Zeitschriftenartikel zum Thema „Guillaume (14..-1558)“

Abstract Purpose/Objective: There is a need for innovative methods to provide high-quality care to vulnerable populations in an effort to reduce disparities. This is particularly important for women facing a new breast cancer diagnosis. The purpose of the present research was to investigate what social determinants may play a role in the existing disparities impacting adherence to care recommendations in communities with poor access to primary care and higher rates of morbidity and mortality. This study characterized adherence with breast health follow-up care in a diverse population of patients where inequalities exist that may negatively impact adherence to treatment recommendations. The objective of the study was to characterize factors associated with adherence to treatment among women with newly diagnosed breast cancer. Materials/Methods: Women diagnosed with stage I through IV breast cancer treated at the University of Florida College of Medicine-Jacksonville between 01/01/2014 and 12/31/2019 were included in the sample. Patterns of adherence were categorized using machine learning methods with data derived from the electronic medical record and the UF Health Jacksonville Tumor registry. Age, race, and Area Deprivation Index (ADI) state rank in 2019 as a disparity proxy were used to build a machine learning model and classify compliance to treatment. Included patients had a diagnostic procedure that identified breast cancer. Compliance to treatment was fulfilled if the patient received surgery following diagnostic confirmation. A machine learning model was used to stratify patients by risk of non-adherence to treatment following a diagnostic procedure. The models were evaluated using their area under the curve (AUC). Results: A total of 6,951 women were included, 629 who were adherent and 6322 non-adherent patients with breast cancer. The average age of the participants was 61.4 years, (Standard Deviation = 12.8 years). The majority of patients were Black (48%) or Caucasian (45%), 2% were Asian, and 5% were Other races. Payer type at diagnosis showed 45% had Medicare, 30% had commercial insurance, 17% were covered by Medicaid, 7% were charity, and 1% had other sources of pay. Most women were diagnosed with stage III breast cancer. Of 346 patients who received surgery that data was available, 127 (36.7%) had surgery within 30 days of diagnosis, 102 (29.5%) between 31 and 60 days, and 37 (10.7%) between 61 and 90 days. Fifteen models were compared using the PyCaret Python library. The ADI appeared as the most important factor to predict adherence in the model, followed by race and characterized by an AUC of 0.63. Conclusion: Our clinic treats predominantly more women diagnosed with biologically aggressive and advanced breast cancer especially in young African American population. The role social conditions play that precipitate and perpetuate health care disparities were investigated to determine their impact on adherence to treatment. At our safety net hospital, over one third were able to undergo surgery within 30 days of diagnosis. The ADI appeared as the most important feature to predict adherence, followed by race. This demonstrated the necessity to better understand the relation between socio-economical determinants and care received by patients. A more detailed description of the patients’ circumstances, such as access to transport, proximity of the hospital, and insurance status may further improve the model. There is a need for innovative methods of providing quality health care to vulnerable populations. Machine learning models can be used to stratify patients by risk of non-adherence to diagnostic follow-up and treatment following a diagnosis of breast cancer. Future research needs to move from identification of non-adherence risk factors to implementation of interventions to improve breast cancer outcomes. Citation Format: Guillaume Labilloy, Brian Celso, Bharti Jasra, Leigh Neumayer, Erin Mobley, Carmen Smotherman, Jennifer Brailsford. Risk factors for lack of adherence with diagnostic follow-up care in breast cancer patients [abstract]. In: Proceedings of the 2021 San Antonio Breast Cancer Symposium; 2021 Dec 7-10; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2022;82(4 Suppl):Abstract nr P5-14-14.

Abstract Large chromosomal alterations are common in cancer and often show preferential gain or loss across many cancer types indicating their selective advantage. Triple negative breast cancer (TNBC) exhibits complex mutational spectrum without common oncogenic drivers yet displays consistent loss of large chromosomal regions. Here, we characterize selection pressures that maintain a recurrently deleted region of chromosome 4p in TNBC. We used bulk WGS phylogenetic analysis of TNCB PT/PDX panel to show that chr4p deletion is an early event in tumor evolution. We used scRNAseq gene expression and inferred copy number analysis to show that chr4p loss is associated with a proliferative state. This finding was confirmed by a combination of RNA in situ hybridization and immunofluorescence. We then tested the dosage sensitivity of genes residing within this region by individual and dual overexpression in TNBC PDX-derived cell lines and control normal cell line by assessing their effect on cell proliferation. The overexpression of genes within chr4p elicited a strong cell proliferation defect in cancer but not normal cell line models. We also characterized an unknown gene within chr4p region as a novel member of the STRIPAK complex. Genome-wide pooled ORFeome library screens identified a global pattern of background-specific dosage sensitive regions. Our study shows that large chromosomal deletions are maintained due to evolutionary early genetic network rewiring rendering multiple genes within such regions to be dosage sensitive. Ultimately, this work enhances our understanding of genetic events that modulate TNBC. Citation Format: Elena Kuzmin, Jean Monlong, Mathieu Bourgey, Tom Lesluyes, Toby Barker, Genevieve Morin, Dongmei Zou, Michael Schwartz, Yang Yang, Alain Pacis, Constanza Martinez, Hellen Kuasne, Anne-Marie Fortier, Rui Li, Claudia Kleinman, Sidong Huang, Peter van Loo, Jiannis Ragoussis, Guillaume Bourque, Morag Park. Evolution of large copy number variants in breast cancer through genetic network rewiring [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 1512.

Abstract Purpose: Immune checkpoint inhibitors (ICI) represent the cornerstone for treatment of patients with metastatic clear-cell renal cell carcinoma (ccRCC). Despite a favourable response for a subset of patients, others present primary progressive disease highlighting the need to better understand plasticity of cancer cells and their crosstalk with the microenvironment to better predict therapeutic response and personalize treatment. Experimental design: We performed single-cell RNA sequencing on 56,421 cells from tumour and normal adjacent tissue of patients with ccRCC at different disease stages. Identification of cancer and microenvironment cell populations was validated by deconvolution analyses of public datasets and their localizations within tumours were determined by spatial transcriptomic analyses. We also assessed association of tumour and microenvironment populations with response of ICI treated patients using data from the BIONIKK clinical trial (NCT02960906). Results: We identify 46 cell populations including 5 tumour subpopulations characterized by distinct transcriptional signatures representing an epithelial to mesenchymal transition gradient as well as novel inflamed state. Deconvolution of the tumour and microenvironment signatures revealed strong correlation between mesenchymal-like ccRCC cancer cells and myofibroblastic cancer-associated fibroblasts (myCAFs) that associate with metastasis and tumour aggressiveness. Spatial transcriptomics revealed their co-localization and we identified therapeutically targetable ligand-receptor interactions underlying their crosstalk. Finally, we show that enrichment in myCAFs was associated with primary resistance to ICI therapy in the BIONIKK clinical trial. Conclusions: We describe epithelial-mesenchymal plasticity of ccRCC cancer cells and identify potentially targetable pathways involved in tumour cell-myCAFs crosstalk opening the way for personalized treatment of ccRCC patients. Citation Format: Alexandra Helleux, Guillaume Davidson, Yann A. Vano, Véronique Lindner, Antonin Fattori, Marie Cerciat, Reza T. Elaidi, Virginie Verkarre, Cheng-Ming Sun, Christine Chevreau, Mostefa Bennamoun, Hervé Lang, Thibault Tricard, Wolf H. Fridman, Catherine Sautes-Fridman, Xiaoping Su, Damien Plassard, Céline Keime, Christelle Thibault-Carpentier, Philippe Barthelemy, Stéphane Oudard, Irwin Davidson, Gabriel G. Malouf. Roles of mesenchymal-like tumour cells and myofibroblastic cancer-associated fibroblasts in progression and therapeutic response of clear-cell renal cell carcinoma [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 1342.

Abstract Patients with Lynch Syndrome, an inherited mismatch repair deficiency, have an increased risk for developing microsatellite unstable (MSI-H) cancers. Our team recently identified shared immunogenic frameshift peptides in patient tumors that can be targets of T cell surveillance and preventative MSI-H cancer vaccines. We hypothesize that in Lynch Syndrome some premalignant lesions are capable of evading immune surveillance from cytotoxic T lymphocytes due to immune checkpoint expression and immunosuppression from tumor, myeloid and stromal cell populations. Immune checkpoint blockade (ICB) showed promising results in the treatment of MSI-H tumors and are being evaluated in the adjuvant setting of patients with Lynch syndrome post surgical resection. However, a significant percentage of advanced MSI-H tumors resist ICB suggesting evolving mechanisms of immune resistance. We will use a MSI-H in vivo mouse model and additionally develop 3D spheroids cocultured with immune cells to characterize the underlying immune resistance mechanisms. The MSI-H mouse model will first be applied to identify shared frameshifts in MSI-H tumors by WES to develop vaccination approaches preventing tumor growth. Then, we will quantify MSI-H tumor growth and immune infiltration in the MSI-H mouse model, with or without ICB. We will analyze the presence of immunosuppressive pathways and myeloid subsets in high growth resisting tumors by immunohistochemistry, scRNAseq, spatial transcriptomics and flow cytometry. In addition, we will perform short-term in vitro spheroid-splenic cell co-cultures to better characterize the early immune resistance mechanisms. We will also perform these spheroid-immune cell co-cultures using the immune cells taken from mice after vaccination or ICB to identify the early myeloid immune resistance mechanisms. Overall, the identification of shared immune frameshifts in MSI-H tumors will allow developing peptide vaccination approaches to prevent tumor growth. Further, the identification of myeloid resistance pathways will guide therapeutic strategies against tumor resisting to ICB and/or vaccination. Citation Format: Guillaume Mestrallet. Overcoming immune resistance in DNA mismatch repair deficient tumors [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 6652.

Abstract Metastatic relapse after treatment is the primary cause of cancer morbidity and mortality. While genetic mechanisms of primary tumors and, to a lesser extent, metastatic cancers have been studied in large cohorts, refractory metastatic tumors are not yet sufficiently characterized. Markers of aggressiveness and resistance that molecular profiling can extract from these tumors have yet to be identified and incorporated into clinical care. In this study, we present a pan-cancer cohort of 1,031 metastatic tumors (which we refer to as META-PRISM) that are resistant to at least one systemic therapy or with no approved treatment options. We retrieved the complete clinical history of patients and performed whole-exome (n=571) and transcriptome sequencing (n=947) for this cohort. The prevalence of detected cancer biomarkers was assessed and compared to an external cohort of primary tumors. In the META-PRISM cohort, we observed an increase in (i) whole-genome duplication frequency, (ii) tumor mutational burden, (iii) germline cancer-predisposing variants, and (iv) somatic alterations in cancer genes, including KRAS, EGFR, CCND1, MYC, and TP53, as compared to the tumor type-matched primary tumors. The most extensive increase in genomic variation at metastatic stage was observed in prostate cancer. We also identified enrichment of standard-of-care resistance biomarkers in most cancer types. However, only 7.6% of tumors harbored at least one such biomarker, indicating that the current understanding of resistance mechanisms remains insufficient. Our cohort demonstrated a significantly improved 6-month survival prediction from models incorporating molecular markers over models with only clinical markers for breast cancer patients and to a lesser extent for other studied tumor types. Overall, our data establish a unique resource for investigating treatment resistance mechanisms and performing predictive analyses in cancer. Citation Format: Yoann Pradat, Julien Viot, Konstantin Gunbin, Andrei Iurchenko, Marc Deloger, Luigi Cerbone, Guillaume Grisay, Loic Verlingue, Veronique Scott, Stefan Michiels, Antoine Hollebecque, Gerome Jules-Clement, Antoine Laine, Luc Friboulet, Laura Mezquita, Yohann Loriot, Benjamin Besse, Fabrice Andre, Paul-Henry Cournede, Daniel Gautheret, Sergey Nikolaev. Integrative pan-cancer genomic and transcriptomic analyses of refractory metastatic cancer [abstract]. In: Proceedings of the AACR Special Conference: Cancer Metastasis; 2022 Nov 14-17; Portland, OR. Philadelphia (PA): AACR; Cancer Res 2022;83(2 Suppl_2):Abstract nr PR009.

Abstract Folate receptor alpha (FOLR1) is a clinically validated target, that is overexpressed extracellularly in numerous solid malignancies with a high unmet medical need. Ovarian, non-small cell lung and breast adenocarcinomas are among indications with the highest frequency of FOLR1-positive patients with one out of two patients showing upregulated topoisomerase I expression. MBK-103, an antibody-drug conjugate targeting FOLR1, is based on Mablink’s novel proprietary hydrophilic drug-linker platform. It is comprised of: (i) an Fc-attenuated humanized IgG1 monoclonal antibody, that selectively binds to FOLR1; (ii) a polysarcosine hydrophobicity masking entity allowing for a high drug-to-antibody ratio (DAR) of 8, while improving the pharmacokinetics and tolerability of the drug; (iii) a proprietary dipeptide cleavable unit and iv) exatecan, a potent topoisomerase I inhibitor. MBK-103 showed excellent ex vivo stability in human plasma enabling a prolonged half-life, similar to that of the unconjugated monoclonal antibody. This led to an increased drug exposure, resulting in a potent antitumor efficacy observed in a collection of more than ten in vivo mouse tumor models with a diverse FOLR1 expression, already at doses as low as 1-3 mg/kg. In addition, the distinct mode of action of exatecan along with the unique characteristics of the proprietary drug-linker platform resulted in the in vivo antitumor potency obtained also in colorectal cancer models, in which tubulin-inhibiting agents exhibit only limited therapeutic effects. The compound was also very well tolerated in cynomolgus monkeys with HNSTD at 50 mg/kg and even when administered repeatedly. High conjugation yields along with 100% homogeneity are major drivers of the successful ongoing manufacturing process with planned IND submission in Q4/2023. Citation Format: Warren Viricel, Louise Conilh, Edouard Leroy, Jean-Guillaume Lafay, Frederique Brune, Lenka Kyrych Sadilkova, Charles Dumontet. MBK-103, a potent novel conjugation platform-based antibody-drug conjugate, changing therapeutic options in folate receptor alpha positive cancer patients [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 1544.

Abstract Background: Over 4,300 children, adolescents, and young adults (CAYA) are diagnosed with cancer each year in Canada, 1/3 of whom have refractory/metastatic disease or will relapse. The PRecision Oncology For Young peopLE (PROFYLE) national, collaborative program, was created to provide equitable access to molecular profiling to identify novel targeted treatment options in a clinically relevant timeframe for all CAYA with hard-to-cure cancers in Canada. Design: Building upon 3 pre-existing regional precision oncology programs, PROFYLE now includes >20 institutions and has united an interdisciplinary team of experts, leaders, research teams, end-users and advocates from across Canada. The program has 14 domain specific nodes that are unified by a shared governance structure, and has harmonized biobanking, genomics, bioinformatics and reporting procedures. PROFYLE includes genomic and transcriptomic sequencing of paired germline and cancer fresh/frozen samples, proteomic analysis, and cancer modelling. Inclusion criteria: ≤29y; treatment at a Canadian center; diagnosis of a hard-to-cure cancer. Profiling results are reviewed by multidisciplinary Molecular Tumor Boards. A report including a results/recommendations summary of actionable findings (therapeutic, diagnostic, prognostic, cancer predisposition), potential targeted therapy options including available clinical trials, clarification of diagnosis, and genetic counseling recommendations is provided to the treating oncologist. Results: >1,000 CAYA are included from all of the provinces. Cancer diagnoses: 34% sarcoma, 16% leukemia/lymphoma, 16% CNS tumor, 11% neuroblastoma, 23% other. 17% of participants had a cancer-predisposing pathogenic/likely pathogenic germline variant, 45% had ≥1 potentially actionable somatic alteration, 22.6% had a therapeutically targetable somatic alteration. The most frequent classes of therapeutic alterations were RAS/MAPK (15%), cell cycle (14%), epigenetic (13%), RTK (12%), PI3K/AKT/mTOR (11%), DNA repair (9%), immune checkpoint (8%). Of clinicians who reported the utility of results, 55% indicated the findings were useful for clinical management. Future Directions: Collaborations with other national and international initiatives and data from this interdisciplinary, multi-institutional research program will inform the development of a framework to innovatively link research, clinical and system considerations with Canadian values relevant to multi-omic profiling and drug access for CAYA. In addition, we believe that with a comprehensive molecular view of cancer, PROFYLE will transform our understanding of underlying disease mechanisms, facilitate and improve diagnostic and prognostic indicators, and identify new therapeutic strategies and targets for CAYA patients with cancer. Citation Format: Stephanie A. Grover, Lesleigh Abbott, Jason N. Berman, Guillaume Bourque, Jennifer A. Chan, Avram E. Denburg, Rebecca J. Deyell, Conrad V. Fernandez, Cynthia Hawkins, Jan-Willem Henning, Meredith S. Irwin, Nada Jabado, Steven J. Jones, Philipp F. Lange, Paul Moorehead, Michael F. Moran, Daniel A. Morgenstern, Sapna Oberoi, Antonia Palmer, Shahrad R. Rassekh, Donna L. Senger, Adam Shlien, Daniel Sinnett, Caron Strahlendorf, Patrick J. Sullivan, Michael D. Taylor, Suzanne Vercauteren, Anita Villani, Stephanie Villeneuve, James A. Whitlock, David Malkin, on behalf of the PROFYLE Consortium. A pan-Canadian precision oncology program for children, adolescents and young adults with hard-to-cure cancer: The PRecision Oncology For Young peopLE (PROFYLE) Program. [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 4509.

Abstract Triple-negative breast cancer (TNBC) is an aggressive subtype of breast cancer. It has no approved targeted therapies due to the lack of the expression of key biomarkers, namely Estrogen and Progesterone receptors, and HER2 amplification. As a result, TNBC has the worst prognosis compared to other breast cancer subtypes. We previously showed that chromosome 4p (chr4p) loss is recurrent, correlates with poor prognosis, is an early event in tumor evolution, and confers a proliferative advantage. Here, we propose to leverage chr4p loss as a genetic vulnerability of TNBC by identifying its genetic interactions (GIs) with genes whose inactivation leads to synthetic lethality i.e. loss of viability of chr4p copy loss cells, but not of the chr4p copy neutral cells, or suppression i.e., positive GIs. We first classified breast cancer cell lines with TNBC enriched molecular subtype called basal-like, as chr4p copy loss (9) and chr4p copy neutral (7), using copy number data from CCLE. Associated with chr4p copy loss, the differential expression analysis carried out using CCLE data revealed transcriptomic changes that distinctly cluster the chr4p copy loss and chr4p copy neutral cell lines. We then used the publicly available CRISPR-based genome-wide gene inactivation data from the DepMap project and quantified chr4p loss-associated GIs using multiple scoring methods such as drugZ and MAGeCK. In total, we identified ~200 negative and ~70 positive statistically significant GIs involving genes annotated to biological processes such as translation, metabolism, and others. We have prioritized a subset of them for subsequent experimental validation using CRISPR-Cas9-based gene editing in TNBC cancer cell line models. Alongside, using the publicly available drug sensitivity data obtained from the PRISM project, we identified compounds that impair cell growth in a chr4p loss-specific manner, the strongest of which targeted redox balance. Collectively, the set of GIs, the compound sensitivities, and the computational methods we have generated are unique resources for developing precision oncology therapeutic strategies for TNBC, as well as for other cancers harboring chr4p loss. Citation Format: Rohan Dandage, Michael Schwartz, Lynn Karam, Alain Pacis, Traver Hart, Guillaume Bourque, Morag Park, Elena Kuzmin. Chromosome arm aneuploidies as genetic vulnerabilities of triple-negative breast cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 1413.

Abstract SAR444245 (formerly THOR-707) is a site-specific pegylated recombinant human IL-2 molecule which blocks IL-2R alpha-binding while retains near-native affinity for beta/gamma IL-2 receptor subunits. SAR444245 reduces B16-F10 tumor proliferation in C57BL/6 mice and induces a short transient migration of CD8+ T and NK cells from peripheral blood to lymphoid organs before a strong proliferation (Ptacin et al, Nat Commun 2021). Here we report CD8+ T cells imaging post SAR444245 as monotherapy in naive and tumor bearing mice using nuclear imaging. An anti-murine CD8+ PET probe was injected in mice 24 hours post SAR444245 administration and nuclear imaging was performed at different times over 6 days. In C57BL/6 naive and in poorly immunogenic B16-F10 tumor bearing animals, images reveal a decreased Standardized Uptake Value (SUV) within blood in treated animals at 3mg/kg compared to control ones. In parallel, blood SUV Ratio (SUVR) increases within spleen, lymph nodes and thymus after SAR444245 injection compared to control mice. On tumor site, a stronger SUVR is measured at 144 hours post injection in SAR444245 treated group than in the control group. A dose escalation ranging from 1 to 6 mg/kg of SAR444245 was performed in highly immunogenic CT26 tumor bearing animal. Blood SUV decreases in the treated groups compared to the control group. In lymphoid organs, all SUVR increase at 144 hours post injection in the treated groups compared to the control one. In CT26 tumors, a slight increase of the SUVR is observed for groups treated at 3 and 6 mg/kg. In parallel, a pronounced reduction of tumor growth is observed in the treated group at 6mg/kg. These studies non-invasively confirm the mode of action of SAR444245 with a rapid relocation of CD8+ T cells from peripheral blood to lymphoid organs and tumor site. Citation Format: Sebastion d'Heilly, Fabrice Tirode, Maxime Gaulin, Dudzicki Anne, Guillaume Bluet, Stéphane Guerif, Francis Descamps, Peter Casteels, Xiangming Li, Rui Wang, Robin Meng, Erwan Jouannot. Noninvasive immuno-PET imaging of CD8+T cell behavior in tumor bearing mice models treated with SAR444245. [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 3580.

Abstract METTL3 is the RNA methyltransferase predominantly responsible for the addition of N-6-methyladenosine (m6A), the most abundant modification to mRNA. The prevalence of m6A and the activity and expression of METTL3 have been linked to the appearance and progression of acute myeloid leukemia (AML)1. EPICS has discovered and optimized a small molecule inhibitor of METTL3 (“M3i”). M3i was shown to inhibit the enzymatic activity of METTL3 (IC50 = 2 nM, SPA assay) and inhibit the presence of intracellular m6A in a cell-based assay (IC50 = 8 nM, Calu6). The anti-proliferative effects of METTL3 was demonstrated in a spheroid model (NCIH-560, IC50 = 100 nM) as well as cell viability assays in various AML (Kasumi-1, IC50 = 30 nM; LEXF 41283, IC50 = 46 nM; MV-4-11, IC50 = 248 nM) and solid tumor (Calu6, IC50= 6 nM; Caov3, IC50 = 27 nM) cell lines. In vivo efficacy was evaluated in a disseminated xenograft model using established systemic MV-4-11-Luc-mCh-Puro2 in female NSG mice where M3i (30 mg/kg, i.p., QDx31 days) significantly (p<0.01, relative to vehicle control) inhibited cancer progression as measured by in-life imaging in addition to flow cytometry of hCD45+ cells in bone marrow, blood and spleen; M3i had minimal effects on hematopoiesis in this model with no significant changes in RBCs, WBCs and neutrophils, but an increase in platelets (measured by IDEXX). In an orthotopic, patient-derived xenograft model of AML (LEXF 41283, intratibial implantation, female NSG mice), M3i (30 mg/kg, i.p., QDx91 days) significantly prolonged survival (p<0.01, relative to vehicle control) with a commensurate absence of hCD45+ cells following compound treatment. These results confirm that the pharmacological inhibition of METTL3 is a viable strategy for the treatment of AML and support the additional exploration of the role of METTL3 inhibitors in alternate blood cancers as well as solid tumors. Citation Format: Graeme Fraser, Catherine Sorlet, Nicolas Parmentier, Elodie Brunel, Anne-France Hartiel, Killian Oukoloff, Guillaume Dutheuil. EP102: Pharmacological inhibition of METTL3 arrests tumor progression and prolongs survival in CDX and PDX models of AML [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 6282.

Abstract Metastatic relapse after treatment is the primary cause of cancer morbidity and mortality. While genetic mechanisms of primary tumors and, to a lesser extent, metastatic cancers have been studied in large cohorts, refractory metastatic tumors are not yet sufficiently characterized. Markers of aggressiveness and resistance that molecular profiling can extract from these tumors have yet to be identified and incorporated into clinical care. In this study, we present a pan-cancer cohort of 1,031 metastatic tumors (which we refer to as META-PRISM) that are resistant to at least one systemic therapy or with no approved treatment options. We retrieved the complete clinical history of patients and performed whole-exome (n=571) and transcriptome sequencing (n=947) for this cohort. The prevalence of detected cancer biomarkers was assessed and compared to an external cohort of primary tumors. In the META-PRISM cohort, we observed an increase in (i) whole-genome duplication frequency, (ii) tumor mutational burden, (iii) germline cancer-predisposing variants, and (iv) somatic alterations in cancer genes, including KRAS, EGFR, CCND1, MYC, and TP53, as compared to the tumor type-matched primary tumors. The most extensive increase in genomic variation at metastatic stage was observed in prostate bladder, and pancreatic cancer types. We also identified enrichment of standard-of-care resistance biomarkers in most cancer types. However, only 9.3% of tumors harbored at least one such biomarker, indicating that the current understanding of resistance mechanisms remains insufficient. Our cohort demonstrated a significantly improved 6-month survival prediction from models incorporating molecular markers over models with only clinical markers for breast cancer patients and to a lesser extent for other studied tumor types. Overall, our data establish a unique resource for investigating treatment resistance mechanisms and performing predictive analyses in cancer. Citation Format: Yoann Pradat, Julien Viot, Konstantin Gunbin, Andrey Yurchenko, Luigi Cerbone, Marc Deloger, Guillaume Grisay, Loic Verlingue, Véronique Scott, Ismael Padioleau, Leonardo Panunzi, Stefan Michiels, Antoine Hollebecque, Gerome Jules-Clément, Laura Mezquita, Antoine Lainé, Yohann Loriot, Benjamin Besse, Luc Friboulet, Fabrice André, Paul-Henry Cournède, Daniel Gautheret, Sergey Nikolaev. Integrative pan-cancer genomic and transcriptomic analyses of refractory metastatic cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 6570.

Abstract METTL3 is the RNA methyltransferase predominantly responsible for the addition of N-6-methyladenosine (m6A), the most abundant epigenetic modification to mRNA. The prevalence of m6A and the activity and expression of METTL3 have been linked to the appearance and progression of numerous hematological and solid tumor cancers. EPICS has discovered and optimized small molecule inhibitors of METTL3 (“M3i”). The aim of the current study is to evaluate the efficacy of M3i in various CDX mouse tumor models. M3i compound potency was evaluated in vitro by measuring enzymatic inhibition of METTL3/14 by scintillation proximity assay (IC50 = 2 nM). The anti-proliferative effects of M3i were measured in cell viability assays following a 72h treatment of A549 (NSCLC, IC50 = 190 nM), SKOV3 (ovarian, IC50 = 9 nM) and FaDu (hypopharyngeal squamous cell carcinoma, IC50 = 24 nM) cells. Complementary data in all three cell lines demonstrates a clear concentration-response for a target engagement biomarker (m6A; IC50 = 2-8 nM, across cell lines) as well as M3i-induced changes in biomarkers of oncogenic drivers. M3i (30 mg/kg, IP, QD) was tested in vivo in mouse models using FaDu (subcutaneous implant) in addition to A549 and SKOV3 (both orthotopic implants); all animal models were performed by LabCorp (MI, USA). Daily M3i treatment significantly prolonged survival in all three disease models consistent with efficacy on tumor growth inhibition as measured by imaging (A549, SKOV3) or direct measurement of tumor volume in the case of subcutaneous implant (FaDu). M3i has previously been shown to be effective in mouse CDX and PDX models of AML. The current results extend the application of METTL3 inhibitors to include the treatment of various types of solid tumors. The efficacy of M3i as a single-agent on tumor growth inhibition and survival advances the validation of METTL3 as a cancer target and provides additional evidence for the applicability of mRNA epigenetic mechanisms to the treatment of solid tumors. Citation Format: Graeme Fraser, Catherine Sorlet, Nicolas Parmentier, Elodie Brunel, Christian Coremans, Sarah Rorive, Anne-France Hartiel, Killian Oukoloff, Guillaume Dutheuil. EP102: Pharmacological Inhibition of METTL3 causes tumor growth inhibition and prolongs survival in preclinical models of NSCLC, ovarian and squamous cell carcinoma [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 3233.

Abstract Background: In patients with advanced lung adenocarcinoma cancers, a subset can be cured by radiotherapy, and/or combined regimens, including chemotherapy, immunotherapy, or targeted therapies based on somatic molecular profiling. Nucleosomes are basic elements of chromatin, composed of 147bp DNA wrapped around an octamer of histones. Cell-free DNA (cfDNA) and nucleosomes are released into the bloodstream upon cell death. In addition to genetic somatic alterations, epigenetic modifications are found to play a key role in tumorigenesis of different cancers. The trimethylated histone H3 at lysine 27 (H3K27Me3) is well known as a transcription-repressive mark and proven to be involved in tumorigenesis, cell cycle progression and proliferation dysregulation. However, the concentration of circulating nucleosomes, as a biomarker of the contributive value of circulating tumor DNA (ctDNA) molecular profiling in patient management at diagnosis has not previously been investigated. Patients and Methods: Plasmas were retrospectively collected from patients with NSCLC at initial diagnosis before treatment (CIRCAN’s cohort, N= 198) and from healthy donors (N=100). We carried out targeted next-generation sequencing (NGS) on paired plasmas. Samples were divided in two sub-groups based on the genetic results: ctDNA negative or positive for presence of somatic alterations. The concentration of global circulating nucleosomes as well as specific methylated marks: H3K4Me2-, H3K9Me3-, H3K27Me3- or H3K36Me3-nucleosomes were measured using chemiluminescent Nu.Q® immunoassays (Belgian Volition SRL, Belgium). Results: Among all the different nucleosome forms evaluated, the highest coefficient correlations with cfDNA concentration were observed for H3K27Me3-nucleosomes (r=0.72, p<0.0001). At a cut-off of 14 ng/ml of H3K27Me3-nucleosomes Nu.Q® assay showed a sensitivity of 69.2% (n=137) at 95% specificity for lung cancers vs healthy donors (AUC= 0.92). Interestingly, by combining H3K37Me3 level and ctDNA molecular profile, we found a contributive diagnostic value of molecular profiling results in 77% of the plasmas versus only 40% using NGS alone as per current practice. 32% of plasmas were positive for both H3K27Me3 and ctDNA; 37% are H3K27Me3-positive but ctDNA-negative; 8% of the samples were ctDNA-positive/H3K27Me3-negative and 23% are negative for both H3K27Me3 and ctDNA. Conclusions: High levels of Nu.Q® H3K27Me3 and the absence of detecting somatic alterations strongly support the presence of non-mutated ctDNA in the corresponding plasma. This greatly improves the confidence in the negative molecular results in cfDNA in LC. This may allow the indication of individualized therapeutic regimens with a short-time medical decision and may reduce invasive tissue re-biopsies. Citation Format: Arnaud Gauthier, Julie Candiracci, Emmanuel Grolleau, Gaelle Lescuyer, David Barthelemy, Christine Haon, Florence Geiduer, Margaux Raffin, Nathalie Hardat, Julie Balandier, Rémi Rabeuf, Anne-Sophie Wozny, Claire Rodriguez-Lafrasse, Guillaume Rommelaere, Fabien Subtil, Sébastien Couraud, Marielle Herzog, Lea Payen-Gay. Circulating H3K27 methylated nucleosome concentration in lung cancer improves the contributive value of ctDNA molecular profiling result at diagnosis [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 1013.

Abstract Background: Elevated levels of CD8+ tumor-infiltrating lymphocytes are associated with response to immunotherapy (IoT)1 in many indications. While immunohistochemistry (IHC) has been the method of choice to evaluate CD8+ tumor infiltration level, the utility of the CD8-PET/CT imaging method offers an attractive non-invasive alternative to biopsies, with the potential to account for both intra-patient tumor heterogeneity and lesion-specific response. Methods: Using the data from ImaginAb’s phase II trial2, we predict response to standard of care (SOC) IoT in patients with advanced solid malignancies. Solely based on CD8-PET/CT scan data, the novel method scores each identifiable lesion by using its volume as measured in the CT data, and mean CD8-PET uptake values before and 4 to 6 weeks after treatment start. All lesion scores are aggregated to compute the patient score, to be subsequently compared to the best standard RECIST evaluation. This methodology was defined and developed using data from 14 patients, and later evaluated using the whole cohort of 40 patients. Results: Despite the multiplicity of indications and SOC IoT in the patient cohort, the computed scores significantly identify and stratify responders (Complete or Partial Response, mean score = -0.78, 10 subjects) from non-responders (Stable or Progressive Disease, mean score = 0.088, 25 subjects): two sample t-test value = 5.378, p-value = 1e-05. RECIST evaluations remain too preliminary to report for 4 subjects. Furthermore, CD8-PET outcomes are available much earlier (median 35 days, median best of RECIST 105 days), which may facilitate important therapy decisions. In addition, the discriminative power between the response groups is reduced when restricting the method to either of the infiltration or lesion size measurements. This result highlights the complementarity of the two measurements. Conclusions: CD8-PET offers new non-invasive opportunities in the early discrimination of responders and non-responders to IoT as well as visualization of the heterogeneity of tumor responses to IoT, supporting its use pre-IoT and early on IoT for adult solid tumors. Acknowledgement: All the data used in this study come from clinical study 2. These data were acquired and discussed within the frame of a pre-competitive collaboration between AstraZeneca, Takeda, Pfizer and imaginAb. We want to thank the respective parties for fruitful discussions which helped us refine our analysis methodology. References: 1 Li F, et al. The association between CD8+ tumor-infiltrating lymphocytes and the clinical outcome of cancer immunotherapy: A systematic review and meta-analysis. EClinicalMedicine. 2021 Sep 16;41:101134. doi: 10.1016/j.eclinm.2021.101134. PMID: 34585125; PMCID: PMC8452798 2 ClinicalTrials.gov Id: NCT03802123 https://clinicaltrials.gov/ct2/show/NCT03802123 Citation Format: Guillaume Potdevin, Isabelle Ayx, Laura Dillon, Emilie Mahieu, Hadassah Sade, Günter Schmidt. A first assessment of CD8-PET/CT with 89Zr-Crefmirlimab as predictive biomarker for response to standard of care immunotherapy in patients with solid tumors. [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 3577.

Abstract Introduction: Treatment options of lung cancer (LC) comprise radiotherapy, and/or combined treatment approaches, including chemotherapy, immunotherapy and targeted therapies based on the tumoral molecular profile. Following curative-intent first-line therapies, clinical surveillance involves serial CT imaging. However, such surveillance can detect only macroscopic disease recurrence and is frequently inconclusive. NGS has been utilized to help identify and monitor treatment plans. Nucleosomes, complexes of DNA and histones proteins, are released during cell death into blood circulation. Trimethylation of lysine 27 on histone H3 (H3K27Me3), catalyzed by enhancer of zeste homolog 2 (EZH2), is a crucial epigenetic process in tumorigenesis. We investigated if H3K27Me3-nucleosome concentration could be a biomarker for molecular residual disease (MRD). Patients and Methods: Plasmas were retrospectively collected from patients with advanced LC during treatment (CIRCAN’s cohort, n= 200) and from healthy donors (n=100). We carried out standard targeted NGS on paired plasmas. Samples were divided in two sub-groups based on genetical results: ctDNA negative (n=120) or positive (n=80) for presence of somatic alterations. Concentration of circulating H3K27Me3-nucleosome was measured using chemiluminescent Nu.Q® immunoassay (Belgian Volition SRL, Belgium). Results: Significantly elevated concentrations of H3K27Me3-nucleosomes were found in LC plasmas during the follow-up of patients compared to healthy donors (median 14.9 ng/ml vs 6.15 ng/ml, respectively, p<0.001). In addition, H3K27Me3 levels is lower in the ctDNA-negative group compared to ctDNA-positive group (median 12.1 vs 24.8 ng/ml, respectively, p<0.001). At a clinical cut-off of 14 ng/ml, 62% of samples were positive for either H3K27Me3 or ctDNA, or for both, suggesting an active disease progression compared to only 40% detection using the NGS assay alone. 38% of the patients have low levels of H3K27Me3 and were ctDNA-negative strongly suggesting a therapeutic response under treatment. Conclusions: High levels of Nu.Q® H3K27Me3 could allow physicians to detect MRD in LC patients following treatment with curative intent. This could be achieved by monitoring testing of patients at defined intervals of treatment and recovery, alongside imaging, to incorporate analyses of evolving molecular landscapes during treatment. In this setting, the H3K27Me3-nucleosome quantification, to complete the molecular exploration of cfDNA is highly encouraging, especially in advanced NSCLC, where re-tissue biopsies are impractical, expensive, and may cause undue harm. H3K27Me3-nucleosome quantification may also be useful in patient identification for specific treatments such as EZH2 inhibitor, but this requires investigation. Citation Format: Emmanuel Grolleau, Julie Candiracci, Arnaud Gauthier, Gaelle Lescuyer, David Barthelemy, Christine Haon, Florence Geiguer, Margaux Raffin, Nathalie Hardat, Julie Balandier, Rémi Rabeuf, Anne-Sophie Wozny, Guillaume Rommelaere, Claire Rodriguez-Lafrasse, Fabien Subtil, Sébastien Couraud, Marielle Herzog, Lea Payen-Gay. Circulating H3K27 nucleosomes to monitor lung cancer patients during treatment, a universal biomarker quantifying the molecular residual disease (MRD) in plasma samples [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 2163.

Abstract The goal of this study is to target pre-existing heterogeneity in gene expression to ultimately overcome resistance to targeted therapy in melanoma. Despite significant advances in the treatment of melanoma, many patients still fail to achieve durable responses to therapy due to the development of drug resistance. Our lab recently discovered that in drug-naive melanoma rare cells transiently enter distinct gene expression states that allows them to ultimately survive treatment with targeted therapy (we refer to these cells as “primed for drug resistance”). The goal of this work is to develop a strategy to overcome this form of resistance by driving cells out of states primed for resistance and into drug-susceptible states. To identify the pathways that regulate the transition to the primed state we combined single-cell RNA sequencing with DNA barcoding to extract the transcriptional profiles and lineage information of over 7,000 drug-naive melanoma cells. We found that cells in lineages that transitioned into states primed for drug resistance express high levels of genes associated with TGFβ and PI3K signaling compared to lineages that stayed in a drug-susceptible state. To test whether TGFβ can directly cause cells to enter the primed state, we treated cells with recombinant TGFβ which led to a significant increase in the number of primed cells. Intriguingly, we also found that treating with TGFB receptor inhibitor or PI3K inhibitor reduced the number of cells in the primed cell state. Further, we sequentially treated these cells first with either the TGFB or PI3K inhibitor followed by targeted therapy and ultimately reduced the number of resistant colonies, demonstrating the potential for this therapeutic paradigm. To better understand how targeting the TGFB and PI3K pathways shift the population dynamics, we used single-cell RNA-seq and cellular barcoding to track lineages following treatment with TGFB, TGFB inhibitor, and PI3K inhibitor. Our analysis reveals the population changes following each therapy and directly demonstrates that treating with PI3K inhibitor decreases the number of primed cells, shifting the population into a drug-susceptible expression state. Ultimately, this work defines two key pathways underlying the primed cell state in melanoma and presents a therapeutic paradigm to eliminate this form of therapy resistance. Citation Format: Guillaume Harmange, Ben Emert, Dylan Schaff, Raul Reyes-Hueros, Sydney Shaffer. Overcoming resistance by driving melanoma cells into drug-susceptible states [abstract]. In: Proceedings of the AACR Special Conference on the Evolutionary Dynamics in Carcinogenesis and Response to Therapy; 2022 Mar 14-17. Philadelphia (PA): AACR; Cancer Res 2022;82(10 Suppl):Abstract nr B028.

Abstract Extensive trials of PARP inhibitors (PARPi) have shown that they improve progression-free survival in patients with various cancers, including ovarian cancers. This improvement is marked in patients carrying BRCA mutations, and in Homologous Recombination Deficiency (HRD) subgroups. To date, identifying patients eligible for PARPi has been a challenge for scientific and clinical teams using next-generation sequencing (NGS) analysis and the persistence of genomic scars in tumors after restoration of proficient HR or epigenetic changes is a limitation of NGS. Functional assays could be used to improve the profiling of HR status and faithfully identify HRD tumors. The Repair Capacity Test assesses the formation of RAD51 foci in proliferating cells after irradiation and can be used on fresh primary cancer tissues irradiated ex vivo as well as on Patient-Derived Tumor Organoids (PDTO). However, RAD51 foci scoring is often performed manually without any possibility for the standardization of techniques. By contrast, recent progress in whole slide imaging referring to scan a complete microscope slide and creating a single high-resolution digital file, could represent an opportunity for automatizing the evaluation of HR. The purpose of this translational study was to develop an automated tool for scoring RAD51-mediated homologous recombination, to use this tool on ovarian PDTO and to compare the result to the sensitivity to olaparib (PARPi), determined by direct exposure of PDTO to the drug. We show that immunofluorescence of RAD51 foci can be automatically detected and quantified in Cyclin A2 positive nuclei in all the cells of each PDTO slice thus offering a new opportunity for the routine management and standardization of HR assessment in PDTO that goes beyond the widely used manual estimation. Finally, this automated scoring will be optimized to directly assess HR on tumor slices to make the Repair Capacity Test a highly relevant functional precision oncology tool to identify patients who will benefit from PARPi. Citation Format: Lucie Thorel, Pierre-Marie Morice, Nicolas Elie, Louis-Bastien Weiswald, Romane Florent, Florence Giffard, Margaux Jacobs, Agathe Ricou, Raphaël Leman, Guillaume Babin, Jean-François Lebrun, Sandrine Martin, Mélanie Briand, Benoit Goudergues, Bernard Lambert, Cécile Blanc-Fournier, Dominique Vaur, Benoit Plancoulaine, Laurent Poulain. Automated scoring to assess RAD51-mediated homologous recombination in patient-derived tumor organoids of ovarian cancers [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 6112.

Abstract Background: It’s now increasingly recognized that ovarian cancer (OC) ascites play a significant role in OC progression - behavior of tumor cells is influenced by the nature of their surrounding microenvironment. Thus, characterization of ascites composition is essential to understand how this milieu affects tumor progression and particularly the immunosuppressive pathways that would underlie immune response dysfunctions and, in turn, how biological ascites effects can be influenced by that composition. Methods: Ascites samples were collected from advanced OC patients. Through respective multiplexed approaches of flow cytometry-based marker expression analysis and quantitation of mediators, immune context was profiled by investigating immune cell composition and levels of a plethora of soluble factors, including cytokines/chemokines, and metabolic pathways of some amino acids known to be involved in immunosuppression. Furthermore, ascites fluids were functionally screened for their biological effects on healthy monocytes, either undifferentiated or undergoing M1 polarization. Results: Unlike healthy PBMCs, OC ascites were mostly “enriched” in regulatory/immunosuppressive immune cell subsets including T and myeloid populations. Also, T cells were shown to highly express immune checkpoints such as PD1 in T cells and TIGIT in Tregs. Intriguingly, a high CD4/CD8 ratio was seen. Also, CD163+ tumor-associated macrophages were shown to express CSF1R, CCR8 and CCR2, and to even display a mixed phenotype since also expressing Arg1, CD80, and iNOS. On acellular fractions, most ascites demonstrated elevated CCL18, IL6, LIF, VEGF, and CCL2 levels, and low IL2, IL4, and IL17 levels. Interestingly, unlike healthy plasmas, these ascites appeared to harbor a metabolically-immunosuppressive profile characterized by high glutaminolysis and tryptophan (Trp) degradation in kynurenine (Kyn). Functionally, we demonstrated that not only ascites basically polarized monocytes into M2 macrophages, but even antagonized with their M1 polarization to ultimately tilt to M2 status. Conclusions: Taken together, our data show that ascites fluids most favorable to the M2 phenotype were associated with high LIF, VEGF, IL6, CCL2, and CCL18 levels, and with elevated Kyn to Trp ratios - Kyn levels being strongly higher than in healthy plasmas. Our results thus highlight a peculiar altered environment of OC ascites where a mixture of suppressive cells and signaling factors mediate extracellular cues leading to immune cell activity dysfunction. Altogether, these translational findings highlight OC ascites as a valuable tool to understand the mechanisms of suppression and develop predictive profiles, and to provide new insights for the identification of new targets and development of targeted-therapies. Citation Format: Assia Chaibi, Coriolan Lebreton, Dominique Bodet, Jean-Philippe Guegan, Guillaume Babin, Antoine Italiano, Alban Bessede, Imane Nafia. Profiling of immune cell components and soluble factors in ovarian cancer ascites highlights impaired immune environment. [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 4705.

Abstract The recent exponential progress of sequencing technologies has dramatically impacted cancer research and paved the way to precision medicine in cancer care. In parallel, light-speed progress in bioinformatics has been essential to allow analysts to embrace the vast amount of data yielded by high-throughput profiling machines, turn this data into cancer biology knowledge, and ultimately develop innovative approaches to cancer care. Still, computational complexity and tools' interoperability remain major challenges for the advancement of -omic data-driven cancer research. Despite the historical prevalence of R, Python is gaining momentum in the bioinformatics landscape. As a general purpose language, it offers numerous advantages: - its overall ecosystem facilitates the integration of bioinformatics tools into large-scale frameworks, increasing their versatility and widening their targeted audience; - its straightforward syntax and user-friendly logic makes it a popular language in education, and explains its wide adoption in various sectors; - its wide adoption favors interdisciplinarity, e.g. it can reduce the learning curve for engineers eager to make an impact in cancer research. These advantages motivate the further development of the Python bioinformatics ecosystem. We thus advocate the necessity to port reference tools from R to Python, with the ambition of shaping a comprehensive ecosystem. Sign of this trend, state-of-the-art tools have been directly developed in Python (e.g. lifelines, a library dedicated to survival analysis) or quickly ported in Python from R (e.g. harmony, an R library for integrating single cell data). We introduce InMoose (Integrated Multi-Omics Open Source Environment), an open source Python unified framework for every -omic data type. It is based on recognized tools and focuses on efficiency and user-friendliness. InMoose is accessible at https://github.com/epigenelabs/inmoose and is released under GPL3 license. The first version of InMoose focuses on bringing to the Python world transcriptomics tools mostly based on the edgeR R package. It features batch-effect correction algorithms, as well as differential expression analysis, for microarray and RNA-seq data. InMoose demonstrates the advantages of our approach: - it is developed with the intent to capitalize on and facilitate interactions with other popular Python data-oriented libraries (e.g. pandas, numpy); - it demonstrates significant computational performance improvements; - our package integrates easily into web-based user-friendly analyses platforms (e.g. Epigene Labs’ proprietary mCUBE platform); - porting existing code from one language to another provides an opportunity window to improve both functionality and performance. We expect that our effort will help foster a larger collaborative effort to build and grow a consistent state-of-the-art Python platform for cancer bioinformatics. Citation Format: Maximilien Colange, Guillaume Appé, Akpéli Nordor, Abdelkader Behdenna. Introducing InMoose, an integrated open source Python package for multi-omic analyses [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 2093.

Abstract Dual chemotherapy (i.e., Pemetrexed and Cisplatin, CT) plus anti-VEGF Bevacizumab could be associated with anti-PD1 such as Pembrolizumab in NSCLC patients with low PDL1 levels. To support this new combination, to what extent those drugs could reshape tumor immunity remains to be evaluated, prior to establishing optimal scheduling or sequencing with Pembrolizumab. Both Pemetrexed and Cisplatin are expected to trigger immunogenic cell death, whereas Bevacizumab could help T lymphocytes to extravasate and better infiltrate tumor micro-environment. Here, we studied the impact of Cisplatin and Pemetrexed at two dose levels (i.e., low dose and Maximal Tolerated Dose, MTD) and fixed flat-dose Bevacizumab, alone or in combination (i.e., Cisplatin, Pemetrexed, Cisplatin + Pemetrexed, with our without Beva), on the immune repertoire of BalbC mice ectopically grafted with KLN-205 murine lung tumors. CD45, CD4, CD8, FoxP3-CD4 (Tregs), and MDSCs were monitored by FACS analysis in blood, spleen, and tumors on D2, D7, D14, and D21 after treatment. Results were normalized by CD45 counts. All treatments led to an increase in CD4+ T cells in blood as compared with control (i.e., untreated) animals. Little increase was observed in tumors, regardless of the treatments but mice treated with the Beva+CT combo (both low dose and MTD CT) showed an increase in CD4+ in the spleen on D14. Regarding CD8+ T cells, all treatments led to an increase from D2 to D7. Beva + CT low dose led to a continuous increase in CD8+ cells, whereas in Beva+CT MTD mice, after an initial increase, a reduction in CD8+ was observed after D7 eventually. In the spleen, CD8+ proved to be stable apart from the Beva + CT low dose group which showed an increase up to D14. In tumors, an increase in CD8+ cells was observed in all the groups. No change in MDSCs levels was observed, regardless of the drugs, the dosing and the organ. Conversely, Tregs decreased in the blood in the Beva + CT low-dose group on D14, and decreased in spleen and tumors as compared with Control mice, regardless of the drugs and the dosing. Overall, our data suggest that the triple combination between Beva, Pemetrexed, and Cisplatin has a stronger impact on immune cells than each drug used as a single agent. MDSCs were the only cells to be unaffected by the treatments. Dosing could be relevant as well since the reduction in CD8+ T cells observed with MTD CT could come from drug-related lymphopenia. The Beva followed by low dose Pemetrexed + Cisplatin group is thus the more likely to harness tumor immunity, with peaks in CD4+ and CD8+ T cells and drop in Tregs observed on D14. This pilot study suggests therefore that sequencing Beva + CT low dose followed by a two-week lag time prior to administrating anti-PD1 could lead to synergism in lung cancer models. Citation Format: Guillaume Sicard, Sarah Giacometti, Fabrice Barlesi, Raphaelle Fanciullino, Joseph Ciccolini. Pemetrexed-cisplatin-bevacizumab combo in lung cancer models: impact on tumor immunity [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 1856.

Abstract Background. Fluorescent imaging technologies now allow for multiplex immunofluorescence (mIF) for up to 100 targets on a single slide. However, the ability to quantitatively analyze the resulting data, especially on whole-slide images (WSI), is limited by scalability and reproducibility. Currently available platforms for segmenting cancer cells and nuclei involve segmentation algorithms that are hand-tuned on individual fields of view, making these methods subjective and difficult to replicate. To this end, we sought to develop an end-to-end workflow for WSI mIF data in cancer, from raw images to cell-level features, using state-of-the-art deep learning models for tissue, cell, and nuclei segmentation. Methods. mIF was performed using the Akoya 6-plex Lung IO panel (CD8, FoxP3, CD68, PD-1, PD-L1, and pan-cytokeratin) with a DAPI counterstain on clinical NSCLC specimens obtained from commercial sources (N=41). Slides were scanned using the Akoya PhenoImager HT. To remove bleed-through between different fluorophores, mIF WSIs were linearly unmixed based on a reference single-stain matrix, generated by selecting pure stain regions in mIF images. Convolutional neural network models previously trained on hematoxylin and eosin (H&E) images for 1) artifact detection, 2) tissue region identification, and 3) nucleus detection and segmentation were then deployed on mIF images that were converted to RGB images using a synthetic H&E transformation. Cell segmentation models were developed and implemented for each marker assessed. For each demixed channel representing a single antibody signal, expert annotations were used to train deep learning models for cell detection and segmentation. Finally, segmentation results from separate channels were aggregated based on spatial colocalization to identify cells and cell phenotypes. Results. We developed a pipeline for mIF image analysis at scale. After unmixing, conversion of an mIF image to a synthetic H&E image allowed for 1) removal of artifact regions, 2) detection of cancer and stroma using tissue segmentation, and 3) nuclei segmentation. Nuclear segmentation using this approach improved upon a widely used commercial segmentation platform when compared to manual expert annotations (Dice scores = 0.85 and 0.52, respectively). Finally, using state-of-the-art instance segmentation models allows a novel approach to cell phenotyping compared to currently available platforms, allowing positive identification of cells lacking a nuclear signal in the sectioned plane (e.g. large cytokeratin-positive cells). Conclusions. We developed a robust workflow to accurately segment cells and nuclei from mIF WSIs at scale. Improving the scalability and reproducibility of complex mIF image analysis may help improve the adoption of this approach for biomarker development and incorporation into clinical trial workflows in oncology. Citation Format: Waleed Tahir, Emma Krause, Jin Li, Howard Mak, Mohammad Mirzadeh, Kevin Rose, Judy Shen, Vignesh Valaboju, Guillaume Chhor, Joseph Lee, Jun Zhang, Jacqueline Brosnan-Cashman, Michael G. Drage, Justin Lee, Carlee Hemphill, Saumya Pant, Robert Egger. Development of a high-throughput image processing pipeline for multiplex immunofluorescence whole slide images at scale [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 6616.

Abstract Background: Cancers of unknown primary (CUP) is a heterogenous group of malignancies characterized by distant metastases in the absence of detectable primary. Specifics subgroups of CUP that share similarities with cancers of known primary beneficiate from tissue-tailored therapeutic strategies. The objective of this study was to characterize the clinical and biological presentation of a cohort of patients with suspected CUP of renal origin (rCUP) and to evaluate their optimal therapeutic management. Methods: All Patients with rCUP identified prospectively between 2020 and 2023 within the French National Multidisciplinary Tumor Board for CUP were included. Centralized pathological review and immunohistochemical stainings were performed. Whole Exome Sequencing (WES), whole transcriptomic sequencing (RNAseq) and application of the deep-learning based algorithm TransCUPtomics for prediction of tissue of origin, and DNA methylation analysis were performed and compared to public renal cell carcinomas (RCC). Progression Free Survival (PFS) and Overall Survival (OS) were calculated using Kaplan-Meier estimates. Results: 23 patients were included. The median age at diagnosis was 57 yo and 17/23 were male. The most common metastatic sites were the bones (N=17) and the lymph nodes (N=14). 19/23 patients presented with at least two distinct distant metastatic sites. Genomic analyses performed in 10 patients led to the identification of recurrent inactivating mutations in genes commonly altered in RCC, including NF2 (N=7), SETD2 (N=3), or VHL (N=1). 10/11 samples analyzed by RNAseq were predicted as RCC by the TransCUPtomics classifier based on their gene expression profiles. DNA methylation profiling performed in 7 samples showed major similarity with known RCC subtypes.Centralized pathological review and integration of molecular characteristics led to the final diagnoses of papillary RCC (N=2), clear cell RCC (N=7), renal medullary carcinoma (N=1), TFEB-amplified RCC (N=1) and unclassified RCC (N=12). 20/23 patients received at least one line of systemic treatment based on the renal origin, including anti-angiogenic TKI, anti-PD1/PDL1 antibody, anti-CTLA4 antibody or mTOR inhibitor. The median PFS under the first renal-oriented systemic treatment was 5 months and 10 patients remained progression-free for more than 6 months under tailored-treatment. The median OS was 31 months, and 11 patients are still alive in October 2023. Conclusions: This study shows that rCUP represent a new entity within CUPs that shares major similarities with classical RCC despite the more frequent presentation of unfavorable prognostic factors. Prolonged survival can be observed with renal-tailored systemic treatment, supporting the individualization of these patients within CUP. Citation Format: Nicolas Jacquin, Julien Masliah-Planchon, Guillaume Grisay, Ronan Flippot, Riwan Brillet, Célia Dupain, Maud Kamal, Isabelle Guillou, Nadège Gruel, Nicolas Servant, Pierre Gestraud, Jennifer Wong, Vincent Cockenpot, Andreia Goncalves, Janick Selves, Hélène Blons, Etienne Rouleau, Oliver Delattre, Christophe Le Tourneau, Ivan Bièche, Yves Allory, Laurence Albigès, Sarah Watson. Metastatic renal cell carcinoma with occult primary: Clinical and biological evidence for a new entity of cancers of unknown primary that beneficiates from tissue-tailored treatment [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 4627.

Abstract Background: In recent years, a relationship between the tumor microenvironment (TME) and patient response to targeted cancer immunotherapy has been suggested. We applied machine-learning algorithms on H&E stained tissue to study the TME in metastatic non-small cell lung cancer (NSCLC) patients. Our goal was to identify digital pathology (DP) features associated with outcome under combination treatment or monotherapy with atezolizumab (atezo), an anti-PD-L1 therapy, and relate those features to other data modalities. We analyzed patient data from two phase 3 clinical trials, OAK (docetaxel versus atezo in 2L+ NSCLC) and IMpower150 (bevacizumab, carboplatin, and paclitaxel (BCP) versus BCP+atezo (ABCP) in advanced 1L non-squamous NSCLC). Methods: As part of our effort to build a DP-based tumor-immune microenvironment atlas, digitized H&E images were registered onto the PathAI research platform. Over 200K annotations from 90 pathologists were used to train convolutional neural networks (CNNs) that classify slide-level human-interpretable features (HIFs) of cells and tissue structures from images and deployed on images from OAK and IMpower150. HIFs and PD-L1 status were associated with outcome in all samples in each arm in OAK and results were validated in IMpower150, using Cox proportional hazard models. Bulk RNAseq was run using samples extracted from the same area as the H&E slide. Results: We identified a composite feature capturing the ratio of immune cells to fibroblasts in the stroma predictive of both overall survival (OS) (HR=0.74 p=0.0046) and progression-free survival (PFS) (HR=0.87 p=0.14). While patients primarily benefit from atezo if they are PD-L1 high, we found that even PD-L1 negative patients benefited from atezo when enriched for this feature (22C3 PD-L1 assay: OS HR=0.59 p=0.015, PFS HR=0.8 p=0.25; SP142 PD-L1 assay: OS HR=0.74 p=0.12, PFS HR=0.88 p=0.45). We thus recognized a DP feature that was predictive for positive outcome with atezo treatment, independent of PD-L1 levels. This association was then validated in IMpower150 comparing ABCP to BCP, both overall (OS HR=0.69 p=0.012) and in PD-L1 negative patients (SP263 assay OS HR=0.56 p=0.034). Integrating with RNAseq, patients enriched for this DP feature showed similar enrichment for B and T gene signatures and depletion in CAF-related gene signatures, thus showing the harmonization of TME between different data modalities. Conclusions: Using a deep learning-based assay for quantifying pathology features of the TME from H&E images in two NSCLC trials, we identified a novel biomarker predictive of outcome to PD-L1 targeting therapy, even in PD-L1 low & negative patients. Importantly, our work shows how different data modalities (DP, gene expression) can be integrated to further our understanding of the TME. Citation Format: Aditi Qamra, Minu K. Srivastava, Eloisa Fuentes, Ben Trotter, Raymond Biju, Guillaume Chhor, James Cowan, Steven Gendreau, Webster Lincoln, Lisa McGinnis, Luciana Molinero, Namrata S. Patil, Amber Schedlbauer, Katja Schulze, Adam Stanford-Moore, Laura Chambre, Ilan Wapinski, David S. Shames, Hartmut Koeppen, Stephanie Hennek, Jane Fridlyand, Jennifer M. Giltnane, Assaf Amitai. Digital pathology based prognostic & predictive biomarkers in metastatic non-small cell lung cancer. [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 5705.

Abstract The effect on the immune system of nanoparticles is poorly described. The aim of this study was to analyze in immuno-competent BalbC mice bearing or not 4T1 murine breast cancers, the impact on immune system of blank liposomes (i.e., Lipo) or blank immunoliposomes grafted with trastuzumab with a maleimide linker (i.e., ANC). Both nanoparticles were 120 nm of diameter and were pegylated. Mice were administered with saline solution (Control) or treated with either Lipo or ANC with single-dosing (SD) or multiple-dosing (MD). Impact on immunity in blood, spleen and tumors was monitored in a longitudinal fashion in blood for up to 8 weeks, and once per animal in spleen and tumors (i.e., D18 or D23). Immune cells were analyzed by flow cytometry. In tumor-free mice, in blood a transient increase in B-cells between D7 and D15 was found (p= 0,003; < 0,001 and 0,009, t test) for ANC-MD, Lipo-SD and Lipo-MD, respectively. Of note, mice treated with ANC-MD exhibited persistent increase in B-cells up to 8 weeks. Regarding T-cells, a transient increase of CD4+ and CD8+ lymphocytes from D0 to D15 was observed. In spleen, more CD4+ T-cells than CD8+ cells were found, both with Lipo and ANC. Blank Lipo or blank ANC did not impact the tumor growth as compared with Control, and tumor sizes were comparable among all groups. In tumor-bearing mice, in blood little difference between B-cells was observed among the groups. Considering T-cells, a significant diminution in CD4+ cells was observed throughout time in mice treated with either ANC or Lipo (p=0.029, Anova). Conversely, a sharp increase in CD8+ cells as compared with Control group was observed between D18 and D23 for all the treatment arms (p<0.05, Anova). Of note, ANC-MD and Lipo-MD showed significantly higher CD8+ as compared with SD treatments. In spleen, more CD8+ cells than CD4+ cells were found. Still, CD4+ T-cells tended to increase, including in the Control group, between D18 and D23, whereas no such increase was observed for CD8+ cells. In treated animals, a sharp increase in FOXP3 CD4+ Tregs was observed in spleen. In tumors, B-cells increased in Control group but decreased in both Lipo and ANC-treated animals. A marked decrease in CD4+ cells was observed in Control group, whereas little change was observed in treated mice. Conversely, a sharp increase in CD8+ was observed, especially in mice treated with Lipo-MD and ANC-SD. Interestingly, no Tregs were found in treated mice (i.e., either Lipo or ANC) on D23, whereas Tregs were still observed in Control group. As a conclusion, our data suggest that nanoparticles can reshape tumor immunity indeed. Little difference was found between ANC and Lipo, suggesting the immunomodulating features we observed are probably supported by the pegylated liposomes, and not the grafted trastuzumab. Increase in CD8+ cells in blood and tumors, associated with decrease in Tregs in tumors, suggest that liposomal nanoparticles could help turning cold tumors into hot ones. Citation Format: Mathilde Dacos, Guillaume Sicard, Benoit Immordino, Sarah Giacometti, Joseph Ciccolini, Anne Rodallec, Raphaelle Fanciullino. Nanoliposomes harness tumor immunity in breast cancer models [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 1857.

Abstract Preclinical models of cancer are essential for understanding cancer biology and developing effective cancer therapeutics, and potentially to guide clinical decision-making in the era of personalized medicine. Therefore, it is required that these preclinical cancer models should closely recapitulate original tumors, including histo- and molecular pathology, and match patient’s drug response. Here we described the establishment and the characterization of 7 models (4 cell lines, 2 PDTO and 1 PDX), all derived from an Ovarian Clear Cell Carcinoma (OCCC) including their responses to treatment and their comparison to the clinical response. This ovarian cancer subtype is considered as a rare tumor, clinically aggressive and chemoresistant, and very few models are yet available. We succeeded in establishing Patient-Derived Xenografts (PDX) from subcutaneous tumor fragment implantation onto nude mice, and a monolayer tumor-derived cell line and Patient-Derived Tumor Organoid (PDTO) after patient tumor dissociation. A PDTO model was derived from the PDX and 3 additional cell lines were established from this PDTO model, from the PDTO derived from the patient tumor and from the PDX model, respectively. In order to determine the relevance of each of these models, comprehensive characterization was performed based on morphological, histological, and transcriptomic analyses as well as on the evaluation of their response to the conventional treatments received by the patient. These results were compared to the features of the original tumor and to the clinical data when it was possible. Only the PDX and PDTO model derived from the patient tumor were able to retain the two components within the patient tumor; one component displaying histological features of OCCC and the other one consisted of undifferentiated cells. The patient from which all these models were derived was refractory to carboplatin as first line chemotherapy and resistant to the next lines of treatments, doxorubicin and gemcitabine, respectively. While tumor cell lines were sensitive to these treatments, PDX and PDTO models displayed resistance to the 3 drugs. The transcriptomic analysis was also consistent with these results since the models recapitulating faithfully the clinical response did cluster together away from the other classical cell culture models. Then we investigated drugs that were not used in the patient clinical management and we identified the HDAC inhibitor Belinostat as a potential effective treatments based on the response of PDTO. In conclusion, PDX and PDTO appear thus to be the most relevant models, but PDTO seem to present all the necessary prerequisites for predictive purposes and could constitute relevant tools for therapeutic decision support in the context of these particularly aggressive cancers refractory to conventional treatments. Citation Format: Lucie Thorel, Pierre-Marie Morice, Hippolyte Paysant, Romane Florent, Guillaume Babin, Cecilia Thomine, Marion Perreard, Edwige Abeilard, Florence Giffard, Emilie Brotin, Christophe Denoyelle, Celine Villenet, Melanie Briand, Alexandra Leconte, Florence Joly, Enora Dolivet, Didier Goux, Cecile Blanc-Fournier, Corinne Jeanne, Marie Villedieu, Matthieu Meryet-Figuiere, Martin Figeac, Laurent Poulain, Louis-Bastien Weiswald. Comparative analysis of response to treatments and molecular features of patient-derived tumor organoids versus cell lines and PDX derived from the same ovarian clear cell carcinoma [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 166.

Abstract Background: There are a growing number of targeted agents that have the potential to greatly improve the clinical outcomes of patients with PDAC beyond current standard of care chemotherapy. However, there is a lack in our understanding of which patients and intrinsic PDAC tumor types will best respond to a given targeted agent(s). In order to optimizing their clinical utility, it is crucial that we a deep understanding of the biological impact that these targeted agents (alone or combination) have within the human system. As such, we designed and implemented the WOO trial to provide a highly adaptable discovery platform for the preliminary assessment of biological activity of one or more targeted agents in patients with PDAC. Utilizing paired, pre- and on-treatment biopsies, along with a robust multiomics pipeline, the WOO trial enables independent assessment of the biological impact of targeted agent(s) on key signaling pathways that can be used to explore novel therapeutic strategies. Methods: WOO is a multi-arm early phase I trial platform designed to assess the pharmacodynamic (PD) effects of one or more study agent(s) alone or in combination in patients with PDAC. A Master Protocol is used to describe overarching design and logistics, and sub-protocols separately describe each study arm comprising the different investigational agent(s). Each participants undergoes a baseline tumor biopsy, then receives their assigned study agent(s) for a specified timeframe (not exceeding 30 days), before undergoing a repeated tumor biopsy, after which they continue to receive therapy per standard of care or clinical trials. Spatially-resolved, single cell multiplex assays are used to compare the effects of the study agent(s) on tumor cell state and heterogeneity between the paired samples. The primary study objective is to independently assess the PD feasibility of detecting a measurable change in tumor biology at post-treatment from baseline for participants within a study arm. The WOO trial uses a 2-stage Bayesian efficacy monitoring approach with a futility-stopping rule for each study arm. In stage 1 of each study arm, if 6 or more of the first 10 participants have a detectable change in tumor biology measurements (i.e., pre vs. post-treatment), then the study arm may continue to enroll an additional 10 participants. To date, 4 study arms are being evaluated: 1) poly (ADP-ribose) polymerase inhibitor (PARPi), olaparib (300 mg PO BID for 10 days), 2) MEK inhibitor (MEKi), cobimetinib (60 mg PO QD for 10 days), 3) ERK inhibitor, LY3214996 (400 mg PO QD for 10 days), and 4) PLK1 inhibitor, onvansertib (12 mg/m2 PO QD for 10 days). To date, a total of 34 participants have been enrolled and treated: n = 14 cobimetinib, n= 15 olaparib, n=2 onvansertib. Future arms to be added. Trial ID: NCT04005690. Citation Format: Charles D. Lopez, Adel Kardosh, Emerson Chen, Guillame Pegna, Alexander Guimaraes, Bryan Foster, Brian Brinkerhoff, Shaun M. Goodyear, Erin Taber, Brindha Rajagopalan, Johnson Vo, Kiara Siex, Brett Johnson, Danielle Galipeau, Dove Keith, Brett Sheppard, Brody Jonathan, Rosalie Sears, Gordon Mills. Advancing biomarker discovery using a novel window of opportunity (WOO) trial for pancreatic ductal adenocarcinoma: Trial updates [abstract]. In: Proceedings of the AACR Special Conference in Cancer Research: Pancreatic Cancer; 2023 Sep 27-30; Boston, Massachusetts. Philadelphia (PA): AACR; Cancer Res 2024;84(2 Suppl):Abstract nr B003.

Abstract To enhance efficacy of anti-PD-1/PD-L1 antibodies, many combinations with various therapeutic agents are being investigated. Blocking the CD47/SIRPα myeloid checkpoint with monoclonal antibodies (mAbs) or decoy receptors is emerging as an effective approach to mobilize dendritic cells and macrophages to support T-cell mediated antitumor responses. The benefit of combining CD47/SIRPα and PD-1/PD-L1 blockade to improve tumor control has been convincingly demonstrated in preclinical models and is now being explored in patients. However, CD47 mAbs are hindered by ubiquitous CD47 expression, leading to pharmacokinetic (PK) and safety issues.NI-2901, an IgG4 CD47xPD-L1 bispecific antibody (bsAb), was generated using the κλ-body platform. In vitro assays were used to characterize its binding profile and checkpoint inhibition as well as its capacity to enhance T-cell activation and macrophage-mediated phagocytosis of tumor cells. PD-L1-independent CD47 antitumor activity was assessed in vivo in a PD-L1-negative xenograft model and compared to the anti-CD47 magrolimab analog. PK and tolerability of NI-2901 were evaluated in non-human primates (NHP), allowing for translational modeling to predict PK and dosing regimens in humans. Consistent with its intermediate affinity to CD47, NI-2901 shows lower binding to RBC as compared to magrolimab analog and is still able to induce CD47/SIRPα blockade on PD-L1-negative tumor cells, that is significantly enhanced once PD-L1 is expressed. As a result, the bsAb is able to enhance the phagocytosis of PD-L1-negative and -positive tumor cell lines induced by mAbs targeting tumor-associated antigens (e.g. rituximab, trastuzumab and anti-CD19) and demonstrates in vivo activity in the Raji B-cell lymphoma xenograft model. Given its high affinity for PD-L1, NI-2901 triggers an effective blockade of the PD-1/PD-L1 interaction, inducing T-cell activation in vitro to a degree similar to anti-PD-L1 benchmark antibodies atezolizumab and avelumab. In immunocompetent huCD47/huSIRPα-transgenic mice engrafted with MC38 cells engineered to express human PD-L1 and CD47, NI-2901 displayed significant anti-tumor activity. In a NHP study, NI-2901 was well-tolerated after four weekly injections at 30mg/kg, showing no signs of hemotoxicity. In contrast, the magrolimab analog induced a significant drop in RBC already after a single injection at 10mg/kg. PK modeling and simulations in humans suggest a more favorable dosing regimen as compared to CD47 targeted approaches. In conclusion, NI-2901, a dual immune checkpoint inhibitor, triggered effective T-cell activation and enhanced phagocytosis of tumor cells. Also, NI-2901 demonstrated significant antitumor activity in vivo and is therefore expected to show improved clinical efficacy over PD-1/PD-L1 blockade alone. The bsAb was well-tolerated in NHP without inducing RBC or platelet depletion. Citation Format: Xavier Chauchet, Sebastien Calloud, Pauline LLoveras, Nicolas Bosson, Margaux Legrand, Laurence Chatel, Laura Cons, Adeline Lesnier, Pauline Malinge, Guillemette Pontini, Christophe Guillamo, Dmitry Shchelokov, Oleg Demin, Ulla Ravn, Valéry Moine, Bruno Daubeuf, Giovanni Magistrelli, Yves Poitevin, Susana Salgado-Pires, Limin Shang, Nicolas Fischer, Walter Ferlin, Krzysztof Masternak. NI-2901, an affinity-optimized CD47xPD-L1 bispecific antibody for dual immune checkpoint blockade [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 2951.

Abstract Purpose: In this paper, we describe the methodology used, and provide a first characterization of the study population and radiotherapy (RT) data in CANTO-RT (CANcer TOxicities Radiotherapy), the largest available multicenter prospective cohort of early breast cancer (BC) patients treated with RT that aims to identify predictors of development, and persistence of long-term toxicities. Methods: CANTO (NCT01993498) is a French prospective clinical cohort study of 10 150 patients with stage I-III BC from 26 cancer centers. Patients matching all CANTO inclusion and exclusion criteria, who received RT and were still in follow up, in the 10 top recruiting CANTO centers, with a minimum follow up of 3 years, were selected for CANTO-RT. Eligible patients had breast/chest wall +/- lymph node RT with curative intent. Individual full DICOM RT files (CT, RT Structure, RT Dose, RT Plan) were collected, anonymized, structured and analyzed on the CANTO-RT/UNITRAD web platform using AQUILAB Share Place™ and Analytics Dose module. Characteristics of the patients and tumors (including TNM, histology, HER2, estrogen and progesterone receptor) were recorded at baseline. Characteristics of the treatments, skin, lung, cardiovascular, neurological, musculoskeletal toxicities (CTCAE v4.0), QOL (BR23, QLQC30), cosmetic, and oncological outcomes were assessed at diagnosis (baseline), 3-6 (M0), 12 (M12), 36 (M36) and 60 (M60) months after completion of primary surgery, chemotherapy or radiotherapy whichever came last together, with blood, plasma and serum tests. Results: CANTO-RT enrolled 3875 BC patients between June 2012 and February 2017 with a median follow-up of 64 months :1947 (50.2%) left side, 1850 (47.8%) right side and 78 (2%) bilateral BC. The vast majority of patients had hormone receptor-positive tumors 3321 (85.7%) and 553 (14.3%) had human epidermal growth factor 2 (HER2) positive tumors; 2586 (66.7%) had stage pT1 and 2525 (65.2%) pN0 disease; 2087 (53.8%) neoadjuvant or adjuvant chemotherapy, 477 (12.3%) adjuvant trastuzumab and 3138 (81%) adjuvant endocrine therapy. Among 3797 patients with unilateral RT, 3065 (80.4%) had breast conserving surgery, 747 (19.6%) total mastectomy; 2712 (71.5%) sentinel node and 1080 (28.5%) axillary dissection. Tumor bed boost was delivered in 2658 patients (68.5%) and lymph node RT in 1356 patients (35%) including internal mammary chain in 844 patients (21.8%). Most patients 3691 (95.3%) were treated with 3D conformal RT and 184 (4.7%) with intensity-modulated RT. Normofractionated RT (2Gy/fraction) was mostly used (69.9%). Clinical target (breast, chest wall, lymph nodes) and contoured organs at risk (heart, left anterior descending coronary, lung, spinal cord, esophagus, thyroid, brachial plexus, contralateral breast, humeral head) contours and dose/volume histograms were automatically extracted after quality control procedure excluding corrupted files and inconsistencies 36 (1%) (Table 1). Conclusion: CANTO-RT is the largest early breast cancer prospective cohort with full individual clinical and DICOM RT data available. CANTO-RT is a valuable resource, open for collaborative projects, for identification and validation of clinical and dosimetric predictive factors of RT related toxicities. Further long term follow up is ongoing. Table 1.Baseline characteristics of the CANTO RT breast cancer patients.CharacteristicsBreast Cancer Patients [N(%) or Mean (range)]Age at enrolmentMean (range), years56.5 (23.3-85.8)Tumour size (pT)T037 (1)T12586 (66.7)T21058 (27.3)T3177 (4.6)Missing17 (0.4)Nodal status (pN)02525 (65.2)11035 (26.7)2223 (5.8)379 (2)Missing13 (0.3)Tumour histologyInfiltrating Ductal3011 (77.7)Lobular473 (12.2)Others (including mixed)381 (9.8)Missing10 (0.3)Hormone Receptors positiveNegative541 (14)Positive3321 (85.7)Missing13 (0.3)HER2Negative3305 (85.3)Positive553 (14.3)Missing17 (0.4)Type of chemotherapyNo chemotherapy1788 (46.1)Neoadjuvant chemotherapy450 (11.6)Adjuvant chemotherapy1629 (42)Peri-adjuvant chemotherapy (neo + adjuvant)8 (0.2)Hormonal therapyNo730 (18.8)Yes3138 (81)Missing7 (0.2)Herceptin treatmentNo or Not applicable3378 (87.2)Yes477 (12.3)Missing20 (0.5)Type of breast surgerylumpectomy3113 (80.3)Mastectomy734 (18.9)Right lumpectomy and Left mastectomy13 (0.3)Right mastectomy and Left lumpectomy9 (0.2)None6 (0.2)Type of lymph node surgerySentinel node2746 (70.9)Axillary dissection1086 (28)Right sentinel node, Left axillary dissection20 (0.5)Right axillary dissection, left sentinel node12 (0.3)None11 (0.3)Radiation therapyRight Side1850 (47.8)Left Side1947 (50.2)Bilateral78 (2.0)Patients with boostNo or Not applicable1217 (31.4)Yes2658 (68.6)Lymph node levels treatedNone2519 (65)Yes1356 (35)Level 1284 (20.9)Level 2340 (25.1)Level 31072 (79.1)Level 41348 (99.4)Internal mammary chain844 (62.2)Irradiation techniques3D3691 (95.3)IMRT184 (4.7)Fractionation regimensNormofractionation 25-fractions2707 (69.9)Hypofractionation 15-16 fractions166 (4.3)Hypofractionation and Partial breast irradiation51 (1.3)Unspecified fractionation - CTV breast or chest wall not delineated951 (24.5) Citation Format: Thomas Sarrade, Rodrigue Allodji, Youssef Ghannam, Guillaume Auzac, Sibille Everhard, Ophélie Querel, Youlia Kirova, Karine Peignaux, Philippe Guilbert, Claire Charra-Brunaud, Julien Blanchecotte, Rezart Belshi, David Pasquier, Séverine Racadot, Céline Bourgier, Sandrine Ducornet, David Gibon, Fabrice André, Florent De Vathaire, Sofia Rivera. CANTO RT: The largest prospective multicenter cohort of early breast cancer patients treated with radiotherapy including full DICOM RT data [abstract]. In: Proceedings of the 2021 San Antonio Breast Cancer Symposium; 2021 Dec 7-10; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2022;82(4 Suppl):Abstract nr P3-19-01.

O tórus mandibular é considerado uma protuberância óssea (exostose) de etiologia desconhecida classificada como não-patológica, comumente vista em região lingual dos incisivos até pré-molares inferiores. Sua característica histopatológica é semelhante ao osso normal descrevendo-se osso cortical lamelar com a presença de canais de Havers, osteócitos normais e áreas dispersas de tecido conjuntivo. Normalmente se apresentam bilateralmente e indolores. Sem causar prejuízo na vida da pessoa que apresenta esta condição, devido estes aspectos, na maioria dos casos, não há intervenção cirúrgica, sendo o caso apenas acompanhado clinicamente. Neste caso apresentado, o objetivo é expor uma condição extrema desta exostose mandibular, tendo como finalidade a melhora na qualidade de vida do paciente.Descritores: Cirurgia Bucal; Exostose; Qualidade de Vida.ReferênciasKumar Singh A, Sulugodu Ramachandra S, Arora S, Dicksit DD, Kalyan CG, Singh P. Prevalence of oral tori and exostosis in Malaysian population - A cross-sectional study. J Oral Biol Craniofac Res. 2017;7(3):158-60.Hiremath VK, Husein A, Mishra N. Prevalence of torus palatinus and torus mandibularis among Malay population. J Int Soc Prev Community Dent. 2011;1(2):60-4.Sathya K, Kanneppady SK, Arishiya T. Prevalence and clinical characteristics of oral tori among outpatients in Northern Malaysia. J Oral Biol Craniofac Res. 2012;2(1):15-19.Jainkittivong A, Apinhasmit W, Swasdison S. Prevalence and clinical characteristics of oral tori in 1,520 Chulalongkorn University Dental School patients. Surg Radiol Anat. 2007;29(2):125-31.Kim YS. Pathogenetic growth potential in the central area of oral exostosis. Korean J Oral Maxillofac Pathol. 2013;37(5):201-10.Morrison MD, Tamimi F. Oral tori are associated with local mechanical and systemic factors: A case-control study. J Oral. Maxillofac Surg. 2013;71(1):14-22.García-García AS, Martínez-González JM, Gómez-Font R, Soto-Rivadeneira A, Oviedo-Roldán L. Current status of the torus palatinus and torus mandibularis. Med Oral Patol Oral Cir Bucal. 2010;15(2):e353-60.Ladizinski B, Lee KC. A nodular protuberance on the hard palate. JAMA. 2014;311(15):1558-59.Khan S, Shah SAH, Ali F, Rasheed D. Concurrence of torus palatinus, torus mandibularis and buccal exostosis. J Coll Physicians Surg Pak. 2016;26(11):111-13.Kün-Darbois JD, Guillaume B, Chappard D. Asymmetric bone remodeling in mandibular and maxillary tori. Clin Oral Investig. 2017;21(9):2781-88.Riley DS, Barber MS, Kienle GS, Aronson JK, von Schoen-Angerer T, Tugwell P et al. CARE guidelines for case reports: explanation and elaboration document. J Clin Epidemiol. 2017;89:218-35.Kucukkurt S, Özle M, Baris E. Peripheral osteoma in an unusual location on the mandible. BMJ Case Rep. 2016;2016. pii: bcr2016216554.

Füssel, Marian / Antje Kuhle / Michael Stolz (Hrsg.), Höfe und Experten. Relationen von Macht und Wissen in Mittelalter und Früher Neuzeit, Göttingen 2018, Vandenhoeck & Ruprecht, 228 S. / Abb., € 55,00. (Alexander Querengässer, Leipzig) Fertig, Christine / Margareth Lanzinger (Hrsg.), Beziehungen – Vernetzungen – Konflikte. Perspektiven Historischer Verwandtschaftsforschung, Köln / Weimar / Wien 2016, Böhlau, 286 S. / Abb., € 35,00. (Simon Teuscher, Zürich) Geest, Paul van/ Marcel Poorthuis / Els Rose (Hrsg.), Sanctifying Texts, Transforming Rituals. Encounters in Liturgical Studies. Essays in Honour of Gerard A. M. Rouwhorst (Brill’s Studies in Catholic Theology, 5), Leiden / Boston 2017, Brill, XL u. 489 S. / Abb., € 145,00. (Martin Lüstraeten, Mainz) Kallestrup, Louise Nyholm / Raisa M. Toivo (Hrsg.), Contesting Orthodoxy in Medieval and Early Modern Europe. 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(Angela Huang, Lübeck) Kirsch, Mona, Das allgemeine Konzil im Spätmittelalter. Organisation – Verhandlungen – Rituale (Heidelberger Abhandlungen zur Mittleren und Neueren Geschichte, 21), Heidelberg 2016, Universitätsverlag Winter, 655 S., € 68,00. (Johannes Helmrath, Berlin) Burton, Janet / Karen Stöber (Hrsg.), Women in the Medieval Monastic World (Medieval Monastic Studies, 1), Turnhout 2015, Brepols, VIII u. 377 S. / Abb., € 90,00. (Cristina Andenna, Dresden) Baker, John, The Reinvention of Magna Carta 1216 – 1616 (Cambridge Studies in English Legal History), Cambridge [u. a.] 2017, Cambridge University Press, XLIX u. 570 S., £ 120,00. (Andreas Pečar, Halle a. d. Saale) Bünz, Enno (Hrsg.), Geschichte der Stadt Leipzig, Bd. 1: Von den Anfängen bis zur Reformation, Leipzig 2015, Leipziger Universitätsverlag, 1055 S. / Abb., € 49,00. (Christian Speer, Halle a. d. S.) Kinne, Hermann, Das (exemte) Bistum Meißen 1: Das Kollegiatstift St. Petri zu Bautzen von der Gründung bis 1569 (Germania Sacra. Dritte Folge, 7), Berlin / Boston 2014, de Gruyter, XII u. 1062 S., € 169,95. (Ulrike Siewert, Chemnitz) Bauch, Martin / Julia Burkhardt / Tomáš Gaudek / Václav Žůrek (Hrsg.), Heilige, Helden, Wüteriche. Herrschaftsstile der Luxemburger (1308 – 1437) (Forschungen zur Kaiser- und Papstgeschichte des Mittelalters, 41), Köln / Weimar / Wien 2017, Böhlau, 449 S. / Abb., € 55,00. (Lenka Bobkova, Prag) Voigt, Dieter, Die Augsburger Baumeisterbücher des 14. Jahrhunderts, 2 Bde., Bd. 1: Darstellung; Bd. 2: Transkriptionen (Veröffentlichungen der Schwäbischen Forschungsgemeinschaft. Reihe 1: Studien zur Geschichte des Bayerischen Schwabens, 43), Augsburg 2017, Wißner, XII u. 228 S. / Abb. / CD-ROM (Bd. 1); X u. 906 S. (Bd. 2), € 65,00. 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(Christina Vanja, Kassel) Mączak, Antoni, Eine Kutsche ist wie eine Straßendirne … Reisekultur im Alten Europa. Aus dem Polnischen von Reinhard Fischer und Peter O. Loew (Polen in Europa), Paderborn 2017, Schöningh, 237 S. / Abb., € 29,90. (Benjamin Müsegades, Heidelberg) Garner, Guillaume (Hrsg.), Die Ökonomie des Privilegs, Westeuropa 16.–19. Jahrhundert / Lʼéconomie du privilège, Europe occidentale XVIe-XIXe siècles (Studien zu Policey, Kriminalitätsgeschichte und Konfliktregulierung), Frankfurt a. M. 2016, Klostermann, VII u. 523 S. / graph. Darst., € 79,00. (Rachel Renault, Le Mans) Gemeine Bescheide, Teil 1: Reichskammergericht 1497 – 1805, hrsg. v. Peter Oestmann (Quellen und Forschungen zur höchsten Gerichtsbarkeit im Alten Reich, 63.1), Köln / Weimar / Wien 2013, Böhlau, VI u. 802 S., € 79,90. (Ralf-Peter Fuchs, Essen) Gemeine Bescheide, Teil 2: Reichshofrat 1613 – 1798, hrsg. v. Peter Oestmann (Quellen und Forschungen zur höchsten Gerichtsbarkeit im Alten Reich, 63.2), Köln / Weimar / Wien 2017, Böhlau, 480 S., € 60,00. (Ralf-Peter Fuchs, Essen) Süß, Thorsten, Partikularer Zivilprozess und territoriale Gerichtsverfassung. Das weltliche Hofgericht in Paderborn und seine Ordnungen 1587 – 1720 (Quellen und Forschungen zur höchsten Gerichtsbarkeit im Alten Reich, 69), Köln / Weimar / Wien 2017, Böhlau, 570 S., € 90,00. (Michael Ströhmer, Paderborn) Luebke, David M., Hometown Religion. Regimes of Coexistence in Early Modern Westphalia (Studies in Early Modern German History), Charlottesville / London 2016, University of Virginia Press, XI u. 312 S. / Abb., $ 45,00. (Alexander Schunka, Berlin) Blum, Daniela, Multikonfessionalität im Alltag. Speyer zwischen politischem Frieden und Bekenntnisernst (1555 – 1618) (Reformationsgeschichtliche Studien und Texte, 162), Münster 2015, Aschendorff, X u. 411 S., € 56,00. 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Kulturgeschichte der evangelischen Geistlichkeit des 17. Jahrhunderts, Berlin / Boston 2017, de Gruyter Oldenbourg, VI u. 234 S. / Abb., € 29,95. (Cornel Zwierlein, Bamberg / Erfurt) Hennings, Werner / Uwe Horst / Jürgen Kramer, Die Stadt als Bühne. Macht und Herrschaft im öffentlichen Raum von Rom, Paris und London im 17. Jahrhundert (Edition Kulturwissenschaft, 63), Bielefeld 2016, transcript, 421 S. / Abb., € 39,99. (Susanne Rau, Erfurt) „Das Beispiel der Obrigkeit ist der Spiegel des Unterthans“. Instruktionen und andere normative Quellen zur Verwaltung der liechtensteinischen Herrschaften Feldsberg und Wilfersdorf in Niederösterreich (1600 – 1815), hrsg. v. Anita Hipfinger (Fontes Rerum Austriacarum. Abt. 3: Fontes Iuris, 24), Wien / Köln / Weimar 2016, Böhlau, 875 S. / Abb., € 97,00. (Alexander Denzler, Eichstätt) Roper, Louis H., Advancing Empire. English Interests and Overseas Expansion, 1613 – 1688, New York 2017, Cambridge University Press, XI u. 302 S., £ 25,99. 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(Pauline Puppel, Berlin) Onnekink, David, Reinterpreting the Dutch Forty Years War, 1672 – 1713, Palgrave Pivot 2016, London, VIII u. 138 S., £ 37,99. (Johannes Arndt, Münster) Froide, Amy M., Silent Partners. Women as Public Investors during Britainʼs Financial Revolution, 1690 – 1750, Oxford / New York 2017, Oxford University Press, VI u. 225 S. / Abb., £ 60,00. (Philipp R. Rössner, Manchester) Mulsow, Martin / Kasper Risbjerg Eskildsen / Helmut Zedelmaier (Hrsg.), Christoph August Heumann (1681 – 1764). Gelehrte Praxis zwischen christlichem Humanismus und Aufklärung (Gothaer Forschungen zur Frühen Neuzeit, 12), Stuttgart 2017, Steiner, XVI u. 265 S. / Abb., € 54,00. (Claire Gantet, Fribourg/Freiburg) Harding, Elizabeth (Hrsg.), Kalkulierte Gelehrsamkeit. Zur Ökonomisierung der Universitäten im 18. Jahrhundert (Wolfenbütteler Forschungen, 148), Wiesbaden 2016, Harrassowitz in Kommission, 300 S. / Abb., € 62,00. (Andrea Thiele, Halle a. d. S.) Fulda, Daniel, „Die Geschichte trägt der Aufklärung die Fackel vor“. Eine deutsch-französische Bild-Geschichte (IZEA. Kleine Schriften, 7/2016), Halle a. d. S. 2017, Mitteldeutscher Verlag, 213 S. / Abb., € 16,00. (Kai Bremer, Kiel) Suitner, Riccarda, Die philosophischen Totengespräche der Frühaufklärung (Studien zum achtzehnten Jahrhundert, 37), Hamburg 2016, Meiner, 276 S. / Abb., € 78,00. (Helmut Zedelmaier, München / Halle a. d. S.) Mintzker, Yair, The Many Deaths of Jew Süss. The Notorious Trial and Execution of an Eighteenth-Century Court Jew, Princeton / Oxford 2017, Princeton University Press, X u. 330 S. / Abb., £ 27,95. (Gudrun Emberger, Berlin) Zedler, Andrea / Jörg Zedler (Hrsg.), Prinzen auf Reisen. Die Italienreise von Kurprinz Karl Albrecht 1715/16 im politisch-kulturellen Kontext (Beihefte zum Archiv für Kulturgeschichte, 86), Köln / Weimar / Wien 2017, Böhlau, 364 S. / Abb., € 50,00. (Michael Maurer, Jena) Streminger, Gerhard, Adam Smith. Wohlstand und Moral. Eine Biographie, Beck 2017, München, 253 S. / Abb., € 24,95. (Georg Eckert, Wuppertal) Home, Roderick W. / Isabel M. Malaquias / Manuel F. Thomaz (Hrsg.), For the Love of Science. The Correspondence of J. H. de Magellan (1722 – 1790), 2 Bde., Bern [u. a.] 2017, Lang, 2002 S. / Abb., € 228,95. (Lisa Dannenberg-Markel, Aachen) Wendt-Sellin, Ulrike, Herzogin Luise Friederike von Mecklenburg-Schwerin (1722 – 1791). Ein Leben zwischen Pflicht, Pläsir und Pragmatismus (Quellen und Studien aus den Landesarchiven Mecklenburg-Vorpommerns, 19), Köln / Weimar / Wien 2017, Böhlau, 468 S. / Abb., € 60,00. (Britta Kägler, Trondheim) Oehler, Johanna, „Abroad at Göttingen“. Britische Studenten als Akteure des Kultur- und Wissenstransfers 1735 bis 1806 (Veröffentlichungen der Historischen Kommission für Niedersachsen und Bremen, 289), Göttingen 2016, Wallstein, 478 S. / graph. Darst., € 39,90. (Michael Schaich, London) Düwel, Sven, Ad bellum Sacri Romano-Germanici Imperii solenne decernendum: Die Reichskriegserklärung gegen Brandenburg-Preußen im Jahr 1757. Das Verfahren der „preußischen Befehdungssache“ 1756/57 zwischen Immerwährendem Reichstag und Wiener Reichsbehörden, 2 Teilbde., Münster 2016, Lit, 985 S. / Abb., € 79,90 (Bd. 3 als Download beim Verlag erhältlich). (Martin Fimpel, Wolfenbüttel) Pufelska, Agnieszka, Der bessere Nachbar? Das polnische Preußenbild zwischen Politik und Kulturtransfer (1765 – 1795), Berlin / Boston 2017, de Gruyter Oldenbourg, VIII u. 439 S., € 74,95. (Maciej Ptaszyński, Warschau) Herfurth, Stefan, Freiheit in Schwedisch-Pommern. Entwicklung, Verbreitung und Rezeption des Freiheitsbegriffs im südlichen Ostseeraum zum Ende des 18. Jahrhunderts (Moderne europäische Geschichte, 14), Göttingen 2017, Wallstein, 262 S. / Abb., € 29,90. (Axel Flügel, Bielefeld) Boie, Heinrich Christian / Luise Justine Mejer, Briefwechsel 1776 – 1786, hrsg. v. Regina Nörtemann in Zusammenarbeit mit Johanna Egger, 4 Bde. im Schuber, Bd. 1: Juni 1776 – Juni 1782; Bd. 2: Juli 1782 – Juni 1784; Bd. 3: Juli 1784 – Juli 1786; Bd. 4: Kommentar, Göttingen 2016, Wallstein, 612 S. (Bd. 1); 608 S. (Bd. 2); 571 S. (Bd. 3); 846 S. / Abb. (Bd. 4), € 149,00. (Barbara Stollberg-Rilinger, Berlin / Münster) Poniatowski, Fürst Stanisław, Tagebuch einer Reise durch die deutschen Länder im Jahre 1784. Aus dem Manuskript übers. u. hrsg. v. Ingo Pfeifer, Halle a. d. S. 2017, Mitteldeutscher Verlag, 269 S., € 24,95. (Michael Maurer, Jena) Blaufarb, Rafe, The Great Demarcation. The French Revolution and the Invention of Modern Property, New York 2016, Oxford University Press, XIV u. 282 S., £ 47,99. (Moritz Isenmann, Köln) Behringer, Wolfgang, Tambora und das Jahr ohne Sommer. Wie ein Vulkan die Welt in die Krise stürzte, 4. Aufl., München 2016, Beck, 398 S. / Abb., € 24,95. (Wolfgang Reinhard, Freiburg i. Br.) Die Tagebücher des Ludwig Freiherrn Vincke 1789 – 1844, Bd. 10: 1830 – 1839, bearb. v. Heide Barmeyer-Hartlieb (Veröffentlichungen des Vereins für Geschichte und Altertumskunde Westfalens, Abt. Münster, 10; Veröffentlichungen der Historischen Kommission für Westfalen. Neue Folge, 45; Veröffentlichungen des Landesarchivs Nordrhein-Westfalen, 69), Münster 2018, Aschendorff, 949 S. / Abb., € 88,00. (Heinz Duchhardt, Mainz)