Acute myeloid leukaemia (AML) is a disease with high intra- and interpatient heterogeneity. Current treatments are still based on cytarabine combined with an anthracycline, with 5-year overall survival rates below 30%. Functionalist oncology utilises edge-weighted digraphs as a representational tool to model the mathematical functional interdependencies of disease factors. This methodology enables a conceptual and mechanistic analysis of key factors influencing therapeutic success, providing a framework that can be parameterised to explore patient-specific treatment responses. It incorporates the oligoclonal nature of AML to perform, in principle, comprehensive cost-benefit analyses regarding the addition of individual drugs to existing treatment regimens and the number and type of chemotherapy courses to be given. Extrapolating from experimental data, we investigate the therapeutic potential and risks of SAMHD1 inhibitors in AML treatment as a proof-of-concept. We also provide a web-based interactive application to visualise hypothetical AML treatment scenarios, which can be combined with ex vivo single-cell phenotypic (gene expression), genotypic (somatic mutations) and functional (drug responses) analyses. Functionalist oncology can thus be used to generate testable hypotheses that might contribute to improving oncological decision-making, for example, by identifying the optimal number, nature and sequence of chemotherapy courses, including both existing and novel drugs, such as SAMHD1 inhibitors.
Despite advances in biomedical research, human psychopathology remains underserved by experimental model systems, limiting therapeutic innovation. Alternative approaches are needed to investigate the mechanisms underlying mental health conditions. We aimed to assess whether large language models (LLMs) could serve as experimental systems to model affective processes relevant to human psychopathology. Using standard psychological induction protocols, including imagery vignettes, we tested whether seven affective states (fear, anxiety, anger, disgust, sadness, worry, and stress) could be systematically induced in six state-of-the-art LLMs (including GPT-4o and several Llama variants) and subsequently reversed with regulation strategies. Separate prompt sequences were created for each affective state, and an additional neutral control condition was included in which no affective stimulation was applied after the initial general prompt. All affective states followed static vignette protocols without interaction, consistent with human studies. The only exception was stress, which, in line with the original Trier Social Stress Test (TSST) protocol, followed a dynamic interactive prompting procedure. The LLMs were intermittently prompted to self-assess their current affective state via visual analogue scales with a fixed numerical range from 0 to 100 (except for anxiety, which was measured with the State version of the State-Trait Anxiety Inventory). To reverse the induction of affective states, a mindfulness-based relaxation technique was used for all conditions except stress, which was followed by a standardised debriefing procedure consistent with the original TSST protocol. Each condition was repeated in five independent runs to ensure reliability. A sentence completion test was used to test for cognitive bias after induction of sadness, with responses rated for emotional valence by three independent human raters. Inter-rater reliability (Cohen's κ) and negativity scores were calculated, and conditions were compared using t tests (Cohen's d). Across all affective states and over five runs, mean scores for GPT-4o increased by 52·83 points (201·20%) from baseline to post affect induction, and downregulation prompts reduced scores by 48·23 points (60·98%). These patterns were broadly replicated across five additional open-weight LLMs, with significant between-model differences for all affective states (p values between 0·045 and <0·0001) except stress (p=0·063). GPT-4o and Llama 4 Maverick showed the strongest effects, whereas Llama 4 Scout showed the weakest responses, indicating that model architecture and scale influence susceptibility to affect induction. In the test for cognitive bias, sadness-related prompts elicited a consistent negativity bias in sentence completions by GPT-4o compared with neutral prompts (mean 15·00 [SD 4·26] vs 8·67 [2·66]; Cohen's d=1·87). Our findings establish LLMs as promising tools for modelling affective processes relevant to human psychopathology. By reproducing key psychological phenomena, LLMs might enable the experimental investigation of mechanisms underlying mental disorders and facilitate the preliminary screening of novel therapeutic interventions, potentially accelerating progress in a field historically constrained by the scarcity of effective model systems. None.
Cisplatin (CIS) and cyclophosphamide (CTX) are commonly used in clinical oncology, often in combination to improve therapeutic outcomes. However, most preclinical studies have examined these agents individually, overlooking the potential synergistic and immunomodulatory effects of combined treatment. This study aimed to compare the antitumor and immune-modulating effects of single versus combined CTX and CIS chemotherapy, and to determine the treatment sequence that achieves maximum efficacy with minimal toxicity. Adult female mice (n = 5/group) were inoculated intraperitoneal (i.p.) with 0.5 × 10⁶ fresh viable Ehrlich ascites carcinoma (EAC) cells. Twenty-four hours' post-inoculation, animals received a single administration of phosphate-buffered saline (PBS), CIS, CTX, or various combinatorial CIS-CTX regimens. Antitumor activity and alterations in myeloid cell subsets, including dendritic cells (DCs), monocytes, and neutrophils, were evaluated in peripheral blood and spleen by flow cytometry. EAC-bearing mice showed changes in splenic and circulating immune cell populations based on CD11c and CD11b expression profiles, suggesting tumor-associated immunomodulatory alterations. CIS and CTX monotherapy partially restored myeloid cell populations. Notably, sequential regimens, particularly CIS followed by escalating doses of CTX-led to substantial increases in both mature neutrophils and plasmacytoid DCs (pDCs), while reducing CD11b+ myeloid cell population associated with immunosuppressive phenotypes. These changes suggest enhanced myelopoiesis and innate immune activation in treated mice. A regimen consisting of high-dose CTX combined with low-dose CIS demonstrated robust anti-tumor activity while limiting leukopenia and systemic toxicity. This protocol may represent an optimal therapeutic strategy, balancing efficacy with safety.
Acute myeloid leukemia (AML) is a hematologic malignancy characterized by heterogeneity, poor prognosis, and limited biomarkers for risk prediction. Mitochondria pathway related genes (MPRGs), as central regulators of cellular metabolism and immune microenvironment dynamics, may provide useful information for prognostic assessment and biological characterization in AML. MPRGs were obtained from the MitoCarta3.0 database. Univariate Cox and Kaplan-Meier methods were conducted to analyze their prognostic relevance. LASSO penalized regression followed by stepwise multivariate Cox analysis yielded an optimal gene panel in the TCGA-LAML dataset. External validation was performed across three GEO datasets (GSE10358, GSE106291, GSE71014). Finally, the role of UCP2 was examined in vitro by assessing UCP2 knockdown effects on MOLM-13 cell behavior. A prognostic signature comprising seven MPRGs (UCP2, FAM162A, ACCS, HSD1, ACSF2, PPIF, and SDHA) was established. High-risk patients exhibited significantly shorter survival. The MPRGs risk score served as an independent predictor of prognosis. Moreover, elevated risk scores correlated with heightened immune checkpoint molecule expression and an immunosuppressive tumor microenvironment. UCP2 knockdown attenuated both proliferative capacity and migratory potential in MOLM-13 AML cells. In summary, the MPRG-based signature provides independent prognostic value in AML and reflects its association with an immunosuppressive microenvironment. These findings provide additional evidence that mitochondrial pathway-related genes are associated with AML prognosis and immune microenvironment features. UCP2 may represent a biologically relevant candidate gene in AML, although further mechanistic and clinical validation is required.
Cleavage under target and tagmentation (CUT&Tag) is a widely used method for profiling chromatin occupancy; however, its reproducibility is impacted by a lack of standardization in experimental and analytical procedures. This study identifies four key parameters critical for optimizing CUT&Tag performance. First, optimal cell number requirements are target and species specific: H3K27me3 detection requires ≥10K cells in both mouse and rat cells, whereas CTCF mapping needs 50K cells in mouse embryonic stem cells and 100K in rat C6 cells, reflecting potential differences in protein abundance and antibody affinity. Second, the peak-calling methodology is crucial; default model-based analysis of chromatin immunoprecipitation followed by sequencing (MACS2) scaling causes a paradoxical decrease in peak numbers with decreasing IgG control size, a limitation resolved by the "scale-to-large" option. Third, duplicate removal strategies differentially affect peak callers, with MACS2 performing best using biological reads and sparse enrichment analysis for CUT&RUN (SEACR) relying on technical duplicates for accurate calling. Finally, mild crosslinking with 0.2 mM ethylene glycol bis (succinimidyl succinate) (EGS) for 5 min enhances CTCF detection and reduces variability. Together, these optimizations establish practical guidelines for reliable CUT&Tag experimental design and analysis.
The development of effective antibody therapeutics has been hampered by a lack of methods to measure drug delivery and activity within tumors at single-cell resolution. Here we introduce single-cell spatial pharmacobiology (SSP), an experimental and analytical framework that integrates in situ imaging of a systemically infused, fluorescently labeled therapeutic antibody with high-plex spatial proteomics to quantify antibody distribution, target engagement and tumor microenvironment (TME) architecture. We applied SSP to tumor tissues from participants with head and neck squamous cell carcinoma and pancreatic ductal adenocarcinoma who received the antibody panitumumab-IRDye800 in phase 1 trials. SSP identified pronounced spatial heterogeneity in single-cell drug delivery and target engagement, shaped by conserved stromal barriers, including periostin-rich extracellular matrix assemblies and fibroblast-activation-protein-positive cancer-associated fibroblast neighborhoods, which were associated with reduced antibody delivery in both tumor types. SSP measures drug-target-TME interactions in human tumors and can support studies of resistance mechanisms, dosing strategies and discovery of spatial biomarkers for precision oncology.
Exercise improves physical function and health-related quality of life (HRQoL) in patients with cancer; however, nationwide structured programmes remain limited. A hospital in Germany implemented a donation-funded Haematological and Oncological Training Therapy with Stationary Strength and Cardio Machines (HOT), and a 3-year real-world data analysis aimed to evaluate its acceptance, feasibility, safety, and effects. HOT comprised 24 sessions of combined resistance and aerobic training, complemented by a sensorimotor module for patients with chemotherapy-induced peripheral neuropathy (CIPN), with a target frequency of two sessions per week. Patients with all cancer types and treatment phases were eligible. The study evaluated acceptance (e.g., participants' clinical and sociodemographic characteristics, reasons for premature discontinuation), feasibility (e.g., completion rate, training density), safety (adverse events), and effects (changes in strength [1-repetition maximum, 1-RM], cardiopulmonary performance [maximum power on a cycle ergometer], and patient-reported outcomes [HRQoL, fatigue, CIPN]) within a quasi-experimental pre-post design. Between April 2021 and March 2024, 70 patients (60% female; mean age, 63 ± 12 years; range, 31-91 years) participated; 61% had solid tumours and 39% haematological malignancies, and 57% were undergoing active cancer treatment. Most participants (94%) had a history of sports participation. A total of 53 participants (76%) completed HOT, while 17 (24%) discontinued prematurely, mainly for health-related reasons. The training density among completers was 1.6 sessions per week. One adverse event occurred during a 1-RM test. Significant improvements were observed in strength and cardiopulmonary performance (p < 0.05), as well as in global HRQoL (p < 0.001), fatigue (p < 0.001), and CIPN (p = 0.003). HOT appeared to primarily attract sports-experienced patients, with haematological malignancies overrepresented. HOT proved to be safe and feasible across diverse cancer entities and treatment phases and was associated with improvement in physical function and HRQoL. The participants' high baseline motivation for exercise may limit generalisability.
We report outcomes of the phase 2 study combining carfilzomib (CFZ) plus vincristine, dexamethasone, asparaginase, and daunorubicin (CFZ-VXLD) in heavily pre-treated, pediatric patients with relapsed/refractory (R/R) acute lymphoblastic leukemia (ALL). We investigated whether post-induction complete remission (CR) rates after CFZ-VXLD induction are superior to an external real-world control (EC). Patients aged ≥1 month to <21 years with R/R ALL received CFZ-VXLD induction, followed by optional consolidation (experimental arm). EC arm included similar patients receiving real-world, curative-intent regimens at Therapeutic Advances in Childhood Leukemia sites between 2000-2023. Primary endpoint was CR after induction; overall response rate (ORR; CR/CR with partial/incomplete hematologic recovery/CR without platelet recovery) was a secondary endpoint. Comparative analyses used inverse probability of treatment weighting. Experimental and EC arms included 105 and 140 patients, respectively. CR rates after induction were 14.8% vs 7.8% (odds ratio [OR] 2.04; 95% CI, 0.54-7.66) in patients with B-ALL and 13.6% vs 9.1% (OR 1.58; 95% CI, 0.47-5.31) in patients with T-ALL. There was no statistically significant increase in post-induction CR for the CFZ-VXLD arm vs the EC arm and the study did not meet its primary endpoint. In patients with B-ALL, ORR was 42.6% vs 26.3% favoring the CFZ-VXLD arm vs the EC arm. Adverse event rates with CFZ were consistent with previous reports. Taken together, the CFZ-VXLD regimen may be an option for some selected pediatric patients with R/R ALL and this study highlights that novel treatments for R/R ALL remain an area of unmet medical need. NCT02303821.
The incidence of early onset (age < 50 years at diagnosis) appendiceal adenocarcinoma (EOAA) is rising alarmingly. Reported data on etiology, treatment, and outcomes is scarce. In this study, we report clinical outcomes for patients with EOAA. Patients diagnosed with appendiceal adenocarcinoma between 2013 and 24 were stratified on the basis of age at diagnosis as having either EOAA (< 50 years) or average-age onset appendiceal adenocarcinoma (AOAA; ≥ 50 years). Clinicopathological and genomic data were abstracted from electronic medical records. Baseline clinicopathologic features and survival outcomes were compared between patients with EOAA and AOAA. Of 181 eligible patients, 49 (27.1%) had EOAA. Median age at diagnosis for patients with EOAA was 42.1 years (interquartile range [IQR], 35.7-46). Of the 76 patients (EOAA = 23 [30.3%] + AOAA = 53 (69.7%)] who had locoregional disease at presentation, 44 (57.9%) remained disease-free for at least 2 years. Within these 44 (conditional survival), the EOAA group showed numerically shorter recurrence-free survival (RFS) and higher risk of recurrence (median RFS, EOAA = 49.5 months versus AOAA = 83.3 months; hazard ratio [HR], 4.07; 95% confidence interval [CI], 1.49-11.18; p = 0.06). Long-term recurrences (≥ 5 years) were seen in 9/15 (60%) patients during the 5-year follow-up period. Of 139 evaluable patients (EOAA = 36 [25.9%] + AOAA = 103 [74.1%]) with metastatic/recurrent disease, patients with EOAA received more non-fluorouracil (FU)-based systemic treatments, including experimental agents, in any line of treatment (8/36, 22.2% versus 6/103, 5.8%; p = 0.009). A higher proportion of patients in the EOAA group received bevacizumab in any line of treatment (24/36, 66.7% versus 49/103, 47.6%; p = 0.055) and three or more lines of systemic therapy (10/36, 27.8% versus 14/103, 13.6%; p = 0.07). There was no significant difference in OS between the EOAA and AOAA groups despite more aggressive therapy in EOAA (median OS, EOAA = 35.2 months versus AOAA = 40.6 months; HR, 0.90; 95% CI, 0.57-1.44; p = 0.66). Among patients who remained disease-free after initial surgical resection for locoregional disease for at least 2 years, recurrence was more frequent in patients with EOAA. Aggressive systemic therapy, including trials and non-FU based therapies were more frequent in patients with EOAA but were not associated with improved survival. EOAA may represent a unique entity within this rare disease, which warrants further exploration.
Creatine supplementation is widely used for its ergogenic benefits but has recently garnered interest for its immunomodulatory properties, particularly its capacity to enhance CD8+ T-cell bioenergetics through SLC6A8-mediated transport and phosphocreatine-dependent ATP (adenosine triphosphate) buffering. Emerging preclinical evidence suggests that creatine may potentiate antitumor immunity and augment the efficacy of immune checkpoint inhibitor (ICI) therapies. Conflicting reports of creatine-associated metastasis in select tumor models have raised uncertainty regarding its role in oncologic settings. This review synthesizes current preclinical evidence to assess whether creatine exerts beneficial, neutral, or detrimental effects across experimental oncologic models treated with immune checkpoint therapy. A total of 230 articles were screened, and 5 studies were included within this systematic review. Current preclinical evidence suggests that creatine supplementation may exert beneficial immunomodulatory and antitumor effects when combined with PD-1 blockade, largely by enhancing T-cell metabolic fitness and macrophage-driven inflammatory responses. However, the potential for metastasis in select tumor types poses a risk to the administration of a creatine supplement. This systematic review emphasizes the absence of human data, highlighting a critical knowledge gap. Rigorous mechanistic studies and early-phase clinical trials are essential to determine whether creatine can be safely and effectively integrated into ICI-based treatment strategies.
Activating mutations in NOTCH1 are frequent in T-cell acute lymphoblastic leukemia (T-ALL) and, in the absence of alterations in RAS or PTEN, are associated with favorable prognosis. Besides classical heterodimerization and PEST domain mutations, juxtamembrane internal tandem duplications (JME-ITDs) represent a third class of activating variants. Their intermediate size and sequence composition can challenge short-read next-generation sequencing and variant calling algorithms, leading to underdetection in routine diagnostics. An index case of newly diagnosed T-ALL harboring a NOTCH1 juxtamembrane insertion that was discordantly detected by variant callers during routine targeted NGS (Hematology OncoKitDx v2) prompted further analysis. The variant was evaluated using VarDict and VarScan2, followed by manual inspection with IGV and orthogonal confirmation by Sanger sequencing. To further assess variability in the detection of JME-ITDs, six additional BAM files from patients with previously confirmed variants were reanalyzed using VarDict, VarScan2, Mutect2, and Pindel. In the index case, VarDict detected a 51-bp NOTCH1 JME-ITD (c.5168-2_5216dup) that was missed by VarScan2 and confirmed by Sanger sequencing. Reanalysis of the seven cases showed that Mutect2 and VarDict detected all JME-ITDs (100%), VarScan2 identified only three (42.8%), and Pindel detected six mutations (85.7%). These results reflect differences in algorithmic strategies where haplotype-based callers incorporate soft-clipped and split reads, whereas pileup-based discard them. Detection of NOTCH1 JME-ITDs depends strongly on variant-calling strategy. Combining haplotype-based callers with split-read structural variant tools may reduce false negatives and improve detection of clinically relevant insertions in diagnostic NGS pipelines.
Central nervous system (CNS) involvement frequently occurs in acute lymphoblastic leukemia (ALL), but many questions remain about how leukemia cells access, persist in, and exploit the CNS. The CNS is protected by the meninges, which are fluid-filled membranes that surround the brain. Within the meninges, meningeal lymphatic vessels, located within the dura mater, drain cerebrospinal fluid to the deep cervical lymph nodes for immunosurveillance, providing a potential pathway for leukemia cells to enter or exit the CNS. Here, we utilized tissue engineered models of the meningeal lymphatics and lymph node stroma to probe how leukemia cells interact with these key microenvironments. We first demonstrated that standard-of-care chemotherapeutics can perturb the meningeal lymphatics, particularly lymphatic endothelial cells. Next, we showed that soluble factors from the meningeal lymphatics conferred a modest upward trend in leukemia cell growth, while soluble factors from the lymph node model under flow significantly promoted leukemia cell migration. Finally, we showed that leukemia cells migrate through the lymph node model under both static and flow conditions. Overall, we have demonstrated the feasibility of using engineered lymphatic models to study leukemia cell behavior in the CNS with the goal of expanding the available experimental platforms for understanding CNS metastasis and relapse. Insight Box Leukemia commonly infiltrates the central nervous system (CNS), requiring intensive CNS-directed therapies that are often ineffective and cause both acute and long-term toxicities, especially in pediatric patients. The meningeal lymphatics and the deep cervical lymph nodes constitute a pivotal axis in CNS immunity, facilitating drainage of fluid and waste, and enabling peripheral immune surveillance in response to CNS-derived signals. Here, we employ in vitro models of the meningeal lymphatics and lymph node stroma to examine their crosstalk in influencing leukemia cell growth and migration. These engineered platforms serve as valuable tools for uncovering mechanistic insights into the meningeal-lymph node axis in the context of CNS-leukemia relapse.
While immune repertoire (IR) dynamics encode critical disease signatures, existing models fail to capture the non-equilibrium processes that govern clonal selection and somatic hypermutation (SHM). We develop a biophysical framework that mathematically reconstructs IR evolution through energy landscape optimization, where 1) clonal emergence probabilities map to metastable states, 2) repertoire transitions obey non-equilibrium dynamics, and 3) inter-IR distances quantify the distribution transformation costs via optimal transport theory. This model enables macroscopic immune state detection from 10k cells by resolving critical fluctuations in sparse sampling regimes. Experimental validation across murine/human cohorts demonstrates precise unsupervised stratification of immune stages and disease states without prior clinical annotations. By bridging stochastic SHM kinetics with deterministic repertoire shifts, our approach establishes quantitative metrics for tracking immunological trajectories and pathological progression. This paradigm shift from static pattern recognition to dynamic process decoding empowers sensitive immunomonitoring and personalized therapeutic design through ultra-low-input clinical sampling.
Immune checkpoint inhibitors (ICIs) have revolutionised the treatment landscape of non-small cell lung cancer (NSCLC) without actionable genomic alterations by opening new possibilities for treatment. However, the predictive value of Programmed Death-Ligand 1 (PD-L1) expression assessed by immunohistochemistry (IHC) is quite modest, leaving many patients without optimal benefit from ICIs. To address this gap, liquid biopsy is gaining momentum as a promising tool to complement tissue-based PD-L1 assessment. Liquid biopsy, which entails the analysis of circulating tumour cells (CTCs), extracellular vesicles (EVs), circulating tumour DNA (ctDNA), and other biomarkers, offers a non-invasive approach for tracking disease evolution and interactions with the immune system. We conducted a narrative review to explore the potential of liquid biopsy in enhancing patient selection for ICIs, its predictive and prognostic value in advanced NSCLC, and the associated technical and clinical implications. By highlighting recent advancements and ongoing research efforts, we underscore the critical role of liquid biopsy in driving precision immuno-oncology. Establishing robust, reproducible methods for PD-L1 assessment through liquid biopsy is essential for translating this approach into clinical practice, with the potential to overcome the limitations of tissue-based assays and better individualise immunotherapy strategies in NSCLC.
Children and adolescents with serious illness or medical complexity are often referred to highly specialized children's hospitals that offer experimental interventions, novel management, or treatments that cannot be provided at local health care centers. Referral to a geographically distant, highly specialized health center presents new complexity, with a distinct set of ethical issues, for patients, families, and clinicians. We describe ethical considerations that occur at the initiation of referral, during the process of referral, and after a referral for pediatric patients receiving hospital-based care. Using illustrative cases, we highlight salient themes surrounding these challenges that span many tenets of pediatric bioethics, including beneficence (ie, weighing benefits against harms), imposing values to impact shared decision-making, and justice (ie, lack of standardization in processes creating risk for discrimination). Pediatric clinicians and ethicists must be aware of the potential challenges arising in referring and transferring patients for higher levels of specialized intervention and treatment. When referral is indicated, institutions should facilitate collaborative care as much as possible by sharing information early, frequently, transparently, and in a standardized manner.
To characterize relapse patterns, identify relapse-related risk factors, and evaluate prognostic determinants of post-relapse outcomes in pediatric acute lymphoblastic leukemia (ALL) patients treated with the Chinese Children's Cancer Group (CCCG)-ALL-2015 protocol. This retrospective study included 380 children diagnosed with ALL between 2016 and 2019. We analyzed relapse timing (very early, early, late), relapse sites (bone marrow (BM) vs. extramedullary), early treatment response (morphology and minimal residual disease (MRD)), and post-relapse outcomes. Survival was estimated using Kaplan-Meier methods. Relapse rate was 11.1% (42/380), predominantly isolated BM (69.0%) and very early timing (47.6%). Relapse varied significantly by risk group (low-risk: 5.9%; intermediate-risk: 10.4%; high-risk: 22.4%; P < 0.001), immunophenotype (B-ALL: 9.8% vs. T-ALL: 23.5%; P = 0.015), Day 19 morphology (M1:9.9% vs. M3:20.0%; P < 0.001), Day 46 morphology (M1:10.5% vs. M3:21.4%; P = 0.025), and Day 46 MRD status (positive:21.4% vs. negative:8.7%; P = 0.002). Fusion gene positivity was significantly associated with earlier relapse timing (P < 0.05). Post-relapse survival differed by relapse timing (very early: 7.2 months; early: 23.2 months; late: 37.8 months; P < 0.001) and initial risk group (low-risk: 38.2 months; high-risk: 18.1 months; P = 0.006). Non-remission on Days 19/46, Day 46 MRD positivity, and high-risk status predict ALL relapse. High-risk patients experience earlier relapses and poorer survival. Fusion gene positivity is associated with earlier relapse timing.
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In addition to well-established immune checkpoints (ICs), such as CTLA-4, PD-1, PD-L1, increasing attention is being directed toward next-generation ICs, including TIM-3, Gal-9, LAG-3, BTLA, HVEM, and CD160. Single nucleotide polymorphisms (SNPs) within IC-related genes may contribute to dysregulation of inhibitory pathways and impair anti-tumor immune responses. This study aimed to evaluate the association between selected IC gene variants and susceptibility to bladder cancer (BC). A total of twelve SNPs located in TIM-3, LGALS9, BTLA, HVEM, and CD160 genes were genotyped using TaqMan assays in 314 BC patients and over 520 healthy controls (HC). Genotype distributions were analyzed under multiple genetic models, and associations with clinicopathological parameters were assessed using multivariate logistic regression. Genotype distributions of BTLA polymorphisms (rs2705511, rs1982809, rs9288953) differed between BC patients and HC, suggesting potential associations with BC risk. Stratified analyses revealed sex-specific effects, with variants in BTLA (rs1982809), HVEM (rs1886730, rs2234167, rs8725), and CD160 (rs231375) showing potential associations with susceptibility among women. Additionally, SNPs in BTLA and HVEM were nominally associated with recurrence and high-grade tumors, while CD160 and LGALS9 variants were potentially linked to primary tumor occurrence. However, these associations lost statistical significance after correction for multiple comparisons. Although the observed associations did not remain significant after multiple testing correction, the results suggest that genetic variation within BTLA, HVEM, and CD160 genes may still play a biologically relevant role in BC susceptibility and disease progression. These findings underscore the potential importance of IC pathways in BC pathogenesis and warrant further investigation in larger, well-powered studies.
Hematological patients are at higher risk of severe SARS-CoV-2 infection and exhibit impaired vaccine responses due to disease and therapy. Treatments like Rituximab are known to compromise immune function, reducing the generation of protective antibodies. In the CORSA trial, we evaluated humoral and cellular responses to messenger RNA vaccines against SARS-CoV-2 in 77 patients with hematological malignancies receiving active treatment and in matched healthy controls. Peripheral blood samples were processed to assess Spike-specific immunoglobulin G titers, T-cell responses by enzyme-linked immunospot assay, and T-cell receptor repertoire sequencing from baseline to six months after vaccination to determine clonal breadth and depth of Spike-specific T-cell receptor clones. Seroconversion occurred in only 8% and 21% of patients after the first and second dose, compared to 93% and 100% of healthy controls. Despite poor antibody responses, 69% of seronegative patients showed measurable T-cell activity, suggesting some level of vaccine-induced protection. Spike-specific TCR repertoire analysis in lymphoma patients (LP) and HC revealed significantly broader clonal breadth (p = 0.0013) and higher clonal depth (p = 0.0265) in HC at day 50 versus baseline, while blunted diversification in LP (p = 0.0407). Lymphoma patients showed significantly weaker Spike-specific clonal responses and reduced diversity compared with healthy controls, supporting the association with increased patient vulnerability.
Humoral immunodeficiency is frequently observed in patients affected by chronic lymphocytic leukemia (CLL). The reasons for the compromised B-cell responses are only partly understood. We hypothesized that the normal residual B cells in CLL patients are negatively influenced in their functionality by soluble factors produced by CLL cells. We performed functional B-cell assays to analyze the influence of serum from CLL patients or conditioned medium from CLL cells cultivated with stromal cells on B cells from healthy donors. Serum from healthy individuals or conditioned medium from normal B cells was used as controls, respectively. We stimulated the B cells in a T-cell-dependent fashion and incubated them in the respective serum or conditioned medium for 5 days. We measured plasma cell differentiation, proliferation, cell death, and B-cell activation, the latter using the markers CD80, CD86, and CD25. We did not detect significant differences supporting the hypothesis that CLL-derived soluble factors negatively influence the proliferation, activation, or cell death levels of normal B cells from healthy individuals in in vitro coculture. Our analysis suggests no major direct inhibitory effects of CLL-derived soluble factors on normal mature B cells. Thus, it is likely that other factors lead to the known B-cell dysfunction in CLL.