Supplementary Figure S4: A. Venn diagram showed that among the 112 genes higher expressed in patients with shorter PFS, 15 overlapped with genes also higher expressed in patients without DCB, 7 were specific to absence of DCB, and 97 were specific to shorter PFS. B. Venn diagram showed that among the 37 genes higher expressed in patients with longer PFS, 14 overlapped with genes also higher in patients with DCB, 35 were specific to DCB, and 23 were specific to longer PFS.

Immune checkpoint blockade treatment is highly effective in microsatellite-instable (MSI) colorectal cancer. However, microsatellite-stable (MSS) tumors, which represent 95% of metastatic colorectal cancer, are intrinsically resistant to immunotherapy. In this study, we sought to better understand the mechanisms of resistance to anti-PD-L1 therapy in colorectal cancer by characterizing the immune profiles of MSS and MSI tumor models. Although both tumor types presented intratumoral CD8+ T-cell responses and PD-L1 expression, the exhausted CD8+ T-cell phenotypes differed. In MSS tumors, exhausted CD8+ T cells coexpressed PD-1 and T-cell immunoglobulin and ITIM domain (TIGIT) and exhibited a terminal exhausted profile with low cytokine secretion and limited cytotoxic function. In contrast, PD-1+ CD8+ T cells in MSI tumors did not express TIGIT and displayed higher cytokine and cytotoxic activities. Interestingly, immunosuppressive M2-like tumor-associated macrophages (TAM) accumulated in MSI tumors and positively correlated with PD-1+ TIGIT+ CD8+ T-cell frequency. M2-like TAM depletion reduced TIGIT expression, increased CD8+ T-cell function, and improved efficacy of PD-L1 blockade. Transcriptomic analysis revealed elevated COX1/2 expression in TAMs in MSS tumors compared with MSI tumors. COX2 and prostaglandin E2 (PGE2) receptor inhibition impeded TIGIT expression and restored CD8+ T-cell activity, whereas PGE2 triggered TIGIT upregulation in CD8+ T cells. Single-cell, spatial, and bulk transcriptomic data from patients with colorectal cancer substantiated the correlation between elevated TIGIT in CD8+ T-cell and COX1/2 in TAMs. Together, these data uncover the role of the TAM axis in inhibiting PD-L1 efficacy in MSS colorectal cancer and support the utility of combining anti-PD-L1 therapy with TIGIT blockade, PGE2 treatment, or M2-like TAM inhibition in colorectal cancer.Significance: Targeting PGE2 signaling activated by tumor-associated macrophages sensitizes microsatellite-stable colon tumors to immune checkpoint blockade by limiting the emergence of an exhausted PD-1+ TIGIT+ CD8+ T population.

PURPOSE:There is no standard second-line therapy for gastroenteropancreatic (GEP) and lung large-cell neuroendocrine carcinoma (NEC) after the failure of platinum-based chemotherapy. This study aimed to investigate the efficacy of nivolumab ± ipilimumab. METHODS:The GCO-001-NIPINEC (ClinicalTrials.gov identifier: NCT03591731) trial was a noncomparative, open-label, phase II trial. The main inclusion criteria were age ≥18 years, performance status (PS) ≤2, advanced large- and small-cell GEP-NEC and large-cell lung NEC, and second- or third-line treatment for NECs refractory to platinum-based chemotherapy. Patients were randomly assigned (1:1) and stratified by age and PS to receive nivolumab (3 mg/kg/once every 2 weeks) ± ipilimumab (1 mg/kg/once every 6 weeks) for 2 years or until progression or unacceptable toxicity. The primary end point was objective response rate (ORR) at 8 weeks, assessed by investigators. RESULTS:A total of 185 patients (91 in the nivolumab arm and 94 in the nivolumab-ipilimumab arm) were enrolled between December 2018 and March 2021; 169 were analyzed (median age of 64.5 years, 71% male, 91% PS 0-1). The main primary tumor locations were lungs (50%), colorectal (15%), gastroesophageal (14%), and pancreatic (13%) regions. The ORR at 8 weeks was 7.2% (95% CI, 2.7 to 15.1]) in the nivolumab arm and 14.0% (95% CI, 7.4 to 23.1) in the nivolumab-ipilimumab arm. The best ORR was 9.6% and 20.9%, respectively, whereas the median progression-free and overall survival were approximately 2 months and 6 months in both arms. One treatment-related death occurred, in the nivolumab arm. The grade 3-4 adverse events (≥5%) were asthenia (13%), gamma-glutamyl transferase increase (10%), alkaline phosphatase increase (9%), dyspnea (7%), and anemia (6%) in the nivolumab-ipilimumab arm. CONCLUSION:Nivolumab-ipilimumab could be a second-/third-line treatment option for patients with NECs. However, given the limited magnitude of benefit, studies are warranted to evaluate its use earlier and/or associated with chemotherapy.

Supplementary Table S1. Representativeness of Study Participants

Managing unresectable metastatic colorectal cancer (mCRC) requires a comprehensive strategy. While chemotherapy, anti-angiogenic, and anti-epidermal growth factor receptor (EGFR) agents are available, strategy trials are needed to optimize their use and sequencing. The STRATEGIC-1 phase III trial (NCT01910610) was designed to determine the optimal treatment sequence in patients with untreated, unresectable wild-type RAS/BRAFV600E mCRC. Patients were randomized (1:1) to FOLFIRI-cetuximab then mFOLFOX6-bevacizumab (arm A) or OPTIMOX-bevacizumab then FOLFIRI-bevacizumab followed by EGFR monoclonal antibody +/- irinotecan (arm B). The primary endpoint was the duration of disease control (DDC). Secondary endpoints were overall survival (OS), time to failure of strategy (TFS), progression-free survival (PFS), overall response rate (ORR), salvage surgery rate, safety, and health-related quality of life (HRQoL). Overall, 263 patients (arm A:131, arm B:132) were randomized. After 68.4 months of median follow-up (95% CI, 76.5-98.0), the median DDC was 22.8 months (95% CI, 20.4-28.8) in arm A and 23.5 months (95% CI, 17.9-26.3) in arm B (HR = 1.01, 95% CI, 0.76-1.34; log-rank P = 0.945). The median OS was 40.4 months (95% CI, 32.4-51.1) in arm A and 34.4 months (95% CI, 27.5-42.2) in arm B (HR = 1.30, 95% CI, 0.99-1.72). The ORR was higher in arm A (82.4% versus 65.4%) in the first-line group but not in the second-line group (20.7% versus 16.4%). Adverse events were consistent with the well-known safety profiles. STRATEGIC-1 did not meet its primary endpoint and was inconclusive in identifying the optimal treatment strategy in wild-type RAS/BRAFV600E mCRC.

Supplementary Figure S4. Patients with focal MET amplification had increased responses to treatment with Temab-A.

Abstract Purpose: Gastrointestinal tumors, including esophageal and gastric/gastroesophageal junction adenocarcinoma (GEA), have a high mortality rate and present significant treatment challenges. Telisotuzumab adizutecan (Temab-A, ABBV-400), a novel antibody–drug conjugate targeting the c-Met protein (also known as MET protein), has shown encouraging results in patients with advanced GEA. Patients and Methods: This phase I, open-label, multicenter study assessed the safety, efficacy, and pharmacokinetics (PK) of Temab-A monotherapy (3 mg/kg every 3 weeks intravenously) in patients with advanced GEA. Patients ≥18 years of age with advanced/metastatic GEA who had received 1 to 2 prior systemic therapies were included. The primary objectives were the evaluation of safety, PK, and efficacy; efficacy endpoints included objective response rate (ORR), duration of response (DOR), progression-free survival (PFS), and overall survival (OS). c-Met expression and MET amplification were retrospectively assessed. Results: Forty-two patients with advanced GEA were enrolled; the median age was 60 years. The median follow-up duration was 12.6 months. All patients had one or more treatment-emergent adverse events (TEAE), with 88% experiencing grade ≥3 TEAEs. The most common hematologic TEAEs were anemia (67%), nausea (52%), and decreased appetite (36%). The ORR was 29%, the clinical benefit rate was 71%, and the median DOR was 4.2 months. The median PFS was 4 months, and the median OS was 5.8 months. Exploratory biomarker analyses showed ORR enrichment in patients with higher c-Met protein expression and MET focal amplification. Conclusions: Temab-A monotherapy demonstrated antitumor activity and a manageable safety profile in patients with advanced GEA. The findings support further clinical development of Temab-A, particularly in combination with other agents to improve outcomes for patients with 2L+ GEA.

PURPOSE:The antibody-drug conjugate Temab-A comprises the c-Met-targeting antibody telisotuzumab conjugated to a novel topoisomerase 1 inhibitor payload, adizutecan. A first-in-human phase I study (ClinicalTrials.gov identifier: NCT05029882) of Temab-A in patients with advanced solid tumors whose disease has progressed is currently ongoing. We report results from all patients in the dose escalation and the monotherapy metastatic colorectal cancer (mCRC) dose expansion. METHODS:Temab-A was administered intravenously once every 3 weeks as a monotherapy starting at 1.6 mg/kg in dose escalation. In mCRC dose expansion, patients with confirmed BRAF wild-type, microsatellite stable/mismatch repair-proficient mCRC were randomly assigned to 1.6 mg/kg, 2.4 mg/kg, or 3.0 mg/kg Temab-A once every 3 weeks. Primary end points included safety, pharmacokinetics, recommended phase II dose of Temab-A monotherapy, and Temab-A efficacy in patients with mCRC. RESULTS:In total, 57 patients received ≥1 dose of Temab-A in dose escalation; 3.0 mg/kg once every 3 weeks was established as the maximum tolerated dose. Collectively, in dose escalation and dose expansion, 122 patients with mCRC received Temab-A (dose escalation, N = 29; randomized dose optimization expansion, N = 93). All patients experienced ≥1 treatment-emergent adverse event; the most frequent were gastrointestinal (78%) and hematologic (71%) toxicities. Treatment-related discontinuations and deaths were infrequent (10% and 3%, respectively). Across all doses in patients with mCRC, overall response rate was 15.6% (95% CI, 9.6 to 23.2), disease control rate was 74.6% (95% CI, 65.9 to 82.0), and duration of response was 5.9 months (95% CI, 4.1 to 10.5); responses were more frequent at doses of 2.4 mg/kg and 3.0 mg/kg once every 3 weeks. Median progression-free survival was 4.6 months (95% CI: 4.0, 5.4), and median overall survival was 10.4 months (95% CI, 8.9 to 13.1). CONCLUSION:Temab-A at 2.4 mg/kg once every 3 weeks has a tolerable and manageable safety profile, with promising antitumor activity.

Abstract Background: Colorectal cancer (CRC) is the third most common cancer worldwide and the second leading cause of cancer death. It comprises two biological subtypes: microsatellite-stable (MSS, 85-95%) and microsatellite-instable (MSI-H) tumors. While MSI-H tumors respond to immunotherapy, MSS tumors remain “cold” and rely on chemotherapy. STC-1010 is a first-in-class, allogeneic antigen-induced immunotherapy derived from the Stimulated Ghost Cells (SGC) platform. SGC are produced by applying controlled physical or chemical stress to CRC cell lines, followed by haptenation to boost immune recognition; the cells are then inactivated. BreAK CRC 001 is a first-in-human trial evaluating STC-1010 with standard chemotherapy in unresectable advanced CRC. This study aims to translate preclinical findings into the clinical setting by evaluating the immunogenicity and antitumor activity of STC-1010 in combination with chemotherapy. Methods: Multi-omics analyses were performed to identify STC-1010 antigens relevance to CRC heterogeneity. The anti-tumor activity of a murine surrogate was evaluated in syngeneic CRC models. Mechanistic studies were conducted ex vivo to evaluate STC-1010-primed CD8⁺ T cells killing potency against CRC cell lines. BreAK CRC 001 is a first-in-human, open-label phase I/IIa study of STC-1010 combined with low-dose of cyclophosphamide, GM-CSF (STC-1010 regimen), and associated to standard chemotherapy for unresectable locally advanced or metastatic CRC. Phase I evaluated safety, dose-limiting toxicities, and recommended phase II dose (RP2D); phase IIa will assess safety, clinical activity and immune responses. Results: STC-1010 provides a broad repertoire of immunogenic antigens capturing CRC heterogeneity. In murine models, mSTC-1010 reduced tumor burden, improved survival, and, when combined with FOLFOX chemotherapy, significantly decreased tumor volumes compared with control. These effects were associated with increased CD8⁺ T cell infiltration. Ex vivo, CD8⁺ T cells primed by STC-1010-treated dendritic cells induced robust apoptosis in multiple CRC cell lines (HCT116, HT29, SW620). As of January 2026, 6 patients with unresectable metastatic CRC received STC-1010 regimen with SOC chemotherapy in the phase I dose-escalation part. Treatment was well tolerated, with predominantly grade 1 infusion-related reactions and no dose-limiting toxicities at the first two dose levels. Despite variable treatment duration, preliminary clinical activity was observed, including partial responses and stable disease per RECIST v1.1. Conclusion: STC-1010 regimen restores anti-tumor immunity and enhances immune cell activation in CRC models, supporting its continued clinical development. Combined with chemotherapy, it showed a manageable safety profile and early signs of clinical activity, consistent with the antitumor effects observed in preclinical models. Citation Format: Diego Tosi, Antoine Italiano, Antoine Hollebecque, Benoît You, Philippe Cassier, Francesco Sclafani, David Tougeron, Eric Christenson, Céline Gongora, Iseulys Richert, George Alzeeb, Paul Marteau, Marion Brun, Leila Feki, Lionel Chalus, Benoit Pinteur, Paul Bravetti, Sarah Kerbouche, Corinne Tortorelli, François Ghiringhelli. From preclinical models to first-in-human evaluation of STC-1010 immunotherapy in unresectable advanced colorectal cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2026; Part 2 (Late-Breaking, Clinical Trial, and Invited Abstracts); 2026 Apr 17-22; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2026;86(8_Suppl):Abstract nr CT051.

Abstract Purpose: Targeting the adenosine pathway may enhance the efficacy of chemo/immunotherapy regimens in patients with heavily pretreated advanced metastatic colorectal cancer (mCRC), for whom treatment options are limited. Patients and Methods: The phase II ARC-9 study, Cohort B (NCT04660812), evaluated the efficacy and safety of etrumadenant (A2a and A2b receptor antagonist), zimberelimab (anti–PD-1 mAb), FOLFOX, and bevacizumab (EZFB) versus regorafenib in patients with third-line mCRC who previously progressed on oxaliplatin- and irinotecan-containing regimens. Results: From September 21, 2021, to September 12, 2022, 112 patients were randomized 2:1 to EZFB (n = 75) or regorafenib (n = 37). As of November 13, 2023, the median survival follow-up was 20.4 months. The primary endpoint of progression-free survival (PFS) was improved with EZFB (6.2 months) versus regorafenib [2.1 months; hazard ratio (HR), 0.27; 95% confidence interval (CI), 0.17–0.43; nominal P < 0.0001], as was the secondary endpoint of overall survival (OS; EZFB, 19.7 months; regorafenib, 9.5 months; HR, 0.37; 95% CI, 0.22–0.63; nominal P = 0.0003). The confirmed overall response rate was 17% (90% CI, 10.6%–26.1%) with EZFB and 3% (90% CI, 0.1%–12.2%) with regorafenib. Treatment-emergent adverse events (TEAE), grade ≥3 TEAEs, and TEAEs leading to discontinuation of all study treatments were reported in 99%, 82%, and 5% of the EZFB arm and in 87%, 49%, and 17% of the regorafenib arm, respectively. Conclusions: EZFB significantly improved survival outcomes compared with regorafenib in patients with mCRC as a third-line treatment, with a manageable safety profile. Further investigation is warranted, given the clinically meaningful improvements in PFS and OS.

Emerging preclinical evidence challenges the long-standing assumption that Interleukin-1β (IL-1β) uniformly promotes non–small cell lung cancer (NSCLC). We show that, in the context of chemo-immunotherapy, IL-1β enhances anti-tumor immunity by inducing tumor-cell CXCL10 expression and recruiting CD8+ T cells, thereby sensitizing “cold” tumors to treatment. These findings contrast sharply with the failure of multiple CANOPY trials targeting IL-1β, suggesting that blockade may be effective only in prevention or early carcinogenesis. Instead, controlled IL-1β activation, guided by biomarkers and combined with chemotherapy plus PD-1 blockade, may represent a promising strategy to overcome resistance in established NSCLC.

Supplementary Figure S9: Deconvolution analysis on 20 immune cell types did not reveal any significant differences in activated NK cell levels between the two groups. No differences in T cell populations were observed between both groups.

List of material used in our study

Supplementary Figure S2: Principal Component Analysis revealed that the center of origin was a significant source of variance in the data. Consequently, the center of origin was included as a covariate in all subsequent differential expression analyses to mitigate its impact.

Patient dispositions. EZFB, etrumadenant, and zimberelimab with mFOLFOX-6 and bevacizumab; mFOLFOX-6, modified FOLFOX-6.

Ethics committee approval information

PFS subgroups analysis. 1L, first-line therapy; BOR, best overall response; CR, complete response; ECOG PS, Eastern Cooperative Oncology Group performance status; EZFB, etrumadenant and zimberelimab with mFOLFOX-6 and bevacizumab; HR, hazard ratio; mFOLFOX-6, modified FOLFOX-6; MSI, microsatellite instability; MSI-L, MSI low; MSS, microsatellite stable; NGS, next-generation sequencing; PCR, polymerase chain reaction; PD, progressive disease; PFS, progression-free survival; PR, partial response; rego, regorafenib; SD, stable disease. aDisease progression occurred >2 months after the last prior oxaliplatin dose in the metastatic setting (n = 11), or inclusion was approved by the medical monitor (n = 1)

Supplementary Table S3. Patients with c-Met expression above the median showed a trend toward better clinical outcomes