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US-20260125701-A1 - COMPOSITIONS AND METHODS FOR CHIMERIC LIGAND RECEPTOR (CLR)-MEDIATED CONDITIONAL GENE EXPRESSION

US20260125701A1US 20260125701 A1US20260125701 A1US 20260125701A1US-20260125701-A1

Abstract

Disclosed are composition comprising (a) an inducible transgene construct, comprising a sequence encoding an inducible promoter and a sequence encoding a transgene, and (b) a receptor construct, comprising a sequence encoding a constitutive promoter and a sequence encoding an exogenous receptor, wherein, upon integration of the construct of (a) and the construct of (b) into a genomic sequence of a cell, the exogenous reporter is expressed, and wherein the exogenous reporter, upon binding a ligand, transduces an intracellular signal that targets the inducible promoter of (a) to modify gene expression. Methods for introducing compositions into cells and the use of the resultant cells in adoptive cell therapies are also provided.

Inventors

  • Eric M. Ostertag
  • Devon Shedlock

Assignees

  • POSEIDA THERAPEUTICS, INC.

Dates

Publication Date
20260507
Application Date
20250701

Claims (12)

  1. 1 .- 132 . (canceled)
  2. 133 . A nucleic acid sequence comprising a) a receptor sequence comprising a constitutive promoter and a sequence encoding at least one chimeric ligand receptor (CLR); and b) a inducible transgene sequence comprising an inducible promoter and a sequence encoding a transgene.
  3. 134 . The nucleic acid of claim 133 , wherein the constitutive promoter is a CMV promoter, a U6 promoter, a SV40 promoter, a PGK1 promoter, a Ubc promoter, a human beta actin promoter, a CAG promoter, or an EF1α promoter.
  4. 135 . The nucleic acid of claim 134 , wherein the constitutive promoter is an EF1α promoter.
  5. 136 . The nucleic acid of claim 133 , wherein the inducible promoter is an NFκB promoter, an NR4A1 promoter, a CD5 promoter, an interferon (IFN) promoter or an interleukin-2 promoter.
  6. 137 . The nucleic acid of claim 136 , wherein the IFN promoter is an IFNγ promoter.
  7. 138 . A vector comprising the nucleic acid of claim 133 .
  8. 139 . The vector of claim 138 , wherein the inducible transgene sequence and the receptor sequence are oriented in the same direction.
  9. 140 . The vector of claim 148 , wherein the inducible transgene sequence and the receptor sequence are oriented in the opposite direction.
  10. 141 . A method of treating a disease or disorder in a subject in need thereof, comprising administering to the subject: a) a population of T-cells wherein a plurality of T-cells in the population comprise at least one chimeric ligand receptor (CLR) and at least one inducible transgene construct, wherein the CLR is a transmembrane protein comprises (i) an ectodomain comprising a ligand recognition region, wherein the ligand recognition region comprises a signal peptide and at least one scaffold protein; (ii) a transmembrane domain; and (iii) an endodomain comprising at least one costimulatory domain, wherein the at least one inducible transgene construct comprises a sequence encoding an inducible promoter and a sequence encoding a transgene; and b) a ligand that binds to the ligand recognition region of the at least one CLR, wherein upon binding of the ligand to the ligand recognition region, the endodomain of the at least one CLR transduces an intracellular signal that targets the inducible promoter and results in expression of the transgene within the plurality of T-cells, thereby treating the disease or disorder in the subject.
  11. 142 . The method of claim 141 , wherein the disease or disorder is cancer.
  12. 143 . The method of claim 141 , wherein the disease or disorder is Hemophilia B.

Description

CROSS-REFERENCE TO RELATED APPLICATIONS This application is a divisional of U.S. patent application Ser. No. 16/640,788, filed Feb. 21, 2020, which is a U.S. National Phase Application, filed under 35 U.S.C. § 371, of International Patent Application No. PCT/2018/050288, filed Sep. 10, 2018, which claims the benefit of provisional application U.S. Ser. No. 62/556,310, filed Sep. 8, 2017, the contents of each of these applications are herein incorporated by reference in their entireties. INCORPORATION OF SEQUENCE LISTING The instant application contains a Sequence Listing, which has been submitted electronically in XML file format, and is hereby incorporated by reference into the specification in its entirety. The XML file containing the Sequence Listing XML is named “000218-0106-303-SL.xml,” was created on Jul. 1, 2025, and is 21,480,788 bytes in size. FIELD OF THE DISCLOSURE The disclosure is directed to molecular biology, and more, specifically, to compositions and methods for use in a conditional gene expression system responsive to a chimeric ligand receptor (CLR)-mediated signal. BACKGROUND There has been a long-felt but unmet need in the art for a method of controlling gene expression in genetically modified cells for the long-term delivery of therapeutic agents. The disclosure provides a solution by genetically modified cells that conditionally express genes upon activation of a cell-surface receptor. SUMMARY The disclosure provides a composition comprising (a) an inducible transgene construct, comprising a sequence encoding an inducible promoter and a sequence encoding a transgene, and (b) a receptor construct, comprising a sequence encoding a constitutive promoter and a sequence encoding an exogenous receptor, wherein, upon integration of the construct of (a) and the construct of (b) into a genomic sequence of a cell, the exogenous reporter is expressed, and wherein the exogenous reporter, upon binding a ligand, transduces an intracellular signal that targets the inducible promoter of (a) to modify gene expression. In certain embodiments, the composition modifies gene expression by increasing gene expression. In certain embodiments, the composition modifies gene expression by decreasing gene expression. In certain embodiments, the composition modifies gene expression by transiently modifying gene expression (e.g. for the duration of binding of the ligand to the exogenous receptor). In certain embodiments, the composition modifies gene expression acutely (e.g. the ligand reversibly binds to the exogenous receptor). In certain embodiments, the composition modifies gene expression chronically (e.g. the ligand irreversibly binds to the exogenous receptor). In certain embodiments of the compositions of the disclosure, the cell may be a prokaryotic cell. Prokaryotic cells of the disclosure include, but are not limited to, bacteria and archaea. For example, bacteria of the disclosure include, but are not limited to, Listeria monocytogenes. In certain embodiments of the compositions of the disclosure, the cell may be a eukaryotic cell. Eukaryotic cells of the disclosure include, but are not limited to, yeast, plants, algae, insects, mammals, amphibians, birds, reptiles, marsupials, rodents, and humans. Preferred eukaryotic cells of the disclosure include, but are not limited to, human cells. Exemplary human cells of the disclosure include but are not limited to, immune cells (e.g. T cells), myeloid cells and bone marrow cells (e.g. hematopoietic stem cells (HSCs)). In certain embodiments of the compositions of the disclosure, the exogenous receptor of (b) comprises an endogenous receptor with respect to the genomic sequence of the cell. Exemplary receptors include, but are not limited to, intracellular receptors, cell-surface receptors, transmembrane receptors, ligand-gated ion channels, and G-protein coupled receptors. In certain embodiments of the compositions of the disclosure, the exogenous receptor of (b) comprises a non-naturally occurring receptor. In certain embodiments, the non-naturally occurring receptor is a synthetic, modified, recombinant, mutant or chimeric receptor. In certain embodiments, the non-naturally occurring receptor comprises one or more sequences isolated or derived from a T-cell receptor (TCR). In certain embodiments, the non-naturally occurring receptor comprises one or more sequences isolated or derived from a scaffold protein. In certain embodiments, including those wherein the non-naturally occurring receptor does not comprise a transmembrane domain, the non-naturally occurring receptor interacts with a second transmembrane, membrane-bound and/or an intracellular receptor that, following contact with the non-naturally occurring receptor, transduces an intracellular signal. In certain embodiments of the compositions of the disclosure, the exogenous receptor of (b) comprises a non-naturally occurring receptor. In certain embodiments, the non-naturally occurring receptor is a s