Publications

Degraders with dual activity against BRD4 and CBP/EP300 were designed. A structure-guided design approach was taken to assess and test potential exit vectors on the dual BRD4 and CBP/EP300 inhibitor, ISOX-DUAL. Candidate degrader panels revealed that VHL-recruiting moieties could mediate dose-responsive ubiquitination of BRD4. A panel of CRBN-recruiting thalidomide-based degraders was unable to induce ubiquitination or degradation of target proteins. High-resolution protein cocrystal structures revealed an unexpected interaction between the thalidomide moiety and Trp81 on the first bromodomain of BRD4. The inability to form a ternary complex provides a potential rationale for the lack of degrader activity with these compounds, some of which have remarkable affinities close to those of (+)-JQ1, as low as 65 nM in a biochemical assay, vs 1.5 μM for their POI ligand, ISOX-DUAL. Such a “degrader collapse” may represent an under-reported mechanism by which some putative degrader molecules are inactive with respect to target protein degradation.

NF-κB signalling is frequently dysregulated in human cancers making it an attractive therapeutic target. Despite concerted efforts to generate NF-κB inhibitors, direct pharmacological inhibition of the kinases mediating canonical NF-κB has failed due to on-target toxicities in normal tissues. So, alternative strategies, designed to target specific components of the NF-κB signalling machinery, have the potential to selectively inhibit tumour cells whilst reducing the toxicities associated with broad inhibition of NF-κB in non-malignant cells. Here we present evidence that a C8-linked pyrrolobenzodiazepine (PBD) containing proteolysis-targeting chimera (PROTAC) selectively degrades the NF-κB subunit, RelA/p65, in a proteasome-dependent manner. Our lead PROTAC (JP-163-16, 15d) showed cytotoxicity with mean LC50 values of 2.9 μM in MDA-MB-231 cells, 0.14 μM in MEC-1 cells and 0.23 μM in primary chronic lymphocytic leukaemia cells. In contrast, 15d was two-logs less toxic in primary B- and T-lymphocytes (mean LD50 19.1 μM and 36.4 μM, respectively). Importantly, the development of 15d, by conjugating the C8-linked PBD with a cereblon-targeting ligand using a five-carbon linker, abolished the ability of the C8-linked PBD to bind to DNA, whilst demonstrating cytotoxicity in cancer cells associated with the degradation of RelA/p65. Mechanistically, 15d displayed PROTAC credentials through the selective degradation of NF-κB RelA/p65 in a proteasome-dependent manner and showed a five-fold reduction in potency in the cereblon deficient, lenalidomide resistant, myeloma cell line, RPMI-8226. To our knowledge, this work describes the first PROTAC capable of selective degradation of a single NF-κB subunit and highlights the therapeutic potential of our strategy for the treatment of RelA/p65-dependent tumours.

Abstract unavailable.

Upon infection, human papillomavirus (HPV) manipulates host cell gene expression to create an environment that is supportive of a productive and persistent infection. The virus-induced changes to the host cell’s transcriptome are thought to contribute to carcinogenesis. Here, we show by RNA-sequencing that oncogenic HPV18 episome replication in primary human foreskin keratinocytes (HFKs) drives host transcriptional changes that are consistent between multiple HFK donors. We have previously shown that HPV18 recruits the host protein CTCF to viral episomes to control the differentiation-dependent viral transcriptional programme. Since CTCF is an important regulator of host cell transcription via coordination of epigenetic boundaries and long-range chromosomal interactions, we hypothesised that HPV18 may also manipulate CTCF to contribute to host transcription reprogramming. Analysis of CTCF binding in the host cell genome by ChIP-Seq revealed that while the total number of CTCF binding sites is not altered by the virus, there are a sub-set of CTCF binding sites that are either enriched or depleted of CTCF. Many of these altered sites are clustered within regulatory elements of differentially expressed genes, including the tumour suppressor gene cell adhesion molecule 1 (CADM1), which supresses epithelial cell growth and invasion. We show that HPV18 establishment results in reduced CTCF binding at the CADM1 promoter and upstream enhancer. Loss of CTCF binding is coincident with epigenetic repression of CADM1, in the absence of CpG hypermethylation, while adjacent genes including the transcriptional regulator ZBTB16 are activated. These data indicate that the CADM1 locus is subject to topological rearrangement following HPV18 establishment. We tested this hypothesis using 4C-Seq (circular chromosome confirmation capture-sequencing) and show that HPV18 establishment causes a loss of long-range chromosomal interactions between the CADM1 transcriptional start site and the upstream transcriptional enhancer. These data show that HPV18 manipulates host cell promoter-enhancer interactions to drive transcriptional reprogramming that may contribute to HPV-induced disease progression.

Maintaining genome integrity is an essential and challenging process. RAD51 recombinase, the central component of several crucial processes in repairing DNA and protecting genome integrity, forms filaments on DNA, which are tightly regulated. One of these RAD51 regulators is FIGNL1 (fidgetin-like 1), which prevents RAD51 genotoxic chromatin association in normal cells and persistent RAD51 foci upon DNA damage. The cryogenic electron microscopy–imaged structure of FIGNL1 in complex with RAD51 reveals that FIGNL1 forms a nonplanar hexamer and encloses RAD51 N terminus in the FIGNL1 hexamer pore. Mutations in pore loop or catalytic residues of FIGNL1 render it defective in filament disassembly and are lethal in mouse embryonic stem cells. Our study reveals a distinct mechanism for removing RAD51 from bound substrates and provides the molecular basis for FIGNL1 in maintaining genome stability.

The P2X purinergic receptor 7 (P2X7) has an essential role in inflammation, innate immunity, tumor progression, neurodegenerative diseases, and several other diseases, leading subsequently to the development of P2X7 modulators. AZ11645373 is a frequently studied P2X7 antagonist tool compound but always used as a racemic mixture. Racemic AZ11645373 can be separated into its respective enantiomers by chiral chromatography, albeit in small batches, and these were stereochemically intact over two years later, by chiral high-performance liquid chromatography (HPLC) analysis. On a higher scale, significant decomposition is observed during purification. One of the enantiomers was crystallized as a palladium complex, and its (R)-configuration was determined by single-crystal X-ray diffraction, further confirmed, in solution, by vibrational circular dichroism. Biological studies demonstrated that both (S)- and (R)-forms were able to fully inhibit human P2X7, but (R)-AZ11645373 was more potent, with an IC50 of 32.9 nM. Contrary to its effect on human P2X7, (S)-AZ11645373 was ineffective on mouse P2X7, while the (R)-AZ11645373 enantiomer was a full antagonist. These results demonstrated that the antagonistic effects of racemic AZ11645373 are mainly due to its (R)-enantiomer. Site-directed mutagenesis and molecular dynamics simulations indicated that the (R)-enantiomer may form specific interactions with Phe95 and the antagonists bound to other P2X7 monomers. Phe95 is situated in the allosteric binding site at the edge of the upper vestibule and appears to be the pivotal molecular gateway between AZ11645373 allosteric binding and locking of the closed state of the P2X7 channel. All together, these structure-function relationships should be helpful for drug design of P2X7 modulators.

Introduction: Physiological age-related bone loss is common, with 50% of women aged ≥80 having osteoporosis. Bone loss is exacerbated in women receiving aromatase inhibitors (AIs) for early breast cancer (EBC), increasing fracture risk. This study explored the management of bone-health in older women (≥70 years) with EBC and factors influencing clinical decision-making. Materials and Methods: This was a sub-study of a larger United Kingdom multicentre observational study into practice variation and outcomes in older women (≥70) with EBC (Age Gap study). Participants were aged ≥70 years with EBC; data were collected on health status, treatments, and outcomes. This sub-study focused on patients recruited at five hospitals, where more detailed data on bone health and management were collected for women with ER + ve (oestrogen receptor positive) cancers who received adjuvant or primary endocrine therapy treatment. We aimed to determine factors influencing treatment selection and outcomes in this age group. Results: The main Age Gap study recruited between 2013 and 2018. In this sub-study, 565 patients had ER + ve cancers, of whom 529 (93.6%) received AIs and 26 (4.6%) tamoxifen. The median age of participants was 77 years (70–98 years). A baseline dual energy x-ray absorptiometry (DEXA) scan was performed in only 354/529 (67%) of the AI group. Bisphosphonates were prescribed for 226/529 (43%). Baseline DEXA scans were more likely to be requested if patients were fit for surgery and were < 80 years old. Of those scanned (n = 354), 148 (42%) were osteopenic and 64 (18%) osteoporotic. Bisphosphonate prescription was associated with younger age (<80 years old) (p = 0.02). From recruitment to 2022, fractures were diagnosed in 23% of participants (122/529), of whom only 38% (46/122) had received prior bisphosphonates. Frailty or prefrailty (Rockwood scale) were present in 94% (431/461), but there was no correlation between frailty and baseline hip (r2 = 0.0098) or spine (r2 = 0.00007) T-scores. Rates of DEXA scanning varied between centres from 36% to 76% (p < 0.001) for unknown reasons. Discussion: Age and general health influenced bone-health management decision-making, but there was considerable variation between centres, highlighting the need for standardised bone-health care for older women with EBC.

Wnt/β-catenin signalling is important for normal hematopoietic stem/progenitor cell (HSPC) biology and heavily implicated in acute and chronic myeloid leukaemia (AML and CML). The central mediator β-catenin is an attractive therapeutic target in myeloid neoplasms however its targeting has been hampered by a poor characterisation of its molecular interactions in haematopoietic cells, which will differ from its network in solid tissues. Our previous β-catenin interactome study identified the significant enrichment of RNA-binding proteins (RBP) implying post-transcriptional roles for β-catenin in myeloid cells. To identify β-catenin interacting mRNAs we performed β-catenin RNA-immunoprecipitation coupled to RNA-sequencing (RIP-seq) and identified significantly enriched Wnt signalling pathway transcripts. Using β-catenin cross-linking immunoprecipitation (CLIP) we demonstrated a limited capacity for β-catenin to bind RNA directly, implying dependence on other RBPs. β-Catenin was found to interact with Musashi-2 (MSI2) in both myeloid cell lines and primary AML patient samples, where expression was significantly correlated. MSI2 knockdown reduced Wnt signalling output (TCF/LEF activity), through suppression of LEF-1 expression and nuclear localisation. Through both RIP and CLIP we demonstrate MSI2 binds LEF1 mRNA in a partly β-catenin dependent fashion, and may impact the post-transcriptional control of LEF-1 expression. Finally, we show that MSI2-mediated expansion of human HSPCs could be partly driven through LEF1 regulation. This is the first study to experimentally demonstrate functional crosstalk between MSI2 and Wnt signalling in human cells, and indicates potential novel post-transcriptional roles for β-catenin in a haematological context.

The Wnt/β-catenin pathway is an evolutionarily conserved signal transduction cascade with critical regulatory roles in cellular proliferation, cell fate determination and tissue homeostasis. Through the regulation of multiple human stem cell systems, canonical Wnt signalling is not only a major contributor to normal development, but also heavily implicated in a multitude of human diseases, including cancer. The central mediator of the pathway β-catenin, first identified as Armadillo (ARM) in Drosophila, has well-defined roles in cell adhesion and transcription within the pathway. However, accumulating evidence suggests β-catenin functionality is more complex than initially anticipated with reported roles beyond those historically characterised, including the regulation of RNA and RNA-binding proteins (RBP). This review will summarise the current understanding around β-catenin as a post-transcriptional regulator in normal and malignant development, drawing particular attention to cell types not traditionally used to characterise Wnt signalling but uniquely placed to reveal novel β-catenin function.

Abstract unavailable.

Objectives: Androgen deprivation therapy (ADT) forms the mainstay of treatment for advanced prostate cancer. Traditionally administered as a luteinising hormone-releasing hormone (LHRH) agonist depot injection, newer options for ADT include transdermal oestradiol patches (tE2) or oral LHRH antagonists. This study aimed to identify whether this is an important choice for men, which treatment men would choose if offered either LHRH agonist injections, tE2 patches or oral LHRH antagonists as ADT, and to explore the factors influencing this decision. Subjects, Patients, and Methods: Five focus groups were conducted. A total of 24 men from around the UK participated in discussions, of whom 10 had never had prostate cancer and 14 had early prostate cancer but had not received ADT. Focus groups were co-facilitated with patient and public involvement representatives. Transcripts were analysed using a critical realist thematic analysis approach. Results: Participants reported that having a choice of ADT is important and being involved in making treatment decisions can help men maintain an element of control. Most men expressed a preference to avoid ADT with LHRH agonist injections; 14 of the 24 men reported they would choose an oral LHRH antagonist, eight reported they would choose tE2 patches, and two that they would choose LHRH agonist injections. Participants reported a large number of factors that influenced their treatment choice that were grouped into: (i) side-effects of treatment, (ii) logistical aspects, and (iii) advice from trusted others. Personal preferences between different types of ADT were based on factors that participants identified as important to them, this prioritisation of factors was influenced by participants’ past experiences, current health beliefs, and future expectations. Conclusions: Men wish to have choice of ADT, and many would not choose LHRH agonists injections if there were other ADT options available. This should be considered, as reimbursement committees and guideline-makers consider the role of alternative ADT strategies.

Copper catalyzed aldehyde-alkyne-amine (A3) couplings lead to multifunctional, racemic, propargylic amines, many on a multigram scale. As part of an industrial collaboration, a selection of linkers was purified by chiral HPLC to afford single enantiomers, the absolute configuration of which was determined by vibrational circular dichroism (vCD). To show medicinal chemistry applications, selected linkers were further derivatized into potential cellular probes and (+)-JQ1 containing PROTACs (proteolysis targeting chimeras), which degraded their target protein BRD4.

Fatigue may affect the decision to deploy effort (cost) for a given rewarded outcome (benefit). However, it remains unclear whether these fatigue-associated changes can be attributed to simply feeling fatigued. To investigate this question, twenty-two healthy males made a series of choices between two rewarded options: a fixed, no effort option, where no physical effort was required to obtain a set, low reward vs. a variable, effortful option, in which both the physical effort requirements (i.e. different durations of a sustained contraction performed in the lower limbs) and rewards (i.e. greater monetary incentives) was varied. Effort-based choices were made under two conditions: (1) a rested (control) state, (2) a pre-induced state of perceived fatigue, evoked through physical exertion in the upper-limbs, thus controlling for the physical manifestation of fatigue (i.e. decline in muscular force production in the lower limbs) on the decision process. Though prior physical exertion increased the perception of fatigue, participants choices did not significantly differ between control and fatigue conditions. Across both conditions, participants demonstrated an anticipated aversion to effort, with greater effort requirements reducing the decision to engage in actions associated with higher rewards. However, in the fatigue state only, decision time was prolonged and self-reported confidence in individuals’ ability to perform high effort actions was reduced. The findings suggest that a perceived state of fatigue does not necessarily alter cost/benefit comparisons within effort-based decisions, but may introduce greater uncertainty within choice and reduce self-confidence. These findings evidence altered evaluative processes during decision making under conditions of fatigue.

This review comprehensively analyzes key parameters that gov-ern the use of luminescent benzoselenadiazole (BSD) derivatives.We examine the main factors affecting their photophysicalproperties, including structural variations, heavy atom effectsand environmental conditions, and discuss their potentialapplications in bioimaging technology. Whenever possible, theproperties were compared to those of 2,1,3-benzoxadiazole and2,1,3-benzothiadiazole analogues. This review also highlightsemerging trends and future directions in the research and devel-opment of BSD compounds, underscoring their significance andpotential impact in advancing phototechnology and relatedfields. We also aim to offer valuable insight into the versatilenature of BSD derivatives and their expanding role in scientificand technological innovations.

DNA damage checkpoints are key regulatory signaling cascades that arrest cell cycle progression upon DNA damage or upon DNA replication stalling and allow time for repair or correction. Failure to elicit these checkpoints can lead to genomic instability that can result in cell death or mutations, ultimately leading to diseases such as cancer. Components of the DNA damage checkpoint are attractive targets for precision medicine to treat cancers. Over the last several years, cutting-edge structural techniques have provided molecular insights into the highly coordinated checkpoint signaling that occurs in response to DNA damage or other obstacles to replication progression. This review summarizes our current mechanistic understanding of the DNA damage checkpoint in eukaryotes, with an emphasis on the sensor kinases ATM (Tel1) and ATR (Mec1), highlighting structure–function and cellular studies.

We report a family of propeller-shaped polynuclear metal complexes whose overall chirality is dictated by a single stereogenic centre within their component amino alcohol-ligand. These topologically intriguing complexes are readily prepared in enantiomerically pure form and are shown here to catalyse the conjugate addition of barbituric acids and their derivatives to nitroalkenes, with a catalyst loading of 1 mol%. Although only low levels of enantioinduction are observed, control experiments indicate that the enantioselectivity is dictated by the overall topology of the complex and not governed by binding to the tetrametallic entity, heralding a potentially new mode of catalysis.

In epidemic outbreak control management, the synergy between healthcare facilities and infrastructure, including road networks and ambulance services, plays a pivotal role. In this article, we formulate and analyze a mathematical model that underscores the significance of infrastructure components along with healthcare facilities to effectively control an epidemic outbreak. By intricately examining the interplay between healthcare accessibility and the swift movement of medical resources, this study contributes valuable insights into optimizing response strategies during such outbreaks. Through qualitative analysis, we establish the model’s susceptibility to a range of bifurcations, including transcritical (forward and backward), saddle-node, Hopf, and codimension-2 Bogdanov–Takens bifurcations. Furthermore, we provide an epidemiological interpretation of the intricate dynamic behaviors observed in the context of disease endemism. The proposed model offers insights that can guide policymakers, healthcare administrators, and urban planners in devising effective strategies for combating epidemic outbreaks while fostering community resilience.

Fragment approaches are long-established in target-based ligand discovery, yet their full transformative potential lies dormant because progressing the initial weakly binding hits to potency remains a formidable challenge. The only credible progression paradigm involves multiple cycles of costly conventional design-make-test-analyse medicinal chemistry. We propose an alternative approach to fragment elaboration, namely performing large numbers of parallel and diverse automated multiple step reactions, and evaluating the binding of the crude reaction products by high-throughput protein X-ray crystallography. We show it is effective and low-cost to perform, in parallel, large numbers of non-uniform multi-step reactions, because, even without compound purification, crystallography provides a high-quality readout of binding. This can detect low-level binding of weakly active compounds, which the target binding site extracts directly from crude reaction mixtures. In this proof-of-concept study, we have expanded a fragment hit, from a crystal-based screen of the second bromodomain of pleckstrin homology domain-interacting protein (PHIP(2)), using array synthesis on low-cost robotics. We were able to implement 6 independent multi-step reaction routes of up to 5 steps, attempting the synthesis of 1876 diverse expansions, designs entirely driven by synthetic tractability. The expected product was present in 1108 (59%) crude reaction mixtures, detected by liquid chromatography mass spectrometry (LCMS). 22 individual products were resolved in the crystal structures of crude reaction mixtures added to crystals, providing an initial structure activity relationship map. 19 of these showed binding pose stability, while, through binding instability in the remaining 3 products, we could resolve a stereochemical preference for mixtures containing racemic compounds. One compound showed biochemical potency (IC50=34 μM) and affinity (Kd=50 μM) after resynthesis. This approach therefore lends itself to routine fragment progression, if coupled with algorithmically guided compound and reaction design and new formalisms for data analysis.

Background: We trialled the first digital pathway (BRCA-DIRECT) aiming to improve capacity for mainstreamed BRCA testing within UK breast oncology services. Patients received standardised digital pretest information, with saliva sampling and consent to testing completed at home. For individualised support, we offered access to a clinical genetics professional via a telephone helpline (TH). Methods: To evaluate the utilisation, uptake and resource requirements for provision of the TH, we analysed data from structured call logs recorded in the BRCA-DIRECT Study. Mixed-methods analysis included combining quantitative data from call logs and patient demographics with thematic analysis of free-text notes establishing reasons for calls. Additional data were analysed from structured telephone interviews. Results: Calls were received from 201/1140 (17.6%) patients. We identified that 84.6% of calls (274 calls, 1097 min) pertained to 'administrative' support needs only. The remaining 15.4% required a clinical genetics professional (50 calls, 344 min). Of the clinical calls received: 26.0% were placed prior to test consent, 36.0% while awaiting results and 38.0% post results, with median (interquartile) call lengths of 8 (4-10) min; 5.5 (4-10) min; and 5 (3-7) min, respectively. Across all 1140 patients, a mean of 0.3 min of clinical time was required per patient. Conclusions: Our findings demonstrate that the 'BRCA-DIRECT' model of standardised information provision served most patients, with a minority using the helpline for supplementary clinical information or support. The modest per-patient requirement for clinical time supports the scalability of this model for expanding mainstream genetic testing within UK oncology services.

A dose optimization trial in oncology may be performed to compare an approved dose level of a given drug with a reduced dose level, testing the hypothesis that efficacy is maintained whilst reducing side effects and consequently improving adherence and quality-of-life. This is particularly relevant with modern therapeutic agents whose mechanisms of action imply that efficacy may not necessarily be linearly related to the dose. Using a conventional non-inferiority framework leads to large sample sizes that are often unfeasible in the phase IV setting. An alternative is to use a margin of practical non-inferiority, which we define in this paper and show how it can be exploited to justify a sample size. Whilst defining the extent of the margin, researchers also pre-specify the other dimensions of interest, such as receptor occupancy and/or side effects and quality-of-life, that will be used to establish practical non-inferiority if the observed efficacy of the reduced dose level lies within the margin. The comparison of efficacy is based on the observed difference between the reduced and the approved levels, instead of the confidence interval of this difference, leading to a reduction in sample size. The reduction in precision due to the smaller sample size is compensated by formally pre-specifying the additional dimensions to the decision process, allowing a more thorough assessment of the opportunity to reduce a dose in practice, with the many advantages that this may involve.

RIF1 is a multifunctional protein that regulates DNA replication and repair. RIF1-deficient cells are hypersensitive to DNA replication stress. Of the two alternatively spliced RIF1 isoforms, called RIF1-Short and RIF1-Long, the RIF1-Long isoform is more capable than RIF1-Short in supporting cell recovery from replication stress. Examining replication stress resistance mechanisms specific to RIF1-Long, we find that prolonged replication stress unexpectedly induces interaction of RIF1-Long with BRCA1. Mechanistically, a phosphorylated SPKF motif unique to the RIF1-Long isoform binds the tandem BRCT domain of BRCA1. BRCA1–RIF1-Long interaction is strongly down-regulated through dephosphorylation by RIF1-associated Protein Phosphatase 1. BRCA1–RIF1-Long interaction requires ATR signaling, and occurs predominantly during S phase. Loss of RIF1-Long impairs the formation of RAD51 foci, and reduces the efficiency of homology-mediated repair at broken replication forks. In summary, our investigation establishes RIF1-Long as a new functional binding partner of the BRCA1-BRCT domain, crucial to protect cells from extended DNA replication stress by enabling RAD51-dependent repair of broken replication forks.

Background: Lemur Tail Kinase 3 (LMTK3) promotes cell proliferation, invasiveness and therapy resistance, and its expression correlates with poor survival in several different malignancies, including breast cancer. Crosstalk through extracellular vesicles (EVs) is an increasingly appreciated mechanism of cell communication within the tumour immune microenvironment, which contributes to different aspects of cancer progression and plays a pivotal role in shaping tumour fate. Methods: Nanoparticle tracking analysis and transmission electron microscopy were used to study the effects of LMTK3 on EV size, while single particle interferometry allowed us to examine LMTK3-dependent effects on the subpopulation distribution of EVs. Quantitative mass spectrometry was used to profile LMTK3-dependent proteomics changes in breast cancer-derived EVs. Bioinformatics analysis of clinical data along with in vitro and cell-based assays were implemented to explore the effects of LMTK3-dependent EV protein cargo on the tumour immune microenvironment. To elucidate the mechanism through which LMTK3 impacts endosomal trafficking and regulates EV biogenesis, we used a variety of approaches, including in vitro kinase assays, confocal and electron microscopy, as well as in vivo subcutaneous and orthotopic breast cancer mouse models. Results: Here, we report that LMTK3 increases the average size of EVs, modulates immunoregulatory EV proteomic cargo and alters the subpopulation distribution of EVs released by breast cancer cells. Mechanistically, we provide evidence that LMTK3 phosphorylates Rab7, a key regulator of multivesicular body (MVB) trafficking, thereby reducing the fusion of MVBs with lysosomes and subsequent degradation of intralumenal vesicles, resulting in altered EV release. Moreover, LMTK3 causes increased packaging of phosphoserine aminotransferase 1 (PSAT1) in EVs, leading to a paracrine upregulation of phosphoglycerate dehydrogenase (PHGDH) in monocytes when these EVs are taken up. PSAT1 and PHGDH play key roles in the serine biosynthesis pathway, which is closely linked to cancer progression and regulation of monocyte behaviour. LMTK3 EV-induced elevated PHGDH expression in monocytes reduces their infiltration into breast cancer 3D spheroids and in vivo breast cancer mouse models. Furthermore, these infiltrating monocytes preferentially differentiate into pro-tumourigenic M2-like macrophages. Additional breast cancer mouse studies highlight the contribution of LMTK3-dependent EVs in the observed immunosuppressive macrophage phenotype. Finally, in vitro experiments show that pharmacological inhibition of LMTK3 reverses the pro-tumourigenic and immunomodulatory effects mediated by EVs derived from LMTK3 overexpressing cells. Conclusion: Overall, this study advances our knowledge on the mechanisms of EV biogenesis and highlights a novel oncogenic role of LMTK3 in the breast TME, further supporting it as a target for cancer therapy.

Cervical cancer is the second most frequent gynaecological malignancy in the world. Human papillomavirus (HPV) infection is the primary etiologic agent of cervical cancer. However, HPV alone is not sufficient for tumor progression. The clinical display of HPV infection also depends on the host’s immune status. Both innate and adaptive immunity recognize and fight foreign pathogens inside the body, but sometimes they prove ineffective against HPV. HPV has several mechanisms for evading the immune system. After infection, HPV multiplies in keratinocytes, which are distant from immune centers and have a naturally short lifespan. The naturally short life cycle of the keratinocyte removes the need for the virus to destroy the cells, which would trigger inflammation and immune response. In addition, HPV downregulates the expression of interferon genes. Despite viral immune evasion, the immune system effectively resists most HPV infections and mounts strong localized cell mediated immune responses. Despite significant progress in observations and clinical practice, many aspects of the complex interactions between HPV and the human immune system remain not fully understood. Langerhans cells (LCs) are known to play a critical role in producing innate and adaptive cellular immune responses against HPV infection. In this paper, we propose and analyze a mathematical of HPV infection with particular focus on the role of Langerhans cells in facilitating immune response, as well as on the treatment of HPV infection by induction of the appropriate virus-specific immune responses in patients. We determine equilibria of the model, analyse their stability, and derive the basic reproduction number. Sensitivity analysis is performed to investigate the effects of individual parameters on system dynamics. We explore the impulsive therapy for controlling HPV infection, and discuss how these findings may be helpful in development of immunotherapy against HPV infection.

Background: Ovarian and tubal cancers are lethal gynaecological cancers, with over 50% of the patients diagnosed at advanced stage. Trial design: Randomised controlled trial involving 27 primary care trusts adjacent to 13 trial centres based at NHS Trusts in England, Wales and Northern Ireland. Methods Participants: Postmenopausal average-risk women, aged 50–74, with intact ovaries and no previous ovarian or current non-ovarian cancer. Interventions: One of two annual screening strategies: (1) multimodal screening (MMS) using a longitudinal CA125 algorithm with repeat CA125 testing and transvaginal scan (TVS) as second line test (2) ultrasound screening (USS) using TVS alone with repeat scan to confirm any abnormality. The control (C) group had no screening. Follow-up was through linkage to national registries, postal follow-up questionnaires and direct communication with trial centres and participants. Objective: To assess comprehensively risks and benefits of ovarian cancer screening in the general population. Outcome: Primary outcome was death due to ovarian or tubal cancer as assigned by an independent outcomes review committee. Secondary outcomes included incidence and stage at diagnosis of ovarian and tubal cancer, compliance, performance characteristics, harms and cost-effectiveness of the two screening strategies and a bioresource for future research. Randomisation: The trial management system confirmed eligibility and randomly allocated participants using computer-generated random numbers to MMS, USS and C groups in a 1:1:2 ratio. Blinding: Investigators and participants were unblinded and outcomes review committee was masked to randomisation group. Analyses: Primary analyses were by intention to screen, comparing separately MMS and USS with C using the Versatile test. Results Recruitment: 1,243,282 women were invited and 205,090 attended for recruitment between April 2001 and September 2005. Randomised: 202,638 women: 50,640 MMS, 50,639 USS and 101,359 C group. Numbers analysed for primary outcome: 202,562 (>99.9%): 50,625 (>99.9%) MMS, 50,623 (>99.9%) USS, and 101,314 (>99.9%) C group. Outcome: Women in MMS and USS groups underwent 345,570 and 327,775 annual screens between randomisation and 31 December 2011. At median follow-up of 16.3 (IQR 15.1–17.3) years, 2055 women developed ovarian or tubal cancer: 522 (1.0% of 50,625) MMS, 517 (1.0% of 50,623) USS, and 1016 (1.0% of 101314) in C group. Compared to the C group, in the MMS group, the incidence of Stage I/II disease was 39.2% (95% CI 16.1 to 66.9) higher and stage III/IV 10.2% (95% CI –21.3 to 2.4) lower. There was no difference in stage in the USS group. 1206 women died of the disease: 296 (0.6%) MMS, 291 (0.6%) USS, and 619 (0.6%) C group. There was no significant reduction in ovarian and tubal cancer deaths in either MMS (p = 0.580) or USS (p = 0.360) groups compared to the C group. Overall compliance with annual screening episode was 80.8% (345,570/420,047) in the MMS and 78.0% (327,775/420,047) in the USS group. For ovarian and tubal cancers diagnosed within one year of the last test in a screening episode, in the MMS group, the sensitivity, specificity and positive predictive values were 83.8% (95% CI 78.7 to 88.1), 99.8% (95% CI 99.8 to 99.9), and 28.8% (95% CI 25.5 to 32.2) and in the USS group, 72.2% (95% CI 65.9 to 78.0), 99.5% (95% CI 99.5 to 99.5), and 9.1% (95% CI 7.8 to 10.5) respectively. The final within-trial cost-effectiveness analysis was not undertaken as there was no mortality reduction. A bioresource (UKCTOCS Longitudinal Women’s Cohort) of longitudinal outcome data and over 0.5 million serum samples including serial annual samples in women in the MMS group was established and to date has been used in many new studies, mainly focused on early detection of cancer. Harms: Both screening tests (venepuncture and TVS) were associated with minor complications with low (8.6/100,000 screens MMS; 18.6/100,000 screens USS) complication rates. Screening itself did not cause anxiety unless more intense repeat testing was required following abnormal screens. In the MMS group, for each screen-detected ovarian or tubal cancer, an additional 2.3 (489 false positives; 212 cancers) women in the MMS group had unnecessary false-positive (benign adnexal pathology or normal adnexa) surgery. Overall, 14 (489/345,572 annual screens) underwent unnecessary surgery per 10,000 screens. In the USS group, for each screen-detected ovarian or tubal cancer, an additional 10 (1630 false positives; 164 cancers) underwent unnecessary false-positive surgery. Overall, 50 (1630/327,775 annual screens) women underwent unnecessary surgery per 10,000 screens. Conclusions: Population screening for ovarian and tubal cancer for average-risk women using these strategies should not be undertaken. Decreased incidence of Stage III/IV cancers during multimodal screening did not translate to mortality reduction. Researchers should be cautious about using early stage as a surrogate outcome in screening trials. Meanwhile the bioresource provides a unique opportunity to evaluate early cancer detection tests.

Postmitotic neurons have high levels of methylated cytosine and its oxidized intermediates such as 5-hydroxymethylcytosine1. However, the functional relevance of these epigenetic modifications of DNA are poorly understood. Here we show that some cytidine analogues, such as cytarabine, cause DNA double-strand breaks during TET-mediated active 5-methylcytosine demethylation by interrupting TDG-dependent base excision repair. These double-strand breaks are frequently converted into deletions and translocations by DNA ligase 4. In vivo, Purkinje and Golgi cells in the cerebellum are the only neuronal populations that exhibit high levels of DNA damage due to cytarabine. In Purkinje cells, TET targets highly expressed gene bodies marked by enhancer-associated histone modifications. Many of these genes control movement coordination, which explains the long-recognized cerebellar neurotoxicity of cytarabine2. We show that other cytidine analogues, such as gemcitabine, cause only single-strand breaks in neurons, which are repaired by DNA ligase 3 with minimal toxicity. Our findings uncover a mechanistic link between TET-mediated DNA demethylation, base excision repair and gene expression in neurons. The results also provide a rational explanation for the different neurotoxicity profiles of an important class of antineoplastic agents.

Optimising the use of approved drugs requires evidence from post-approval trials that investigate variations of their use. Determining optimal drug use goes beyond the dominant, academic effort to conduct trials to identify effective lower doses of new drugs. Other important therapeutic approaches that use either less, similar, or more drug than the standard dose need testing in clinical trials, to get the most out of these drugs. Trial objectives on survival outcomes vary greatly; some aim for superiority, others for equivalent exposure or non-inferiority. This Personal View aims to inform academic trialists in how to conceive and prioritise questions aimed at determining the optimal use of drugs, taking into account the perspectives of patients, clinicians, and trial funders, to maximise the chances of successful delivery and impact for patients globally.

Acute myeloid leukemia (AML) is a challenging blood cancer to treat, with only about 24% of patients surviving for 5 years after diagnosis. A key challenge is that AML cells stick to normal cells in the bone marrow (BM), and these BM cells protect them from chemotherapy. The aim of this project is to find drugs that disrupt AML cell adherence to BM cells and release them into the blood, where chemotherapy will be more effective. To achieve this, we have created a model of adhesive BM and shown that it mimics the drug resistance seen clinically. We have used the model as a testing platform for drugs that disrupt AML cell adhesion. We have shown that the combined targeting of CD44 and FAK, using anti-CD44 and the clinical-grade FAK inhibitor defactinib, inhibits the adhesion of the most primitive AML cells that are associated with drug resistance and disease relapse.

Background: Metformin is a widely used anti-diabetic drug. Several studies have suggested that metformin has anticancer activity in some malignancies, including prostate cancer. Metformin might also mitigate the adverse metabolic effects of androgen-deprivation therapy (ADT). We hypothesised that metformin might improve survival in patients with metastatic hormone-sensitive prostate cancer and reduce metabolic complications associated with ADT. Methods: The STAMPEDE multi-arm, multi-stage, randomised phase 3 trial recruited patients with high-risk locally advanced or metastatic adenocarcinoma of the prostate staged by conventional imaging with isotope bone and CT scanning. This publication reports findings for the most recent STAMPEDE research question, testing the addition of metformin to standard of care for non-diabetic (glycated haemoglobin [HbA1c] <48 mmol/mol [equivalent to <6·5%]) patients with metastatic disease with adequate renal function (glomerular filtration rate ≥45 ml/min/1·73 m2) and WHO performance status 0–2. This trial recruited from 112 hospitals in the UK and Switzerland to the STAMPEDE protocol. Patients were randomly allocated (1:1) to standard of care or standard of care plus metformin 850 mg twice daily. Random assignment was by telephone using minimisation with a random element of 20% (developed and maintained by the MRC Clinical Trials Unit at UCL), stratified for randomising hospital, age (<70 years vs ≥70 years), WHO performance status (0 vs 1 or 2), type of ADT, regular long-term use of aspirin or non-steroidal anti-inflammatory drugs (NSAIDs; yes vs no), pelvic nodal status (positive vs negative), planned radiotherapy (yes vs no), and planned docetaxel or androgen receptor pathway inhibitor (ARPI) use (docetaxel vs abiraterone, enzalutamide, or apalutamide vs none). Standard of care comprised ADT with or without radiotherapy and with or without docetaxel or ARPI. The primary outcome measure was overall survival, defined as the time to death from any cause, assessed in the intention-to-treat population. Safety was assessed in patients who started treatment. The trial is registered with ClinicalTrials.gov, NCT00268476 and ISRCTN, ISRCTN78818544. Findings: Between Sep 5, 2016, and Mar 31, 2023, 1874 patients with metastatic disease were randomly allocated to standard of care (n=938) or standard of care plus metformin (n=936). The median patient age was 69 years (IQR 63-73) and the median PSA was 84 ng/mL (24–352). 1758 (94%) of 1874 patients were newly diagnosed with metastatic disease and 116 (6%) were diagnosed with metachronous relapsing disease. 1543 (82%) of 1874 patients received ADT plus docetaxel and 52 (3%) received abiraterone, enzalutamide, or apalutamide. The median time to most recent case report form follow-up was 60 months (IQR 49–72). 473 deaths were reported in the standard of care group; median survival was 61·8 months (IQR 29·7 to not reached). There were 453 deaths in the metformin group; median survival was 67·4 months (32·5 to not reached; HR 0·91, 95% CI 0·80–1·03; p=0·15). Grade 3 or worse adverse events were reported in 487 (52%) of 938 patients in the standard of care group and 523 (57%) of 921 patients in the standard of care plus metformin group. 61 (7%) patients in the standard of care group and 84 (9%) patients in the standard of care plus metformin group reported at least one grade 3 or worse gastrointestinal adverse event; all other body systems showed no difference in grade 3 adverse events. There were six drug-related deaths in the standard of care group and one in the standard of care plus metformin group. Interpretation: We did not find significant evidence of an overall survival benefit of adding metformin to standard of care in the overall population of patients with metastatic hormone-sensitive prostate cancer. The side-effect profile of metformin was as expected and consisted mainly of diarrhoea. Adverse metabolic side-effects of ADT were significantly reduced in the metformin group compared with the standard of care group. Funding: Cancer Research UK, Prostate Cancer UK, and UK Research and Innovation Medical Research Council.

Most women with ovarian cancer are diagnosed after the disease has spread widely (advanced stage – III and IV) and more than half die within 5 years. We wanted to find out if testing women without symptoms could pick up ovarian cancer at an earlier stage before it has spread beyond the ovaries and tubes and reduce deaths. We also wanted to assess the risks and benefits of such screening. We invited over 1.2 million women living near 13 centres in England, Wales and Northern Ireland. Of them, 202,638 joined the trial. All women were between 50 and 74 and were no longer having periods. They had never been diagnosed with ovarian cancer or were not having treatment for any other cancer. They did not have many relatives with ovarian or breast cancer. The volunteers were placed into one of three groups at random: The blood test group contained 50,640 women who had yearly CA125 blood tests. If these showed a moderate or high chance of ovarian cancer, they had repeat CA125 tests and a scan. The scan group contained 50,639 women who had yearly internal scans of their ovaries and tubes which were repeated if they showed an abnormality. The no-screening group contained 101,359 women. Those in the blood and scan groups had screening every year until December 2011. We sent all women health questionnaires and also, with their permission, received information about them from the national cancer and death registries till mid-2020. Women in the screened groups had an average of eight years of screening. We followed them for approximately 16 years after they had joined the trial. During this period, 2055 women were diagnosed with ovarian and tubal cancer. It was about 1 in 100 women (1%) in all three groups: 522 of 50,625 in the blood group 517 of 50,623 in the scan group 1016 of 101,314 in the no-screening group More women were diagnosed with early-stage cancer and fewer were diagnosed with advanced cancer in the blood group compared to the no-screening group. There was no difference in the number diagnosed with early or advanced disease between the scan and no-screening group. Despite this difference, the number of women in each group who died from ovarian and tubal cancer was similar in all three groups: 296 of 50,625 (0.6%) in the blood group, 291 of 50,623 (0.6%) in the scan group and 619 of 101,314 (0.6%) in the no-screening group. Other results showed: Overall, 81% women in the blood group and 78% in the scan group attended all of their annual screening appointments. In the blood group, screening detected 84% of ovarian and tubal cancers diagnosed within one year of the test and correctly classified as normal 99.8% of women who did not have ovarian and tubal cancer. In the scan group, screening detected 72% of ovarian and tubal cancers diagnosed within one year of the last test and correctly classified 99.5% of those who did not have ovarian and tubal cancer. Both screening tests were associated with minor complications. While screening did not increase anxiety, there was slightly increased worry in women who were asked to return for more intense repeat testing. Both screening methods picked up changes that were in fact not ovarian cancer. This meant that women had unnecessary surgery together with the worry and risk of complications that go with it. In the blood group 14 women had unnecessary surgery for every 10,000 women screened annually. This means that for each woman found to have ovarian cancer, an additional 2 women had unnecessary surgery. In the scan group 50 women had unnecessary surgery for every 10,000 women screened annually. This means that for each woman found to have ovarian cancer, an additional 10 women had unnecessary surgery. A biobank with all the donated data and over 0.5 million serum samples, including yearly samples from women in the blood group, was built and continues to be used in many new studies, mainly focused on early detection of cancer. Screening using the CA125 blood test or transvaginal ultrasound scan to test for ovarian cancer did not save lives. Additionally, it was associated with some harm. Therefore, an ovarian cancer screening programme for most women cannot be currently recommended. The trial also showed for the first time that ovarian cancer can be detected earlier through screening. However, for screening to save lives, the test needs to pick up many more women earlier in the course of the disease so that available treatments are effective. The biobank provides an opportunity for scientists to see if newer tests for cancer can detect the disease earlier.

Abstract unavailable.

Flap endonuclease 1 (FEN1)-dependent long-patch repair has been considered a minor sub-pathway of DNA single-strand break repair (SSBR), activated only when short-patch repair is not feasible. However, the significance of long-patch repair in living cells remains unclear. Here, we employed human RPE-1 cells with FEN1 deletion to compare the requirements for short- and long-patch pathways for the rapid repair of various types of DNA single-strand breaks (SSBs). We found that SSBs arising from abortive topoisomerase 1 activity are repaired efficiently without FEN1. In contrast, the rapid repair of SSBs arising during base excision repair following treatment with methyl methanesulphonate (MMS) or following treatment with hydrogen peroxide (H2O2) exhibits an unexpectedly high dependence on FEN1. Indeed, in G1 phase, FEN1 deletion slows the rate of SSBR to a similar or even greater extent than deletion of the short-patch repair proteins XRCC1 or POLβ. As expected, the combined deletion of FEN1 with XRCC1 or POLβ has an additive or synergistic effect, severely attenuating SSBR rates after MMS or H2O2 exposure. These data highlight an unanticipated requirement for FEN1 in the rapid repair of SSBs in human cells, challenging the prevailing view that long-patch repair is a minor sub-pathway of SSBR.

Background: People living with metastatic breast cancer (MBC) may have different support requirements to those with early stage breast cancer (EBC). These differences can be substantial, particularly as care pathways and information are often designed around the latter. There is limited understanding of how these discrepancies impact patients with MBC. Aims: In the LIMBER study (Living with Metastatic Breast Cancer), we explored the distinct and unmet needs of people living with MBC. Methods: In collaboration with people living with MBC and healthcare professionals (HCPs), we developed an online survey comprising fixed and free text responses. Fixed responses and overall study demographics from the main LIMBER study have been published elsewhere. A framework analysis of the free text comments is reported here. Results: The resulting thematic map has seven main themes - friends and family, reactions of others, healthcare professionals, systems & processes, knowledge & information, outlook & goals and wellbeing. Participants reflected that comments made by friends and family were often well-meaning but showed misunderstanding of the disease. This was particularly noticeable in understanding the difference between MBC and EBC. There were references to the lack of support and information from HCPs. Conclusions: The analysis of free text comments from this survey demonstrates the impact that MBC can have, particularly without robust support or accessible information. Understanding areas where patients have outstanding needs provides insight into how best to promote coping strategies and improved quality of life, while informing those who provide informal and formal care.

The emergence of CRISPR-Cas9 technology has transformed functional genomics, offering unmatched opportunities to dissect and understand biological pathways and identify novel therapeutic targets in cancer. Breast cancer is a complex, heterogeneous disease and remains a major cause of morbidity and mortality in women, particularly when diagnosed at advanced or metastatic stages where effective treatments are limited. High-throughput CRISPR screening is undoubtedly a powerful tool to discover novel drug targets, uncover synthetic lethal interactions, and identify vulnerabilities in cancer. This review focuses on advances in our understanding of breast cancer developed through CRISPR-based screening technology, particularly in identifying drivers of breast cancer progression, growth, and metastasis, as well as in identifying potential new therapeutic targets and combination therapies. We discuss recent discoveries, current challenges, and limitations of this approach and explore how advancements in CRISPR technology could have a profound impact on the future of breast cancer treatment.

Background: Gynaecological oncology (GO) surgery involves a wide range of procedures, from minor diagnostic interventions to highly complex cytoreductive operations. Accurate perioperative diagnostics—particularly in major surgery—are critical to optimise patient care, predict morbidity, and facilitate shared decision-making. This study aimed to evaluate current practices in perioperative risk assessment amongst UK GO specialists, focusing on the use, perception, and applicability of diagnostic risk prediction tools. Methods: A national multicentre survey was distributed via the British Gynaecological Cancer Society (BGCS) to consultants, trainees, and nurse specialists. The questionnaire examined clinician familiarity with and use of existing tools such as POSSUM, P-POSSUM, and ACS NSQIP, as well as perceived reliability and areas for improvement. Results: Fifty-four clinicians responded, two-thirds of whom were consultant gynaecological oncologists. While 51.9% used morbidity prediction tools selectively, only 7.4% used them routinely for all major surgeries. The most common models were P-POSSUM (39.6%) and ACS NSQIP (25%), though over 20% did not use any formal tool. Despite this, 80% of respondents expressed a desire for more accurate, GO-specific models. Conclusions: This study reveals a gap between available perioperative diagnostics and real-world clinical use in GO surgical planning. There is an urgent need for validated, user-friendly, and GO-specific risk prediction tools—particularly for high-risk, complex surgical cases. Further research should focus on prospective validation of tools such as ACS NSQIP and their integration into routine practice to improve outcomes in gynaecological oncology.

Temozolomide (TMZ) resistance is one of the critical factors contributing to the poor prognosis of glioblastoma (GBM). As a first-line chemotherapeutic agent for GBM, TMZ exerts its cytotoxic effects through DNA alkylation. However, its therapeutic efficacy is significantly compromised by enhanced DNA damage repair (DDR) mechanisms in GBM cells. Although several DDR-targeting drugs have been developed, their clinical outcomes remain suboptimal. Post-translational modifications (PTMs) in GBM cells play a pivotal role in maintaining the genomic stability of DDR mechanisms, including methylguanine-DNA methyltransferase-mediated repair, DNA mismatch repair dysfunction, base excision repair, and double-strand break repair. This review focuses on elucidating the regulatory roles of PTMs in the intrinsic mechanisms underlying TMZ resistance in GBM. Furthermore, we explore the feasibility of enhancing TMZ-induced cytotoxicity by targeting PTM-related enzymatic to disrupt key steps in PTM-mediated DDR pathways. By integrating current preclinical insights and clinical challenges, this work highlights the potential of modulating PTM-driven networks as a novel therapeutic strategy to overcome TMZ resistance and improve treatment outcomes for GBM patients.

Background: The use of technology in medical education has been increasing with more students exposed to some form of online learning or tutorials, under the umbrella of virtual learning (VL). Many programmes, particularly those involving virtual reality, have centred on practical skills, such as surgical techniques or anatomical knowledge, rather than communication. The study presented here examined the feasibility and acceptability of a VL module developed to aid communication when handling angry patients and their relatives. Methods: Participants were 4th and 5th year medical students at the Brighton and Sussex Medical School. Students were randomly allocated to receive training about having angry conversations in a clinical setting via virtual reality headset or desktop application. Prior to the intervention, everyone completed the SE12 self-efficacy questionnaire, a 5-item confidence measure, and free-response study specific survey. Following the module, they completed another study specific survey, with fixed and free responses, the confidence measure, along with the UTAUT2 questionnaire on acceptance and use of technology. Quantitative data was analysed descriptively, conceptual content analysis was applied to free responses. Participants received a £25 voucher for their time. Results: Twenty students took part in the project. Scores on the SE12 did not differ significantly between intervention arms. Confidence improved across all five categories - recognising responses that diffuse or exacerbate anger, identifying anger signals, remaining calm in hostile situations, moving forward with empathy, and applying techniques to different situations. Responses to the UTAUT2 indicated acceptance of VL, including the psychological safety it provides. Nineteen categories for free text responses were developed via content analysis. Participants spoke frequently about the challenges of navigating anger. There was initial apprehension VL would not feel realistic, though this was largely reversed post-intervention. Students expressed preference for a combination of VL, whichever modality, and face-to-face teaching, recognising benefits of both. Conclusion: Students found the training to be acceptable, providing them with tangible skills. There should be a consideration as to how to incorporate VL, with a mix of face-to-face practice for added realism. Trial Registration: Clinical trial number not applicable.

The time of day of administration (chronotherapy) of certain medications can affect both their toxicity and efficacy. In this pragmatic, multicenter trial, women starting adjuvant endocrine therapy (ET) for hormone receptor-positive early-stage breast cancer were randomized (1:1) to either morning or evening administration. The primary endpoint was endocrine toxicity/tolerability measured by the change in total Functional Assessment of Cancer Therapy-Endocrine Subscale (FACT-ES) score from baseline to 12-weeks. Secondary endpoints included: endocrine toxicity/tolerability and quality of life (FACT-ES and FACT-B) from baseline to 4, 8, 12, and 52 weeks, non-persistence or non-adherence, and patient preference for timing of ET. Between June 30, 2021, and March 18, 2022, 245 eligible participants were randomized to either morning (122/245, 49.8%) or evening ET (123/245, 50.2%). In the overall population, there was no statistical difference in the change in total FACT-ES score from baseline to 12 weeks (p = 0.086). There were no statistically significant differences for any of the secondary endpoints between the two groups. The study provides evidence for the enthusiasm of patients and investigators to take part in chronotherapy studies. Additional prospective studies should be performed to assess how the timing of ET affects survival outcomes to ensure optimal patient care.

Cognitive impairment is common in people diagnosed with breast cancer, but the molecular mechanisms that underlie maladaptive changes in the brain are unknown. The psychological stress of a cancer diagnosis is certainly a contributing factor. Here, we investigated alterations in the hippocampal proteome in response to both cancer and psychological stress using label-free quantitative mass spectrometry techniques. An orthotopic syngeneic model of triple-negative breast cancer (TNBC) was established by injecting Py230 cells into the mammary fat pads of female C57Bl/6 mice. Half of the mice were subjected to a daily restraint stress paradigm. Mice that experienced both cancer and restraint stress lost weight and displayed larger tumours compared to non-stressed mice. Their urinary corticosterone levels were also elevated, as measured by enzyme-linked immunosorbent assay. Non-stressed tumour-bearing mice displayed higher levels of TNFα in the prefrontal cortex (PFC) compared to stressed mice with cancer. Flow cytometry results suggested that the CD4+/CD8+ T cell ratios were also raised in non-stressed tumour-bearing mice compared to both controls and stressed mice with TNBC. Bioinformatic analysis of hippocampal proteomes indicated that cancer alone causes reduced mitochondrial respiration and ATP synthesis, as well as impaired glutamate recycling and synaptic plasticity. Moreover, daily stress in TNBC mice caused further mitochondrial dysfunction, increased oxidative phosphorylation, and altered lipid metabolism. Importantly, over half of the mammary tumours that initially developed spontaneously regressed after 7–9 weeks in these young immunocompetent mice. Tumour regression inhibited TNFα increases in the PFC. However, the hippocampal proteomes of tumour-bearing mice were largely similar to mice in which tumours regressed, suggesting that spontaneous regression of breast cancer confers lasting physiological dysregulations that impact hippocampal protein expression. This study in mice may help to identify molecular mechanisms responsible for long-term memory impairments in cancer survivors and reveal novel drug targets for cancer-related cognitive impairment.

The advent and refinement of state-of-the-art spatial biology technologies have facilitated analysis that combines the advantages of high-throughput single cell analysis with techniques that preserve tissue architecture. This combination of cellular phenotyping with retained spatial context provides a much greater understanding of cellular interactions within the tumour microenvironment (TME). For glioblastoma, with its significant intra-tumoural heterogeneity, cellular plasticity, and complex TME, appreciating and understanding these spatial patterns may prove key to improving patient outcomes. This review examines the advances in spatial biology techniques, discusses how these methodologies are being applied to study glioblastoma, and explores how spatial information improves understanding of the TME. Ultimately, it is this spatial context that will accelerate the identification of more effective treatments for glioblastoma.

Background and objective: Outcomes after nephrectomy for intermediate- and high-risk renal cell carcinoma (RCC) according to histological subtype are poorly characterised. This study aims to determine the value of RCC histology in predicting survival and to inform on surveillance strategies in relation to patterns of first recurrence. Methods: We pooled data from phase 3 trials: SORCE (n = 1689) and ASSURE (n = 1853). Of 3542 patients, 2881 had clear-cell RCC (ccRCC), 269 had papillary RCC (pRCC), 201 had chromophobe RCC (chRCC), and 191 had sarcomatoid RCC (sRCC). Relapse rates, median time to relapse (TTR), and first relapse sites were reported. Multivariable Cox regression models evaluated overall survival by histology, adjusting for initial relapse location and other important clinical factors. Key findings and limitations: Patients with pRCC and ccRCC had similar overall survival (log-rank p = 0.1). The median TTR for those with pRCC was 1.34 yr (interquartile range [IQR] 0.76, 2.59) compared with 1.78 yr (IQR 0.96, 3.38) for ccRCC patients (p = 0.012). Patients with chRCC had a median TTR of 2.72 yr (IQR 0.91, 4.11), and those with sRCC had a median TTR of 0.74 yr (IQR 0.50, 1.55). For sRCC patients, relapsing in the chest was associated with a lower risk of death than those relapsing in the abdomen (hazard ratio [HR] 0.5, confidence interval [CI]: 0.3, 0.88; p = 0.06). A similar trend was shown for pRCC (HR 0.5, CI: 0.2, 1.3; p = 0.1). Recurrence patterns for World Health Organization 2020 molecularly classified RCCs were not included. Despite pooling phase three datasets, small event numbers led to imprecise estimates, particularly for chRCC. Conclusions and clinical implications: Patients with intermediate and high-risk pRCC relapse earlier than those with ccRCC. Papillary RCC and sRCC first recurring in the abdomen exhibit poor prognosis, warranting consideration of additional abdominal imaging to enhance early relapse detection. ChRCC exhibits favourable prognosis and could avoid image-based surveillance until year 2. Patient summary: This study evaluates pooled data from large phase 3 trials to precisely delineate relapse patterns for patients with intermediate- and high-risk cell renal cell carcinoma (RCC) according to their histology. The site and timing of first relapse provide useful information to support histology-specific RCC surveillance after nephrectomy. Development of genetic and molecular signatures corresponding to relapses at poor prognosis sites for each histology will individualise follow-up and is the next step.

Aims: To investigate physical and psychological sexual dysfunction (SD) in prostate cancer (PCa) patients, according to disease stage and treatment modality. Materials and methods: Participants diagnosed with PCa completed an online survey reporting sexual side effects across 13 domains, the importance of sexual function, and their support needs. Disease stage and treatment data were collected to identify variations in experience. Results were analysed descriptively and with chi-squared significance testing. Results: Six hundred fifty-four participants diagnosed with localised (66.1%), locally advanced (25.1%), and advanced (8.9%) PCa responded to the survey. Their disease management included radical prostatectomy (RP; 49.7%), radiotherapy (RT; 45.9%), and androgen deprivation therapy (ADT; 43.6%). More than 98% reported new-onset post-treatment sexual problems. The most common physical dysfunctions were erectile dysfunction (ED; 91.0%), ejaculatory disturbance (82.9%), and anatomical penile change (70.0%). The most common psychosexual dysfunctions were loss of sexual confidence (76.2%), loss of sex drive (67.1%), and loss of self-esteem (57.1%). Participants diagnosed with advanced disease were significantly more likely to report SD than participants with localised or locally advanced disease in 5 of 13 domains (p < .05). Participants whose treatment included a combination of RP, RT, and ADT were most likely to report SD in 7 of 13 domains. Overall, 78.3% of participants said sexual activity was important to them, with 61.8% placing sexual problems in their top three current concerns. Furthermore, 78.3% wanted to discuss sexual problems with a healthcare professional, with most wishing to focus on ED, loss of sexual confidence, and low libido. Conclusion: SD is a common, wide-ranging, and distressing side effect of treatment, and PCa survivors place a high level of importance on sexual recovery. Those with advanced disease are among the worst affected and report high levels of psychosexual problems. Holistic rehabilitation strategies addressing a broad range of side effects would benefit all, but particularly those treated with permanent ADT.

Upregulation of programmed death ligand-1 (PD-L1) has been observed in patients with MDS, and its expression on myeloblasts is associated with progression to AML. This open-label, phase 1 study evaluated the safety and tolerability of the PD-L1 antibody durvalumab as monotherapy (part 1) and in combination with tremelimumab, with or without azacitidine (part 2), in patients with MDS who progressed following hypomethylating agent treatment. Sixty-seven adults with MDS were enrolled (part 1, 40 with low/intermediate-1 or intermediate-2/high IPSS risk status; part 2, 27 with intermediate-2/high IPSS risk status). Primary safety endpoints included dose-limiting toxicities (DLTs) and treatment-emergent adverse events (TEAEs). Secondary endpoints included evaluation of clinical outcomes, survival, and pharmacokinetics. Dose-limiting toxicities were experienced by no patients in part 1 and 3 patients (11%) in part 2. The most common treatment-emergent adverse events were diarrhea and fatigue (40% each) in part 1 and fatigue (44%) and anemia (37%) in part 2. In parts 1 and 2, 15% of patients experienced marrow complete response as their best overall response, according to IWG criteria. Hematologic improvement was observed in 35% and 30% of patients respectively in part 1 and part 2. The study was terminated early due to limited efficacy.

Background: Optimal therapy following breast-conserving surgery in older adults with low-risk, early-stage breast cancer remains uncertain. The EUROPA trial aims to compare the effects of radiotherapy and endocrine therapy as single-modality treatments on health-related quality of life (HRQOL) and ipsilateral breast tumour recurrence (IBTR) outcomes in this population. Methods: This non-inferiority, phase 3, randomised study was conducted at 18 academic hospitals across Italy (17 centres) and Slovenia (one centre). Eligible patients were women aged 70 years or older with histologically confirmed, stage I, luminal A-like breast cancer, who had undergone breast-conserving surgery and had an Eastern Cooperative Oncology Group performance status of 0 or 1. Patients were randomly assigned (1:1) to receive single-modality endocrine therapy or radiotherapy. Endocrine therapy consisted of daily oral aromatase inhibitors or tamoxifen, for a total planned duration of 5–10 years as per clinical discretion, while radiotherapy was administered as either whole breast or partial breast irradiation, delivered in 5–15 fractions. Randomisation was stratified by health status according to the Geriatric 8 (G8) screening tool and by age, with allocation concealed and no blinding. The co-primary endpoints were the change in HRQOL, assessed by the global health status (GHS) scale of the European Organisation for Research and Treatment of Cancer Quality of Life Questionnaire 30-item core module at 24 months, and 5-year IBTR rates (not reported here). This preplanned interim analysis was performed once at least 152 patients completed the 24-month GHS HRQOL assessment. The safety population comprised patients who received the study intervention at least once after randomisation. The study is registered with ClinicalTrials.gov, NCT04134598, and is ongoing and actively recruiting. Findings: Between March 4, 2021, and June 14, 2024, 731 women were randomly assigned to receive radiotherapy (n=365) or endocrine therapy (n=366). This analysis included 104 patients in the radiotherapy group and 103 in the endocrine therapy group, with a median follow-up of 23·9 months (IQR 22·9–24·2). Patients were predominantly White (204 [99%] of 207) and the median age was 75·0 years (IQR 73·0–80·0) in the radiotherapy group and 74·0 years (72·0–80·0) in the endocrine therapy group. 86 patients in the radiotherapy group and 75 in the endocrine therapy group completed the 24-month HRQOL assessment. The mean baseline GHS score was 71·9 (SD 19·1) in the radiotherapy group and 75·5 (19·3) in the endocrine therapy group. At 24 months, the age-adjusted, G8 score-adjusted mean change from baseline in GHS was –3·40 (95% CI –7·82 to 1·03; p=0·13) in the radiotherapy group and –9·79 (–14·45 to –5·13; p<0·0001) in the endocrine therapy group, with an adjusted mean difference of 6·39 (0·14 to 12·65; p=0·045) favouring radiotherapy. Treatment-related adverse events were less frequent in the radiotherapy group (65 [67%] of 97 patients) compared with the endocrine therapy group (76 [85%] of 89). The most common grade 3–4 adverse events were arthralgia (six [7%] of 89 in the endocrine therapy group vs 0 of 97 in the radiotherapy group), pelvic organ prolapse (three [3%] vs 0), fatigue, hot flashes, myalgia, bone pain, and fractures (two [2%] vs 0 for each). Serious adverse events were reported in 15 (15%) patients in the radiotherapy group and 13 (15%) in the endocrine therapy group. There were no treatment-related deaths in either group. Interpretation: Endocrine therapy was associated with a greater reduction in HRQOL, as measured by GHS, compared with radiotherapy at 24 months. While these interim results suggest radiotherapy might better preserve HRQOL in older women with low-risk early breast cancer, further data on disease control outcomes and final patient accrual are needed to draw definitive conclusions. Funding: Fondazione Radioterapia Oncologica.

Abstract unavailable.

Purpose: The IMPACTOR study (IMPact of AbemaCiclib on patienTs’ rOles and Responsibilities–ISRCTN17281696) was developed to capture experiences of women with MBC being treated with abemaciclib in a real-world setting. The primary aim was to explore changes to quality of life over time and our secondary aim was to understand these changes in detail via qualitative interviews, as presented here. Methods: A singular interview was offered to participants who had expressed an interest at the point of consent. These were all conducted remotely using a semi-structured interview topic guide. Results: Twenty interviews were completed and analysed using a framework approach to thematic analysis. Eight themes were developed—COVID-19, experience of MBC, side effects, side effect management, treatment information and support, relationship impacts, impact on daily life, and finances and employment. Conclusions: It was apparent that participants faced side effects from treatment but undertook steps to manage these as much as possible. Adaptations were often led by a belief about the benefits of remaining on treatment. Adjustments ranged from modifying routines to carrying personal hygiene supplies when out in public in case of diarrhoea. While this was anticipated, other side effects were less well known with variable clinical support and available information. Family support was raised frequently, predominantly in relation to the impact MBC had on roles and relationships. Themes from this work can be thought of via theories about treatment belief and adherence, such as the common-sense and self-regulation models, as participants reflected on both emotional and cognitive coping strategies.

Defects in DNA single-strand break repair are associated with neurodevelopmental and neurodegenerative disorders. One such disorder is that resulting from mutations in XRCC1, a scaffold protein that plays a central role in DNA single-strand base repair. XRCC1 is recruited at sites of single-strand breaks by PARP1, a protein that detects and is activated by such breaks and is negatively regulated by XRCC1 to prevent excessive PARP binding and activity. Loss of XRCC1 leads to the toxic accumulation and activity of PARP1 at single-strand breaks leading to base excision repair defects, a mechanism that may underlie pathological changes in patients carrying deleterious XRCC1 mutations. Here, we demonstrate that xrcc1 knockdown impairs development of the cerebellar plate in zebrafish. In contrast, parp1 knockdown alone does not significantly affect neural development, and instead rescues the cerebellar defects observed in xrcc1 mutant larvae. These findings support the notion that PARP1 inhibition may be a viable therapeutic candidate in neurological disorders.

The alternative lengthening of telomeres (ALT) pathway is a telomerase-independent mechanism for immortalization in cancer cells and is commonly activated in low-grade and high-grade glioma, as well as osteosarcoma. The ALT pathway can be activated under various conditions and has often been shown to include mutational loss of ATRX. However, this is insufficient in isolation and so other cellular event must also be implicated. It has been shown that excessive accumulation of DNA:RNA hybrid structures (R-loops) and/or formation of DNA–protein crosslinks (DPCs) can be other important driving factors. The underlying cellular events leading to R-loop and DPC formation in ALT cancer cells to date remain unclear. Here, we demonstrate that excessive cellular reactive oxygen species (ROS) is an important causative factor in the evolution of ALT-telomere maintenance in ATRX-deficient glioma. We identified three sources of elevated ROS in ALT-positive gliomas: co-mutation of SETD2, downregulation of DRG2, and hypoxic tumour microenvironment. We demonstrate that elevated ROS leads to accumulation of R-loops and, crucially, resolution of R-loops by the enzyme RNase H1 prevents ALT pathway activity in cells exposed to elevated ROS. Further, we found a possible causal link between the formation of R-loops and the accumulation of DPCs, in particular, formation of TOP1 complexes covalently linked to DNA (Top1cc). We also demonstrate that elevation of ROS can trigger over-activity of the ALT pathway in osteosarcoma and glioma cell lines, resulting in excessive DNA damage and cell death. This work presents important mechanistic insights into the endogenous origin of excessive R-loops and DPCs in ALT-positive cancers, as well as highlighting potential novel therapeutic approaches in these difficult-to-treat cancer types.

Purpose: Addition of a CDK4/6 inhibitor to endocrine therapy (ET) prolongs survival in HR + /HER2-metastatic breast cancer (MBC). Gastrointestinal side effects, predominantly diarrhoea and abdominal pain, are common in patients receiving abemaciclib. This can potentially increase symptom burden, reduce quality of life (QoL) and affect treatment adherence. This longitudinal mixed-methods study with a 6-month follow-up explored patients’ outcomes and experiences. Methods: Participants (n = 44) completed validated QoL measures at study-entry and at 1, 3 and 6 months. Weekly diarrhoea diaries with free-text response options assessed bowel movements and self-management strategies. Optional interviews gathered insight in patients’ experiences. Results: Forty-two participants completed study measures at study-entry and 24 at 6 months. 17/42 reported no gastrointestinal side-effects. Above threshold diarrhoea (≥ 3 loose/liquid stools daily) was reported at least once by 25/42, with 3/42 having persistent symptoms. Strategies to control diarrhoea, employed by 28/42, included dietary modifications, non-prescribed medication-use and nonadherence (dose interruption or reduction). Meaningful decline on the QoL diarrhoea subscale was observed in 12/37 at 1 month, 13/28 at 3 months and 8/23 at 6 months. Free-text analysis showed that diarrhoea disrupted everyday life in those affected. Conclusion: A proportion of this small sample of MBC patients treated with abemaciclib and ET-reported diarrhoea which affected symptom burden and QoL. Close symptom monitoring alongside targeted supportive/educational interventions should be introduced to reduce the negative impact on patients’ lives.

Mutation of some genes drives uncontrolled cell proliferation and cancer. The Philadelphia chromosome in chronic myeloid leukaemia (CML) provided the very first such genetic link to cancer1,2. However, little is known about the trajectory to CML, the rate of BCR::ABL1 clonal expansion and how this affects disease. Using whole-genome sequencing of 1,013 haematopoietic colonies from nine patients with CML aged 22 to 81 years, we reconstruct phylogenetic trees of haematopoiesis. Intronic breaks in BCR and ABL1 were not always observed, and out-of-frame exonic breakpoints in BCR, requiring exon skipping to derive BCR::ABL1, were also noted. Apart from ASXL1 and RUNX1 mutations, extra myeloid gene mutations were mostly present in wild-type cells. We inferred explosive growth attributed to BCR::ABL1 commencing 3–14 years (confidence interval 2–16 years) before diagnosis, with annual growth rates exceeding 70,000% per year. Mutation accumulation was higher in BCR::ABL1 cells with shorter telomere lengths, reflecting their excessive cell divisions. Clonal expansion rates inversely correlated with the time to diagnosis. BCR::ABL1 in the general population mirrored CML incidence, and advanced and/or blast phase CML was characterized by subsequent genomic evolution. These data highlight the oncogenic potency of BCR::ABL1 fusion and contrast with the slow and sequential clonal trajectories of most cancers.

Background: Localised squamous cell carcinoma of the anus is treated with radical chemoradiotherapy. Cure rates are high, but treatment can result in substantial acute and long-term morbidity. We aimed to assess whether lower dose chemoradiotherapy maintains high local control rates in patients with early-stage disease, with the secondary aim of reducing toxicity. Methods: ACT4 is a phase 2, prospective, multicentre, open-label, two-arm non-comparative, randomised, controlled trial, investigating reduced-dose intensity-modulated radiotherapy (rd-IMRT: 41·4 Gy in 23 fractions) in patients with early-stage anal cancer; T1–2 (≤4 cm) N0–NxM0. Eligible patients were at least 16 years of age, with an Eastern Cooperative Oncology Group performance status of 0–1. The primary outcome is 3-year loco-regional failure rates. Patients were randomly assigned 1:2 (with stratification by T stage, N stage, gender, HIV status, and randomising site) to standard-dose IMRT (sd-IMRT: 50·4 Gy in 28 fractions) or rd-IMRT with concurrent mitomycin and capecitabine chemotherapy. Here, we report the pre-planned, modified intention-to-treat analysis of secondary endpoints 6 months after treatment end—complete clinical response, compliance, patient-reported outcomes (EORTC QLQ-C30 and ANL27), and safety data. The trial is registered at the ISRCTN registry (ISRCTN88455282) and is ongoing but no longer recruiting. Findings: 163 patients were recruited from 28 UK tertiary centres between April 24, 2017, and Dec 1, 2020. 160 patients were included in the primary analysis (sd-IMRT n=55; dr-IMRT n=105). Data on ethnicity were not collected. The median patient age was 66 years (IQR 58–72 years); 117 (73%) were female and 43 (27%) male; and 129 (94%) of 138 evaluable samples were p16 positive. Complete clinical responses at 6 months were 87% (46 of 53) for sd-IMRT and 92% (89 of 97) for rd-IMRT. Radiotherapy interruptions of 3 days or more occurred in 14 (26%) of 55 patients in sd-IMRT and 16 (15%) of 105 patients in rd-IMRT. Chemotherapy modifications occurred in 27 (49%) of 55 patients in sd-IMRT and 39 (37%) of 105 patients in rd-IMRT. Grade 3 or worse acute toxicity was reported in 25 (46%) of 55 patients in sd-IMRT and 37 (35%) of 105 patients in rd-IMRT. The most common grade 3 or worse adverse events were radiation dermatitis (seven [13%] of 55 in sd-IMRT and ten [10%] of 105 in rd-IMRT), and diarrhoea (four [7%] of 55 in sd-IMRT and nine [9%] of 105 in rd-IMRT). Serious adverse events occurred in eight (15%) of 55 patients in sd-IMRT and ten (10%) of 105 patients in rd-IMRT. Patient-reported outcomes for most issues deteriorated at the end of treatment and resolved to baseline by 6 weeks in both groups. Poorer sexual function for men and women was observed at 6 months following sd-IMRT. Interpretation: Good 6-month complete clinical responses rates were seen in both groups. Early results suggest rd-IMRT is well tolerated with oncological outcomes maintained. 3-year locoregional failure rates are awaited. Funding: Cancer Research UK and Stand Up to Cancer.

Degraders with dual activity against BRD4 and CBP/EP300 were designed. A structure-guided design approach was taken to assess and test potential exit vectors on the dual BRD4 and CBP/EP300 inhibitor, ISOX-DUAL. Candidate degrader panels revealed that VHL-recruiting moieties could mediate dose-responsive ubiquitination of BRD4. A panel of CRBN-recruiting thalidomide-based degraders was unable to induce ubiquitination or degradation of target proteins. High-resolution protein cocrystal structures revealed an unexpected interaction between the thalidomide moiety and Trp81 on the first bromodomain of BRD4. The inability to form a ternary complex provides a potential rationale for the lack of degrader activity with these compounds, some of which have remarkable affinities close to those of (+)-JQ1, as low as 65 nM in a biochemical assay, vs 1.5 μM for their POI ligand, ISOX-DUAL. Such a “degrader collapse” may represent an under-reported mechanism by which some putative degrader molecules are inactive with respect to target protein degradation.

The development of the first European Organisation for Research and Treatment of Cancer (EORTC) Quality of Life Group (QLG) health-related quality of life (HRQoL) questionnaires contributed to the systematic uptake of HRQoL as an endpoint in cancer clinical trials, and to the measurement of HRQoL for individual assessment in routine care. Following a modular approach, these patient-reported outcome (PRO) measures (PROMs) ensure that both generic and disease-specific issues are assessed, enabling comparison of PROs across groups and studies. The application of a comprehensive and continually refined methodology for developing and updating these PROMs has been crucial in supporting their psychometric and cross-cultural validity, and their continued implementation in clinical research. However, the advancement of measurement science, the more widespread implementation of PROMs, and the significant evolution of anti-cancer therapies over the last decades have highlighted the need to adopt more flexible approaches to PRO assessment to ensure that PROMs remain relevant and fit-for-purpose. The QLG has responded to this call by implementing more tailored PRO measurement approaches through the development and release of the computerised adaptive test (CAT) version of the EORTC QLQ-C30 (i.e., the EORTC CAT Core) and the EORTC Item Library. The EORTC Item Library is an interactive online platform that allows for the creation of customised questionnaires (item lists) from the pool of available items derived from established EORTC QLG PROMs. The aim of this article is to describe the current EORTC QLG approach to PRO measurement in oncology, covering important historical developments and best practice recommendations.

Cytoplasmic dynein 1, a motor protein essential for retrograde axonal transport, is increasingly implicated in the pathogenesis of neurodegenerative diseases such as amyotrophic lateral sclerosis (ALS). In this study, we developed a novel mouse model that combines the Legs at odd angles (Loa, F580Y) point mutation in the dynein heavy chain with a cholinergic neuron-specific knockout of the dynein heavy chain. This model, for the first time, allows us to investigate the impact of Loa allele exclusivity in these neurons into adulthood. Our findings reveal that this selective increase in dynein dysfunction exacerbated the phenotypes observed in heterozygous Loa mice including pre-wean survival, reduced body weight and grip strength. Additionally, it induced ALS-like pathology in neuromuscular junctions (NMJs) not seen in heterozygous Loa mice. Notably, we also found a previously unobserved significant increase in neurons displaying TDP-43 puncta in both Loa mutants, suggesting early TDP-43 mislocalisation – a hallmark of ALS. The novel model also exhibited a concurrent rise in p62 puncta that did not co-localise with TDP-43, indicating broader impairments in autophagic clearance mechanisms. Overall, this new model underscores the fact that dynein impairment alone can induce ALS-like pathology and provides a valuable platform to further explore the role of dynein in ALS.

Squamous cell carcinoma of the anus (SCCA) is a human papillomavirus (HPV)-associated cancer with an increasing incidence. Localized disease is treated curatively with chemoradiotherapy (CRT) but has substantial toxicities and impact on quality of life. Despite increasing research over the past decade, much of our biological understanding of SCCA is extrapolated from other HPV-associated cancers. Recently launched initiatives aim to connect clinicians and basic researchers working in SCCA to foster collaboration and knowledge sharing and facilitate access to SCCA materials. To improve outcomes for SCCA patients, a better understanding of SCCA-specific biology is required. This review will discuss areas of SCCA biology that are important to developing targets for drug development and biomarker-based individualized precision treatment. It will discuss the current understanding of radiosensitivity, differences in HPV-positive versus HPV-negative disease, hypoxia, immune environment, molecular characteristics, circulating tumor HPV-DNA, and the microbiome. It will detail when this understanding has been extrapolated from other HPV-associated cancers. Finally, it will discuss our current and upcoming preclinical biological models and discuss future models needed to further our understanding of SCCA biology.

Objective Explaining gene expression profiling (GEP) test results to patients can be challenging. We examined the utility of two 8 min films about Oncotype DX and Prosigna to aid the knowledge and decision-making of women with early-stage oestrogen receptor positive (ER+) breast cancer. Methods and analysis Patients awaiting GEP test results completed an anxiety questionnaire and the intolerance of uncertainty scale (IUS) before randomisation and divided into Group A (standard verbal and/or written hospital information) or Group B (standard information plus GEP film). Prior to results, they were interviewed about their GEP test knowledge and how the recurrence risk helps determine treatment options. After the results consultation, participants answered two further questionnaires. Participating clinicians completed IUS scales and reported their satisfaction with the results discussions. Results 230/251 patients completed the study (Group A (n=106) and Group B (n=124)). The total knowledge score was higher in Group B (estimated between groups mean difference of 2.5 (95% CI:1.7 to 3.4) p<0.001). Most treatment decisions adhered to recommended risk of recurrence thresholds, although patients with higher trait anxiety were more likely to make less apparently rational decisions OR=0.93 (95%CI 0.88 to 0.97) p=0.002 (163/230; 70.8% received ET alone; 65/230; 28% ET plus chemotherapy, and two sought second opinions). Clinicians reported slightly longer consultations for Group A participants who tended to ask more difficult and unexpected questions. Conclusion Patients who received standard verbal and written information plus film had increased knowledge about GEP tests compared with standard information alone.

Combining mathematical modeling with experiments enables quantitative understanding of cell signaling, transcriptional regulation, and cell fate decisions. Here, we provide a systems biology approach to link signal transduction with B cells fate decisions, to enable quantitative prediction of B-cell proliferation, and differentiation. We describe methodology to run simulations that reveal how signal transduction regulates gene expression and predicts cell fate decision. We describe how to quantitively validate modeling predictions with wet-lab experiments.

The aim of this study was to examine the role of cardiac interoception on self-regulated (Experiment 1) and externally prescribed (Experiment 2) exercises. Cardiac interoception was assessed using heartbeat tracking and discrimination tasks in both experiments. Based on heartbeat discrimination performance, participants were partitioned into groups demonstrating GOOD and POOR cardiac interoceptive accuracy. In Experiment 1, 20 participants completed two self-regulated 20-min cycling tasks at two intensities (light rated physical exertion [RPE on Borg Scale = 10] vs. hard-to-very hard, RPE = 16). During self-regulated exercise, the POOR cardiac interoception group showed lower differences in their exercise work rates and physiological responses between light and hard-to-very hard intensity exercises. These differences were partly attributable to a higher work rate over the first 5 min of light intensity exercise and a higher initial rate of work in the first min of hard-to-very hard intensity exercise. In Experiment 2, 15 participants completed an externally prescribed, constant-load cycling task performed at 80% of the peak power output, to task failure. During externally prescribed exercise, GOOD and POOR groups did not differ in their time-to-task failure nor in their physiological and perceptual responses to the exercise. Together these findings demonstrate that individual differences in interoceptive accuracy influence the regulation of self-paced exercise but do not affect externally prescribed exercise tolerance under constant load.

Randomized controlled trials (RCTs) are the gold standard for assessing healthcare interventions. There is a rich culture of assessing cancer therapy through RCTs. The many challenges include defining the right question to ask, recruiting a diverse population of patients, and avoiding research waste. There are a number of innovative RCT designs that can be used to address these challenges. For example, platform trials test multiple treatments within a single trial, basket trials test treatments across multiple cancer types, and umbrella trials test treatments within a single cancer type but stratified by different molecular profiles. National and international cooperation can also help to avoid research waste and reduce trial management costs. By working together, researchers can ensure that trials are well designed and that the results are used to improve patient care.

Using linked primary care, hospital and cancer registry data, we show that presenting symptom and existence comorbid long-term conditions offering alternative explanations for symptoms, both influence the diagnostic delay for ovarian cancer.

Nuclease–helicase DNA2 is a multifunctional genome caretaker that is essential for cell proliferation in a range of organisms, from yeast to human1, 2, 3–4. Bi-allelic DNA2 mutations that reduce DNA2 concentrations cause a spectrum of primordial dwarfism disorders, including Seckel and Rothmund–Thomson-related syndromes5, 6–7. By contrast, cancer cells frequently express high concentrations of DNA2 (refs. 8, 9, 10–11). The mechanism that precludes cell proliferation in the absence of DNA2 and the molecular aetiology of DNA2-linked diseases remain elusive. Here we used yeast and human cells to demonstrate that DNA2 suppresses homologous recombination-restarted replication and checkpoint activation at stalled DNA replication forks. Loss of this control mechanism upon degradation of DNA2 in human cells causes recombination-dependent DNA synthesis and build-up of RPA-bound single-stranded DNA in the G2 phase of the cell cycle. Consequently, DNA2 deprivation triggers the DNA damage checkpoint and invariably leads to ATR–p21-dependent cell-cycle exit before mitosis. These findings explain why DNA2 is essential for cell proliferation and reveal that replication fork processing to restrict recombination is indispensable for avoiding cellular senescence. Stochastic entry into senescence stifles the proliferative potential of cells following the expression of a Seckel syndrome patient-derived DNA2 hypomorph or partial degradation of DNA2, providing a conceptual framework to explain global growth failure in DNA2-linked primordial dwarfism disorders.

The BTRR (BLM/TOP3A/RMI1/RMI2) complex resolves DNA replication and recombination intermediates to maintain genome stability. Alongside PICH, they target mitotic DNA intertwinements, known as ultrafine DNA bridges, facilitating chromosome segregation. Both BLM and PICH undergo transient mitotic hyper-phosphorylation, but the biological significance of this remains elusive. Here, we uncover that during early mitosis, CDK1 and PLK1 constrain BTRR complex activities at centromeres. CDK1 destabilises the complex, limiting its binding to PICH at specialised chromatin underneath kinetochores. Inactivating the BLM-TOP3A interaction compromises the UFB-binding complex functions and prevents centromere destruction. Different phosphorylation on BLM affects the TRR subcomplex interaction and the mitotic activity, particularly phosphorylation at Ser144 and multiple PLK1-target sites suppresses illegitimate centromeric DNA unwinding. However, unleashing such activity after sister-chromatid cohesion inactivation facilitates the separation of entangled chromosomes. Here, we show a centromere protection pathway in human mitotic cells, heavily reliant on a tight spatiotemporal control of the BTRR complex.

The Greatwall kinase inhibits PP2A-B55 phosphatase activity during mitosis to stabilise critical Cdk1-driven mitotic phosphorylation. Although Greatwall represents a potential oncogene and prospective therapeutic target, our understanding of the cellular and molecular consequences of chemical Greatwall inactivation remains limited. To address this, we introduce C-604, a highly selective Greatwall inhibitor, and characterise both immediate and long-term cellular responses to the chemical attenuation of Greatwall activity. We demonstrate that Greatwall inhibition causes systemic destabilisation of the mitotic phosphoproteome, premature mitotic exit and pleiotropic cellular pathologies. Importantly, we show that the cellular and molecular abnormalities associated with reduced Greatwall activity are specifically dependent on the B55α isoform, rather than other B55 variants, underscoring PP2A-B55α phosphatases as key mediators of the cytotoxic effects of Greatwall-targeting agents in human cells. Additionally, we establish that sensitivity to Greatwall inhibition varies in different cell line models and that dependency on Greatwall activity reflects the balance between Greatwall and B55α expression levels. Our findings highlight Greatwall dependency as a cell-specific vulnerability and propose the B55α-to-Greatwall expression ratio as a predictive biomarker of cellular responses to Greatwall-targeted therapeutics.

In Diffuse Large B-cell Lymphoma (DLBCL), elevated anti-apoptotic BCL2-family proteins (e.g., MCL1, BCL2, BCLXL) and NF-κB subunits (RelA, RelB, cRel) confer poor prognosis. Heterogeneous expression, regulatory complexity, and redundancy offsetting the inhibition of individual proteins, complicate the assignment of targeted therapy. We combined flow cytometry ‘fingerprinting’, immunofluorescence imaging, and computational modeling to identify therapeutic vulnerabilities in DLBCL. The combined workflow predicted selective responses to BCL2 inhibition (venetoclax) and non-canonical NF-κB inhibition (Amgen16). Within the U2932 cell line we identified distinct resistance mechanisms to BCL2 inhibition in cellular sub-populations recapitulating intratumoral heterogeneity. Co-cultures with CD40L-expressing stromal cells, mimicking the tumor microenvironment (TME), induced resistance to BCL2 and BCLXL targeting BH3-mimetics via cell-type specific upregulation of BCLXL or MCL1. Computational models, validated experimentally, showed that basal NF-κB activation determined whether CD40 activation drove BH3-mimetic resistance through upregulation of RelB and BCLXL, or cRel and MCL1. High basal NF-κB activity could be overcome by inhibiting BTK to resensitize cells to BH3-mimetics in CD40L co-culture. Importantly, non-canonical NF-κB inhibition overcame heterogeneous compensatory BCL2 upregulation, restoring sensitivity to both BCL2- and BCLXL-targeting BH3-mimetics. Combined molecular fingerprinting and computational modelling provides a strategy for the precision use of BH3-mimetics and NF-κB inhibitors in DLBCL.

Purpose: Navigating prognosis and end-of-life (EoL) communications in advanced breast cancer (ABC) is difficult for healthcare professionals, people with the disease, and their caregivers. Greater multidisciplinary guidance is needed to optimise the effectiveness of these discussions. Methods: In November 2023, the ABC Global Alliance held a multidisciplinary workshop at their annual meeting in Lisbon, Portugal, exploring practices in communicating prognosis and discussing EoL with people with ABC. Seventy-one participants from 27 countries participated in the workshop. Reference and insights from secondary research were reviewed and incorporated into the workshop findings. Results: Workshop participants proposed a number of best practices for communicating prognosis and discussing EoL with people living with ABC, which aligned with four key topics: (1) When EoL discussions should happen - although individual preferences might differ, most participants felt this should be when life expectancy was within 6–12 months. (2) Who should be involved – if acceptable to the patient, presence of a care-giver or other family member should be encouraged. (3) What should be discussed- there was broad agreement for honesty with a realistic approximation of prognosis providing best and worst case scenarios all the time helping patients to maintain hope for plausible outcomes. (4) How EoL such conversations should be conducted- these sensitive conversations need to be conducted preferably by a Multidisciplinary Team Member (MDT) member who knows the patient and is trusted by them. Provision of a safe environment and plenty of time for discussion were seen as pre-requisites. Conclusion: The workshop highlighted the complexity of EoL communication in ABC. Participants identified challenges and opportunities, and suggested best practices. While these approaches should be universally adopted, they must always be guided by patient preferences, considering individual and cultural differences to ensure compassionate and effective communication.

Background: Epidemiological studies have linked psychological stress with an increased risk of breast cancer, however few studies have linked stress hormone signalling to cancer initiation mechanistically. This may be particularly pertinent in populations already at risk due to mutations in the cancer predisposition genes BRCA1/2. Methods: Here we employ BRCA1/2 knockdown in breast and prostate epithelial cells to examine the effects of the stress hormone cortisol on DNA damage and repair. We perform a retrospective analysis of plasma cortisol and urinary 8-OHdG in a female BRCA-mutation carriers cohort (n = 62) and validate our findings in a male cohort (n = 70). Results: Cortisol promotes DNA damage in normal mammary epithelial cells, and in a BRCA-deficient setting, delays DNA repair. In female BRCA-mutation carriers higher plasma cortisol levels are associated with an increased risk of cancer. In a male BRCA-mutation cohort risk of prostate cancer was also significantly increased in those with higher cortisol levels. Urinary 8-OHdG, a biomarker of oxidative DNA damage, was also correlated with a risk of breast cancer and prostate cancer. Conclusion: Taken together these findings demonstrate that psychological stress, through the induction of DNA damage by cortisol, may increase the cumulative risk of cancer in BRCA-mutation carriers.

Decades characterising the genetic drivers of lymphoma, has led to a detailed understanding of B cell malignancies but also revealed daunting heterogeneity. While current therapies for diffuse large B-cell lymphoma are effective for some patients, they are largely agnostic to the biology of each individual’s disease, and approximately one third of patients experience relapsed/refractory disease. Consequently, the challenge is to understand how each patient’s mutational burden and tumour microenvironment combine to determine their response to treatment; overcoming this challenge will improve outcomes in lymphoma. This mini review highlights how data-driven modelling, statistical approaches and machine learning are being used to unravel the heterogeneity of lymphoma. We review how mechanistic computational models provide a framework to embed patient data within knowledge of signalling. Focusing on recurrently dysregulated signalling networks in lymphoma (including NF-κB, apoptosis and the cell cycle), we discuss the application of state-of-the-art mechanistic models to lymphoma. We review recent advances in which computational models have demonstrated the power to predict prognosis, identify promising combination therapies and develop digital twins that can recapitulate clinical trial results. With the future of treatment for lymphoma poised to transition from one-size-fits-all towards personalised therapies, computational models are well-placed to identify the right treatments to the right patients, improving outcomes for all lymphoma patients.

There have been considerable therapeutic advances within the last decade in the field of multiple myeloma (MM) which have translated into superior outcomes for the majority of patients living with this condition. Concurrently, there has been increasing focus on frailty in MM, appreciating the importance of balancing overall survival with healthy life years through minimising treatment-related toxicities and maintaining quality of life. This guideline explores how to best identify and assess frailer patients living with MM. We outline the current therapeutic landscape in this patient population in addition to exploring emerging areas of interest in the field of frailty.

Background: The in-depth understanding of the impact of a hereditary cancer predisposition syndrome (HCPS) on the health-related quality of life (HRQOL) of individuals with a hereditary cancer burden contributes to the improvement of counselling strategies as well as care planning and informs the development of patient-reported outcome measures (PROMs) for standardised HRQOL assessment. This is the first review to systematically identify and synthesise the evidence from qualitative literature on HRQOL issues relevant for adult individuals living with (the risk of) HCPS between 1991 and 2024. Methods: Eligible studies were qualitative studies of adult individuals' experiences, including direct quotes and studies on the development or validation of health outcome measures. The literature was searched from 1991 to 2024 using the databases PubMed, CINAHL, Embase, and PsycINFO. Results: We screened 13,410 references for study inclusion by title and abstract, resulting in the retrieval of 606 full papers. More than 6800 qualitative patient quotes were extracted and coded by four raters. Reviewed literature provided a comprehensive picture of the experience of individuals living with (the risk of) HCPS in nine identified HRQOL domains (decision-making, impact on family and social relationships, emotional response to test result, living with HCPS, perspective on life and self, HCPS-related symptoms, taking measures to prevent the development or progression of cancer, issues related to the health care system, practical issues of life). Conclusion: Results contribute to insight on how individuals at risk cope with genetic testing and will inform the development of a PROM on their HRQOL that will be applicable for individualised patient management and service evaluation.

Whilst T-cell directed therapies have revolutionized the therapeutic landscape in triple-class refractory multiple myeloma, ongoing long-term safety concerns remain, including the risk of second primary malignancy (SPM) development. We systematically evaluated the incidence and distribution of SPMs in R/R MM patients post T-cell-directed therapy. MEDLINE, EMBASE, and Cochrane CENTRAL databases were searched for clinical trial and real-world studies reporting outcomes for patients infused with either chimeric antigen receptor (CAR) T-cell or bispecific antibody therapies reported until March 2025. Patient-specific characteristics and SPM outcomes were extracted from eligible studies with calculation of point estimate confidence intervals (CIs) achieved via the Clopper–Pearson Exact Method. A total of 12 studies (7 RCTs and 5 RWS) were eligible for analysis, encompassing a total of 2743 adult R/R MM patients. Eleven studies were related to CAR T-cell therapy, with only 1 study reporting on bispecific antibody therapy. The pooled SPM point estimate for CAR T-cell therapy was 6.3%, with hematological malignancies representing the most common subtype. This highlights the potential risk of SPMs in patients eligible for T-cell directed therapy. Further robust, prospective clinical trial and pharmacovigilance data will continue to inform the true level of risk in this cohort of patients.

Background: The rising cancer burden in low- and middle-income countries (LMICs) has been accompanied by an increase in clinical trials. However, there is a paucity of research from LMICs on patient preferences for trial participation. Methods: We undertook a cross-sectional qualitative study using in-depth interviewing to explore the views of Indian cancer patients (n = 11), caregivers(n = 10) and public (n = 10), regarding clinical trials. Clinical researchers (n = 10) were also interviewed. Data were analysed using the framework of qualitative content analysis. Results: Five themes were identified regarding clinical trials: a) Perception: Only a minority had a prior understanding; when explained, most were willing to be randomised and attend additional monitoring visits. b) Recruitment: Consensus that trial discussions should be with the patient, with caregivers and family included where appropriate, variability in when a patient should be first approached. c) Patient information: Need for both written and audio-visual information material using simple local language. d) Benefits/ adverse effects: Discussion of all pros and cons, including the possibility of dying was preferred. There were divided views regarding disclosure of all versus common risks. Challenges in understanding quantitative risks/benefits were voiced. e) Consent: Honesty and transparency, imbalance of power/trust between trialists and participants and financial vulnerability of patients were voiced by participants. Conclusion: Cancer clinical trials in LMICs can be enriched by patient and public involvement during planning research and conduct of the clinical trial. The financial vulnerability of patients and the power imbalance between them and researchers need to be addressed, especially in international multiregional clinical trials.