European Commission Initiative on Breast Cancer (ECIBC): Plenary 2016

The European Commission Initiative on Breast Cancer (ECIBC) Plenaries are an opportunity to inform representatives from the 28 EU Member States and 7 other countries participating in the ECIBC, as well as patients and other stakeholders, policymakers, and the scientific and health policy communities, about the aims, activities and achievements of the ECIBC. They also provide a platform for the exchange of ideas, feedback and input into the ECIBC. The 2016 ECIBC Plenary, entitled “When science and policy collaborate for health”, took place on 24-25 November in Varese, Italy. Its main focus was the implementation of both the voluntary European Quality Assurance scheme for Breast Cancer Services (European QA scheme) and the European guidelines for breast cancer screening and diagnosis (European Breast Guidelines). In this context, the first concrete results were presented, with the launch of the first four European Breast Guidelines recommendations on screening. The first day of the Plenary was dedicated to the JRC informing the audience about the various tools that ECIBC is developing. The second day instead, gave the floor to the audience, who informed the JRC of their views in terms of the challenges and opportunities related to implementing the ECIBC in the respective European countries. The event opened with welcome speeches from the European Commission’s Joint Research Centre (JRC), a moving presentation from a breast cancer survivor and reflections on how to ensure science makes its way into policy. The JRC and ECIBC working group members then brought the audience up to date with progress on the European QA scheme, the European Breast Guidelines, as well as the Guidelines Platform, the template for training on digital mammography, as well as about how ECIBC plans to monitor its impact. Participants also received in-depth explanations of the accreditation framework selected for the European QA scheme, as well as two countries’ experiences of using the ISO 15189 standard for accreditation, which is foreseen for the European QA scheme. The second day saw a focus on the individual countries represented at the Plenary. Presentations assessed how the European QA scheme could potentially fit into three different health systems (Scotland, the Netherlands, Romania), while a special breakout session gave national representatives from the 27 countries present (out of the 35 countries participating in the ECIBC) the chance to discuss implementation of the European Breast Guidelines and the European QA scheme themselves. The results, collected through questionnaires, fed into a roundtable debate on what needs to be done at European and national level to ensure ECIBC implementation. The meeting was closed by Member of the European Parliament and President of MEPs Against Cancer (MACs), Alojz Peterle. An evaluation of the event revealed that the third ECIBC Plenary met its aims to inform stakeholders: all responding participants felt that the event succeeded in providing a comprehensive overview of how the ECIBC is progressing, and what the challenges are. Discussions also provided the JRC with valuable information and feedback. The fourth ECIBC Plenary will take place once the results from piloting the European QA scheme are available.JRC.F.1-Health in Societ

PO-332 Genomic landscapes, neoantigen profiles and biological impact of MLH1 inactivation in cancer cells

Introduction Alterations in DNA repair pathways are thought to fuel tumour progression. Mismatch Repair (MMR) deficient cancers show peculiar biological features such as an indolent progression and a resolute therapeutic response to checkpoint inhibitors. The genomic and biological bases of the peculiar clinical features are poorly understood. Further progress in this area is limited by the paucity of models to study the impact of MMR genes inactivation at the genomic and biological levels. To address this issue we developed a bioinformatic workflow to monitor the neoantigen repertoire induced by inactivation of the Mlh1 gene (a key player of the MMR machinery), in murine cell lines. Material and methods We inactivated Mlh1 throughout the CRISPR-Cas9 technology in CT26 (colon cancer), PDAC (pancreatic cancer) and TSA (breast cancer) murine cell lines. We performed whole exome sequencing (WES) at different time points and then we quantified the amount of mutations (SNVs and indels). We generated a pipeline that characterises the neoantigen repertoire, starting from annotated alterations and the HLA of specific murine strain. In parallel, we inoculated MMR-proficient and -deficient cells in immuno-compromised and -competent mice and monitored their growth. Results and discussions In all pre-clinical models analysed we found a massive increment in the number of non-synonymous alterations (up to 100% increase respect to basal population) after Mlh1 inactivation. Notably, analysis of MMR deficient mouse cells at different time points showed a renewal of mutational profile and consequently an accumulation of predicted neoantigens. We further characterised the SNVs and frameshifts acquired by Mlh1-knockout cells. In agreement with data in human tumours, the fraction of predicted neoantigens derived from frameshifts was higher than the SNV-derived neoantigens. When injected in immuno-compromised mice the Mlh1-knockout cells and their wild type counterpart showed comparable growth. On the contrary, MMR-deficient cells but not their control counterpart grew poorly in immuno-competent mice and responded promptly to treatment with checkpoint inhibitors. Conclusion We find that Mlh1 gene inactivation drives dynamic neoantigen profiles, which can be monitored with an ad hoc bioinformatic pipeline. These analyses provide mechanistic support to understand why MMR deficient cells engage the immune system of the host, foster immune surveillance and tumour control

Report on the call for feedback about The Scope of the European guidelines for breast cancer screening and diagnosis: European Commission Initiative on Breast Cancer

In 2015, the European Commission Initiative on Breast Cancer (ECIBC) started the development of the European guidelines for breast cancer screening and diagnosis (henceforth the European Breast Guidelines) under the auspices of the Directorate-General for Health and Food Safety (DG SANTE) and the technical and scientific coordination of the Directorate-General Joint Research Centre (JRC). To support the JRC in this task, a Guidelines Development Group (GDG), consisting of independent experts and individuals, was established. The European Breast Guidelines’ scope (The Scope) represented the first output of the development process of the European Breast Guidelines. Via a public call for feedback, stakeholders and individual citizens were invited to provide their feedback on The Scope. The call for feedback was open from 18 December 2015 to 17 January 2016 and an online questionnaire was made available on the ECIBC web hub via the EU Survey platform. The JRC received a total of 82 valid responses, from 40 individuals from 18 different countries and from 42 organisations from 20 different countries. During a meeting held in Varese (Italy) in March 2016, the GDG discussed the new version of The Scope which was prepared taking into account the results of the call for feedback. The Scope was finalised and approved by the GDG after some minor editing on 6 September 2016 and was later made publicly available together with this report.JRC.F.1-Health in Societ

European Commission Initiative on Breast Cancer: Concept document

The European Commission Initiative on Breast Cancer (ECIBC) is aimed at ensuring and harmonising breast cancer services quality across European countries. It is coordinated by Commission’s Joint Research Centre, under the supervision of the Directorate-General Health and Food Safety. This document describes the background of the initiative, its general goals and objectives, and its foreseen outcomes.JRC.F.1-Health in Societ

Nailfold Capillaroscopy Analysis Can Add a New Perspective to Biomarker Research in Antineutrophil Cytoplasmic Antibody-Associated Vasculitis

Background: Antineutrophil cytoplasmic antibody (ANCA)-associated vasculitis (AAV) includes granulomatosis with polyangiitis (GPA), microscopic polyangiitis (MPA), and eosinophilic granulomatosis with polyangiitis (EGPA), all of which are characterised by inflammation of small-medium-sized vessels. Progressive understanding of these diseases has allowed researchers and clinicians to start discussing nailfold video capillaroscopy (NVC) as a future tool for many applications in daily practice. Today, NVC plays a well-established and validated role in differentiating primary from secondary Raynaud's phenomenon correlated with scleroderma. Nevertheless, there has not been sufficient attention paid to its real potential in the ANCA-associated vasculitis. In fact, the role of NVC in vasculitis has never been defined and studied in a multicentre and multinational study. In this review, we carried out a literature analysis to identify and synthesise the possible role of capillaroscopy for patients with ANCA-associated vasculitis. Methods: Critical research was performed in the electronic archive (PUBMED, UpToDate, Google Scholar, ResearchGate), supplemented with manual research. We searched in these databases for articles published until November 2023. The following search words were searched in the databases in all possible combinations: capillaroscopy, video capillaroscopy, nailfold-video capillaroscopy, ANCA-associated vasculitis, vasculitis, granulomatosis with polyangiitis, EGPA, and microscopic polyangiitis. Results: The search identified 102 unique search results. After the evaluation, eight articles were selected for further study. The literature reported that capillaroscopy investigations documented non-specific abnormalities in 70-80% of AAV patients. Several patients showed neoangiogenesis, capillary loss, microhaemorrhages, and bushy and enlarged capillaries as the most frequent findings. Furthermore, the difference between active phase and non-active phase in AAV patients was clearly discernible. The non-active phase showed similar rates of capillaroscopy alterations compared to the healthy subjects, but the active phase had higher rates in almost all common abnormalities instead. Conclusions: Microvascular nailfold changes, observed in patients affected by vasculitis, may correlate with the outcome of these patients. However, these non-specific abnormalities may help in the diagnosis of vasculitis. As such, new analysis analyses are necessary to confirm our results

Landscapes of cellular phenotypic diversity in breast cancer xenografts and their impact on drug response

Funder: Cancer Research UK (CRUK); doi: https://doi.org/10.13039/501100000289Funder: AstraZeneca; doi: https://doi.org/10.13039/100004325Abstract: The heterogeneity of breast cancer plays a major role in drug response and resistance and has been extensively characterized at the genomic level. Here, a single-cell breast cancer mass cytometry (BCMC) panel is optimized to identify cell phenotypes and their oncogenic signalling states in a biobank of patient-derived tumour xenograft (PDTX) models representing the diversity of human breast cancer. The BCMC panel identifies 13 cellular phenotypes (11 human and 2 murine), associated with both breast cancer subtypes and specific genomic features. Pre-treatment cellular phenotypic composition is a determinant of response to anticancer therapies. Single-cell profiling also reveals drug-induced cellular phenotypic dynamics, unravelling previously unnoticed intra-tumour response diversity. The comprehensive view of the landscapes of cellular phenotypic heterogeneity in PDTXs uncovered by the BCMC panel, which is mirrored in primary human tumours, has profound implications for understanding and predicting therapy response and resistance

Tumour ecology of breast cancer patient-derived xenografts

Tumours are complex ecosystems where several neoplastic and non-transformed cell populations (the tumour microenvironment – TME) interact in heterogeneous and dynamic ways. Descriptive studies in patient material, although extremely informative, often lack the temporal and spatial resolution needed to fully recapitulate this variability. Mouse models represent a natural complement to these correlative analyses, allowing longitudinal and spatially-extensive studies as well as perturbation experiments. Among pre-clinical models, patient-derived tumour xenografts (PDTXs) have been largely excluded from ecological descriptions due to the absence of a human TME and to the immune-compromised status of the mouse host. Evidence however suggests that PDTXs might be good candidates for ecosystem analyses because they contain composite tumour and TME compartments where cells may form functionally relevant interactions. To demonstrate this, we used the extensive collection of breast cancer PDTXs established in our laboratory and performed the first in-depth description of ecosystem complexity in these model systems, confirming their potential as study tools for tumour ecology. After a global survey of all 100 available models, we thoroughly analysed a set of 15 xenografts using single cell RNA- and DNA-sequencing (scRNA/DNA-seq), three-dimensional imaging with serial two-photon tomography (STPT) and two-dimensional transcriptomic and proteomic profiling (the latter using imaging mass cytometry - IMC). In this thesis, we will present part of this work, focusing in particular on the phenotypic and spatial complexity revealed by scRNA-seq, STPT, IMC and *in situ* transcriptomics. These analyses detail for the first time the features of the PDTX TME, which contains multiple cell types that significantly overlap with phenotypes described in other model systems and clinical samples. These cells have varying frequencies in different PDTXs and include multiple innate immune populations with tumour-reactive phenotypes. Expanding previous descriptions, we also show pervasive inter- and intra-model transcriptional heterogeneity in the PDTX tumour compartment. By applying ligand-receptor analysis, we study how different populations interact with each other and generate complex ecological dynamics. Using STPT, IMC and *in situ* transcriptomics we show that the geographical distribution of different neoplastic and TME cells is largely model-specific, indicating a concerted organisation of PDTX spatial ecosystems by the malignant cell-autonomous compartment. In agreement with this, we predict multiple exclusive interactions between cancer and stroma and show that regional variations in tumour phenotypes result in heterogeneous TME features within specific samples. Overall, this thesis unveils the phenotypic and spatial heterogeneity of PDTX ecosystems, showing their variable organisation between and within models. By providing evidence of potential interactions between different cell populations, our study suggests the functional relevance of this variation. In general, these findings support a wider application of PDTXs for the study of tumour ecology and have strong implications for the interpretation of xenograft studies and for the characterisation of tumour-host interactions *in vivo*. The future integration of other data modalities (scDNA-seq) and extension to patient data will likely confirm these observations and offer complementary information on the processes controlling phenotypic and spatial cellular variation in cancer

Cytokine Profiles as Potential Prognostic and Therapeutic Markers in SARS-CoV-2-Induced ARDS

Background. Glucocorticoids (GCs) have been shown to reduce mortality and the need for invasive mechanical ventilation (IMV) in SARS-CoV-2-induced acute respiratory distress syndrome (ARDS). It has been suggested that serum cytokines levels are markers of disease severity in ARDS, although there is only limited evidence of a relationship between the longitudinal cytokine profile and clinical outcomes in patients with SARS-CoV-2-induced ARDS treated with GC. Methods. We conducted a single-center observational study to investigate serial plasma cytokine levels in 17 patients supported with non-invasive ventilation (NIV) in order to compare the response in five patients who progressed to IMV versus 12 patients who continued with NIV alone. All patients received methylprednisolone 80 mg/day continuous infusion until clinical improvement. Results. The study groups were comparable at baseline. All patients survived. Although IL-6 was higher in the NIV group at baseline, several cytokines were significantly higher in the IMV group on day 7 (IL-6, IL-8, IL-9, G-CSF, IP-10, MCP-1, MIP-1α) and 14 (IL-6, IL-8, IL-17, G-CSF, MIP-1α, RANTES). No significant differences were observed between groups on day 28. Conclusions. Patients in the IMV group had higher inflammation levels at intubation than the NIV group, which may indicate a higher resistance to glucocorticoids. Higher GC doses or a longer treatment duration in these patients might have allowed for a better control of inflammation and a better outcome. Further studies are required to define the prognostic value of cytokine patterns, in terms of both GC treatment tailoring and timely initiation of IMV