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A figure showing the process of generating and using PDX models.

Patient-derived tumor xenograft (PDX) mouse models have emerged as an important oncology research platform to study tumor evolution, mechanisms of drug response and resistance, and tailoring chemotherapeutic approaches for individual patients. The lack of robust standards for reporting on PDX models has hampered the ability of researchers to find relevant PDX models and associated data. Here we present the PDX models minimal information standard (PDX-MI) for reporting on the generation, quality assurance, and use of PDX models. PDX-MI defines the minimal information for describing the clinical attributes of a patient's tumor, the processes of implantation and passaging of tumors in a host mouse strain, quality assurance methods, and the use of PDX models in cancer research. Adherence to PDX-MI standards will facilitate accurate search results for oncology models and their associated data across distributed repository databases and promote reproducibility in research studies using these models. Cancer Res; 77(21); e62–66. ©2017 AACR.

Counteracting high failure rates in oncology drug development and improving therapeutic management of cancer patients requires preclinical models that can account for the complexity and heterogeneity of human tumors. Patient-derived cancer xenografts (PDXs) maintain histopathological features and genetic profiles of the original patient tumors and are increasingly recognized as reliable models to predict treatment efficacy and discover sensitivity and resistance biomarkers with immediate clinical relevance.Launched in 2013, the EurOPDX Consortium now gathers 18 academic research institutions throughout Europe and in the US (www.europdx.eu). The goal of the Consortium is to maximize exploitation of PDXs and other patient-derived models for cancer research by: (i) integrating institutional collections into a multicentre repository; (ii) defining common standards to improve the quality and reproducibility of oncology preclinical data; (iii) sharing models within and outside the consortium to perform collaborative precision oncology “xenopatient” trials. Building on its first successes, EurOPDX is now teaming up with other key academic and SME partners in a four-year project to build the “EurOPDX Distributed Infrastructure for Research on patient-derived Xenografts" (EDIReX project, Horizon 2020 grant no. 731105).This new cutting-edge European infrastructure offers access to PDX resources for academic and industrial cancer researchers through 6 state-of-the-art installations or “nodes”. We will present the specific objectives of the project, including our work towards standardization and optimization of biobanking, quality control and data tracking, and the performance of in vivo drug efficacy experiments. Access to the resource, including the distribution of cryopreserved samples from established models, the structured biobanking of user-developed models and the performance of drug efficacy studies, is offered through a grant application system which last deadline is planned mid-June 2020. Selection of the models by users and browsing of PDXs annotation data is made possible thanks to the newly-developed EurOPDX Data Portal (dataportal.europdx.eu), which will display approximately 1,000 models by April 2020 (including 700+ models of colorectal cancer, 80+ gastric and 80+ breast cancer models).We aim to improve preclinical and translational cancer research and promote innovation in oncology by integrating a European PDX repository and facilitating access to this much-needed resource for European and worldwide researchers. Citation Format: Emilie Vinolo, Joaquin Arribas, Andrea Bertotti, Alejandra Bruna, Annette T. Byrne, Robert B. Clarke, Nathalie Conte, Steven de Jong, Didier Decaudin, Zdenka Dudova, Jos Jonkers, Daniela Krasser, Ales Krenek, Luisa Lanfrancone, Eleonora Leucci, Elisabetta Marangoni, Gunhild Mari Maelandsmo, Michaela Th. Mayrhofer, Terrence F. Meehan, Jens Henrik Norum, Hector G. Palmer, Alejandro Piris Gimenez, Leo Price, Sergio Roman-Roman, Francesca Sarno, Violeta Serra, Laura Soucek, Livio Trusolino, Marieke van de Ven, Luca Vezzadini, Alberto Villanueva, Andrea Wutte, Enzo Medico, on behalf of the EurOPDX Research Infrastructure. The EurOPDX Research Infrastructure: Supporting European and worldwide cancer research with patient-derived xenografts [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 1685.

Nature Reviews Cancer 17, 254–268 (2017) In the online html version of this article, Joan Seoane's affiliations were not correct. He is also a member of the EurOPDX Consortium and is at the Vall d'Hebron Institute of Oncology, 08035 Barcelona, the Universitat Autonoma de Barcelona, 08193 Bellaterra,and the Institucio Catalana de Recerca i Estudis Avancats (ICREA), 08010 Barcelona, Spain.

Patient-derived xenografts (PDXs) have emerged as an important platform to elucidate new treatments and biomarkers in oncology. PDX models are used to address clinically relevant questions, including the contribution of tumour heterogeneity to therapeutic responsiveness, the patterns of cancer evolutionary dynamics during tumour progression and under drug pressure, and the mechanisms of resistance to treatment. The ability of PDX models to predict clinical outcomes is being improved through mouse humanization strategies and the implementation of co-clinical trials, within which patients and PDXs reciprocally inform therapeutic decisions. This Opinion article discusses aspects of PDX modelling that are relevant to these questions and highlights the merits of shared PDX resources to advance cancer medicine from the perspective of EurOPDX, an international initiative devoted to PDX-based research.

Counteracting high attrition rates in anticancer drug development and providing optimal therapeutic management of cancer patients require preclinical models that properly recapitulate the complexity and diversity of human tumours. Patient-derived cancer xenografts (PDXs), developed by transplanting human tumor fragments into immunodeficient mice, retain the idiosyncratic characteristics of different tumors from different patients. The possibility of population-scale correlations between therapeutic response in PDXs and extensive, multidimensional molecular annotation has enabled the identification of several sensitivity and resistance biomarkers in a number of different tumor types, with immediate clinical relevance. To effectively recapitulate and therapeutically interrogate through patient-derived models the heterogeneity that typifies human cancer, 13 European cancer centres and university hospitals joined forces in 2013 to start EurOPDX, an academic research consortium that now gathers 19 institutions throughout Europe and in the US. EurOPDX includes world-renowned experts at the forefront of research in basic, preclinical, translational and clinical oncology across multiple pathologies, and displays a wide range of expertise in technological platforms. The primary goal of the Consortium is to maximize exploitation of PDXs and other patient-derived cancer models for cancer research by: (i) integrating institutional collections into an organic, multicentre collection of patient-derived models, now reaching 2,000 subcutaneous and orthotopic models for 30+ different pathologies (www.europdx.eu); (ii) defining common operating procedures to improve and standardize molecular and pharmacologic characterization of the models; (iii) sharing models within and outside the consortium to perform collaborative precision oncology “xenopatient” trials, to discover predictive biomarkers, new targets, and new strategies to overcome drug resistance. Towards these objectives, the EC granted 5 million euros under the H2020 programme for a “EurOPDX Distributed Infrastructure for Research on patient-derived Xenografts" - EDIReX EU project n. 731105, starting on February 2018. By teaming up with other key academic, technological and SME partners, our goal with EDIReX is to establish a cutting-edge European infrastructure offering transnational access to PDX resources to academic and industrial cancer researchers, including the distribution of cryopreserved samples to third parties, the structured biobanking of user-developed models, and the performance of drug efficacy studies. We will provide an overview of the current achievements of the Consortium and the objectives of the EDIReX project. Citation Format: Enzo Medico, Mohamed Bentires-Alj, Andrew V. Biankin, Alejandra Bruna, Annette T. Byrne, Carlos Caldas, Robert B. Clarke, Georges Coukos, Olivier Elemento, Manuel Hidalgo, Giorgio Inghirami, Steven de Jong, Jos Jonkers, Ales Krenek, Eleonora Leucci, Gunhild Mari Maelandsmo, Michaela Th. Mayrhofer, Terrence F. Meehan, Simone P. Niclou, Pier Giuseppe Pelicci, Alejandro Piris-Gimenez, Leo Price, Sergio Roman-Roman, Livio Trusolino, Luca Vezzadini, Alberto Villanueva, Emilie Vinolo, on behalf of the EurOPDX Consortium. The EurOPDX EDIReX project: Towards a European research infrastructure on patient-derived cancer models [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 985.

The NEMO (NF-kappaB essential modulator) protein plays a crucial role in the canonical NF-kappaB pathway as the regulatory component of the IKK (IkappaB kinase) complex. The human disease anhidrotic ectodermal dysplasia with immunodeficiency (EDA-ID) has been recently linked to mutations in NEMO. We investigated the effect of an alanine to glycine substitution found in the NEMO polypeptide of an EDA-ID patient. This pathogenic mutation is located within the minimal oligomerization domain of the protein, which is required for the IKK activation in response to diverse stimuli. The mutation does not dramatically change the native-like state of the trimer, but temperature-induced unfolding studied by circular dichroism showed that it leads to an important loss in the oligomer stability. Furthermore, fluorescence studies showed that the tyrosine located in the adjacent zinc finger domain, which is possibly required for NEMO ubiquitination, exhibits an alteration in its spectral properties. This is probably due to a conformational change of this domain, providing evidence for a close interaction between the oligomerization domain and the zinc finger. In addition, functional complementation assays using NEMO-deficient pre-B and T lymphocytes showed that the pathogenic mutation reduced TNF-alpha and LPS-induced NF-kappaB activation by altering the assembly of the IKK complex. Altogether, our findings provide understanding as to how a single point mutation in NEMO leads to the observed EDA-ID phenotype in relation to the NEMO-dependent mechanism of IKK activation.

International audience; Two types of polymeric post-translational modifications of alpha/beta-tubulin, glycylation and glutamylation, occur widely in cilia and flagella. Their respective cellular functions are poorly understood. Mass spectrometry and immunoblotting showed that two closely related species, the ciliates Tetrahymena and Paramecium, have dramatically different compositions of tubulin post-translational modifications in structurally identical axonemes. Whereas the axonemal tubulin of Paramecium is highly glycylated and has a very low glutamylation content, the axonemal tubulin of Tetrahymena is glycylated and extensively glutamylated. In addition, only the alpha-tubulin of Tetrahymena undergoes detyrosination. Mutations of the known glycylation sites in Tetrahymena tubulin affected the level of each polymeric modification type in both the mutated and nonmutated subunits, revealing cross-talk between alpha- and beta-tubulin. Ultrastructural analyses of glycylation site mutants uncovered defects in the doublet B-subfiber of axonemes and revealed an accumulation of dense material in the ciliary matrix, reminiscent of intraflagellar transport particles seen by others in Chlamydomonas. We propose that polyglycylation and/or polyglutamylation stabilize the B-subfiber of outer doublets and regulate the intraflagellar transport.

International audience; NEMO (NF-κB essential modulator) plays a key role in the canonical NF-κB pathway as the scaffold/regulatory component of the IκB kinase (IKK) complex. The self-association of NEMO involves the C-terminal halves of the polypeptide chains containing two putative coiled-coil motifs (a CC2 and a LZ leucine zipper), a proline-rich region, and a ZF zinc finger motif. Using purified truncation mutants, we showed that the minimal oligomerization domain of NEMO is the CC2-LZ segment and that both CC2 and LZ subdomains are necessary to restore the LPS-dependent activation of the NF-κB pathway in a NEMO-deficient cell line. We confirmed the association of the oligomerization domain in a trimer and investigated the specific role of CC2 and LZ subdomains in the building of the oligomer. Whereas a recombinant CC2-LZ polypeptide self-associated into a trimer with an association constant close to that of the wild-type protein, the isolated CC2 and LZ peptides, respectively, formed trimers and dimers with weaker association constants. Upon mixing, isolated CC2 and LZ peptides associated to form a stable hetero-hexamer as shown by gel filtration and fluorescence anisotropy experiments. We propose a structural model for the organization of the oligomerization domain of activated NEMO in which three C-terminal domains associate into a pseudo-hexamer forming a six-helix bundle. This model is discussed in relation to the mechanism of activation of the IKK complex by upstream activators.

International audience; The regulatory NEMO (NF-kappaB essential modulator) protein has a crucial role in the canonical NF-kappaB signaling pathway notably involved in immune and inflammatory responses, apoptosis and oncogenesis. The regulatory domain is located in the C-terminal half of NEMO and contains a classical CCHC-type zinc finger (ZF). We have investigated the structural and functional effects of a cysteine to phenylalanine point mutation (C417F) in the ZF motif, identified in patients with anhidrotic ectodermal dysplasia with immunodeficiency. The solution structures of the wild type and mutant ZF were determined by NMR. Remarkably, the mutant adopts a global betabetaalpha fold similar to that of the wild type and retains thermodynamic stability, i.e., the ability to bind zinc with a native-like affinity, although the last zinc-chelating residue is missing. However, the mutation induces enhanced dynamics in the motif and leads to an important loss of stability. A detailed analysis of the wild type solution structure and experimental evidences led to the identification of two possible protein-binding surfaces that are largely destabilized in the mutant. This is sufficient to alter NEMO function, since functional complementation assays using NEMO-deficient pre-B and T lymphocytes show that full-length C417F pathogenic NEMO leads to a partial to strong defect in LPS, IL-1beta and TNF-alpha-induced NF-kappaB activation, respectively, as compared to wild type NEMO. Altogether, these results shed light onto the role of NEMO ZF as a protein-binding motif and show that a precise structural integrity of the ZF should be preserved to lead to a functional protein-recognition motif triggering full NF-kappaB activation.

WOS: 000238940500017, PubMed ID: 16818673, Germline mutations in five autosomal genes involved in interleukin (IL)-12-dependent, interferon (IFN)-gamma-mediated immunity cause Mendelian susceptibility to mycobacterial diseases (MSMD). The molecular basis of X-linked recessive (XR)-MSMD remains unknown. We report here mutations in the leucine zipper (LZ) domain of the NF-kappa B essential modulator (NEMO) gene in three unrelated kindreds with XR-MSMD. The mutant proteins were produced in normal amounts in blood and fibroblastic cells. However, the patients' monocytes presented an intrinsic defect in T cell-dependent IL-12 production, resulting in defective IFN-gamma secretion by T cells. IL-12 production was also impaired as the result of a specific defect in NEMO- and NF-kappa B/c-Rel-mediated CD40 signaling after the stimulation of monocytes and dendritic cells by CD40L-expressing T cells and fibroblasts, respectively. However, the CD40-dependent up-regulation of costimulatory molecules of dendritic cells and the proliferation and immunoglobulin class switch of B cells were normal. Moreover, the patients' blood and fibroblastic cells responded to other NF-kappa B activators, such as tumor necrosis factor-alpha, IL-beta, and lipopolysaccharide. These two mutations in the NEMO LZ domain provide the first genetic etiology of XR-MSMD. They also demonstrate the importance of the T cell- and CD40L-triggered, CD40-, and NEMO/NF-kappa B/c-Rel-mediated induction of IL-12 by monocyte-derived cells for protective immunity to mycobacteria in humans., Intramural NIH HHSUnited States Department of Health & Human ServicesNational Institutes of Health (NIH) - USA; Medical Research CouncilMedical Research Council UK (MRC) [G0900867]

e22056 Background: Triple negative breast cancer (TNBC) is associated with aggressive histological features, high relapse rates, and poor overall survival. To improve the management of TNBC patient...