Junko Matsuzaki

Adoptive chimeric antigen receptor (CAR)-T cell therapies have limited efficacy in solid tumors because of poor infiltration and impaired effector function, whereas radiotherapy is constrained by radioresistance and metastatic spread. Here we show that integrating the spatial precision of ionizing radiation with the biological specificity of CAR-T cells enables localized delivery of the inflammatory cytokine tumor necrosis factor (TNF). We identified radiation-responsive genes in tumor-infiltrating T cells and engineered synthetic radioinducible promoters to drive TNF expression. Human CAR-T cells carrying this construct (riTNF CAR-T cells) secreted high levels of TNF upon irradiation, independently of antigen engagement. In human xenograft and murine tumor models, the combination of riTNF CAR-T cells and local irradiation produced greater tumor regression than either treatment alone or standard CAR-T cells with irradiation. This response was associated with high intratumoral, but not systemic, TNF levels, increased tumor necrosis and apoptosis, vascular disruption and greater CAR-T cell infiltration. These findings establish a platform for spatially and temporally controlled cytokine delivery from engineered CAR-T cells.

Human peripheral blood CD8+ T cells require NKR-mediated co-stimulation for optimal effector response to weak TCR stimulation.

NKRs assist CTLs to recognize and conjugate cancer cells expressing low-level MHCI/peptide complexes.

DNAM-1 and NKG2D expression is correlated with long term survival and cytotoxic genes in tumor samples from melanoma patients (TCGA).

Expression of DNAM-1 and DNAM-1-competing (inhibitory) receptors reflects CTL cytotoxic function.

DNAM-1 signaling assists TCR signaling in CTLs.

Chemotherapeutic drugs elevate NKR-L expression on cancer cells.

DC-primed MART-1-specific CTLs express high levels of NKRs, but recognize and kill cancer cells in a strictly TCR-dependent manner.

Developing molecules that selectively bind targets of interest remains a critical bottleneck in biological research and biotechnology. Here, we present a workflow that leverages the Phage-Assisted Non-Continuous Selection for Binders (PANCS-Binders) technology for rapid de novo binder discovery. In a blinded testcase, we pursued three cancer-related targets: NSD3, NMNAT2, and CSF1R. Within 26 days, we obtained sequence- and function-verified binders for all three targets with nano-to-micromolar affinities. By incorporating an NSD3 binder into an engineered E3 ligase, RNF8, we developed a potent NSD3 degrader that potently depleted endogenous NSD3 and inhibited colorectal cancer cell proliferation. We then applied this degrader to reveal previously unknown NSD3 dependencies in ovarian cancer cell lines, uncovering new therapeutic vulnerabilities. Together, our work establishes a rapid, robust workflow for accelerated binder discovery and demonstrates how binders can expedite biotechnology development.

The use of corticosteroids at the initiation of ICI therapy correlates with decreased ORR and worse prognosis in patients with NSCLC. A–D, ORR, PFS, and OS of patients at RPCCC (A and C) and USC (B and D) treated with ICI therapy on the basis of baseline steroid use. The ORR was analyzed by the Fisher exact test. P values for PFF and OS were calculated by a log-rank (Mantel–Cox) test. mOS, median OS; mPFS, median PFS.