Abstract The Trimethyl guanosine Synthase (TGS1) is a highly conserved enzyme mediating di-methylation of the 5’-cap 7-methylguanosine of RNA to generate 2,2,7-trimethylguanosine (m 2,2,7 G). Known TGS1 targets include snRNAs, snoRNAs, the telomeric RNA component and a limited number of mRNAs encoding selenoproteins. TGS1 is highly expressed in acute myeloid leukaemia (AML) cells, and its expression correlates with poor prognosis. Here, we report that TGS1 directly methylates the cap of more than 500 mRNAs in AML cells. Specifically, we demonstrate that TGS1 modifies nuclear mRNAs encoding mitochondrial proteins, including critical components of complexes involved in both the TCA cycle and oxidative phosphorylation, promoting their translation mediated by mitochondria associated cytosolic ribosomes. Functionally, we report that TGS1 depletion impairs mitochondrial respiration and increases oxidative stress. This, in turn, impairs the growth of AML cells causing differentiation and cell cycle arrest in vitro and in vivo . Finally, we demonstrate that TGS1 depletion sensitizes AML cells to RSL3, a small molecule promoting ferroptosis. Taken together, our findings establish TGS1 as a key regulator of oxidative phosphorylation and mitochondrial redox status of AML cells and highlight its potential as a therapeutic target in leukaemia.

Immunotherapy has revolutionized the treatment of solid cancers in recent years. However, T-cell lymphomas (T-NHLs) originate from immune cells themselves and are biologically heterogeneous, rendering investigations of immune checkpoint inhibitor (ICI) mechanisms of action complex. While case reports and individual Anaplastic Large Cell Lymphoma (ALCL) cases enrolled in T-NHL trials demonstrated favourable responses to ICI, hyperprogression was observed in other T-NHL subtypes. We therefore utilized a syngeneic mouse model of ALK+ ALCL to investigate immune surveillance and ICI-induced immune response. Transplantation experiments combined with depletion of relevant immune axes revealed that ALCL immune surveillance is mediated by CD4+ T cells and NK cells. Innate and adaptive immune cell infiltration was confirmed on a large series of primary human ALK+ ALCL samples. ICI monotherapy demonstrated robust efficacy in murine ALCL, inducing complete remissions in approximately 50% of treated animals. Mechanistically, PD-L1 blockade reversed Treg-mediated immunosuppression and increased the frequency of circulating effector CD8+ T lymphocytes, thereby prolonging survival significantly. Importantly, CD4+ T cells proved indispensable for driving and sustaining immunotherapy-induced anti-tumour responses in murine ALCL. CD4+ T cells of non-responder animals exhibited an exhausted phenotype and a transcriptomic Th22-like signature, implicating persistent T-cell exhaustion and polarization as a meaningful immune-evasion mechanism. Our findings uncover CD4+ T cells as key players in spontaneous and immunotherapy-mediated anti T-cell lymphoma immunity, which demonstrates the critically needed preclinical proof-of-concept for the safe and effective use of immunotherapy for ALCL.

Burkitt lymphoma (BL) is an aggressive B-cell lymphoma historically classified into endemic, sporadic, or immunodeficiency-associated BL based on clinical and epidemiological features. Recent genomic and epigenomic studies have refined this framework, distinguishing EBV-positive and EBV-negative BL molecular subtypes that cut across traditional clinical and epidemiological categories and exhibit distinct biological profiles. In high-income countries, BL is highly curable with intensive multiagent chemotherapy and antibody-based therapy. In contrast, survival in sub-Saharan Africa (SSA) rarely exceeds 50%, reflecting diagnostic delays, limited access to effective treatment, and challenges in supportive care. This review summarizes advances in BL epidemiology, molecular biology, diagnosis, and treatment, highlighting how genomic insights have identified novel biomarkers for earlier detection and monitoring, as well as potential targeted therapies with reduced toxicity. These advances present opportunities to develop scalable diagnostic tools and tailored therapeutic strategies to narrow survival disparities, particularly in resource-limited settings, where most cases occur, and among adults and patients with relapsed or refractory BL in high income countries. We also outline key unanswered questions in BL epidemiology and pathogenesis, discuss opportunities for translational research, and emphasize the need for improved model systems to study therapeutic resistance to close the survival gap.

Abstract T cell lymphomas (TCL) are a heterogeneous collection of malignancies whose origins and pathogenesis are poorly understood and for which few efficacious therapeutic options exist. Here, we conduct single-cell transcriptomic profiling spanning eight TCL entities and describe entity-associated programmes. We predict the cell of origin for these tumours through an integrative analysis of transcriptome and T cell receptor (TCR) maturation states. By identifying tumours with TCR states ranging from the pre-TCR through non-productive and productive TCR alpha and beta chain rearrangements we shed new light on their developmental origins. Furthermore, we apply our drug2cell computational drug target predictions with drug screens using patient-derived cell models, systematically benchmarking the performance of drug2cell and validating compounds and targets. This process identifies SYK inhibitors as a therapeutic opportunity and prioritises TIM3 for immunotherapy based on combined spatial transcriptomics analysis. Overall, our data provide a resource for diagnostics and therapies for tumours of critical unmet need.

This current framework (version 3) updates the previous one published within the WCRF Gateway on Health Open Research (publication data of version 2: 23 September 2025). The purpose of the Framework is to provide guidance on how to: identify, review, and summarise the biological mechanisms that underpin the associations between diet, nutrition, body weight, and physical activity and cancer incidence found within CUP Global assess the strength of evidence that a given biological mechanism operates between an exposure and outcome of interest These biological plausibility investigations apply to 1) exposures with an established link to cancer (strong evidence in CUP/CUP Global) and for which there are strong mechanistic data, but further work is needed (for example, a poorer understanding of the evidence for certain cancer sites); 2) exposures with a strong epidemiological link to cancer (strong evidence in CUP/CUP Global) but with limited mechanistic understanding; 3) exposures with a putative epidemiological link to cancer (limited suggestive evidence in the CUP/CUP Global) for which more robust mechanistic data would strengthen conclusions. Note that mechanisms evaluated using this Framework are defined as biological processes linking diet, nutrition, body weight, and physical activity and cancer incidence. The focus is primarily on molecular mechanisms but may expand to physiological mechanisms depending on the research question.

Patient-derived xenograft (PDX) models of lymphoma typically involve the injection of human tumor cells into an immunocompromised murine host. PDXs have the advantage that the tumor cells grow in a 3D environment within the mouse, meaning the selection pressure of in vitro establishment is avoided and the tumor cells better maintain their genetic heterogeneity. Here, we outline a method for producing and maintaining a PDX model of lymphoma. We describe three different methods to isolate a single cell suspension of the primary patient tumor, followed by either subcutaneous or intraperitoneal injection into an immunocompromised mouse. We then detail how to monitor tumor growth and how to harvest, passage, and store the tumors once they have grown. We highlight and discuss important protocol considerations including technical hints as well as the advantages and disadvantages of the methods described.

The risk of ALK-ve ALCL associated with textured breast implants (BIA-ALCL) is a notable example of lymphomas associated with medical devices. We describe 2 cases of ALK-ve ALCL associated with meshes, commonly used devices in surgical procedures such as herniorrhaphy. Our cases, each presenting some 10 years following mesh insertions, show anatomical and causal association to index tumours. In addition, we show that the genetic profile has significant overlaps with BIA-ALCL, invoking similar etiopathogenesis. Hence, we propose that mesh implant associated-ALCL (MIA-ALCL) is an unrecognised association and unreported disease entity that merits recognition, broader investigation, and surveillance. ### Competing Interest Statement The authors have declared no competing interest. ### Funding Statement We are grateful to the two patients for their consent and involvement. SDT is supported by research funding from a Cancer Research UK Cambridge Centre [C9685/A25117] award, a European Union Horizon 2020 Marie Skł Odowska-Curie Doctoral Training Network grant (no. 101072735) and an European Union National Institute for Cancer Research Programme (EXCELES, no. LX22NPO5102). ERJ is supported by a UK-MRC doctoral training award. AK was supported by funding from a NIHR Clinical Research Fellowship. AR and IK are funded by Clatterbridge Research Doctoral Fellowships. NK and JS receive funding support from Merseyside against blood cancers charity (MABC, The Bloom appeal, UK Registered Charity no. 1157459). ### Author Declarations I confirm all relevant ethical guidelines have been followed, and any necessary IRB and/or ethics committee approvals have been obtained. Yes The details of the IRB/oversight body that provided approval or exemption for the research described are given below: The collection of data and samples as well processing for the studies within the manuscript were approved and covered by the following: 1. Research Ethics Committee approval for the Clatterbridge Cancer Centre Biobank (Integrated Research Application System reference no. 305466 and identification number: 152025; Approval reference: 14/Northwest/1054 (Haydock, United Kingdom) and Human Tissue Authority License 16929). Granted 2014. 2. ARROVEN: Post-authorisation safety study (Brentuximab vedotin) study reference no. PASS: MA25101 granted by the European Network of Centres for Pharmacoepidemiology and Pharmacovigilance Reference number: EUPAS3583. 3. Human tissue authority ethics approval no. 07-Q0104-16, United Kingdom and Research Ethics Committee (Huntington, United Kingdom) approval number 07-Q0104-16 for sample handling and processing. I confirm that all necessary patient/participant consent has been obtained and the appropriate institutional forms have been archived, and that any patient/participant/sample identifiers included were not known to anyone (e.g., hospital staff, patients or participants themselves) outside the research group so cannot be used to identify individuals. Yes I understand that all clinical trials and any other prospective interventional studies must be registered with an ICMJE-approved registry, such as ClinicalTrials.gov. I confirm that any such study reported in the manuscript has been registered and the trial registration ID is provided (note: if posting a prospective study registered retrospectively, please provide a statement in the trial ID field explaining why the study was not registered in advance). Yes I have followed all appropriate research reporting guidelines, such as any relevant EQUATOR Network research reporting checklist(s) and other pertinent material, if applicable. Yes Reasonable requests for sequencing and FISH and any other data should be directed to the corresponding author.

Dose–response curves (DRCs) are a standard protocol in experimental biology to assess the effect of anticancer agents in vitro. Yet, therapy itself can reshape tumor composition, driving the gradual evolution of the DRC. Monitoring such changes can reveal the emergence of resistance, as well as evolutionary trade-offs that may be exploited to enhance therapeutic efficacy. Here, we investigate the evolution of DRCs in response to three different ALK inhibitors (ALKi) in ALK-positive anaplastic large cell lymphoma (ALCL) using patient-derived cell lines. Over the course of six months, ALKi concentrations were gradually escalated, allowing ALCL cells to adapt and survive at higher drug levels. Strikingly, at the same time, these resistant populations exhibited reduced fitness in the absence of the drug, indicating an evolutionary cost. This phenomenon, known as “drug addiction” or dependence, has been previously observed in melanoma following resistance to MAPK inhibitors and suggests that treatment holidays could optimize tumor control in the long term. Guided by these findings, we developed a mathematical model to capture the temporal evolution of DRCs and to test how therapy design might exploit addiction in ALCL. Calibrated with experimental data, the model simulated tumor growth under alternative dosing regimens. Simulations revealed that continuous therapy rapidly selects for resistance, leading to tumor regrowth within ∼150 days. In contrast, intermittent dosing schedules maintained long-term control for more than 300 days. Moreover, the model identified an adaptive strategy: switching from continuous to intermittent dosing precisely when tumor burden reaches its minimum. This evolutionarily informed approach prevents resistant clones from dominating while leveraging the fitness disadvantage associated with drug addiction. Preliminary experiments with naïve and resistant ALCL cell lines, cultured under intermittent dosing in both monoculture and co-culture systems, support the model’s predictions. These experiments also enable the refinement of the model's parameters to incorporate tumor heterogeneity. While additional validation is required, our findings demonstrate that integrating mathematical modeling with experimental DRCs provides a quantitative framework for predicting the evolutionary dynamics of resistance and addiction. Collectively, this study highlights the potential of adaptive and evolution-based therapy in ALCL. Optimized treatment schedules informed by evolutionary trade-offs can reduce tumor burden, delay progression, and exploit vulnerabilities generated by resistance mechanisms. Franco Pradelli, Lucy Hare, Jamie Matthews, Roxane Chen, Aiindrila Dhara, G. A. Amos Burke, Suzanne D. Turner, Jeffrey West. Capturing dose-response adaptation to identify evolutionarily-informed treatment strategies [abstract]. In: Proceedings of the AACR Special Conference in Cancer Research: Cancer Evolution: The Dynamics of Progression and Persistence; 2025 Dec 4-6; Albuquerque, NM. Philadelphia (PA): AACR; Cancer Res 2025;85(23_Suppl):Abstract nr B037.

Anaplastic Large Cell Lymphoma (ALCL) is an aggressive T-cell lymphoma affecting children and young adults. About 30% of patients develop therapy resistance therefore new precision medicine drugs are highly warranted. Multiple rounds of structure-activity optimization of Caffeic Acid Phenethyl Ester have resulted in CM14. CM14 causes upregulation of genes involved in oxidative stress response and downregulation of DNA replication genes leading to G2/M arrest and subsequent apoptosis induction. In accordance with this, an unbiased proteomics approach, confocal microscopy and molecular modeling showed that TUBGCP2, member of the centrosomal γ-TuRC complex, is a direct interaction partner of CM14. CM14 overcomes ALK inhibitor resistance in ALCL and is also active in T-cell Acute Lymphoblastic Leukemia and Acute Myeloid Leukemia. Interestingly, CM14 also induced cell death in docetaxel-resistant prostate cancer cells thus suggesting an unexpected role in solid cancers. Thus, we synthesized and thoroughly characterized a novel TUBGCP2 targeting drug that is active in ALCL but has also potential for other malignancies.

Anaplastic Large Cell Lymphoma (ALCL) is a CD30+ T-cell lymphoma accounting for ~10% of pediatric non-Hodgkin Lymphoma which is often driven by fusions resulting from translocations involving the anaplastic lymphoma kinase (ALK) and nucleophosmin (NPM1) genes. The COG-sponsored ANHL12P1 trial assessed brentuximab vedotin (BV) or crizotinib (CZ) combined with standard chemotherapy in newly diagnosed pediatric ALK+ALCL. It has previously been reported that levels of NPM::ALK fusion transcript and immune response to the NPM::ALK antigen, as determined by ALK autoantibody (Ab) titers, are predictive of outcomes, but the significance of these biomarkers following either BV or CZ is unknown. Methods: Patients <22 years of age with newly diagnosed ALCL were randomly assigned to receive BV or CZ in combination with an ALCL-99 chemotherapy backbone. Serial assessments for NPM::ALK fusion transcript and Ab to ALK were performed using peripheral blood samples. NPM::ALK transcript levels were measured by quantitative reverse transcription polymerase chain reaction (qRT-PCR) and normalized to the copy numbers (NCN) per 104 copies of the ABL gene. Based on prior studies >10 NCNs were considered a positive result and an ALK Ab titer of >1:750 was classified as a high-titer. Results: In total, 137 patients were enrolled with 68 and 69 patients randomized to the BV and CZ arms, respectively. Minimal residual disease (MRD), defined as detectable NPM::ALK transcript post-cycle 1, was associated with an inferior 2-year EFS overall (20.0% vs. 85.8% MRD-; p<0.0001), and by treatment arm (BV: 28.6 vs 91.1%, p<0.0001; CZ: 81.4 vs. 0.0%, p<0.0001). The evolution of transcript levels from baseline (minimal detectable disease, MDD) to post-cycle 1 (MRD) enabled the stratification of patients into three risk groups based on their associated 2-year EFS: low (MDD-/MRD-): 92.2%, and high (MDD+/MRD+): 12.5% (p<0.0001). High vs. low anti-ALK Ab titers at baseline did not impact EFS overall (80.8% vs. 73.9%, p=0.15) or by treatment arm (BV: 79.3% vs. 81.0%, p=1.0; CZ: 82.4% vs. 68.7%, p=0.08). Titers post-cycle 1 were not associated with outcome in the CZ arm (EFS 82.6% vs 74.2% for high-titer vs low-titer, p=0.42), but high-titer in the BV arm and in the entire population overall resulted in superior 2-year EFS of 89.9% vs. 68.0% (p=0.03) and 86.7% vs. 71.6% (p=0.04) respectively. Persistence of a high-titer from baseline through post-cycle 1 resulted in a superior 2-year EFS of 88.2%, compared to 73.1% and 68.4% for patients with persistent low titers and those transitioning from high to low-titers, respectively (p=0.01). Conclusion: In this study of pediatric ALK+ ALCL, NPM::ALK transcript levels proved to be a critical prognostic marker, with detectable MRD significantly associated with poor outcomes. Conversely, following the initiation of treatment the presence and persistence of high anti-ALK Ab titers showed promise as a favorable prognostic marker. These findings underscore the importance of serial monitoring of NPM::ALK transcript levels and anti-ALK Ab titers in guiding therapeutic strategies and predicting outcomes in pediatric ALK+ ALCL. Further studies are warranted to validate these biomarkers and explore their potential for personalized treatment approaches.

Prostate cancer (PCa) and Type 2 diabetes (T2D) often co-occur, yet their relationship remains elusive. While some studies suggest that T2D lowers PCa risk, others report conflicting data. This study investigates the effects of peroxisome proliferator-activated receptor (PPAR) agonists Bezafibrate, Tesaglitazar, and Pioglitazone on PCa tumorigenesis. Analysis of patient datasets revealed that high PPARG expression correlates with advanced PCa and poor survival. The PPARγ agonists Pioglitazone and Tesaglitazar notably reduced cell proliferation and PPARγ protein levels in primary and metastatic PCa-derived cells. Proteomic analysis identified intrinsic differences in mTORC1 and mitochondrial fatty acid oxidation (FAO) pathways between primary and metastatic PCa cells, which were further disrupted by Tesaglitazar and Pioglitazone. Moreover, metabolomics, Seahorse Assay-based metabolic profiling, and radiotracer uptake assays revealed that Pioglitazone shifted primary PCa cells' metabolism towards glycolysis and increased FAO in metastatic cells, reducing mitochondrial ATP production. Furthermore, Pioglitazone suppressed cell migration in primary and metastatic PCa cells and induced the epithelial marker E-Cadherin in primary PCa cells. In vivo , Pioglitazone reduced tumor growth in a metastatic PC3 xenograft model, increased phosho AMPKα and decreased phospho mTOR levels. In addition, diabetic PCa patients treated with PPAR agonists post-radical prostatectomy implied no biochemical recurrence over five to ten years compared to non-diabetic PCa patients. Our findings suggest that Pioglitazone reduces PCa cell proliferation and induces metabolic and epithelial changes, highlighting the potential of repurposing metabolic drugs for PCa therapy. Graphical Abstract

This current framework (version 2) updates the previous one published within the WCRF Gateway on Health Open Research (publication data of version 1: 12th February 2025). The purpose of the Framework is to provide guidance on how to: identify, review, and summarise the biological mechanisms that underpin the associations between diet, nutrition, body weight, and physical activity and cancer incidence found within CUP Global assess the strength of evidence that a given biological mechanism operates between an exposure and outcome of interest These biological plausibility investigations apply to 1) exposures with an established link to cancer (strong evidence in CUP/CUP Global) and for which there are strong mechanistic data, but further work is needed (for example, a poorer understanding of the evidence for certain cancer sites); 2) exposures with a strong epidemiological link to cancer (strong evidence in CUP/CUP Global) but with limited mechanistic understanding; 3) exposures with a putative epidemiological link to cancer (limited–suggestive evidence in the CUP/CUP Global) for which more robust mechanistic data would strengthen conclusions. Note that mechanisms evaluated using this Framework are defined as biological processes linking diet, nutrition, body weight, and physical activity and cancer incidence. The focus is primarily on molecular mechanisms but may expand to physiological mechanisms depending on the research question. Other mechanisms, for example those involving the social determinants of health, are likely to be at play in cancer risk, but have historically fallen outside of WCRF International’s research. WCRF International is seeking to clarify how these other mechanisms might be investigated within CUP Global in the future.