Methods and compositions for transducing lymphocytes and regulating the activity thereof

Inventors

Frost, Gregory Ian • Onuffer, Jr., James Joseph • Guibinga, Ghiabe H. • Haerizadeh, Farzad • Kundu, Anirban

Assignees

Exuma Biotech Corp

Abstract

The present disclosure provides methods for genetically modifying lymphocytes and methods for performing adoptive cellular therapy that include transducing T cells and/or NK cells. The methods can include inhibitory RNA molecule(s) and/or engineered signaling polypeptides that can include a lymphoproliferative element, and/or a chimeric antigen receptor (CAR), for example a microenvironment restricted biologic CAR (MRB-CAR). Additional elements of such engineered signaling polypeptides are provided herein, such as those that drive proliferation and regulatory elements therefor, as well as replication incompetent recombinant retroviral particles and packaging cell lines and methods of making the same. Numerous elements and methods for regulating transduced and/or genetically modified T cells and/or NK cells are provided, such as, for example, those including riboswitches, MRB-CARs, recognition domains, and/or pH-modulating agents.

Core Innovation

The invention relates to a replication incompetent recombinant retroviral particle, including lentiviral particles, that comprises one or more envelope polypeptides and one or more transcriptional units operatively linked to a promoter active in T cells. The transcriptional unit encodes engineered signaling polypeptides configured to promote proliferation and/or survival of T cells through a cytokine receptor polypeptide signaling domain capable of activating a Jak pathway.

The transcriptional unit further encodes a second engineered signaling polypeptide comprising an antigen specific targeting region (ASTR), a transmembrane domain, and an intracellular activating domain. The engineered signaling polypeptides are expressed from transcriptional units tailored for activity in T cells and/or NK cells, while maintaining replication incompetence for safety.

The particle further includes an activation element on the surface of the replication incompetent recombinant retroviral particle. The activation element is fused to a heterologous membrane attachment sequence and comprises a means for binding to CD3, combining engineered signaling, antigen-specific targeting, and CD3-binding surface activation.

Claims Coverage

The independent claim coverage centers on four inventive features: a replication-incompetent retroviral particle with T-cell-active transcriptional units, engineered signaling polypeptides for Jak-pathway lymphoproliferative signaling and antigen-specific targeting, and a surface activation element that binds CD3 through a heterologous membrane attachment sequence.

Overall claim coverage centers on replication-incompetent retroviral particles that express T-cell-active transcriptional units encoding a lymphoproliferative cytokine receptor signaling polypeptide with Jak-pathway activation, an ASTR-based antigen-targeting signaling polypeptide with transmembrane and intracellular activating domains, and a surface CD3-binding activation element fused to a heterologous membrane attachment sequence.

Stated Advantages

Documented Applications