Search

JP-7855638-B2 - Viral vectors and packaging cell lines

JP7855638B2JP 7855638 B2JP7855638 B2JP 7855638B2JP-7855638-B2

Inventors

  • アンドリュー シャーレンバーグ
  • ローリー バイツ

Assignees

  • ウモジャ バイオファーマ インコーポレイテッド

Dates

Publication Date
20260508
Application Date
20240524
Priority Date
20180412

Claims (19)

  1. A composition comprising lentiviral particles for activating and efficiently transducing T cells, comprising a nucleic acid sequence encoding a small molecule controllable T cell/NK cell activating receptor, wherein the nucleic acid sequence is operably linked to a promoter, and the small molecule controllable T cell/NK cell activating receptor is (i) A first polypeptide chain comprising an extracellular FK506-binding protein (FKBP) domain bound to an intracellular IL2Rβ signaling domain via a transmembrane domain; and (ii) A second polypeptide chain containing an extracellular FKBP12-rapamycin-binding protein (FRB) domain bound to the intracellular IL2Rγ signaling domain via a transmembrane domain. Includes, The T cell/NK cell activation receptor can be activated by a small molecule, and the small molecule includes rapamycin or rapalog. The composition further comprises a lentiviral particle having a viral surface containing an anti-CD3 antibody or scFv.
  2. The composition according to claim 1, wherein the virus surface further comprises a 41bb ligand.
  3. The composition according to claim 1 or 2 , wherein the lentiviral particles further comprise a nucleic acid sequence encoding a protein that provides resistance to immunosuppressants.
  4. The composition according to claim 3 , wherein the immunosuppressant is selected from the group consisting of methotrexate, rapamycin, rapalog, tacrolimus, and cyclosporine.
  5. The composition according to any one of claims 1 to 4 , wherein the lentiviral particles further comprise a nucleic acid sequence encoding a TGF beta-dominant-negative inhibitory receptor.
  6. A combination for use in a method for treating a subject suffering from cancer, comprising the composition according to any one of claims 1 to 5 and the small molecule, wherein the method is A combination comprising a) administering the composition to the subject, and b) administering the small molecule to the subject, characterized in that the cancer is treated in the subject.
  7. A combination for use in a method for promoting the proliferation of T cells capable of recognizing and killing tumor cells in a target requiring such proliferation, comprising the composition according to any one of claims 1 to 5 and the small molecule, wherein the method is A combination comprising i) administering the composition to the subject, and b) administering the small molecule to the subject, characterized in that T cells capable of recognizing and killing tumor cells in the subject proliferate.
  8. The composition according to any one of claims 1 to 5, or the combination according to claim 6 or 7 , wherein the extracellular FKPB domain shares at least 95% sequence identity with SEQ ID NO: 13, the intracellular IL2Rβ signaling domain shares at least 95% sequence identity with SEQ ID NO: 14 , the extracellular FRB domain shares at least 95% sequence identity with SEQ ID NO: 16 , and the intracellular IL2Rγ signaling domain shares at least 95% sequence identity with SEQ ID NO: 17 .
  9. (a) The first polypeptide chain shares at least 95% sequence identity with SEQ ID NO: 12, (b) The second polypeptide chain shares at least 95% sequence identity with SEQ ID NO: 15 The composition or combination according to claim 8 .
  10. The composition according to claim 1, wherein the viral surface of the lentivirus particle further comprises a CD28 ligand.
  11. The composition according to claim 10 , wherein the CD28 ligand is CD86.
  12. The composition according to claim 1, wherein the binding of the extracellular domain to the small molecule is sufficient for the intracellular cytokine receptor signaling domains of the two polypeptide chains to activate cytokine signaling.
  13. The composition according to claim 1, wherein the T cell/NK cell activating receptor does not contain an antigen-binding domain.
  14. The composition according to claim 1, wherein the lentiviral particle further comprises a nucleic acid sequence encoding a chimeric antigen receptor.
  15. The composition according to claim 1, wherein the small molecule comprises rapamycin.
  16. A composition according to any one of claims 1 to 5 for use in a method for treating a subject suffering from cancer, wherein the method comprises the step of administering the composition to the subject in combination with small molecules, characterized in that the cancer is treated in the subject.
  17. A composition for use in a method for treating a subject suffering from cancer, comprising small molecules, wherein the method comprises the step of administering the composition to the subject in combination with the composition according to any one of claims 1 to 5 , characterized in that the cancer is treated in the subject.
  18. A composition according to any one of claims 1 to 5 for use in a method for increasing T cells capable of recognizing and killing tumor cells in a subject requiring such proliferation, wherein the method comprises the step of administering the composition to the subject in combination with small molecules, characterized in that T cells capable of recognizing and killing tumor cells proliferate in the subject.
  19. A composition for use in a method for increasing T cells capable of recognizing and killing tumor cells, including small molecules, in a subject requiring such proliferation, the method comprising the step of administering the composition to the subject in combination with the composition according to any one of claims 1 to 5 , characterized in that T cells capable of recognizing and killing tumor cells proliferate in the subject.

Description

Cross-reference of related applications This application claims priority to U.S. Provisional Patent Application No. 62/656,823, filed on 12 April 2018, the disclosure of which is incorporated herein by reference in its entirety for all purposes. Description of the Electronically Submitted Text File The sequence listing relating to this application is provided in text format instead of a paper copy and is incorporated herein by reference. The name of the text file containing the sequence listing is VITI_001_01WO_SeqList_ST25.txt. The text file is 94KB in size, was created on April 11, 2019, and submitted electronically via EFS-Web. This disclosure generally relates to viral vectors, packaging cell lines, and related uses, particularly methods for the proliferation of immune cell populations in vivo for the treatment of disease conditions. Cancer immunotherapy is a treatment approach based on the therapeutic induction of an immune response against tumors. Adoptive T-cell therapy (ACT) is a form of cancer immunotherapy. Lymphocytes, particularly tumor-infiltrating lymphocytes (TILs), are isolated from the body, cultured ex vivo, proliferated, and then reinjected. The proliferation process may include antigen-specific proliferation or genetic modification of TILs. ACT is outlined in Rosenberg et al. Adoptive cell transfer as personalized immunotherapy for human cancer. Science. 348:62-8 (2015). The inventors have realized that, as an alternative to ACT, in vivo transduction of TILs or other immune cells can promote cell proliferation in vivo rather than ex vivo. Furthermore, the inventors have realized that in vivo transduction of TILs or other immune cells would enable the treatment of cancer or other disease conditions without the high cost, time-consuming, and risky procedures required by ACT. Therefore, there is a need for means of growing TILs or other immune cell populations in vivo. In particular, there is a need for therapeutic agents capable of selectively growing desired populations of TILs or other immune cells in vivo. This disclosure provides viral vectors, packaging cell lines, and related methods of use for in vivo growth of TILs or other immune cells for the treatment of disease pathologies. Rosenberg et al. Adaptive cell transfer as personalized immunotherapy for human cancer. Science. 348:62-8 (2015) Figures A and B show lentiviral particles. Figure A is a diagram of an embodiment of a surface-modified lentiviral particle containing surface-expressed anti-CD3 and T-cell costimulatory molecules that are not present in the lentiviral particle shown in figure B.This document describes the experimental protocol used to generate the HATSE-293 packaging cell line derived from HEK-293 T cells transduced with lentiviral vectors encoding anti-CD3 scFV, CD86, and CD137L. CD86 + and CD137L + cells were isolated by fluorescence-activated cell sorting, grown, and frozen for long-term storage and use.FACS analysis of parental HEK-293 cells and HATSE-293 packaging cell line for CD46 (expected constitutive expression in both cell lines) and CD86 +/ CD137L + expression (expected expression only in HATSE-293 cell line) is shown. Analysis of parental HEK-293 cell line for CD46 (expected constitutive expression) and CD86/CD137L expression (no expected expression) is shown. No detectable fluorescent labels were observed in unstained (mock) samples (Figure 3A). High expression of CD46 was detected by anti-CD46 antibody staining (Figures 3A-B; PE anti-human CD46), while expression of either CD86 (Figure 3E; Pacific Blue anti-human CD86) or CD137L (Figure 3D; PE anti-human 4-1BB ligand (CD137L)) was not detected by specific staining for CD86 and CD137L, respectively.FACS analysis of parental HEK-293 cells and HATSE-293 packaging cell lines for CD46 (expected constitutive expression in both cell lines) and CD86 +/ CD137L + expression (expected expression only in HATSE-293 cell lines) is shown. Analysis of parental HEK-293 cell lines for CD46 (expected constitutive expression) and CD86/CD137L expression (no expected expression) is shown. High expression of CD46 was detected by anti-CD46 antibody staining (Figure 3A-B; PE anti-human CD46), while expression of either CD86 (Figure 3E; Pacific Blue anti-human CD86) or CD137L (Figure 3D; PE anti-human 4-1BB ligand (CD137L)) was not detected by specific staining for CD86 and CD137L, respectively.The FACS analysis of parental HEK-293 cells and HATSE-293 packaging cell lines for CD46 (expected constitutive expression in both cell lines) and CD86 +/ CD137L + expression (expected expression only in HATSE-293 cell lines) is shown. The analysis of parental HEK-293 cell lines for CD46 (expected constitutive expression) and CD86/CD137L expression (no expected expression) is also shown.FACS analysis of parental HEK-293 cells and HATSE-293 packaging cell lines for CD46 (expected constitutive expression in both cell lines) and CD86 +/ CD137L + expression (expected expression only in