Recombinant Eomes Protein Restores Exhausted NK Cell Activity

The patent applies preliminary action by introducing the NLS-Eomes-PTD polypeptide into NK cells before they become exhausted or at the early stages of exhaustion. This polypeptide directly targets the transcriptional machinery to prevent or reverse exhaustion markers, establishing a more durable functional state before complete exhaustion occurs. The approach of pre-treating or early intervention with the fusion protein creates lasting changes in gene expression patterns that sustain anti-cancer activity longer than checkpoint inhibitors alone.

The patent employs parameter changes by modifying the Eomes protein structure into a fusion construct (NLS-Eomes-PTD) with enhanced cellular uptake and nuclear delivery properties. This structural parameter change enables the protein to efficiently enter cells and reach the nucleus, where it can sustainably alter transcriptional parameters of exhaustion markers. The fusion protein creates lasting changes in gene expression levels of cytotoxic molecules and cytokines, providing prolonged anti-cancer activity beyond the temporary effects of checkpoint blockade.

The patent applies the taking out principle by extracting and delivering only the essential transcriptional activation function of Eomes directly into the nucleus of NK cells, rather than engaging in complex membrane receptor signaling as with CAR technology. The NLS-Eomes-PTD polypeptide bypasses the membrane signaling cascade entirely and delivers the core transcriptional activation capability directly to nuclear targets, avoiding the harmful downstream effects of uncontrolled signaling that lead to exhaustion. This extraction of the essential function from the problematic signaling pathway resolves the contradiction between activity enhancement and exhaustion prevention.

The patent uses the NLS-Eomes-PTD fusion protein as an intermediary that mediates the transfer of transcriptional activation capacity from the extracellular environment directly to nuclear genes. This intermediary approach avoids the need for persistent membrane receptor engagement and downstream signaling cascades that cause exhaustion. The fusion protein serves as a direct mediator between the therapeutic intervention and the target genes, providing controlled up-regulation of anti-cancer signaling molecules without triggering the harmful feedback loops that lead to functional exhaustion.

The patent applies universality by designing the NLS-Eomes-PTD polypeptide to simultaneously address multiple functional deficits in exhausted NK cells. The fusion protein universally targets multiple exhaustion-related genes and pathways, restoring not only cytotoxicity but also cytokine secretion, proliferation, and overall functional capacity in a single intervention. This multi-functional approach overcomes the limitation of partial recovery by concurrently restoring all critical NK cell functions through coordinated transcriptional reprogramming of multiple target genes.

The patent uses preliminary action by introducing the NLS-Eomes-PTD polypeptide to preemptively restore transcriptional programs before complete functional exhaustion sets in, or at early stages when reversal is most effective. This timing strategy allows the fusion protein to re-establish expression of cytotoxic molecules and cytokines before their synthesis machinery is permanently downregulated. The preliminary transcriptional restoration prevents the irreversible loss of productive functions, achieving complete recovery rather than partial adaptation.