DuoBody®-CD40x4-1BB (GEN1042/BNT312) is an investigational bispecific antibody designed to conditionally activate costimulatory receptors CD40 and 4-1BB on immune cells to induce antitumor immunity. Comprehensive in vitro studies demonstrated that dual target engagement by DuoBody-CD40x4-1BB is required to enhance antigen presentation and T-cell priming. As the expected primary sites of action of DuoBody-CD40x4-1BB are the tumor-draining lymph nodes and the tumor microenvironment, we developed a chimeric bispecific surrogate antibody (chi-CD40x4-1BB) containing the VH/VL sequences of DuoBody-CD40x4-1BB in an Fc-inert mouse IgG2a backbone to enable mechanistic studies in vivo. Chi-CD40x4-1BB retained the functional and pharmacokinetic properties of DuoBody-CD40x4-1BB and showed antitumor activity in MC38 and B16F10-OVA syngeneic tumor models in double knock-in mice that express extracellular domains of human CD40 and 4-1BB. In the MC38 tumor model, chi-CD40x4-1BB elicited peripheral immunomodulation, including activation of CD8⁺ and CD4⁺ T cells as well as B cells. Transcriptomic profiling of tumors and lymph nodes revealed induction of gene expression consistent with CD40 and 4-1BB engagement mainly in the secondary lymphoid organs. Combination of chi-CD40x4-1BB with PD-1 blockade resulted in enhanced antitumor activity beyond the single agents. These findings establish chi-CD40x4-1BB as a relevant surrogate to evaluate the activity and mechanism of action of DuoBody-CD40x4-1BB in vivo. Chi-CD40x4-1BB induces CD40 and 4-1BB costimulation, promoting coordinated innate and adaptive immunity, resulting in antitumor activity that can be potentiated in combination with PD-1 blockade. These results may provide context for clinical investigations of DuoBody-CD40x4-1BB, as monotherapy and in combination with PD-1 inhibitors, in solid tumors.
JAK1 signaling supports a suppressive tumor-associated macrophage (TAM) state and may also promote tumor-intrinsic oncogenic signaling. Leiomyosarcoma (LMS) is a soft tissue sarcoma (STS) subtype known to have abundant TAM infiltration, which is often polarized to an immunosuppressive M2 phenotype and contributes to immune evasion. We hypothesized that using a JAK1 inhibitor could alter TAM phenotypes in the LMS tumor microenvironment (TME), helping patients and potentially enabling future combination studies. Here, we present results from a pilot study of itacitinib, a JAK1 inhibitor, for LMS. We analyzed patients in the LMS cohort of a pilot study of the selective JAK1 inhibitor itacitinib (started at 300 mg daily in 28-day cycles) (NCT03670069). Serial tumor biopsies and peripheral blood samples were collected for correlative analysis. Despite the heavily pretreated LMS population, PFS rate at 3 and 6 months for LMS was 75% (95% CI: 50%–100%) and 25% (95% CI: 5%–100%), respectively, and the median OS was 101 weeks (range: 70–256). Six of eight patients with LMS achieved stable disease. Serial biopsies demonstrated a decrease in M2 TAM but without a corresponding inflammatory response and with a decrease of inflammatory infiltrates, suggesting that JAK1 inhibition may have direct anti-tumor effects in this subtype and that the impacts of the TAM reduction may be related to their tumor supportive and angiogenic functions, rather than immune inhibitory functions. These data support further investigation of JAK1 inhibition in LMS as part of rational immunotherapeutic combinations.
The RIG-I pathway, a key component of the innate immune system that classically detects cytosolic viral RNA, has also been implicated in regulating tumor cell viability, radiosensitivity, and patient survival across multiple cancer types. However, these effects appear to be context-dependent and partly contradictory. Whether radiotherapy modulates tumor-intrinsic RIG-I signaling in head and neck squamous cell carcinoma (HNSCC) depending on HPV status, and whether such modulation is associated with changes in the tumor immune phenotype, remains unknown. Four human HNSCC cell lines (two HPV-negative and two HPV-positive), along with siRNA-mediated RIG-I knockdown and a CRISPR/Cas9-generated RIG-I knockout derivative, were treated with radiotherapy. Tumor cells were analyzed for their immune phenotype, including RIG-I expression via Western blot and flow cytometry, IFN-β secretion via ELISA, and PD-L1 and PD-L2 expression via flow cytometry. Radiotherapy, dose- and fraction-dependent, induced the upregulation of RIG-I expression and IFN-β secretion from HPV-negative HNSCC cells. The knockdown and knockout of the RIG-I-encoding gene DDX58 in the HPV-negative cell line HSC4 minimized both IFN-β secretion and PD-L1 and PD-L2 expression. These effects could be replicated by pre-exposing HPV-negative cell lines to the RIG-I inhibitor RIG012 before irradiation. Interestingly, HPV-positive cells showed less radiotherapy-mediated modulation of RIG-I and of the tumor cell phenotype of immune-related surface markers and showed no significant reaction to the inhibitor treatment. The previously unrecognized RIG-I-IFN-β axis is a central regulator of RT-induced immune checkpoint modulation for HPV-negative HNSCC. These results provide new opportunities to reshape the immune response following RT in HNSCC.
Background Rates of cancer cure increase; however, recurrence and refractoriness remain clinical challenges that require alternative therapeutic strategies. Natural killer (NK) cells could play a decisive role if their antitumor reactivity can be enhanced. The interaction between killer-cell immunoglobulin-like receptors (KIRs) and their human leukocyte antigen class-I (HLA-I) ligands significantly influences NK-cell education and may impact clinical outcomes in cancer patients. In this study, we evaluated the impact of KIR/HLA-I interactions on the transcriptomic profile of circulating NK-cells and their association with patient outcome.Methods KIR and HLA-I genotyping, together with KIR expression analysis in circulating NK cells, were performed in patients with solid and hematologic malignancies (n = 1,153). In addition, single-cell transcriptomic analyses (scRNA-seq) of circulating NK and T lymphocytes were carried out in selected donors with KIR/HLA-I genotypes associated with short and long survival.Results Activating KIRs (aKIRs), more than inhibitory KIRs, were associated with cancer patient outcomes. The accumulation of aKIR–HLA interactions was related to shorter overall survival, whereas KIR2DS1/C2 interaction, in the absence of other aKIR interactions, was associated with longer survival. Three major NK-cell transcriptional profiles were identified: mature cytotoxic, immature/regulatory, and adaptive cytotoxic. The adaptive-like transcriptional profile was associated with longer survival.Conclusions In conclusion, aKIR/HLA-I interactions were associated with cancer patient outcomes. In particular, KIR2DS1/C2 interaction in the absence of other activating signals was associated with adaptive NK-cell transcriptomic profiles and improved survival. Understanding the mechanisms underlying effective NK-cell-mediated tumor immunity may provide new opportunities for patients with refractory tumors.
Some patients with metastatic non-small-cell lung cancer (NSCLC) receiving anti-PD-1 or anti-PD-L1 immune checkpoint inhibitors (ICIs) progress at a few sites while disease elsewhere stays controlled, a pattern consistent with focal, oligo-acquired immune escape rather than systemic treatment failure. Ablating the escaping focus may eliminate the resistant clone and preserve an otherwise effective immunotherapy line. We performed a PRISMA 2020-compliant, PROSPERO-registered systematic review and meta-analysis of local ablative treatment (LAT) for oligoprogressive or oligo-resistant metastatic NSCLC during active ICI therapy, with continuation of the same immunotherapy line after LAT. MEDLINE, PubMed, Web of Science, Scopus, and the Cochrane Library were searched from inception to March 8, 2026. From 1270 records, 22 studies were included in the qualitative synthesis. Four studies, one randomized subgroup, and three retrospective cohorts contributed to each primary analysis. LAT-oriented management was associated with improved progression-free survival (PFS; pooled hazard ratio [HR] 0.45, 95% CI 0.33–0.62; 95% prediction interval [PI] 0.28–0.74; I2 = 0%) and overall survival (OS; pooled HR 0.41, 95% CI 0.27–0.63; 95% PI 0.24–0.69; I2 = 0%). PFS and OS estimates were consistent across sensitivity analyses, including the exclusion of the one PFS estimate measured to a second progression. The evidence is predominantly observational and of low certainty, reflecting the current absence of NSCLC-specific randomized trials. On this evidence, LAT, delivered predominantly as stereotactic body radiotherapy, may be considered a continuation-enabling option for carefully selected patients with focal progression on otherwise effective immunotherapy, pending ongoing randomized trials.
