173, 2-174, 2-175, 2-176, 2-177, 2-178, 2-179 or 2-180 of the full-length MyD88, optionally of SEQ ID NO:128; or (ii) wherein the two MyD88 polypeptides each independently is a sequence of amino acids 2-171 or 2-172 of the full-length MyD88, optionally of SEQ ID NO:128.
  8. The chimeric signaling receptor of any of claims 1-7, wherein: (a) the first MyD88 polypeptide is set forth in SEQ ID NO: 2; (b) the second MyD88 polypeptide is set forth in SEQ ID NO: 2; (c) the first MyD88 polypeptide and the second MyD88 polypeptide are identical; and/or (d) the first MyD88 polypeptide and the second MyD88 polypeptide are connected by a peptide linker, optionally wherein the peptide linker is set forth in SEQ ID NO: 47 ((GGGGS) n ), where n is an integer between 1 and 4, inclusive, optionally wherein the peptide linker is selected from the group consisting of SEQ ID NO:48 (GGGGS), SEQ ID NO: 85 (GGGGSGGGGS), SEQ ID NO: 49 (GGGGSGGGGSGGGGS), and SEQ ID NO: 50 (GGGGSGGGGSGGGGSGGGGS).
  9. A polynucleotide comprising a sequence of nucleotides encoding the chimeric signaling receptor of any of claims 1-8, optionally wherein: (a) the sequence of nucleotides is a first sequence of nucleotides and the polynucleotide further comprises a second sequence of nucleotides encoding another recombinant molecule, further optionally wherein the recombinant molecule is a recombinant antigen receptor, a cytokine, or a bispecific antibody, optionally wherein: (i) the recombinant antigen receptor is a recombinant T cell receptor (TCR) or is a chimeric antigen receptor (CAR); (ii) the cytokine is an interleukin or a functional portion thereof, further optionally wherein the interleukin is IL-2, IL-12, or IL-15 or a functional portion thereof; or (iii) the bispecific antibody is a bispecific T cell engager (BiTE).
  10. A vector comprising the polynucleotide of claim 9.
  11. A method of producing an engineered cell, the method comprising introducing the polynucleotide of claim 9 or the vector of any of claim 10 into a cell under conditions for expression of the chimeric signaling receptor on the surface of the cell, wherein the method is not a method for treatment of the human or animal body by surgery or therapy, or a diagnostic method practiced on the human or animal body.
  12. An engineered cell comprising a chimeric signaling receptor of any of claims 1-8 or the polynucleotide of claim 9, optionally wherein the engineered cell is a T cell, a tumor infiltrating lymphocyte (TIL), a B cell, a natural killer (NK) cell, or a macrophage.
  13. The engineered cell of claim 12, wherein the engineered cell further comprises a recombinant molecule, optionally wherein the recombinant molecule is a recombinant antigen receptor, a cytokine, or a bispecific antibody, further optionally wherein: (a) the recombinant antigen receptor is a recombinant T cell receptor (TCR) or is a chimeric antigen receptor (CAR); (b) the cytokine is an interleukin or a functional portion thereof, optionally wherein the interleukin is IL-2, IL-12, or IL-15 or a functional portion thereof; or (c) the bispecific antibody is a bispecific T cell engager (BiTE).
  14. A pharmaceutical composition comprising the engineered cell of claim 12 or claim 13, optionally further comprising a pharmaceutically acceptable excipient.
  15. The engineered cell of any of claims 12 to 13 or the pharmaceutical composition of claim 14 for use in a method of treating cancer in a subject, said method comprising administering the engineered cell of any of claims 12 to 13 or the pharmaceutical composition of claim 14 to the subject having cancer, optionally wherein the engineered cell of the pharmaceutical composition comprises an antigen receptor targeted against an antigen of the cancer.

Description

Cross-Reference to Related Applications This application claims priority from U.S. provisional application No. 63/053,529, filed July 17, 2020, entitled "CHIMERIC MYD88 RECEPTORS FOR REDIRECTING IMMUNOSUPPRESSIVESIGNALING AND RELATED COMPOSITIONS AND METHODS," the contents of which are incorporated by reference in their entirety. Incorporation by Reference of Sequence Listing The present application is being filed along with a Sequence Listing in electronic format. The Sequence Listing is provided as a file entitled 165282000140SeqList.txt, created July 10, 2021, which is 228,223 bytes in size. The information in the electronic format of the Sequence Listing is incorporated by reference in its entirety. Field The present disclosure relates in some aspects to chimeric signaling receptors containing an extracellular domain capable of binding a molecule, such as an immunosuppressive cytokine, and a MyD88-containing intracellular domain capable of engaging a signaling pathway to activate an immune cell. In some aspects, the disclosure further relates to engineered cells, such as T cells, and compositions comprising the chimeric signaling receptors or engineered cells, and methods and uses thereof. In some embodiments, the cells may further express a genetically engineered recombinant antigen receptor directed against an antigen, such as a chimeric antigen receptor (CAR) or recombinant T cell receptor (TCR) or, in some cases, secrete a recombinant molecule, for example, a bispecific antibody. Background Cell therapy strategies for treatment of diseases or conditions, such as cancers or tumors, include engineering immune cells to express genetically engineered recombinant antigen receptors, such as chimeric antigen receptors (CARs) or recombinant T cell receptors (TCRs), and administering compositions containing such cells to subjects. Improved strategies are needed to increase efficacy of the treatments, for example, by preventing and/or reversing immunosuppressive effects of cytokine signaling in tumor microenvironments, to improve the persistence, survival, and cytotoxicity of engineered cells. Provided are compositions, cells, and methods that meet such needs. Summary Provided herein are chimeric signaling receptor constructs that are able to redirect immunosuppressive signals by inducing a positive signal and T cell activation in response to an immunosuppressive cytokine. In provided aspects, the immunosuppressive cytokine is TGFβ and the chimeric signaling receptor contains an extracellular domain of a TGFβ receptor (TGFβR) or a portion thereof that binds TGFβ and MyD88 signaling domain(s) to mediate the positive signal in the cells upon binding of TGFβ to the extracellular domain of the chimeric signaling receptor. Thus, the provided chimeric signaling receptor can provide a positive signal to T cells even when the cells are present in an immunosuppressive environment, such as the TME, thereby improving the T cell activity, proliferation, persistence, or survival. In some embodiments, the chimeric receptor signaling receptor provided herein contains: (a) an extracellular domain of a TGFβ receptor (TGFβR) or a portion thereof that binds TGFβ; (b) a transmembrane domain; (c) a first truncated MyD88 polypeptide that lacks the full-length TIR domain of full-length MyD88; and (d) a second truncated MyD88 domain that lacks the full-length TIR domain of full-length MyD88. In some embodiments, the TGFβR is a TGFβR2. In some embodiments, the extracellular domain or the portion thereof contains: (i) the sequence of amino acids set forth in SEQ ID NO: 20; (ii) a sequence of amino acids that exhibits at least or about 85%, at least or about 90%, at least or about 92%, at least or about 95%, at least or about 97% sequence identity to the sequence set forth in SEQ ID NO: 20; or (iii) a portion of (i) or (ii) that binds TGFβ. In some embodiments, the extracellular domain or the portion thereof is set forth in SEQ ID NO:20. In some embodiments, the transmembrane domain contains the native transmembrane domain of the TGFβR. In some embodiments, the transmembrane domain contains the sequence of amino acids set forth in SEQ ID NO: 22 or a sequence of amino acids that exhibits at least or about 85%, at least or about 90%, at least or about 92%, at least or about 95%, at least or about 97% sequence identity to the sequence set forth in SEQ ID NO: 22. In some embodiments, the transmembrane domain is set forth in SEQ ID NO:22. In some embodiments, the transmembrane domain is a heterologous transmembrane domain from a transmembrane protein other than the TGFβR. In some embodiments, the transmembrane domain and the first truncated MyD88 polypeptide are directly linked. In some embodiments, the transmembrane domain and the first truncated MyD88 polypeptide are indirectly linked by a linker. In some embodiments, the linker is or comprises a peptide linker. In some embodiments, the linker is or contains a partial sequence of N-termin