12141 Background: Current adverse event (AE) reporting standards emphasize peak toxicity, which may inadequately capture the cumulative burden of persistent low-grade AEs characteristic of targeted therapies. We conducted a post-hoc analysis of the LUX-Lung 3 trial to compare longitudinal toxicity patterns of targeted therapy and chemotherapy over time. Methods: This post-hoc analysis used individual, de-identified patient-level data from the phase III LUX-Lung3 clinical trial comparing afatinib with platinum-based chemotherapy as first-line treatment for advanced EGFR-mutant NSCLC, accessed through Vivli. Select treatment-related AEs were evaluated longitudinally by grade. Cumulative toxicity burden was quantified using the area-under-the-curve (AUC) of AE grade prevalence over time. Pearson correlation was used to assess temporal trends in AUC. Results: A total of 340 patients had evaluable AE data, including 229 (67%) treated with afatinib and 111 (33%) treated with chemotherapy. The median onset of grade 1-2 AEs generally occurred earlier with chemotherapy than afatinib. The median onset of chemotherapy-related fatigue was 4 days and nausea 2 days, whereas median onset of afatinib-related diarrhea was 4 days, rash 8 days, and paronychia 37 days. Longitudinal AUC analysis (Table 1) demonstrated a significant improvement in grade 1-2 chemotherapy-related nausea (p=0.003) and stomatitis (p=0.05), and a trend towards improvement in fatigue (p=0.2) by 12-months. In contrast, grade 1-2 afatinib-related rash (p=0.4) and diarrhea (p=0.99), remained stable, while paronychia increased over time (p=0.004). For grade 3+ AEs, the AUC improved over time for both the chemotherapy (p=0.04) and afatinib (p=0.02) groups. Conclusions: Assessment of time to onset and longitudinal AUC reveal distinct temporal AE patterns for targeted therapy and chemotherapy. Longitudinal toxicity reporting is feasible and highlights the sustained burden of chronic AEs associated with targeted therapies that are clinically meaningful yet underrepresented by conventional peak toxicity reporting. Cumulative toxicity of afatinib and chemotherapy was quantified using the area under the curve of grade 1-2 adverse events prevalence at landmark time points.* Adverse event Month 1 Month 3 Month 6 Month 12 Pearson’s coefficient p-value Afatinib (%) Rash 0.24 0.39 0.44 0.45 0.24 0.4 Diarrhea 0.30 0.44 0.47 0.45 -0.002 0.99 Paronychia 0.02 0.20 0.32 0.39 0.74 0.004 Chemotherapy (%) Fatigue 0.05 0.09 0.09 0.07 -0.36 0.2 Nausea 0.07 0.17 0.17 0.10 -0.60 0.03 Stomatitis 0.02 0.02 0.02 0.01 -0.56 0.05 *Trends were assessed using Pearson’s correlation coefficient and corresponding p-value.

Introduction Individual health and life expectancy, rather than age, should drive management decisions for patients with prostate cancer. There is limited available data regarding treatment outcomes and tolerance for older adults where health status warranted treatment with curative intent. Herein, we report treatment outcomes for patients 80 years and older who received definitive radiation therapy (RT) for localized prostate cancer. Methods This retrospective study includes 41 patients who received RT for localized prostate cancer at the age of 80 years or older between 2004 and 2022. The median age was 83. Baseline details and outcomes, including treatment toxicity, symptom scores, biochemical recurrence (BR), distant metastases (DM), and overall survival (OS), were collected in a secure database and analyzed with Graphpad Prism (GraphPad Software, Boston, MA, USA) and R software (version 4.5; R Foundation for Statistical Computing, Vienna, Austria). Results No patients experienced adverse events that were Common Terminology Criteria for Adverse Events (CTCAE) grade 4 or higher. One patient (2.4%) experienced late CTCAE grade 3 proctitis, seven patients (17.1%) experienced acute CTCAE grade 2 genitourinary adverse events (GU AE), and five patients (12.2%) experienced late grade 2 GU AEs. Mean urinary symptom scores improved after RT (with the International Prostate Symptom Score (IPSS) decreasing from 12.1 pre-RT to 8.2 post-RT). The estimated four-year cumulative incidence of metastases was 6.7% (95% CI 0-16.1%). There were trends in outcomes based on risk group, but no statistically significant differences between intermediate risk cancer (IR) and high or very high-risk cancer (HR/VHR) regarding BF (15% vs. 20%, p=0.17), DM (0% vs. 10%, p=0.20), or OS (94% vs. 96%, p=0.38). Conclusions Definitive RT for patients 80 years or older was generally well tolerated and effective in this small cohort. These findings may indicate a favorable risk/benefit ratio for definitive prostate cancer treatment for select patients 80 years or older. Decision-making for prostate cancer treatment should involve a comprehensive approach, accounting for each individual patient's life expectancy and overall health rather than avoidance of treatment based simply on age.

Figure S3. Post-hoc analysis comparing stage 1 and stage 2 clinical outcomes.

PURPOSE:Doxorubicin is standard chemotherapy for metastatic soft tissue sarcomas (STS) but also enhances innate/adaptive immune responses by inducing immunogenic cell death. Most STS are immune "cold" tumors that do not respond to immune checkpoint inhibitors (ICI) blocking PD-1 and cytotoxic T lymphocyte antigen-4. We hypothesized that concurrent doxorubicin would improve tumor immunogenicity and boost the efficacy of ICI in STS. PATIENTS AND METHODS:We conducted a single-arm, phase 2 trial of doxorubicin plus zalifrelimab (anti-cytotoxic T lymphocyte antigen-4 antibody) and balstilimab (anti-PD-1 antibody) for patients with advanced/metastatic STS without prior doxorubicin or ICI (NCT04028063). The study was a Simon minimax two-stage design to accrue 28 patients evaluable for primary endpoint of progression-free survival rate at 6 months (PFS6mo) by RECIST 1.1. The study aimed to improve PFS6mo by 20% over a historic null rate of 43.4% with doxorubicin monotherapy. Secondary endpoints included the objective response rate, disease control rate, overall survival, duration of response, and adverse events (AE). RESULTS:The PFS6mo for 28 evaluable patients was 46.4% [95% confidence interval (CI), 27.5-66.1] and not superior to the null rate, with a median PFS of 25.3 weeks (95% CI, 24.0-42). The best objective response rate was 33.3% (95% CI, 17.3-52.8) with a disease control rate of 80.0% (95% CI, 61.4-92.3), including STS types unlikely to respond to doxorubicin or ICI alone. Grade 3/4 treatment-related AE occurred in 45% of patients, with immune-mediated AE requiring immunosuppression in 9%. CONCLUSIONS:Although the study did not meet the predefined endpoint for PFS improvement, promising signals of efficacy warrant future investigation including response/resistance biomarkers to inform patient selection.

Changes in intratumoral heterogeneity in mHMTA PDXs can be explained by neoAg selection. A, NeoAg distribution across HLA alleles (B) Kullback–Leibler logos show enrichment of hydrophobic amino acids at position 9 in the neoAg pool. The height of each one letter amino acid code is the absolute enrichment relative to unselected peptides. Values above the x axis are positively enriched whereas those below are depleted. C, Distribution of estimated changes in variant allele frequency between the mHMTA and mHM models in all variants identified by exome sequencing. Values to the right of the dotted line represent variants that enriched, or underwent positive selection in mHMTA, whereas values to the left represent variants that underwent negative selection. D, Significant changes between mHM and mHMTA variant allele frequencies are plotted against the predicted HLA binding affinity for nine-mer peptides derived from each variant. The points are colored by which class I HLA molecule the peptide was predicted to bind. All significantly altered variants were found to produce at least two neoAgs. Error bars represent SE. E, Single letter codes for each amino acid are shown with the mutant residue outlined in each. The majority of mutations occurred at TCR contact residue locations (positions 3–8).

Breast cancer is the second most common cancer globally, with most deaths caused by metastatic disease, often following long periods of clinical dormancy1. Understanding the mechanisms that disrupt the quiescence of dormant disseminated cancer cells (DCCs) is crucial for addressing metastatic progression. Infections caused by respiratory viruses such as influenza and SARS-CoV-2 trigger both local and systemic inflammation2,3. Here we demonstrate, in mice, that influenza and SARS-CoV-2 infections lead to loss of the pro-dormancy phenotype in breast DCCs in the lung, causing DCC proliferation within days of infection and a massive expansion of carcinoma cells into metastatic lesions within two weeks. These phenotypic transitions and expansions are interleukin-6 dependent. We show that DCCs impair lung T cell activation and that CD4+ T cells sustain the pulmonary metastatic burden after the influenza infection by inhibiting CD8+ T cell activation and cytotoxicity. Crucially, these experimental findings align with human observational data. Analyses of cancer survivors from the UK Biobank (all cancers) and Flatiron Health (breast cancer) databases reveal that SARS-CoV-2 infection substantially increases the risk of cancer-related mortality and lung metastasis compared with uninfected cancer survivors. These discoveries underscore the huge impact of respiratory viral infections on metastatic cancer resurgence, offering new insights into the connection between infectious diseases and cancer metastasis.

Background: The majority of ER+ breast cancers (BC) express androgen receptors (AR). In a randomized phase II trial for women with ER+/HER2- primary BC T2 or greater, neoadjuvant fulvestrant (Fulv) alone or with enzalutamide (Combo) was given for 4 months prior to surgery. A total of 59 patients were evaluable: 33 on Combo and 26 on Fulv. The addition of AR blockade to Fulv reduced residual tumor at time of surgery as measured by modified preoperative endocrine predictive index (PEPI) score. Fresh tumor biopsies were required at study entry (baseline), after 4 weeks on therapy (W5), and at surgery. The Combo arm achieved PEPI=0 more frequently (24%: 8/33) than Fulv (8%: 2/26). Interestingly, the odds of response were 4.6-fold (95% CI: 0.9-22) higher for patients with invasive lobular cancer (IDC) versus invasive ductal (IDC). Results: When examining all tumors, gene expression analyses showed significantly decreased estrogen response and cell division gene sets in tumors in both arms; however, only Combo treated tumors exhibited significant enrichment of immune activation genes sets, including interferon gamma, complement, inflammation, antigen processing, and B and T cell activation. AR protein was significantly reduced by time of surgery in the Combo arm only (P<0.05) as measured by immunohistochemistry. AR was also significantly lower at time of surgery in the PEPI=0 as compared to PEPI >0 (P<0.05) and in the Ki67 responders versus non-responders (p<0.02). Because of the 4.6-fold higher odds of PEPI=0 response in ILCs versus IDC, we examined phosphoproteins that changed with treatment in these two histologic groups by reverse phase phosphoprotein assay (RPPA) from frozen tumor sections. Both AR and phosphoS650 AR showed significantly more decrease from BL to W5 in ILC versus IDC. Cyclin D1, S6RP, HIF-1 alpha, ATP citrate lyase, were significantly lower in ILC than IDC, while CHK1 and ALK were higher. As would be expected with the higher odds of response in ILC, cell cycle proteins decreased significantly more with treatment in this histologic subtype, as did growth/survival and metabolism proteins. Polaris Multiplex immunofluorescence, used to study the tumor immune microenvironment (TIME), revealed that the number of tertiary lymphoid structures (TLS) per area surrounding resected tumor at time of surgery was higher in the Combo arm and was significantly higher near tumors that achieved PEPI=0 (P<0.03). Additionally, the average number of TLS/mm2 was higher in the ILC versus IDC (P < 0.059). T regulatory cells were reduced in the Combo arm (p<0.0002) and in Ki67 responsive tumors (p<0.004); however, T regs were not significantly different in ILC versus IDC. Tumor-associated macrophages decreased by time of surgery only in the Combo arm (p<0.0001), only in tumors that achieved PEPI=0 (p<0.0005) and trended towards decreased numbers in ILC vs IDC. Conclusions: AR inhibition in combination with a SERD activates the immune system in ER+ BC, particularly in ILC. Citation Format: Jennifer Richer, Anthony D. Elias, Alyse W. Staley, Monica Fornier, Gregory A. Vidal, Vida Alami, Sharon Sams, Nicole S. Spoelstra, Andrew Goodspeed, Peter Kabos, Jennifer R. Diamond, Elena Shagisultanova, Rosa I Gallagher, Julia Wulfkuhle, Emanuel Petricoin, Kathryn Zolman, Tessa McSpadden, Christian Rickert, Kimberly R. Jordon, Jill E. Slansky, Virginia F. Borges, Dexiang Gao. Differential odds of response in ILC versus IDC correlate with changes in the TIME in a phase II trial of pre-operative fulvestrant with or without enzalutamide [abstract]. In: Proceedings of the San Antonio Breast Cancer Symposium 2024; 2024 Dec 10-13; San Antonio, TX. Philadelphia (PA): AACR; Clin Cancer Res 2025;31(12 Suppl):Abstract nr PS18-07.

Establishing recommended phase II dose (RP2D) and dose limiting toxicities (DLTs)

All adverse events experienced by patients, including those unrelated to treatment

PFS before and after palbociclib dose change

Background:Primary analysis of the phase I/II clinical trial combining pembrolizumab with all-trans retinoic acid (ATRA) for patients with metastatic melanoma showed a median progression-free survival (PFS) of 20.3 months and median overall survival (OS) not reached. Objective:Report 5-year OS and PFS rates. Design:In this single-center, single-arm trial (NCT03200847), 24 anti-PD-1 naïve patients with metastatic melanoma were enrolled between 10/2017 and 07/2020. Methods:The Kaplan-Meier method was used to estimate OS and PFS. Results:At data cutoff (May 14, 2024), all patients had completed treatment, and 58% (n = 14) remained alive with a median follow-up of 48 months. The updated 5-year OS rate is 54.7% (95% confidence interval (CI): 36.4-82.1), and the 5-year PFS is 36.1% (95% CI: 21.0-62.2), with 33% (n = 8) having an ongoing complete response. Conclusion:These results underscore the long-term clinical benefit of combining anti-PD-1 with ATRA, with survival rates comparable to anti-PD-1/combinations but with lower toxicity.

Supplementary figure 1. (A) Changes in other circulating cytokines. Colored box denotes the time when the patients were being treated with ATRA. (B) example gating strategy for CD8+ T cell activation.

Grade 3 and 4 AEs - palbociclib 125mg vs 75mg

Examination of T-cell development and activity in the mHMTA. A, Representative cytometry of the human CD45+ cells and HSPCs present in mHM and mHMTA bone marrow. B, Comparison of the average percentage of human CD45+ cells and CD34+ HSPCs within this CD45+ cell population within mHM and mHMTA bone marrow (P = 0.118 and 0.379, respectively, Wilcoxon rank-sum test). C, Comparison of peripheral human CD45+ lymphocytes in mHM and mHMTA (P = 0.728, Wilcoxon rank-sum test) and of CD19+ B cells in mHM and mHMTA (0.224, Wilcoxon rank-sum test). D, Cytometric analysis and a comparison of human CD45+ cells, as well as T-cell and B-cell populations, found in the spleens of mHM and mHMTA. E, Comparison of the average human CD45+ cell populations, human T cells, and human B cells on mHM and mHMTA spleens (P = 0.844, 0.450, and 0.646, Wilcoxon rank-sum test, respectively). F, IFNγ expression was detected in the spleen of both mHM and mHMTA (P < 0.005). G, Gating and representative cytometry of T cells in mHM and mHMTA show relative populations used to analyze T-cell activation. Purple boxes in the first panels show the T-cell populations gated in the remaining scatter plots. The red boxes show the CD8+ T-cell populations gated for subsequent analysis. H, Representative cytometry of the CD8+ T cells, gated from the red boxes in the previous panel. I, Comparisons of the ratio of HLA-DR + PD1– T cells with all HLA-DR+ T cells shows an increasingly activated T-cell population in mHMTA (P = 0.0207, two-group t test) and a nonsignificantly increased population of CD4+ T cells. J, Comparison of the HLA-DR + PD1– population with the total HLA-DR+ T-cell population among CD8+ T cells also reveals a trend toward increased activation in mHMTA. P values: *, ≤0.05.

Trial inclusion/exclusion criteria

Representativeness of study participants