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Group 4 tumours (MBGrp4) represent the majority of non-WNT/non-SHH medulloblastomas. Their clinical course is poorly predicted by current risk-factors. MBGrp4 molecular substructures have been identified (e.g. subgroups/cytogenetics/mutations), however their inter-relationships and potential to improve clinical sub-classification and risk-stratification remain undefined. We comprehensively characterised the paediatric MBGrp4 molecular landscape and determined its utility to improve clinical management. A clinically-annotated discovery cohort (n = 362 MBGrp4) was assembled from UK-CCLG institutions and SIOP-UKCCSG-PNET3, HIT-SIOP-PNET4 and PNET HR + 5 clinical trials. Molecular profiling was undertaken, integrating driver mutations, second-generation non-WNT/non-SHH subgroups (1–8) and whole-chromosome aberrations (WCAs). Survival models were derived for patients ≥ 3 years of age who received contemporary multi-modal therapies (n = 323). We first independently derived and validated a favourable-risk WCA group (WCA-FR) characterised by ≥ 2 features from chromosome 7 gain, 8 loss, and 11 loss. Remaining patients were high-risk (WCA-HR). Subgroups 6 and 7 were enriched for WCA-FR (p Grp4 risk-stratification scheme defines: favourable-risk (non-metastatic disease and (i) subgroup 7 or (ii) WCA-FR (21% of patients, 5-year PFS 97%)), very-high-risk (metastatic disease with WCA-HR (36%, 5-year PFS 49%)) and high-risk (remaining patients; 43%, 5-year PFS 67%). These findings validated in an independent MBGrp4 cohort (n = 668). Importantly, our findings demonstrate that previously established disease-wide risk-features (i.e. LCA histology and MYC(N) amplification) have little prognostic relevance in MBGrp4 disease. Novel validated survival models, integrating clinical features, methylation and WCA groups, improve outcome prediction and re-define risk-status for ~ 80% of MBGrp4. Our MBGrp4 favourable-risk group has MBWNT-like excellent outcomes, thereby doubling the proportion of medulloblastoma patients who could benefit from therapy de-escalation approaches, aimed at reducing treatment induced late-effects while sustaining survival outcomes. Novel approaches are urgently required for the very-high-risk patients.

The fusion gene MLL-AF4 defines a high-risk subtype of pro-B acute lymphoblastic leukaemia. However, relapse can be associated with a switch from acute lymphoblastic to acute myeloid leukaemia. Here we show that these myeloid relapses share oncogene fusion breakpoints with their matched lymphoid presentations and can originate in either early, multipotent progenitors or committed B-cell precursors. Lineage switching is linked to substantial changes in chromatin accessibility and rewiring of transcriptional programmes indicating that the execution and maintenance of lymphoid lineage differentiation is impaired. We show that this subversion is recurrently associated with the dysregulation of repressive chromatin modifiers, notably the nucleosome remodelling and deacetylation complex, NuRD. In addition to mutations, we show differential expression or alternative splicing of NuRD members and other genes is able to reprogram the B lymphoid into a myeloid gene regulatory network. Lineage switching in MLL-AF4 leukaemia is therefore driven and maintained by defunct epigenetic regulation.Statement of SignificanceWe demonstrate diverse cellular origins of lineage switched relapse within MLL-AF4 pro-B acute leukaemia. Irrespective of the developmental origin of relapse, dysregulation of NuRD and/or other epigenetic machinery underpins fundamental lineage reprogramming with profound implications for the increasing use of epitope directed therapies in this high-risk leukaemia.

Deep Gaussian processes (DGPs) provide a rich class of models that can better represent functions with varying regimes or sharp changes, compared to conventional GPs. In this work, we propose a novel inference method for DGPs for computer model emulation. By stochastically imputing the latent layers, our approach transforms a DGP into a linked GP: a novel emulator developed for systems of linked computer models. This transformation permits an efficient DGP training procedure that only involves optimizations of conventional GPs. In addition, predictions from DGP emulators can be made in a fast and analytically tractable manner by naturally utilizing the closed form predictive means and variances of linked GP emulators. We demonstrate the method in a series of synthetic examples and empirical applications, and show that it is a competitive candidate for DGP surrogate inference, combining efficiency that is comparable to doubly stochastic variational inference and uncertainty quantification that is comparable to the fully-Bayesian approach. A $\texttt{Python}$ package $\texttt{dgpsi}$ implementing the method is also produced and available at https://github.com/mingdeyu/DGP., This article has been accepted for publication in Technometrics, published by Taylor & Francis

Supplementary Figure 1 from Role for Amplification and Expression of Glypican-5 in Rhabdomyosarcoma

Overexpression of genes, through genomic amplification and other mechanisms, can critically affect the behavior of tumor cells. Genomic amplification of the 13q31-32 region is reported in many tumors, including rhabdomyosarcomas that are primarily pediatric sarcomas resembling developing skeletal muscle. The minimum overlapping region of amplification at 13q31-32 in rhabdomyosarcomas was defined as containing two genes: Glypican-5 (GPC5) encoding a cell surface proteoglycan and C13orf25 encompassing the miR-17-92 micro-RNA cluster. Genomic copy number and gene expression analyses of rhabdomyosarcomas indicated that GPC5 was the only gene consistently expressed and up-regulated in all cases with amplification. Constitutive overexpression and knockdown of GPC5 expression in rhabdomyosarcoma cell lines increased and decreased cell proliferation, respectively. A correlation between expression levels of nascent pre-rRNA and GPC5 (P = 0.001), but not a C13orf25 transcript containing miR-17-92, in primary samples supports an association of GPC5 with proliferative capacity in vivo. We show that GPC5 increases proliferation through potentiating the action of the growth factors fibroblast growth factor 2 (FGF2), hepatocyte growth factor (HGF), and Wnt1A. GPC5 enhanced the intracellular signaling of FGF2 and HGF and altered the cellular distribution of FGF2. The mesoderm-inducing effect of FGF2 and FGF4 in Xenopus blastocysts was also enhanced. Our data are consistent with a role of GPC5, in the context of sarcomagenesis, in enhancing FGF signaling that leads to mesodermal cell proliferation without induction of myogenic differentiation. Furthermore, the properties of GPC5 make it an attractive target for therapeutic intervention in rhabdomyosarcomas and other tumors that amplify and/or overexpress the gene. [Cancer Res 2007;67(1):57–65]

Shared mobility spaces have become increasingly popular internationally as attempts to increase the uptake of active travel modes (walking, cycling and running) have turned pavements, shopping streets and public spaces into multi-mode mobility spaces. From a sustainability perspective, policy makers in the UK have argued that shared spaces afford greater opportunities for cycling off-road in areas with busy traffic, whilst in public spaces they provide greater accessibility and connectivity to a wider range of users. Yet there has been little conceptual critique and empirical research on the impacts of how individuals and groups negotiate what are new forms of public space in the UK. Accordingly, in this paper we use insights from the new mobilities paradigm and social practice theories to analyse data gathered from qualitative research with different travel mode users in the city of Exeter (South-west England) to demonstrate the complexity of shared spaces, the tensions they produce and the challenges they may pose for promoting sustainable mobility. First, we explore the practices that unfold within shared spaces and demonstrate how researchers need to appreciate the social complexity of negotiating new and conflictual sites of practice. Second, we examine how a fragmented approach to the design of shared spaces may compromise the development of sustainable mobility practices through representing a partial and dysfunctional approach towards sharing space in cities. Third, we demonstrate the problematics of deploying shared spaces as short-term and politically expedient devices for delivering individually-focused behavioural goals instead of radical alternatives that embed sustainable mobility infrastructure into urban fabrics. We conclude by suggesting that to realise the benefits of collectively sharing mobility space in the UK requires long-term changes in urban infrastructure that can embed practices and promote a shift away from the political dominance of the private vehicle as the axis around which urban development pivots.

The smart cities agenda has garnered considerable interest recently as the spread of mobile technologies and notions of ‘big data’ have opened possibilities for promoting greater efficiencies in urban metabolisms. This has been particularly prominent in the realm of environmental sustainability, where smart technologies have been viewed as a way of reducing traffic congestion and delivering energy efficiencies. Key to these aspirations is the way in which technologies are seen to interact with human behaviour and how digital technologies can promote behavioural change through the provision of ‘better’ information. However, smart city programmes adopt a particular intellectual and pragmatic framing of behavioural change that we argue is fundamentally narrow and unambitious, raising concerns about how behavioural science is mobilised, by whom and its potential to promote sustainable urban futures. First, we propose that the focus in smart city narratives on quantitative data and insights from ‘big data’ is methodologically narrow and is representative of a highly individualised, libertarian paternalist perspective that privileges rationalistic and atomised understandings of behaviour. Second, we argue that the logic of smart cities leads city governments towards a focus on superficial change and the language of ‘encouraging’ shifts in individual behaviour that presents a distraction from the urgent need to reconfigure city infrastructures for low carbon forms of living. Third, we explore how such behavioural change approaches are fundamentally didactic and often lapse into assuming that publics are the passive receivers of ‘smarter’ information rather than active citizens who can question, campaign and present alternative visions to those of corporate-government interests. In this way, we argue that the suffusing of the smart cities and behavioural change agendas act as a neo-liberal distraction to the ways in which cities can develop to support the priorities of human and ecological wellbeing.

We reconstructed the natural history and temporal evolution of the most common childhood brain malignancy, medulloblastoma, by single-cell whole-genome sequencing (sc-WGS) of tumours representing its major molecular sub-classes and clinical risk groups. Favourable-risk disease sub-types assessed (MBWNT and infant desmoplastic/nodular MBSHH) typically comprised a single clone with no evidence of further evolution. In contrast, highest risk sub-classes (MYC-amplified MBGroup3 and TP53-mutated MBSHH) were most clonally diverse and displayed gradual evolutionary trajectories. Clinically adopted biomarkers (e.g. chromosome 6/17 aberrations; CTNNB1/TP53 mutations) were typically early-clonal/initiating events, exploitable as targets for early-disease detection; in analyses of spatially distinct tumour regions, a single biopsy was sufficient to assess their status. Importantly, sc-WGS revealed novel events which arise later and/or sub-clonally and more commonly display spatial diversity; their clinical significance and role in disease evolution post-diagnosis now require establishment. These findings reveal diverse modes of tumour initiation and evolution in the major medulloblastoma sub-classes, with pathogenic relevance and clinical potential.

SummaryMedulloblastoma is currently sub-classified into distinct DNA methylation subgroups/subtypes with particular clinico-molecular features. Using RNA-seq in large well annotated cohorts of medulloblastoma we show that transcriptionally Group3 and Group4 medulloblastomas exist not as discrete types but as intermediates on a bipolar continuum between archetypal Group3 and Group4 entities. Continuum position is prognostic, reflects propensity for specific DNA copy-number changes, key switches in isoform/enhancer usage and RNA-editing. Examining scRNA-seq profiles we show intra-tumoral transcriptional heterogeneity along the continuum is limited in a subtype-dependent manner. By integrating with a human scRNA-seq reference atlas we show this continuum is mirrored by an equivalent continuum of transcriptional cell types in early fetal cerebellar development. We identify unique developmental niches for all four major subgroups and link each to a common developmental antecedent. Our findings show a transcriptional continuum arising from oncogenic disruption of highly specific fetal cerebellar cell types, linked to almost every aspect of Group3/Group4 molecular biology and clinico-pathology.

Coupled climate-ice sheet models are crucial to evaluating climate-ice feedbacks' role in future ice sheet evolution. Such models are calibrated to reproduce modern-day ice sheets, but current observations alone are insufficient to constrain the strength of climate-ice feedbacks. The extent of the Northern Hemisphere ice sheets during the last glacial maximum, ~20,000 years ago, is well known and could provide a benchmark for calibrating coupled climate-ice sheet models. We test this with the FAMOUS-ice coupled Climate-Ice Sheet model (Smith et al., 2020), a fast GCM coupled to the Glimmer ice sheet model. We ran Last Glacial Maximum simulations using FAMOUS-ice with interactive North American Ice Sheet, following the PMIP4 protocol (Kageyama et al., 2018). We find that the standard model setup, calibrated to produce a good present-day Greenland (Smith et al., 2020), produced a collapsed North American ice sheet at the Last Glacial Maximum. We ran ensembles of hundreds of simulations to explore the influence of uncertain ice sheet, albedo, atmospheric, and oceanic parameters on the ice sheet extent. The North American continent deglaciated rapidly in most of our simulations, leaving only a handful of useful simulations out of 280. We thus developed a method to efficiently identify regions of the parameter space that can produce a reasonable ice-sheet extent. This involved emulating the equilibrium ice volume and area as a function of the surface mass balance at the start of our simulations. We then ran three waves of short simulations for 20-50 years to identify parameter values and surface mass balance conditions potentially suitable to grow a realistic ice sheet. This enabled us to find ~160 simulations with good ice extent.Through analysis of these simulations, we find that albedo parameters determine the majority of uncertainty when simulating the Last Glacial Maximum North American Ice Sheets. The differences in cloud cover over the ablation zones of the North American and Greenland ice sheet explains why the ice sheets have different sensitivities to surface mass balance parameters. Based on our work, we propose that the Last Glacial Maximum can provide an “out-of-sample” target to avoid over calibrating coupled climate-ice sheet models to the present day.References:Kageyama, M. et al. The PMIP4 contribution to CMIP6 – Part 4: Scientific objectives and experimental design of the PMIP4-CMIP6 Last Glacial Maximum experiments and PMIP4 sensitivity experiments. Geosci. Model Dev. 10, 4035–4055 (2017).Smith, R. S., George, S., and Gregory, J. M.: FAMOUS version xotzt (FAMOUS-ice): a general circulation model (GCM) capable of energy- and water-conserving coupling to an ice sheet model, Geosci. Model Dev., 14, 5769–5787, https://doi.org/10.5194/gmd-14-5769-2021, 2021.

Summary Background Marked variation exists in the use of genomic data in tumour diagnosis, and optimal integration with conventional diagnostic technology remains uncertain despite several studies reporting improved diagnostic accuracy, selection for targeted treatments, and stratification for trials. Our aim was to assess the added value of molecular profiling in routine clinical practice and the impact on conventional and experimental treatments. Methods This population-based study assessed the diagnostic and clinical use of DNA methylation-based profiling in childhood CNS tumours using two large national cohorts in the UK. In the diagnostic cohort—which included routinely diagnosed CNS tumours between Sept 1, 2016, and Sept 1, 2018—we assessed how the methylation profile altered or refined diagnosis in routine clinical practice and estimated how this would affect standard patient management. For the archival cohort of diagnostically difficult cases, we established how many cases could be solved using modern standard pathology, how many could only be solved using the methylation profile, and how many remained unsolvable. Findings Of 484 patients younger than 20 years with CNS tumours, 306 had DNA methylation arrays requested by the neuropathologist and were included in the diagnostic cohort. Molecular profiling added a unique contribution to clinical diagnosis in 107 (35%; 95% CI 30–40) of 306 cases in routine diagnostic practice—providing additional molecular subtyping data in 99 cases, amended the final diagnosis in five cases, and making potentially significant predictions in three cases. We estimated that it could change conventional management in 11 (4%; 95% CI 2–6) of 306 patients. Among 195 historically difficult-to-diagnose tumours in the archival cohort, 99 (51%) could be diagnosed using standard methods, with the addition of methylation profiling solving a further 34 (17%) cases. The remaining 62 (32%) cases were unresolved despite specialist pathology and methylation profiling. Interpretation Together, these data provide estimates of the impact that could be expected from routine implementation of genomic profiling into clinical practice, and indicate limitations where additional techniques will be required. We conclude that DNA methylation arrays are a useful diagnostic adjunct for childhood CNS tumours. Funding The Brain Tumour Charity, Children with Cancer UK, Great Ormond Street Hospital Children's Charity, Olivia Hodson Cancer Fund, Cancer Research UK, and the National Institute of Health Research.

Weakly stationary Gaussian processes (GPs) are the principal tools in the statistical approaches to the design and analysis of computer experiments (or uncertainty quantification). Such processes a...

Background Less than 5% of medulloblastoma (MB) patients survive following failure of contemporary radiation-based therapies. Understanding the molecular drivers of medulloblastoma relapse (rMB) will be essential to improve outcomes. Initial genome-wide investigations have suggested significant genetic divergence of the relapsed disease. Methods We undertook large-scale integrated characterization of the molecular features of rMB—molecular subgroup, novel subtypes, copy number variation (CNV), and driver gene mutation. 119 rMBs were assessed in comparison with their paired diagnostic samples (n = 107), alongside an independent reference cohort sampled at diagnosis (n = 282). rMB events were investigated for association with outcome post-relapse in clinically annotated patients (n = 54). Results Significant genetic evolution occurred over disease-course; 40% of putative rMB drivers emerged at relapse and differed significantly between molecular subgroups. Non-infant MBSHH displayed significantly more chromosomal CNVs at relapse (TP53 mutation-associated). Relapsed MBGroup4 demonstrated the greatest genetic divergence, enriched for targetable (eg, CDK amplifications) and novel (eg, USH2A mutations) events. Importantly, many hallmark features of MB were stable over time; novel subtypes (>90% of tumors) and established genetic drivers (eg, SHH/WNT/P53 mutations; 60% of rMB events) were maintained from diagnosis. Critically, acquired and maintained rMB events converged on targetable pathways which were significantly enriched at relapse (eg, DNA damage signaling) and specific events (eg, 3p loss) predicted survival post-relapse. Conclusions rMB is characterised by the emergence of novel events and pathways, in concert with selective maintenance of established genetic drivers. Together, these define the actionable genetic landscape of rMB and provide a basis for improved clinical management and development of stratified therapeutics, across disease-course.

Aims Application of advanced molecular pathology in rare tumours is hindered by low sample numbers, access to specialised expertise/technologies and tissue/assay QC and rapid reporting requirements. We assessed the feasibility of co‐ordinated real‐time centralised pathology review (CPR), encompassing molecular diagnostics and contemporary genomics (RNA‐seq/DNA methylation‐array). Methods This nationwide trial in medulloblastoma (, Application of advanced molecular pathology in rare tumours is hindered by low sample numbers, access to specialised expertise/technologies, and tissue/assay QC and rapid reporting requirements. We undertook a UK‐wide trial in medulloblastoma (

Background Occasions of self-discharge from health services before being seen by a health profession or against medical advice are often used by health systems as an indicator of quality care. People self-discharge because of factors such as dissatisfaction with care, poor communication, long waiting times, and feeling better in addition to external factors such as family and employment responsibilities. These factors, plus a lack of cultural safety, and interpersonal and institutional racism contribute to the disproportionately higher rates of Indigenous people self-discharging from hospital. This qualitative study aimed to increase understanding about the causative and contextual factors that culminate in people self-discharging and identify opportunities to improve the hospital experience for all. Methods Semi-structured interviews with five Aboriginal and/or Torres Strait Islander (hereafter, respectfully, Indigenous) people and six non-Indigenous people who had self-discharged from a major tertiary hospital in Brisbane, Australia, were audio-recorded, transcribed and thematically analysed. Results Study participants all respected hospitals’ vital role of caring for the sick, but the cumulative impact of unmet needs created a tipping point whereby they concluded that remaining in hospital would compromise their health and wellbeing. Five key categories of unmet needs were identified – the need for information; confidence in the quality of care; respectful treatment; basic comforts; and peace of mind. Although Indigenous and non-Indigenous participants had similar unmet needs, for the former, the deleterious impact of unmet needs was compounded by racist and discriminatory behaviours they experienced while in hospital. Conclusions Respectful, empathetic, person-centred care is likely to result in patients’ needs being met, improve the hospital experience and reduce the risk of people self-discharging. For Indigenous people, the ongoing legacy of white colonisation is embodied in everyday lived experiences of interpersonal and institutional racism. Racist and discriminatory behaviours experienced whilst hospitalised are thus rendered both more visible and more traumatic, and exacerbate the deleterious effect of unmet needs. Decreasing self-discharge events requires a shift of thinking away from perceiving this as the behaviour of a deviant individual, but rather as a quality improvement opportunity to ensure that all patients are cared for in a respectful and person-centred manner.

The rapid emergence of SARS-CoV-2 mutants with new phenotypic properties is a critical challenge to the control of the ongoing pandemic. B.1.1.7 was monitored in the UK through routine testing and S-gene target failures (SGTF), comprising over 90% of cases by March 2021. Now, the reverse is occurring: SGTF cases are being replaced by an S-gene positive variant, which we associate with B.1.617.2. Evidence from the characteristics of S-gene positive cases demonstrates that, following importation, B.1.617.2 is transmitted locally, growing at a rate higher than B.1.1.7 and a doubling time between 5-14 days. S-gene positive cases should be prioritised for sequencing and aggressive control in any countries in which this variant is newly detected.One-Sentence SummaryThe B.1.617.2 variant of SARS-CoV-2 is replacing B.1.1.7 and emerging as the dominant variant in England, evidenced by sustained local transmission.

Purpose Acute rheumatic fever (ARF) and rheumatic heart disease (RHD) persist as public health issues in developing countries and among disadvantaged communities in high-income countries, with rates in Aboriginal and Torres Strait Islander peoples in Australia among the highest recorded globally. A robust evidence base is critical to support policy recommendations for eliminating RHD, but available data are fragmented and incomplete. The End RHD in Australia: Study of Epidemiology (ERASE) Project aims to provide a comprehensive database of ARF and RHD cases in Australia as a basis for improved monitoring and to assess prevention and treatment strategies. The objective of this paper is to describe the process for case ascertainment and profile of the study cohort. Patients and methods The ERASE database has been built using linked administrative data from RHD registers, inpatient hospitalizations, and death registry data from 2001 to 2017 (mid-year). Additional linked datasets are available. The longitudinal nature of the data is harnessed to estimate onset and assess the progression of the disease. To accommodate systematic limitations in diagnostic coding for RHD, hospital-only identified RHD has been determined using a purposefully developed prediction model. Results Of 132,053 patients for whom data were received, 42,064 are considered true cases of ARF or RHD in the study period. The patient population under 60 years in the compiled dataset is more than double the number of patients identified in ARF/RHD registers (12,907 versus 5049). Non-registered patients were more likely to be older, non-Indigenous, and at a later disease stage. Conclusion The ERASE Project has created an unprecedented linked administrative database on ARF and RHD in Australia. These data provide a critical baseline for efforts to end ARF/RHD in Australia. The methodological work conducted to compile this database resulted in significant improvements in the robustness of epidemiological estimates and entails valuable lessons for ARF/RHD research globally.

Background: Prevention of avoidable preterm birth in Aboriginal and Torres Strait Islander (Indigenous) families is a major public health priority in Australia. Evidence about effective, scalable strategies to improve maternal and infant outcomes is urgently needed. In 2013, a multiagency partnership between two Aboriginal Community Controlled Health Organisations and a tertiary maternity hospital co-designed a new service aimed at reducing preterm birth: ‘Birthing in Our Community’. Methods: A prospective interventional cohort study compared outcomes for women with an Indigenous baby receiving care through a new service (n = 461) to women receiving standard care (n = 563), January 2013–December 2017. The primary outcome was preterm birth (

The role of general practitioners in cancer care has expanded in recent years. However, little is known about utilization of primary health care (PHC) services by patients with cancer, particularly among socio-economically disadvantaged groups. We describe utilization of PHC services by patients with cancer, and the nature of the care provided. The study focuses on a disadvantaged group in Australia, namely Indigenous Australians. A retrospective audit of clinical records in ten PHC services in Queensland, Australia. Demographic and clinical data of Indigenous Australians diagnosed with cancer during 2010–2016 were abstracted from patient’s medical records at the PHC services. The rates of cancer-related visits were calculated using person years at risk as a denominator. A total of 138 patients’ records were audited. During 12 months following the cancer diagnosis, patients visited the PHC service on average 5.95 times per year. Frequency of visits were relatively high in remote areas and among socioeconomic disadvantaged patients (IRR = 1.87, 95%CI 1.61–2.17; IRR = 1.79, 95%CI 1.45–2.21, respectively). Over 80% of visits were for seeking attention for symptoms, wound care, and emotional or social support. Patients who did not undergo surgery, had greater comorbidity, received chemotherapy and/or radiotherapy, and male gender had significantly greater rate of visits than their counterparts. The frequency of utilization of PHC services, especially by patients with comorbidities, and the range of reasons for attendance highlights the important role of PHC services in providing cancer care. The reliance on PHC services, particularly by patients in remote and disadvantaged communities, has important implications for appropriate resourcing and support for services in these locations.

Background: Gastrointestinal cancers (GI) are among the most common cancers worldwide. Current treatment options rely on a chemotherapy backbone combined with targeted antibody therapies, but the prognosis generally remains poor. Iksuda has developed a next-generation Antibody Drug Conjugate (ADC) targeting a cancer-specific carbohydrate antigen (CanAg) which is highly overexpressed in GI cancers but has no expression on normal tissues. This CanAg-targeting ADC is comprised of a proprietary PBD prodrug coupled to an anti-CanAg antibody using PermaLink conjugation technology. The PBD prodrug is designed with a tumor-selective trigger to attenuate its potency before entry of the ADC into the target cancer cells. PermaLink conjugation chemistry ensures conjugate stability. Here, the potential of this anti-CanAg ADC was explored in preclinical models as a proof of concept for targeting GI cancers. Methods: The ADC is composed of a PBD prodrug conjugated to an anti-CanAg antibody through PermaLink to generate conjugate with a drug to antibody ratio of approximately 2. This CanAg-targeting ADC was evaluated in vitro in the CanAg-expressing Colo205 cancer cell line. In vivo efficacy studies were conducted in SCID mice with CanAg-expressing human cancer xenografts representing colorectal (Colo 205), gastric (N87) and pancreatic cancers (BxPC3). An exploratory toxicology study was conducted in cynomolgus monkeys. Results: The CanAg-targeting ADC showed highly potent, specific activity in the Colo205 colorectal cancer cell line with IC50 values in the pM range. A single administration of anti-CanAg ADC in CanAg-positive GI xenograft models showed significant dose-dependent antitumor activity and induced complete tumor regressions with no observable toxicity. In the gastric cancer model, the anti-CanAg ADC gave a complete response by day 36 when dosed at 0.3 mg/kg, and 1 and 3 mg/kg doses exhibited long-lasting tumor regression. In the pancreatic cancer model, administration of anti-CanAg ADC at 0.3 mg/kg induced 76% tumor growth inhibition (TGI) on day 21, and at 1 mg/kg, and 3 mg/kg showed complete tumor regression by day 40 with no subsequent tumor recurrence for the 3mg/kg dose. In colorectal cancer, the anti-CanAg ADC exhibited 58% and 98% TGI with 0.3 mg/kg and 1 mg/kg doses respectively, and with the 3 mg/kg dose regressed tumor with no recurrence. The anti-CanAg ADC was well tolerated in cynomolgus monkeys with 1 mg/kg defined as the highest non-severely toxic dose (HNSTD). Conclusions: This anti-CanAg ADC has shown favorable preclinical anti-tumor activity in gastrointestinal cancers. Our findings support the clinical development of the anti-CanAg ADC for the treatment of GI cancers. Citation Format: Justyna H. Mysliwy, Adam Lodge, Daniel Williamson, Jenny Thirlway, Jutta Deckert, Robert J. Lutz. Developing a next generation antibody drug conjugate for treatment of gastrointestinal cancers [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 1751.

Malignant Rhabdoid Tumours (MRT) are aggressive paediatric malignancies seen in the central nervous system (Atypical Teratoid Rhabdoid Tumours (ATRT)), and kidney and other soft tissues (Extra-cranial Rhabdoid Tumours (ECRT)). With current therapies often proving ineffective and a lack of clear prognostic associations with consensus subgroups, we explored the possibility of using prognostic molecular signatures to further identify the biological characteristics of high risk ATRT patients. By employing a cross-validated feature selection method the methylation profiles of 121 MRT patients were analysed with clinical data to obtain meta-CpG signatures associated with prognosis for ATRT, ECRT and MRT. The relationship between these meta-CpG signatures and the consensus subgroups were further explored, along with the correlation of meta-CpGs with gene expression to establish biological significance. By selecting CpGs for their ability to predict survival this method obtained three novel prognostic methylation signatures which predict MRT outcome (ATRT-5, ECRT-14 and MRT-42). These signatures are independent of molecular subgroup and each signature was significantly associated with overall survival (OS) and event free survival (EFS) in their respective cohorts (p