Brad H. Nelson

Tumour-infiltrating lymphocyte (TIL) therapy can be effective against metastatic melanoma, including immune checkpoint-resistant disease, but current protocols require lymphodepletion and high-dose interleukin-2 (IL-2), which can cause substantial toxicity. To overcome this limitation, we engineered a chimeric receptor, G2R, that generates an IL-2-like proliferative signal in response to granulocyte colony-stimulating factor (G-CSF), a well-tolerated cytokine widely used in oncology. Human T cells expressing G2R expanded in vitro in response to G-CSF in place of IL-2 or interleukin-15 (IL-15), and the three cytokines induced broadly similar intracellular signalling and cytokine secretion profiles. In immune-competent mice bearing established B16-F10 melanoma, adoptive transfer of G2R-expressing, tumour-reactive CD8 ⁺ T cells followed by systemic G-CSF treatment induced profound T cell expansion. This was accompanied by complete, durable tumour regression in over 75% of animals without the need for lymphodepletion, exogenous IL-2 or any other therapy. G-CSF treatment induced transient neutrophil expansion in peripheral blood but no increase in intratumoural neutrophils or macrophages. G2R-expressing memory T cells persisted long-term and could be re-expanded upon G-CSF re-treatment, enabling rejection of second tumour challenges. Thus, G-CSF can be repurposed to enable clinician-controlled anti-tumour T cell responses of profound magnitude, efficacy and durability, an approach that is readily translatable to humans.

Breast cancer that arises in the postpartum period often carries a poor prognosis and urgently requires chemoprevention strategies, particularly for women at elevated risk. Here, we report that a single post-lactation dose of cabergoline, a dopaminergic agonist that blocks prolactin secretion, delays the onset and reduces the incidence of postpartum mammary cancer in K14-Cre; Brca1F/F, Trp53F/F mice. In these mice, cabergoline treatment remodels the mammary gland by potentiating involution, reducing ductal area and proliferation, and reversing T cell exhaustion, potentially through modulation of ion channel expression. Notably, independent retrospective studies in two large cohorts of European women showed that cabergoline treatment was associated with a lower incidence of postpartum breast cancer, with a meta-analysis indicating an approximate 69% reduction in risk. Our work underscores the importance of targeting postlactational involution as a strategy for the prevention of postpartum breast cancer and positions cabergoline as a novel, low-risk chemoprevention strategy.

Univariable DSS according to immune biomarkers. Kaplan–Meier curves show DSS stratified by (A) PD-L1 CPS 0 vs. ≥1; (B) PD-L1 CPS <10 vs. ≥10; (C) dMMR vs. pMMR; (D) intraepithelial CD3+ T-cell presence vs. absence; (E) intraepithelial CD8+ T-cell presence vs. absence; (F) stromal CD3+ T-cell counts categorized as low vs. high; (G) intraepithelial CD3+ T-cell counts categorized as low vs. high; (H) stromal CD8+ T-cell counts categorized as low vs. high; and (I) intraepithelial CD8+ T-cell counts categorized as low vs. high. High vs. low count categories were defined using the cohort mean for each marker and compartment (CD3: 56 cells in stroma and 11 cells in epithelium; CD8: 27 cells in stroma and 5 cells in epithelium). Numbers at risk are displayed within each panel. P values are from two-sided log-rank tests. Epi, epithelium; Str, stroma.

Prevalence of descriptive MMR/CD8 phenotypes among 98 tumors with both markers evaluable. dMMR was assigned regardless of CD8 localization; inflamed pMMR denotes pMMR with intraepithelial CD8+ T-cell presence; and noninflamed pMMR denotes pMMR without intraepithelial CD8+ T-cell presence. The categories are descriptive and are not treatment-selection classes.

Table 1 from Compartment-Specific CD8 Infiltration and Disease-Specific Survival in Resected Ampullary Adenocarcinoma

Likelihood-ratio P values for biomarker associations with pathology variables.

Exploratory adjusted associations between TIL measures and DSS.

Ampullary adenocarcinoma is rare, and its immune biomarker landscape is incompletely characterized. This article evaluated mismatch repair (MMR), programmed cell death ligand 1 (PD-L1; CD274), and compartment-specific CD3 (CD3E)/CD8 (CD8A) tumor-infiltrating lymphocytes (TIL) in resected tumors and explored associations with disease-specific survival (DSS). Ninety-nine patients underwent pancreaticoduodenectomy at Vancouver General Hospital (VGH) between 1984 and 2013. Duplicate 0.6-mm tissue microarrays (TMA) were assessed for MMR proteins, PD-L1 combined positive score (CPS), and stromal and intraepithelial CD3 and CD8 counts. DSS was estimated using the Kaplan–Meier method. Exploratory Cox models were adjusted for age, histologic subtype, pathologic tumor (pT) group, pathologic node (pN) status, and adjuvant chemotherapy. There were 44 DSS events. MMR deficiency (dMMR) was present in 37 of 99 (37.4%) tumors. PD-L1 was evaluable in 87 tumors; 57 of 87 (65.5%) tumors had CPS ≥1, and 24 of 87 (27.6%) tumors had CPS ≥10. Intraepithelial CD8+ T cells were present in 23 of 98 (23.5%) tumors. PD-L1 CPS ≥1, PD-L1 CPS ≥10, and MMR status were not associated with DSS by log-rank testing (P = 0.35, P = 0.98, and P = 0.74, respectively). In the pT-adjusted model, intraepithelial CD8 presence was associated with lower disease-specific hazard [hazard ratio (HR), 0.422; 95% confidence interval (CI), 0.144–0.994; likelihood-ratio P = 0.048], although the Wald sensitivity test was not significant (P = 0.074). Continuous log-transformed intraepithelial CD8 counts showed a concordant, method-dependent signal. dMMR and PD-L1 expression were common in this historical resected cohort. Intraepithelial CD8 infiltration was associated with DSS, but the borderline and inferential method–dependent estimates require external validation. These prognostic data do not establish benefit from immune checkpoint blockade. Significance:Ampullary cancer is a rare disease in which treatment is extrapolated from other gastrointestinal cancers. In this cohort, immune features varied among tumors with functioning MMR. CD8+ T cells within the tumor epithelium showed an exploratory association with improved DSS, which supports the prospective evaluation of immune stratification, not immediate treatment selection.

Ampullary adenocarcinoma is rare, and its immune biomarker landscape is incompletely characterized. This article evaluated mismatch repair (MMR), programmed cell death ligand 1 (PD-L1; CD274), and compartment-specific CD3 (CD3E)/CD8 (CD8A) tumor-infiltrating lymphocytes (TIL) in resected tumors and explored associations with disease-specific survival (DSS). Ninety-nine patients underwent pancreaticoduodenectomy at Vancouver General Hospital (VGH) between 1984 and 2013. Duplicate 0.6-mm tissue microarrays (TMA) were assessed for MMR proteins, PD-L1 combined positive score (CPS), and stromal and intraepithelial CD3 and CD8 counts. DSS was estimated using the Kaplan–Meier method. Exploratory Cox models were adjusted for age, histologic subtype, pathologic tumor (pT) group, pathologic node (pN) status, and adjuvant chemotherapy. There were 44 DSS events. MMR deficiency (dMMR) was present in 37 of 99 (37.4%) tumors. PD-L1 was evaluable in 87 tumors; 57 of 87 (65.5%) tumors had CPS ≥1, and 24 of 87 (27.6%) tumors had CPS ≥10. Intraepithelial CD8+ T cells were present in 23 of 98 (23.5%) tumors. PD-L1 CPS ≥1, PD-L1 CPS ≥10, and MMR status were not associated with DSS by log-rank testing (P = 0.35, P = 0.98, and P = 0.74, respectively). In the pT-adjusted model, intraepithelial CD8 presence was associated with lower disease-specific hazard [hazard ratio (HR), 0.422; 95% confidence interval (CI), 0.144–0.994; likelihood-ratio P = 0.048], although the Wald sensitivity test was not significant (P = 0.074). Continuous log-transformed intraepithelial CD8 counts showed a concordant, method-dependent signal. dMMR and PD-L1 expression were common in this historical resected cohort. Intraepithelial CD8 infiltration was associated with DSS, but the borderline and inferential method–dependent estimates require external validation. These prognostic data do not establish benefit from immune checkpoint blockade. Significance: Ampullary cancer is a rare disease in which treatment is extrapolated from other gastrointestinal cancers. In this cohort, immune features varied among tumors with functioning MMR. CD8+ T cells within the tumor epithelium showed an exploratory association with improved DSS, which supports the prospective evaluation of immune stratification, not immediate treatment selection.

TableS1. Patient characteristics of the CHL study training cohorts. Table S2. Gene list for the targeted sequencing panel and HLGen. Table S3 Patient Demographic and molecular alterations. Table S4. HLGen correlation with Age and EBV in Training and Validation cohort. Table S5. Antibodies used for immunohistochemistry staining. Table S6. Differential gene expression analysis followed by gene ontology analysis in L-428 model. Table S7. Differential gene expression analysis followed by gene ontology analysis in U-HO1 model + IL-3. Table S8 Differential gene expression analysis followed by gene ontology analysis in U-HO1 model + IL-5. Table S9 Gene expression changes across all three experimental models. Table S10. Classification methods for HLGen. Table S11 HLGen Prediction in Training cohort. Table S12 HLGen Prediction in Stanford cohort (Alig and colleagues, Nature 2024). Table S13. Antibodies used for immunohistochemistry staining. Table S14. IMC Clustering Strategy. Table S15. Antibodies used for flow cytometry staining. Table S16. Antibodies used for Western blotting.