Engineered trimeric CD70 proteins and uses thereof
Inventors
Jaeger, Carla A. • Correnti, Colin E. • Riddell, Stanley R.
Assignees
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Publication Number
US-12351618-B2
Publication Date
2025-07-08
Expiration Date
Abstract
Engineered trimeric CD70 proteins for use in ex vivo T cell manufacturing are described. Use of the proteins during manufacturing creates expanded T cell populations with enhanced properties such as earlier proliferation in culture; selective expansion of nave and memory T cell subsets; longer persistence in vivo following administration to a subject; and improved therapeutic effect. Use of the proteins as therapeutics provide anti-cancer and anti-viral effects. The proteins can also be used as agonistic cell culture reagents in in vitro uses.
Core Innovation
The invention relates to an ex vivo method of manufacturing an activated, expanded T cell population for administration to a subject. The method includes obtaining a population of T cells and culturing them in culture media comprising a CD3 stimulating molecule and an Fc-dimerized protein. The Fc-dimerized protein comprises two copies of the single chain protein sequence set forth in SEQ ID NO: 5 or a sequence having at least 95% sequence identity to that sequence to increase T cell activation.
After the activation phase, the method further includes culturing the activated T cell population within culture media lacking the CD3 stimulating molecule. The expanded T cell population is then formulated into a pharmaceutically acceptable carrier in a therapeutically effective amount, thereby ex vivo manufacturing the T cell population for administration to the subject.
The document describes engineered trimeric CD70 proteins used to provide CD27 agonist signaling in the context of ex vivo T cell manufacturing. It presents dimer-trimer, trimeric monomer via trimerization domains, and circular-tandem repeat scaffolds as enabling increased CD27-mediated activation during manufacturing, together with improved proliferation and expansion outcomes.
The disclosed manufacturing scope further connects this ex vivo activation and expansion framework with engineered T cell features described in the document, including CAR constructs and related design elements, as well as control features such as tag cassettes and an iCaspase9 suicide switch for depletion, tracking, enrichment, and elimination.
Claims Coverage
The provided material identifies one independent ex vivo manufacturing method claim. The claim includes three inventive features: CD3 stimulation with an Fc-dimerized protein defined by two copies of the SEQ ID NO: 5 single chain sequence (or at least 95% identity), CD3-free expansion of activated T cells, and formulation of the expanded population for administration.
Activation with an Fc-dimerized protein plus CD3 stimulation
Culturing a population of T cells within culture media comprising a CD3 stimulating molecule and an Fc-dimerized protein at a concentration sufficient to increase T cell activation, wherein the Fc-dimerized protein has two copies of the single chain protein sequence set forth in SEQ ID NO: 5 or a sequence having at least 95% sequence identity to the sequence set forth in SEQ ID NO: 5.
CD3-free expansion of activated T cells
Culturing the activated T cell population within culture media lacking the CD3 stimulating molecule.
Formulation into a therapeutically effective dose for administration
Formulating the expanded T cell population into a pharmaceutically acceptable carrier in a therapeutically effective amount, thereby ex vivo manufacturing the T cell population for administration to the subject.
Overall, the independent claim centers on ex vivo T cell activation by combining a CD3 stimulating molecule with an Fc-dimerized protein defined by two copies of the SEQ ID NO: 5 single chain, followed by culturing without the CD3 stimulating molecule and formulating the resulting expanded T cells for administration in a therapeutically effective amount.
Stated Advantages
Increases T cell activation during ex vivo culturing with the CD3 stimulating molecule and the Fc-dimerized protein.
Earlier and greater T cell proliferation and expansion when manufacturing uses the engineered trimeric CD70 proteins, often alongside αCD3/OKT3.
Selective expansion of naïve/memory T cell subsets.
Metabolic/stemness advantages, including reduced effector metabolic differentiation and weaker mTORC1/S6K signaling.
Improved survival/persistence and in vivo engraftment.
Enhanced killing capability, including serial tumor-killing.
Improved anti-cancer and anti-viral therapeutic effects.
Enables ex vivo manufacturing of an expanded T cell population for administration to a subject.
Documented Applications
Ex vivo method of manufacturing a T cell population for administration to a subject.
Use of agonistic culture reagents based on the described engineered trimeric CD70 proteins in cell culture contexts.
Anti-cancer treatment, including tumor control and serial killing as described in the document’s example data.
Anti-pathogen treatment targeting one or more specified viruses, including adenovirus and multiple other virus families listed in the dependent claim context.
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