Abstract Background Combination immune checkpoint inhibition with anti-CTLA-4 and anti-PD-1 antibodies improves outcomes in metastatic renal cell carcinoma (mRCC), yet 40–60% of patients fail to achieve meaningful responses. Robust biomarkers that predict deep and durable responses are lacking. We aimed to define systemic and tumor-associated immune signatures associated with therapeutic efficacy using an integrated multi-omic approach. Methods We analyzed peripheral blood and tumor samples from patients with intermediate- and poor-risk mRCC enrolled in a phase II clinical trial evaluating a response-adapted regimen of nivolumab and ipilimumab (NCT03297593, SAKK07/17). High-dimensional mass cytometry was used to profile immune cell populations. Single-cell proteomic and transcriptomic profiling was performed using CITE-seq, and bulk RNA sequencing was conducted on tumor biopsies. Imaging mass cytometry enabled spatial validation within the tumor microenvironment. Differential abundance, gene expression, and pathway analyses were integrated across datasets to identify correlates of response. Results Among evaluable patients, 16 of 24 achieved partial or complete responses, while 8 exhibited stable or progressive disease. Responders demonstrated a significant expansion of CD4+CCR6+ Th17 cells in peripheral blood following therapy, accompanied by increased CD8 effector memory T cells and dendritic cell subsets. Single-cell multi-omic analysis revealed enrichment of Th17-associated transcriptional programs. Pseudotime analysis showed a delayed transition from naïve CD4+ T cells to Th17 cells in non-responders, associated with anti-inflammatory gene signatures, suggesting impaired immune activation. Spatial profiling using imaging mass cytometry validated increased infiltration of CD4+CCR6+ Th17 cells within responder tumors, alongside enhanced immune cell colocalization and reduced stromal features. In contrast, non-responders exhibited immune-excluded, stroma-rich tumor microenvironments. Notably, tumor mutational features, including PBRM1, BAP1, and VHL alterations, did not correlate with response in this cohort. Conclusions A Th17-centric immune signature is strongly associated with deep and sustained responses to dual checkpoint blockade in mRCC. These findings highlight Th17 cells as key mediators of antitumor immunity and potential predictive biomarkers. Integrating systemic and tumor immune profiling provides a framework for patient stratification and the development of therapeutic strategies to overcome resistance. DOD CDMRP Funding yes

Background Prostate-specific membrane antigen (PSMA)-targeted radioligand therapy is the leading theranostic strategy for metastatic castration-resistant prostate cancer (mCRPC). Randomised trials have shown survival benefit with [177Lu]Lu-PSMA-617, but prospective multicentre real-world data for the widely used alternative ligand [177Lu]Lu-PSMA-I&T remain scarce. In this study, we evaluated the safety and clinical activity of [177Lu]Lu-PSMA-I&T in routine practice. Methods This prospective, multicentre, Swiss registry enrolled consecutive men older than 18 years with histologically confirmed prostate adenocarcinoma undergoing intravenous [177Lu]Lu-PSMA-I&T therapy (7 GBq every 6 weeks for up to six cycles). Eligible patients had Eastern Cooperative Oncology Group performance status of 2 or less and PSMA-positive mCRPC with progressive disease after androgen receptor pathway inhibitor therapy or chemotherapy, or both, unless deemed unfit. The primary outcome was safety, assessed by treatment-emergent adverse events graded according to Common Terminology Criteria for Adverse Events (version 5.0). Assessments were performed at predefined timepoints before each treatment cycle and during the post-therapy follow-up. Secondary outcomes included the best prostate-specific antigen (PSA) response and PSA decline of 50% or more (PSA50) response according to the Prostate Cancer Working Group 3 criteria. All patients who underwent at least one treatment cycle were included in the final analysis of the observational registry. This study is registered at ClinicalTrials.gov, NCT06830408. Findings Between May 25, 2020, and June 4, 2024, 333 patients with mCRPC received [177Lu]Lu-PSMA-I&T therapy and were included in the analysis. The median follow-up was 12·0 months (IQR 6·5–20·0). Grade 3 treatment-emergent haematological toxicities included anaemia in 26 (8·8%; 95% CI 5·8–12·6) of 295, thrombocytopenia in nine (3·1%; 1·4–5·7) of 295, leukopenia in five (1·7%; 0·6–3·9) of 295, lymphopenia in 54 (21·1%; 16·3–26·6) of 256, and neutropenia in one (0·4%; 0·0–2·2) of 256. No grade 3 or worse renal toxicity or grade 4–5 treatment-emergent adverse events were observed. Best PSA response was achieved in 198 (64·3%; 95% CI 58·7–69·6) of 308 patients and PSA50 in 137 (44·5%; 38·8–50·2). Interpretation [177Lu]Lu-PSMA-I&T showed clinically meaningful activity with a favourable safety profile in heavily pretreated mCRPC. These findings provide prospective evidence supporting the safety and activity of PSMA-targeted radioligand therapy with [177Lu]Lu-PSMA-I&T in routine clinical practice. Funding Swiss Hadron Foundation.

4503 Background: Perioperative chemo-IO is a new standard for MIBC but pathological complete remission (pCR) rates remain modest. Intravesical Bacillus Calmette Guérin (BCG) is very effective in non-muscle invasive bladder cancer, inducing a local immune response and likely activation of adaptive immunity, with evidence of systemic immune modulation that may enhance downstream immune-oncological effects. The use of BCG in MIBC has not yet been explored, primarily due to concerns about systemic complications. The recombinant BCG vaccine VPM1002BC (rBCG) is associated with enhanced immunogenicity and an improved safety profile. We hypothesize that the combination of intravesical rBCG and chemo-IO in MIBC may act synergistically, augmenting systemic immune activation and hereby improve antitumor efficacy without compromising safety. Methods: SAKK 06/19 is an open-label single arm phase II trial for cT2-T4a N0-1 MIBC patients (pts) eligible for cisplatin and radical cystectomy with lymphadenectomy (RC). rBCG was instilled weekly x3 (day 1, 8, 15). Atezolizumab (Atezo) 1200 mg was administered on day 1 and then x4 every 3 weeks (q3w). Cisplatin and gemcitabine started on day 22 and were given for 4 cycles q3w followed by RC. Only in case of ≥ ypT2 or ypN+, Atezo q3w was given after surgery for 13 cycles. pCR defined as ypT0 ypN0 and assessed by central pathological review was the primary endpoint. Based on Simon’s minimax 2-stage design with H0 pCR ≤ 35% and H1 pCR ≥ 55%, accrual of 46 pts was needed (including 15% dropouts). Secondary endpoints included event free survival (EFS), overall survival (OS), pathological response (PaR, ≤ ypT1N0), feasibility and safety. NCT04630730. Results: 47 pts were included between 04/22 and 04/25 at 10 Swiss sites. 7 pts did not undergo RC (6 refused, 1 unfit for surgery). Of the 40 resected pts 53% had cT2, 37% cT3, 10% cT4 and 88% cN0, 12% cN1. rBCG was instilled in 95% of pts, 78% had all 3 doses. 98% received 4 doses of Atezo and 95% had 4 cycles of platinum (20% switched to carboplatin). pCR according to central review was 65% (26/40; one-sided 95% CI lower boundary of 51%) and PaR was 80% (32/40). Overall pCR for all included pts was 55% (26/47). TRAEs overall, G3, G4 were 48%, 9%, 0% to rBCG, 55%, 15%, 2% to Atezo and 98%, 38%, 17% to chemotherapy. No treatment-related deaths occurred. At a median follow-up of 11.8 months 1-year overall EFS was 88% (95% CI 71 - 95) and OS 95% (95% CI 81 - 99). Conclusions: This is the first trial to combine intravesical rBCG with chemo-IO in MIBC. The combination was feasible and safe without unexpected toxicities. The centrally assessed pCR rate of 65% and PaR of 80% are among the highest reported in an unselected MIBC population. Integration of intravesical rBCG into novel MIBC treatment regimens appears highly promising, particularly in the context of bladder-preservation approaches. Clinical trial information: NCT04630730 .

PURPOSE:Intravesical Bacillus Calmette-Guérin (BCG) is highly effective in non-muscle-invasive bladder cancer (MIBC) but has not been evaluated in MIBC. The recombinant BCG vaccine VPM1002BC (rBCG) has potentially enhanced immunogenicity and an improved safety profile. We investigated neoadjuvant intravesical rBCG combined with chemoimmunotherapy in MIBC. PATIENTS AND METHODS:SAKK 06/19 was an open-label single-arm phase II trial for cT2-T4a N0-1 MIBC eligible for cisplatin and radical cystectomy with lymph node dissection (RC-LND). rBCG was instilled once per week three times starting on day 1. Atezolizumab was administered on day 1 for a total of four doses, and cisplatin/gemcitabine was started on day 22 for four cycles followed by RC-LND. Adjuvant atezolizumab was only administered in the case of >yT1 ypN0. The primary end point was centrally reviewed pathologic complete response (pCR, ypT0 ypN0). Based on Simon's minimax two-stage design with H0 pCR ≤35%, H1 pCR ≥55%, one-sided alpha 5%, and power 80%, 46 patients were needed. Secondary end points included pathologic overall response (PaR, ≤ypT1 ypN0), event-free survival (EFS), overall survival (OS), and safety. RESULTS:Forty-seven patients were included between April 2022 and April 2025. Seven patients did not undergo RC-LND (six declined, one unfit for surgery). rBCG was instilled in 95%, and 78% had all three doses. Centrally reviewed pCR was 68% (27 of 40; one-sided 95% CI lower boundary 53%), and PaR was 83% (33 of 40; 95% CI, 67 to 93). Treatment-related adverse events (any grade, grade 3, grade 4) were 42%, 9%, and 0% for rBCG; 55%, 15%, and 2% for atezolizumab; and 96%, 38%, and 17% for chemotherapy. CONCLUSION:To our knowledge, this is the first trial combining intravesical rBCG with chemoimmunotherapy in MIBC, demonstrating high pCR and PaR rates that warrant further investigation in prospective randomized trials.

Nivolumab monotherapy has been approved for the adjuvant treatment of adult patients with urothelial carcinoma who are at high risk of recurrence after undergoing radical resection of urothelial carcinoma based on results of the phase 3 CheckMate 274 trial, in which adjuvant nivolumab versus placebo demonstrated improvement in the primary endpoint of disease-free survival (DFS) in high-risk muscle-invasive urothelial carcinoma (MIUC). Identification of biomarkers associated with treatment outcomes can help refine patient selection, and inform on the immunobiology of disease. To assess the relevance of key biomarkers in the adjuvant MIUC setting, extensive exploratory analyses of tumor biomarkers, including associations with DFS, were performed. Differential gene expression and gene signature analysis found that immune-related genes and pathways, in particular a high interferon-γ signature, were predictive of improved DFS in nivolumab-treated patients. Positive predictive and prognostic associations, respectively, were found for CD4 gene expression and measures of CD8 T cell infiltration. A composite predictive model suggested that high tumor cell PD-L1 expression, high CD4 gene expression, high tumor mutational burden score, receipt of neoadjuvant cisplatin and low transforming growth factor-β gene signature score made the greatest contributions to predicting improved outcomes in nivolumab-treated patients. These results reinforce studies establishing the importance of tumor biomarkers of adaptive immunity in influencing response to PD-1-PD-L1 blockade, indicating the potential predictive rather than solely prognostic nature of such findings. ClinicalTrials.gov identifier: NCT02632409 .

INTRODUCTION:The landscape of diagnosis and treatment of locally advanced and metastatic prostate cancer has seen an unprecedented evolution in recent years. Incorporation of results from pivotal studies is crucial for state-of-the-art counselling of patients and to inform treatment recommendations. However, in daily practice ambiguous situations may occur or various options may seem adequate. Furthermore, specific national regulations, e.g. availability of diagnostic tools and drug approvals, may influence treatment choices. This paper presents the voting results of a Swiss expert panel that convened in August 2024 and discussed selected questions addressed by an international panel of experts during the Advanced Prostate Cancer Conference held in Lugano in April 2024. The objective is to facilitate national harmonization of diagnosis and treatment of locally advanced and metastatic prostate cancer and thereby providing a basis for discussion with individuals with prostate cancer.

The infiltration of diffuse large- and other mature B-cell lymphomas with T- and myeloid cells is a key tumor microenvironmental feature but is not currently factored into treatment decisions. Here, we have used multiplex immunofluorescence microscopy to quantify the immune infiltrates of >260 diffuse large B-cell- (DLBCL), follicular- (FL) and mantle cell lymphomas (MCL), and chronic lymphocytic leukemias (CLL) relative to clinical outcomes, mutational landscape and phenotype. MCL were found to be the “coldest” and DLBCL the “hottest” entities. The lymphoma microenvironment of DLBCL featured numerically dominant populations of CD8+ and T-follicular helper (Tfh) T-cells that were indicative of superior prognosis. Mutations in EZH2, PTEN and KMT2D were overrepresented in DLBCL with low CD8+ T-cell infiltration. A unique feature of DLBCL was its infiltration by large numbers of PDL1+ macrophages that constituted up to 70% of total cellularity. PDL1+ macrophage infiltration was mutually exclusive with regulatory T-cell infiltration. The inducible ablation of PDL1 on macrophages was sufficient to improve immune control of MYC-expressing lymphoma in a syngeneic immunocompetent model. These results implicate the macrophage/CD8+ T-cell axis as a key pathogenetic determinant and immunotherapeutic target in a subset of DLBCL patients with poor prognosis.

The advent of combination immune checkpoint inhibitors (cICI), specifically ipilimumab and nivolumab, has transformed the treatment landscape for metastatic renal cell carcinoma (mRCC), offering durable remissions and improved outcomes for intermediate- and poor-risk patients. However, intrinsic resistance remains a significant challenge, with 40-60% of patients failing to achieve meaningful responses. Here, we conducted a comprehensive, multi-omic analysis of systemic and tumor-associated immune responses in mRCC patients enrolled in a clinical trial (CA209-980, SAKK07/17, NCT03297593 ) testing a novel response-adapted cICI regimen. Our study aimed to identify immune correlates of response to cICI therapy. High-dimensional mass cytometry and single-cell proteomic and bulk RNA sequencing of the tumor revealed an enrichment of Th17 CD4+ T cells in responders. These cells exhibited upregulated IL-21-driven pathways, IL-17 signaling, and inflammasome-associated processes, highlighting their central role in therapeutic efficacy. Our integrative analysis underscores the importance of Th17 cells in mediating effective responses to cICI and provides a framework for developing predictive biomarkers and therapeutic strategies to overcome resistance. These findings highlight the translational potential of targeting Th17-centric pathways to enhance immunotherapy outcomes in mRCC and potentially other cancers. This work represents a significant step toward advancing personalized oncology through integrated immune profiling.

658 Background: In the phase 3, randomized, double-blind CheckMate 274 trial, adjuvant NIVO demonstrated statistically significant and clinically meaningful disease-free survival (DFS) benefit vs PBO in pts with high-risk MIUC after radical surgery (RS) ± prior neoadjuvant cisplatin-based chemotherapy (NAC). With extended 3-y median follow-up, continued improvements in DFS were seen with NIVO vs PBO in the primary efficacy populations (intent-to-treat [ITT], tumor programmed death ligand 1 [PD-L1] expression ≥ 1%) and in pts with MIBC. Early trends in interim OS favored NIVO vs PBO in ITT and tumor PD-L1 ≥ 1% pts. Here we report additional efficacy outcomes for pts with MIBC. Methods: Pts were randomized 1:1 to NIVO 240 mg every 2 wk or PBO for ≤ 1 y of adjuvant treatment, stratified by tumor PD-L1 expression, nodal status, and prior NAC. Primary endpoints were DFS in ITT and tumor PD-L1 expression ≥ 1% pts. OS in ITT and PD-L1 ≥ 1% pts was a secondary endpoint. Analysis of MIBC pts was exploratory. MIBC OS data are from preplanned interim analyses of ITT and PD-L1 ≥ 1% pts. OS follow-up is ongoing as the prespecified statistical boundaries for significance in ITT and PD-L1 ≥ 1% pts were not crossed at the time of these analyses. Results: Of 709 randomized pts (ITT), 560 (79%) had MIBC (NIVO, n = 279; PBO, n = 281); 284 (51%) of MIBC pts had prior NAC. With median follow-up of 36.1 mo (ITT), DFS improvement with NIVO vs PBO was consistent between all pts with MIBC (hazard ratio [HR] 0.63) and those with (HR 0.58) and without prior NAC (HR 0.69; Table). For OS, HRs favored NIVO vs PBO in all pts with MIBC (HR 0.70) and the tumor PD-L1 ≥ 1% subgroup (HR 0.48), as well as in pts with MIBC with (HR 0.74) and without prior NAC (HR 0.67). Safety was consistent with previous data in ITT pts; no new safety signals were identified. Conclusions: With 3-y median follow-up, consistent benefit in DFS was observed with NIVO vs PBO in all MIBC pts and across prior NAC subgroups. The HR for OS favored NIVO in all MIBC pts, in those with PD-L1 ≥ 1%, and regardless of prior NAC status. These results continue to support adjuvant NIVO as a standard of care for high-risk MIUC and MIBC, potentially providing an opportunity for a curative outcome. Clinical trial information: NCT02632409 . NIVOn NIVOMedian(95% CI), mo PBOn PBOMedian(95% CI), mo HR (95% CI) DFS All MIBC 279 25.6 (19.2–41.8) 281 8.5 (7.3–13.7) 0.63 (0.51–0.78) With prior NAC 142 19.6 (15.6–48.2) 142 8.3 (5.6–11.2) 0.58 (0.43–0.79) No prior NAC 137 25.9 (19.2–51.5) 139 13.7 (7.8–22.1) 0.69 (0.50–0.94) OS All MIBC 279 NR (45.0–NE) 281 39.9 (29.8–52.1) 0.70 (0.55–0.90) PD-L1 ≥ 1% 113 NR (NE–NE) 117 37.6 (26.9–NE) 0.48 (0.29–0.77) With prior NAC 142 55.2 (41.8–NE) 142 40.2 (28.8–53.7) 0.74 (0.53–1.03) No prior NAC 137 NR (40.7–NE) 139 37.7 (28.7–65.2) 0.67 (0.47–0.95) NE, not estimable; NR, not reached.

ABSTRACT:In the phase 3 POLARIX study, Pola-R-CHP (polatuzumab vedotin plus rituximab, cyclophosphamide, doxorubicin, and prednisone) improved progression-free survival (PFS) vs R-CHOP (rituximab, cyclophosphamide, doxorubicin, vincristine, and prednisone) in patients with previously untreated diffuse large B-cell lymphoma (DLBCL). This post hoc subgroup analysis of POLARIX evaluated the efficacy and safety of Pola-R-CHP vs R-CHOP in older patients aged ≥60, ≥65, ≥70, and ≥75 years. As of 15 June 2022 (median follow-up, 40 months), 629 patients aged ≥60 years were included (Pola-R-CHP, n = 311; R-CHOP, n = 318). Clinically meaningful improvements in PFS with Pola-R-CHP vs R-CHOP were observed across all age groups, particularly in patients aged ≥70 years whereby the risk of disease progression, relapse, or death was reduced by 37% (unstratified hazard ratio [HR], 0.63; 95% confidence interval [CI], 0.41-0.96). In patients aged ≥60 years, overall survival was similar with Pola-R-CHP vs R-CHOP (unstratified HR, 0. 99; 95% CI, 0.67-1.47). Safety profiles were similar for Pola-R-CHP vs R-CHOP among patients aged ≥60 years, including rates of grade 3 to 4 adverse events (AEs; 62.7% vs 61.5%), grade 3 to 5 infections (15.0% vs 12.9%), and grade 5 AEs (3.6% vs 3.2%); no novel toxicities were reported. Incidence of grade 3 to 4 febrile neutropenia was higher with Pola-R-CHP than R-CHOP (16.3% vs 7.6%), highlighting the importance of granulocyte colony-stimulating factor prophylaxis in older patients receiving Pola-R-CHP. The benefit-risk profile favored Pola-R-CHP vs R-CHOP in older patients with previously untreated DLBCL. This trial was registered at www.ClinicalTrials.gov as #NCT03274492.

SummaryPost‐transplant lymphoproliferative disorders (PTLD) and lymphomas in immunocompromised individuals represent significant clinical challenges, with a limited understanding of their pathogenesis. We investigated a PTLD cohort (n = 50) consisting of ‘early lesions’ (infectious mononucleosis‐like PTLD, plasmacytic and follicular hyperplasias), polymorphic PTLD and post‐transplant diffuse large B‐cell lymphomas (PT‐DLBCL). The study also included 15 DLBCL with autoimmune/immunocompromised backgrounds (IS‐DLBCL) and 14 DLBCL, not otherwise specified (DLBCL, NOS), as control. To investigate microarchitectural and genetic changes, immunohistochemistry, multiplex immunofluorescence (mIF), fluorescence in situ hybridisation and high‐throughput sequencing were performed. Scarcity of viral infections other than Epstein–Barr virus (EBV) was observed. mIF revealed lower Treg infiltration in PT‐DLBCL and high CD8+/PD1+ T cells in IS‐DLBCL. MYC rearrangements were most common in PT‐DLBCL, followed by IS‐DLBCL and DLBCL, NOS, all EBV‐negative. TP53 mutations were frequent in EBV‐negative PT‐DLBCL and DLBCL, NOS but absent in ‘early lesions’. NOTCH1 mutations were predominant in PT‐DLBCL (N1 DLBCL‐subgroup). Gene expression profiling showed a significant overlap between ‘early lesions’ and polymorphic PTLD. The presence of clonal haematopoiesis of indeterminate potential (CHIP)‐like mutations and the absence of immune‐escape gene mutations in ‘early lesions’ suggest these disorders may represent clonal expansions driven by exogenic immunosuppression and/or EBV infection ‘substituting’ for mutations of the latter group of genes.

Adoptive cell therapy (ACT) with expanded autologous tumor-infiltrating lymphocytes (TILs) can induce durable responses in metastatic melanoma, yet many patients relapse. We profiled tumor-reactive T cell dynamics during TIL therapy using single-cell RNA and TCR sequencing from seven patients to elucidate underlying reasons. We found that tumor-reactive T cells preferentially expanded early during ex vivo TIL culture, transitioning from exhausted to reinvigorated effector states. Particularly, CD8+ exhausted T cells (Tex) and CD4+ follicular helper T cell (Tfh), but not CD4+ Tex, were efficiently reinvigorated. Further, we resolved the heterogeneity of tumor-reactive CD8+ and CD4+ subsets, defining unique signatures for their identification during TIL expansion. In addition, non-responders (NRs) exhibit increased levels of Type 17 T cells in TIL products, suggesting a potential association with resistance to therapy. After transfer, tumor-reactive clones rapidly extravasated and established a stem-like reservoir. However, in NRs, CD4+ regulatory T cells (Tregs) expanded de novo and tumor-reactive CD8+ T cells reacquired exhaustion markers, limiting their functionality. By contrast, responders (Rs) retained a pool of less differentiated, stem-like cells. Collectively, these data provide a comprehensive analysis of T cell fates during TIL-ACT providing the basis for new approaches to enhance therapeutic strategies. ### Competing Interest Statement D. Koenig reports grants from Geistlich-Stucki-Stiftung, personal fees from Amgen, AstraZeneca, MSD, Novartis, Swiss Oncology in Motion, Mirati, Bristol Myers Squibb, Merck, and PharmaMar, personal fees and nonfinancial support from Amgen, Roche and Sanofi, all: outside the submitted work. A. Zingg reports grants from Bristol Myers Squibb during the conduct of the study. K. Glatz reports grants from Bristol Myers Squibb during the conduct of the study. L.T. Jeker reports grants, nonfinancial support, and other support from Cimeio Therapeutics outside the submitted work. F. Stenner reports personal fees from Bristol Myers Squibb and grants from Bristol Myers Squibb during the conduct of the study, as well as personal fees from Roche, Ipsen, AstraZeneca, MSD, and Takeda and grants from Takeda outside the submitted work. M. Matter reports other support from Thermo Fisher Scientific, Merck, GlaxoSmithKline, Roche, Incyte, and Novartis outside the submitted work. A. Zippelius reports grants from Bristol Myers Squibb and Fondaction during the conduct of the study, as well as grants from Roche, Bright Peak Therapeutics, T3 Pharma, and Astra-Zeneca outside the submitted work. H. Laeubli reports grants from Fondaction and Bristol Myers Squibb during the conduct of the study, as well as grants and nonfinancial support from Bristol Myers Squibb, nonfinancial support from Merck Sharp Dohme, grants and personal fees from GlycoEra, and grants from Palleon Pharmaceuticals outside the submitted work. No disclosures were reported by the other authors. H. Laeubli is a co-founder of Glycocalyx Therapeutics.

Viral vectors have successfully modified T cells to express chimeric antigen receptors (CARs), leading to clinical approvals. However, their high cost and regulatory challenges hinder rapid clinical translation. Here, we demonstrate that our lentivirally (LV) manufactured R110-CAR T cells, targeting a leukemia neoepitope, can also be engineered using the non-viral sleeping beauty (SB) transposition with minimal-sized DNA vectors. Flow cytometry and single-cell sequencing were used to compare the two production modes using healthy donor and chronic lymphocytic leukemia (CLL)-patient-derived T cells and a CD19-CAR T cell control. SB products shifted toward CD8+ subsets with activation/co-inhibition marker expression (CD69, LAG-3, and TIM-3) despite their naïve-like phenotype and lack of antigenic challenge. The CAR binding moiety modulated these patterns, with R110-CAR T cells showing more aberrant phenotypes. Moreover, SB engineering resulted in inflammatory signatures with RIG-I-like and TOLL-like nucleotide sensing potentially due to the transfection procedure. Patient-derived products showed fewer CAR-expressing cells, reduced proliferation clusters, and lower T cell diversity, particularly with SB manufacturing, indicating potential challenges with this method when engineering CLL T cells. Together, our data suggest that the engineering mode may substantially influence T cell properties and that these are further modulated by the CAR binding moiety and the T cell donor.

Abstract Background Tumor lysis syndrome (TLS) is a potentially life‐threatening condition resulting from the rapid destruction of malignant cells, leading to electrolyte imbalances and severe complications, such as acute kidney injury, arrhythmias, and seizures. TLS can be managed through hyperhydration, urate‐lowering treatments, and a steroid prophase strategy. Aims This study aims to explore the impact of fractionated rituximab, an anti‐CD20 antibody, on the occurrence and severity of TLS during the initial cycle in patients with aggressive B‐cell non‐Hodgkin lymphoma (B‐NHL). Methods Data was retrospectively collected from 94 of 186 patients. Results Among the 94 patients included in the analysis, the median age was 70. Histologies were diffuse large B‐cell lymphoma (75%), Burkitt lymphoma (13%) and high‐grade B‐cell lymphoma (8%). The majority were at an advanced stage (93%) with a high IPI score (75%). Most patients received anthracycline‐containing regimens (72%) and prophylactic allopurinol (83%) and/or rasburicase (26%). Steroid prophase was administered to 82% of patients. The study identified one clinical TLS case and six laboratory TLS cases. Significant TLS factors included BL histology, elevated baseline LDH (⟩500 U/l), and rasburicase usage. Infusion reactions were rare (3%). Median progression‐free survival was 2.6 years, and 2‐year overall survival was 33%, irrespective of TLS occurrence. Conclusion In this real‐life study, clinical TLS occurrence was low (1%). TLS appeared more frequent in BL but did not impact overall survival. Fractionated initial rituximab dosing in addition to preventive strategies is a feasible approach in preventing clinical TLS, warranting further prospective investigation.

Clinical trials frequently include multiple end points that mature at different times. The initial report, typically based on the primary end point, may be published when key planned co-primary or secondary analyses are not yet available. Clinical Trial Updates provide an opportunity to disseminate additional results from studies, published in JCO or elsewhere, for which the primary end point has already been reported . CheckMate 274 is a phase III, randomized, double-blind trial of adjuvant nivolumab versus placebo for muscle-invasive urothelial carcinoma (MIUC) at high risk of recurrence after radical resection. The primary end points of disease-free survival (DFS) in intent-to-treat (ITT) and tumor PD-L1 expression ≥1% populations were met. We report results at an extended median follow-up of 36.1 months in the ITT population. In addition, we report interim overall survival (OS) data for the first time and an exploratory analysis among patients with bladder primary tumors (muscle-invasive bladder cancer [MIBC]). Consistent DFS benefit with nivolumab versus placebo was observed in both the ITT (hazard ratio [HR], 0.71 [95% CI, 0.58 to 0.86]) and PD-L1 ≥1% (HR, 0.52 [95% CI, 0.37 to 0.72]) patients. The HR for OS with nivolumab versus placebo was 0.76 (95% CI, 0.61 to 0.96) in the ITT population and 0.56 (95% CI, 0.36 to 0.86) in the PD-L1 ≥1 population. Continuous benefit in nonurothelial tract recurrence-free survival and distant metastasis-free survival was also observed in both patient populations. The exploratory analysis of patients with MIBC also showed continued efficacy benefits, irrespective of PD-L1 status. No new safety signals were reported. Overall, these results further support adjuvant nivolumab as a standard of care for high-risk MIUC after radical resection.