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US-12624083-B2 - Chimeric antigen receptors against human cytomegalovirus

US12624083B2US 12624083 B2US12624083 B2US 12624083B2US-12624083-B2

Abstract

Disclosed herein are CMV-specific CARs. In some embodiments. the present invention is directed to a method of treating, reducing, or inhibiting an infection by a cytomegalovirus in a subject, which comprises administering to the subject (a) an expression vector that encodes a CMV-specific CAR as described herein, or (b) one or more cells that are transduced with the expression vector.

Inventors

  • Otto O. Yang
  • Ayub Ali
  • Don J. Diamond
  • Flavia CHIUPPESI
  • Felix WUSSOW

Assignees

  • THE REGENTS OF THE UNIVERSITY OF CALIFORNIA
  • CITY OF HOPE

Dates

Publication Date
20260512
Application Date
20201012

Claims (17)

  1. 1 . A cytomegalovirus specific chimeric antigen receptor (CMV-specific CAR), which comprises a single chain antibody (scFv) sequence having the following CDR sequences: VH CDR1-SEQ ID NO: 30, VH CDR2-SEQ ID NO: 39, VH CDR3-SEQ ID NO: 46, VL CDR1-SEQ ID NO: 59, VL CDR2-SEQ ID NO: 66, and VL CDR3-SEQ ID NO: 71.
  2. 2 . The CMV-specific CAR according to claim 1 , wherein the single chain antibody comprises SEQ ID NO: 11 and SEQ ID NO: 12.
  3. 3 . An expression vector comprising a nucleic acid sequence encoding a CMV-specific CAR according to claim 1 .
  4. 4 . A host cell comprising one or more expression vectors according to claim 3 .
  5. 5 . The host cell according to claim 4 , wherein the host cell is a CD8+ T lymphocyte, hematopoietic stem cell, or a hematopoietic progenitor cell.
  6. 6 . A cell that is the progeny of the host cell of claim 4 .
  7. 7 . The cell according to claim 4 , wherein the cell expresses one or more chimeric antigen receptors encoded by the one or more expression vectors.
  8. 8 . A method of treating, reducing, or inhibiting an infection by a cytomegalovirus in a subject, which comprises transplanting one or more cells according to claim 4 , in the subject.
  9. 9 . The method according to claim 8 , wherein the subject is human and/or the cytomegalovirus is a human cytomegalovirus.
  10. 10 . The method according to claim 8 , wherein the subject has an immunodeficiency.
  11. 11 . A method of killing cells infected with a cytomegalovirus, which comprises contacting the infected cells with one or more cells (a) that express one or more CMV-specific CARs according to claim 1 , or (b) comprise an expression vector that encodes the one or more CMV-specific CARs.
  12. 12 . A method of reducing replication of a cytomegalovirus in a cell or a subject, which comprises contacting the cell with or administering to the subject one or more cells (a) that express one or more CMV-specific CARs according to claim 1 , or (b) comprise an expression vector that encodes the one or more CMV-specific CARs.
  13. 13 . A method of treating, reducing, or inhibiting an infection by a cytomegalovirus in a subject, which comprises administering to the subject (a) an expression vector that encodes the CMV-specific CAR according to claim 1 , or (b) one or more cells that are transduced with the expression vector.
  14. 14 . A method of treating, reducing, or inhibiting an infection by a cytomegalovirus in a subject, which comprises transplanting one or more cells that express one or more CMV-specific CARs according to claim 1 to the subject.
  15. 15 . A cytomegalovirus specific chimeric antigen receptor (CMV-specific CAR), which comprises or consists of SEQ ID NO: 89.
  16. 16 . A method of treating, reducing, or inhibiting an infection by a cytomegalovirus in a subject, which comprises administering to the subject (a) an expression vector that encodes the CMV-specific CAR according to claim 15 , or (b) one or more cells that are transduced with the expression vector.
  17. 17 . The method according to claim 16 , wherein the subject is human and/or the cytomegalovirus is a human cytomegalovirus.

Description

CROSS-REFERENCE TO RELATED APPLICATIONS This application claims the benefit of U.S. Patent Application No. 62/914,408, filed Oct. 11, 2019, which is herein incorporated by reference in its entirety. ACKNOWLEDGEMENT OF GOVERNMENT SUPPORT This invention was made with Government support under Grant Number AI103960, awarded by the National Institutes of Health. The Government has certain rights in the invention. REFERENCE TO A SEQUENCE LISTING SUBMITTED VIA EFS-WEB The content of the ASCII text file of the sequence listing named “20201012_034044_209WO1_ST25” which is 85.9 kb in size was created on Oct. 7, 2020 and electronically submitted via EFS-Web herewith the application is incorporated herein by reference in its entirety. BACKGROUND OF THE INVENTION 1. Field of the Invention Compositions and methods for treating cytomegalovirus infections. 2. Description of the Related Art Human cytomegalovirus (CMV) infection is widespread, ranging from about 60% of adults in the US to nearly 100% in other parts of the world. In most healthy persons, infection is lifelong but immunologically contained and asymptomatic. Some perinatally infected persons, and persons who are significantly immunosuppressed (due to AIDS or iatrogenic/therapeutic immunosuppression for conditions such as bone marrow or organ transplantation) can develop disseminated infection with significant morbidity and mortality due to end organ damage. The major arm of immunity controlling CMV infection in healthy hosts is cellular immunity, particularly CD8+ T lymphocytes (C8TLs). Autologous immunotherapy using expanded CMV-specific C8TLs has provided proof-of-concept that C8TLs can treat CMV in immunocompromised hosts, but this approach is not generally applicable due to various technical limitations. On the other hand, gene therapy with a chimeric antigen receptor (CAR) targeted against CMV could be readily applied to generate CMV-specific C8TLs in patients, analogous to the growing use of CAR T-cell gene therapy for various cancers. To date only one CMV-specific CAR has been reported in the literature, and it has not been advanced to clinical trials to our knowledge. SUMMARY OF THE INVENTION “CMV-specific CARs”: In some embodiments, the present invention is directed to a cytomegalovirus specific chimeric antigen receptor (CMV-specific CAR), which comprises a single chain antibody sequence or fragment thereof having SEQ ID NO: 21, SEQ ID NO: 22, SEQ ID NO: 23, SEQ ID NO: 24, SEQ ID NO: 25, and SEQ ID NO: 26 as follows: G-X1-X2-X3-X4-X5-X6-X7-X8-X9 Formula H1 (SEQ ID NO: 21), wherein, X1 is F or Y, X2 is S or T, X3 is L or F, X4 is S or T, X5 is D, I, N, S, or T, X6 is F, Y, or S, X7 is G, Y, or W, X8 is present or absent and if present X8 is L or I, and, X9 is present or absent and if present X9 is G; I-X10-X11-X12-X13-X14-X15-X16 Formula H2 (SEQ ID NO: 22), wherein, X10 is D, N, S, or W, X11 is D, N, P, T, or W, X12 is D, G, N, Y, or S, X13 is D, G, or T, X14 is D, G, N, or S, X15 is E, K, S, or Y, and, X16 is present or absent and if present X16 is P or T; X17-X18-X19-X20-X21-X22-X23-X24-X25-X26-X27-X28-X29-X30-X31 Formula H3 (SEQ ID NO: 23), wherein, X17 is A, S, or V, X18 is R, N, or S, X19 is E, G, K, P, R, or S, X20 is G, H, K, L, Y, or W, X21 is D, L, R, S, or Y, X22 is D, F, G, L, S, or Y, X23 is A, D, F, G, L, P, or Y, X24 is A, D, I, Q, S, V, or Y, X25 is present or absent and if present X25 is E, F, N, S, or Y, X26 is present or absent and if present X26 is A, G, M, L, or P is, X27 present or absent and if present X27 is D, L, or Y, X28 is present or absent and if present X28 is A, F, L, or Y, X29 is present or absent and if present X29 is D, F, G, or M, X30 is present or absent and if present X30 is C, D, or Y, and, X31 is present or absent and if present X31 is S or Y; X32-X33-X34-X35-X36-X37-X38-X39-X40-X41-X42 Formula L1 (SEQ ID NO: 24), wherein, X32 is E, K, or Q, X33 is G or S, X34 is I, L, or V, X35 is D, G, S, or V, X36 is D, H, N, S, or T, X37 is D, N, S, or Y, X38 is present or absent and if present X38 is D, G, or N, X39 is present or absent and if present X39 is G, N, or Y, X40 is present or absent and if present X40 is K, N, or S, X41 is present or absent and if present X41 is F, Y, or T, and, X42 is present or absent and if present X42 is Y; X43-X44-S Formula L2 (SEQ ID NO: 25), wherein, X43 is D, L, R, T, or Y, and, X44 is A, T, or V; X45-X46-X47-X48-X49-X50-P-X51-T Formula L3 (SEQ ID NO: 26), wherein, X45 is S, Q, or W, X46 is H, N, or Q, X47 is D, G, S, or Y, X48 is H, N, R, S, T, or Y, X49 is E, H, K, R, S, or T, X50 is D, F, L, S, V, or W, and, X51 is L, P, W, or Y. In some embodiments, a) Formula H1 (SEQ ID NO: 21) is GFSLSTYGIG (SEQ ID NO: 27), GFSLTTSGLG (SEQ ID NO: 28), GFTFSDYY (SEQ ID NO: 29), GYTFTIYG (SEQ ID NO: 30), GYTFTNFG (SEQ ID NO: 31), GYTFTSYG (SEQ ID NO: 32), GYTFTSYW (SEQ ID NO: 33), GYTFTIYW (SEQ ID NO: 34), or GYTFTSYW (SEQ ID NO: 35); b) Formula H2 (SEQ ID NO: 22) is IDPSDSET (SEQ ID NO: 36), IDPSDSET (