All treatment-emergent adverse events

Abstract Background Previous retrospective work found an association between earlier immune-checkpoint inhibitors (ICI) infusion times and survival outcomes in renal cell carcinoma (RCC). A recent randomized trial in non-small cell lung cancer (NSCLC) also reported benefit, however, a recent analysis of several randomized trials in NSCLC found no association. Prior studies have used heterogeneous exposure definitions. Whether time-of-day (ToD) of ICI infusion impacts survival in real-world RCC patients remains unclear. We evaluated the association between ToD of ICI infusion and overall survival (OS) in RCC patients treated at Dana-Farber Cancer Institute (DFCI), using the first infusion (C1D1) as the primary exposure on biological grounds. Methods We conducted a retrospective cohort study of RCC patients receiving first- or second-line ICI-based therapy at DFCI (June 2015-December 2024). OS was assessed considering C1D1 infusion time (before vs after 12 PM; before vs after 3 PM; and three categories (<12 PM, 12-4 PM, >4 PM)); and a cumulative exposure definition matching prior mRCC studies (≥25% of infusions after 4:30 PM vs < 25%). Cox models were adjusted for age, sex , season, tumor mutational burden and line of therapy. 3-month landmark analyses addressed immortal time bias. Results 117 RCC patients received first- or second-line ICI-based therapy at DFCI between June 2015 and January 2024 (median follow-up 33 months, IQR 16.3–62.3; 84 deaths). C1D1 infusion time was not associated with OS at any cutoff, before vs after 12 PM (HR 0.77, 95% CI 0.47–1.27, p = 0.31), before vs after 3 PM (HR 0.98, 95% CI 0.59–1.62, p = 0.93) or three-category comparisons (morning vs late afternoon/evening: HR 0.64, 95% CI 0.31–1.33, p = 0.23; midday/afternoon vs late afternoon/evening: HR 0.80, 95% CI 0.41–1.54, p = 0.50). However, the cumulative exposure definition (≥25% (n = 15) of infusions after 4:30 PM vs < 25% (n = 102)) showed statistically significant worse OS for the group that received late infusions (HR 1.93, 95% CI 1.01–3.67, p = 0.046). This signal persisted after a 3-month landmark analysis (HR 2.14, 95% CI 1.11–4.12, p = 0.023), arguing against immortal time bias. Conclusions In this real-world cohort, C1D1 timing of ICI infusion was not associated with OS. An exploratory cumulative-exposure analysis (≥25% of all infusions after 4:30 PM) was associated with worse OS, persisting after landmark analysis. Biologically, any effect of infusion timing would plausibly be largest at the first infusion; we therefore consider the exploratory cumulative-exposure signal difficult to reconcile with a direct circadian effect on ICI activity. Whether this signal reflects a true biological effect or unmeasured confounding factors needs further investigation.

Patients with resectable stage III-IV melanoma are at high risk of recurrence following surgical excision. Neoadjuvant therapy (NEO) with immune checkpoint inhibition (ICI) improves event-free survival and may allow for pathologic treatment response assessment with potential adjuvant therapy de-escalation. Identification of a biomarker correlated with tumor response could facilitate individualized therapeutic decision-making. This study investigates the utility of circulating tumor DNA (ctDNA) as a predictive biomarker of pathologic response in melanoma treated with NEO ICI. We identified melanoma patients treated with NEO ICI at Dana-Farber Cancer Institute and measured personalized ctDNA plasma levels pretreatment, before each NEO ICI cycle, and postoperatively. We evaluated whether ctDNA levels during the NEO course correlated with graded pathologic response. Of 18 patients who underwent NEO ICI, there were 10 pathologic complete responses (pCR; 0% viable tumor), 1 pathologic near-complete response (near pCR; >0% but ≤10% viable tumor), 1 partial response (pPR; >10%-≤50% viable tumor), and 6 nonresponses (pNR; >50% viable tumor). Responders (pCR, near pCR, or pPR) were more likely to have negative presurgical ctDNA levels (100% (12/12) vs. 33% (2/6), P =0.005). In addition to absolute levels, ctDNA kinetics during NEO ICI were also predictive of pathologic response, with nonresponders more likely to show an increase in ctDNA levels between the first and final ICI cycles compared with responders (mean change ctDNA (SD), 0.09 (0.15) vs. -1.16 (3.31), P =0.004). These findings underscore the potential of ctDNA as a dynamic biomarker to inform response-driven, personalized treatment strategies following NEO ICI.

Taxonomic differences from baseline between treatment arms at after antibiotic preconditioning and at C1D1

CD8 intra-tumoral infiltration percentage by IHC

Tertiary lymphoid structures (TLSs) are associated with improved responses to immune checkpoint blockade across solid tumours1,2, but how they impact the phenotypic properties of tumour-specific T cells remains unclear. Here we found, across 24 treatment-naive renal cell carcinoma (RCC) tumours, that TLS-containing tumours are more heavily infiltrated by exhausted CD8+ T cells and have a reduced terminal exhaustion transcriptional program compared with TLS- tumours. Specificity screening of 554 T cell clonotypes expanded within the microenvironment of 6 RCC tumours revealed 82 TCRs that were reactive against tumour cells and/or RCC antigens. A subset of tumour-specific T cell clonotypes (12%) was enriched within TLSs, and these expressed an increased program of stem-like progenitor exhaustion, associated with favourable anti-tumour immunity. However, in 60 independent RCC tumours, macrophages within tumour margins of TLS-containing tumours had an inferred immunosuppressive phenotype and were colocalized with exhausted putative tumour-reactive T cells in a subgroup that was further analysed, therefore supporting this mode of immune evasion as a counterbalance to T cell immune pressure. Our data reveal that TLSs are reservoirs of tumour-specific T cells with stem-like progenitor features that could be leveraged by T cell immunotherapies.

Peripheral Treg populations of interest by baseline Ruminococcaceae status prior to initiating anti-PD1 therapy

The NCCN Guidelines for Melanoma: Cutaneous provide strategies for the diagnosis, treatment, and follow-up of patients with cutaneous melanoma, which is among the most common cancers in the United States. The outcome of cutaneous melanoma is significantly influenced by stage at initial presentation. Generally, localized disease and smaller primary tumors have been associated with more favorable prognosis. Thus, the select content featured in this issue focuses on risk factors associated with cutaneous melanoma development and which should be considered during an initial assessment. These include (1) phenotypic predispositions, (2) personal medical history or comorbidities, (3) genetic predispositions, and (4) environmental factors.

The NCCN Guidelines for Melanoma: Cutaneous provide strategies for the diagnosis, treatment, and follow-up of patients with cutaneous melanoma, which is among the most common cancers in the United States. The outcome of cutaneous melanoma is significantly influenced by stage at initial presentation. Generally, localized disease and smaller primary tumors have been associated with more favorable prognosis. Thus, the select content featured in this issue focuses on risk factors associated with cutaneous melanoma development and which should be considered during an initial assessment. These include (1) phenotypic predispositions, (2) personal medical history or comorbidities, (3) genetic predispositions, and (4) environmental factors.

Assessment of gut microbiome networks by treatment arm and across timepoints

Improved methods to identify therapeutically relevant tumor neoantigens and their cognate T cells would aid the development of precision medicines for cancer. Here, we developed Slide-GoTags, a droplet-based single-nucleus spatial transcriptomics approach that characterizes neoantigen-specific immunity by integrating targeted transcript genotyping and T cell receptor (TCR) sequencing with single-nucleus RNA sequencing from the same slice of frozen tissue. Application of Slide-GoTags to mouse and human tumors revealed colocalization of clonally expanded, neoantigen-specific T cells with tumor cells expressing their cognate neoantigen. We also identified distinct spatial immune landscapes shaped by anti-PD1 or anti-CTLA4 blockade in mouse colorectal tumors. Across human tumor types, Slide-GoTags detected TCR–neoantigen interactions through spatial proximity and identified an enrichment of interferon-driven immunogenicity niches in immunologically ‘hot’ tumors compared to ‘cold’ tumors. These niches harbored three T cell clonotypes that colocalized with genotyped neoantigens, highlighting a spatially organized antitumor immune response. Collectively, Slide-GoTags establishes a framework for in situ mapping of T cell–tumor interactions directly from individual tissue. Spatial relationships between tumor neoantigens and cognate T cells are studied.

Progression-free survival in a retrospective melanoma immunotherapy cohort by baseline Ruminococcaceae status

Summary of clinical response by treatment arm and baseline Ruminococcaceae group

Disease control and objective response rates by Ruminococcaceae status

Overview of exploratory correlative specimen availability

Although immunotherapy has revolutionized cancer treatment, many patients still experience limited benefit, highlighting the urgent need for improved biomarkers1. Although immunotherapy is founded on unleashing T cells2, most existing biomarkers remain tumour-centric and mainly overlook host immune competence. The thymus is a key immune organ that is crucial for T cell maturation, and we hypothesized that thymic functionality is associated with immunotherapy outcomes3. Here we show that thymic health, a radiographic measure of thymic functionality, is strongly associated with immunotherapy outcomes across several cancer types. Using a deep-learning framework applied to routine computed tomography images, we quantified thymic health in a pan-cancer cohort of 3,476 patients receiving immune checkpoint inhibitors. In patients with non-small cell lung cancer, higher thymic health was associated with reduced risks of progression and all-cause mortality. These associations remained significant across clinically relevant levels of programmed death ligand 1 (PD-L1) and tumour mutation burden. In the prospective TRACERx lung cancer study, thymic health was positively associated with T cell receptor diversity and T cell receptor excision circles, and correlated with immune-system signalling pathways, supporting radiographic thymic health as a proxy for thymic activity and adaptive immune competence. Analysis across patients with melanoma, breast cancer or renal cancer demonstrated pan-cancer relevance. Together, these findings identify thymic health as a previously unrecognized, tumour-agnostic determinant of immunotherapy efficacy, with potential implications for patient stratification, treatment timing and the development of immune-rejuvenating strategies in precision immuno-oncology.

Best objective response by baseline Ruminococcaceae abundance levels

Longitudinal serum proteomics analysis

Patient characteristics by baseline Ruminococcaceae status

Differences in baseline peripheral immune cell populations by response and baseline Ruminococcaceae levels