Cancer Research Unit Research Articles

Interview with Journal of Cell Science Editor Renata Basto

ABSTRACT Renata Basto is a Centre National de la Recherche Scientifique (CNRS) Team Leader at the Institut Curie and Deputy Director of the Cell Biology and Cancer Research Unit in Paris, France. Renata completed her undergraduate degree at the University of Lisbon, Portugal, before moving abroad to Gif-sur-Yvette, France for her doctoral studies on mitotic spindle assembly checkpoint proteins in Drosophila, mentored by Prof. Rui Gomes and Prof. Roger Karess. She then received a postdoctoral fellowship with Prof. Jordan Raff at the University of Cambridge, UK, where her work helped identify centrosome amplification as a tumour-initiating mechanism. In 2008, she joined the Department of Cell Biology at CNRS and the Institut Curie, where her laboratory studies the roles of centrosomes, and mechanisms of mitotic spindle assembly, polyploidy and genetic instability in development, health and disease. Renata joined Journal of Cell Science as an Editor in August 2023, bringing to the journal her significant expertise in the cell biology of centrosomes, spindle proteins, microtubules and mitosis. We spoke with Renata over Zoom to learn about her fascinating career, her advice for young scientists and the impact she hopes to make as an Editor.

The International Society for Biological and Environmental Repositories Presents Abstracts from Its Annual Meeting

The International Society for Biological and Environmental Repositories Presents Abstracts from Its Annual Meeting

Investigation on Circulating Neonatal Nav1.5 (nNav1.5) and Its Antibodies in the Blood and Serum of 4T1 Orthotopic Mice Model and Breast Cancer Patients from Hospital Universiti Sains Malaysia (HUSM)

Investigation on Circulating Neonatal Nav1.5 (nNav1.5) and Its Antibodies in the Blood and Serum of 4T1 Orthotopic Mice Model and Breast Cancer Patients from Hospital Universiti Sains Malaysia (HUSM)

Epstein barr virus associated Hodgkin and non-Hodgkin lymphoma

Background: The Epstein-Barr virus (EBV) has been linked to a variety of B-cell lymphomas, including Burkitt lymphoma (BL), classical Hodgkin lymphoma (cHL), and diffuse large B-cell lymphoma (DLBCL), at rates ranging from 5 to 10% of DLBCL cases to >95% in endemic BL. Methods: The Present study conducted in AL-Najaf Holy City; it is a cross sectional study. All patients were recruited from Middle Euphrates Cancer Research Unit Laboratory, from pre-existing samples. Tissues collected from the previously mentioned laboratory were fixed in 10% formalin and placed in a cassette before being embedded in paraffine. The tissues were examined and diagnosed as with Hodgkin disease. Eighty samples were subjected in the present study. All study group was diagnosed formerly as paraffin imbedding blocks. Molecular analysis also used to detection of EBV-LMP-1 Gene. Histopathological examination using hematoxylin and eosine stain (H&E) then examining under light microscope to check for EBV-LMP-1 protein in twenty block samples of tissues with lymphoma. Results: The result of the current study revealed that EBV-LMP-1 marker found in 12.5% of cases that had been diagnosed by PCR and IHC.

Effect of age and gender on EBV associated-Hodgkin lymphoma among sample of Iraqi patients

Background: Data on the age-specific prevalence of Epstein–Barr virus (EBV) infection. According to most studies, by the age of 35, more than 90% of adults are EBV antibody positive. Comparison of demographic groups could identify factors associated with its acquisition. Methods: The Present study conducted in AL-Najaf Holy City; it is a cross sectional study. All patients were recruited from Middle Euphrates Cancer Research Unit Laboratory, from pre-existing samples. Tissues collected from the previously mentioned laboratory were fixed in 10% formalin and placed in a cassette before being embedded in paraffine. The tissues were examined and diagnosed as with Hodgkin disease. forty samples were subjected in the present study. All study group was diagnosed formerly as paraffin imbedding blocks. Histopathological examination using hematoxylin and eosine stain (H&E) then examining under light microscope to check for twenty block samples of tissues with lymphoma. Results: From a total of 40 FFPET blocks were tested. The mean age was 34.92 years. The age group prevalence was as follows: ≤10 years, 2.5%; 10–20 years, 25%; 21–30 years, 22.5%; 31–40 years, 10%; and 41–50 years, 20%; ≥ 50, 20%.

Genomic Profiling In Fit, Elderly Patients Receiving Oral Fludarabine, Oral Cyclophosphamide and Intravenous Rituximab (ALLG CLL5 OFOCIR trial) As Initial Treatment Of Chronic Lymphocytic Leukemia (CLL)

BackgroundAnalysis for genomic alterations by fluorescence in-situ hybridization (FISH) allows prognostic stratification in chronic lymphocytic leukemia (CLL). Recurrent mutations in genes including NOTCH1, SF3B1, BIRC3 and others have recently been identified in patients with CLL by next-generation sequencing and may provide independent prognostic information (Rossi D et al. Blood 2013). The prognostic impact of these mutations following first-line therapy chemotherapy has not been consistent in the literature (Oscier D et al. Blood 2012; Ysebaert L et al. ASH Abstract 2012; Stilgenbauer S et al. ASH Abstracts 2012) and the prognostic significance of these mutations in elderly patients requiring therapy is not clearly defined (Chiaretti S et al. ASH Abstracts 2012). AimWe aimed to describe the incidence of genomic alterations by FISH and multiplexed targeted resequencing of genes with potential prognostic importance in an elderly population of patients requiring frontline treatment for CLL. MethodsPre-treatment peripheral blood and bone marrow aspirate samples were obtained from patients enrolled on a phase II randomized clinical trial investigating oral fludarabine, oral cyclophosphamide and i.v. rituximab (poFCivR) tolerance in previously untreated fit elderly patients with CLL (Australasian Leukemia and Lymphoma Group [ALLG] CLL5, ACTRN12608000404325). Peripheral blood lymphocytes were purified by Ficoll gradient separation. Bone marrow aspirate samples were analyzed for CLL-associated genomic changes with a Vysis CLL FISH probe kit (Abbott, Des Moines, IL). DNA was extracted from peripheral blood samples and we performed targeted genome sequencing of genes including TP53, ATM, NOTCH1, SF3B1, BIRC3, MYD88, XPO1 and FBXW7 using a TruSeq Custom Amplicon Design Panel on a MiSeq Sequencer as per manufacturer's protocol (Illumina, San Diego, CA). DNA mutation data was analyzed and compared to reported mutations in the literature and COSMIC database. Non-synonymous mutations with a mutation frequency >10% are reported. Results120 patients were enrolled into the CLL5 study and 78 samples were available for analysis. In these patients, FISH analysis identified 17p deletion in 10 pts (13%), 11q deletion in 6 pts (8%), trisomy 12 in 15 pts (19%), deletion 13q in 28 pts (36%), no abnormality (n=14, 18%), and not performed (n=5, 6%). By targeted sequencing, mutations were identified in TP53 (n=7, 9%), ATM (n=7, 9%), NOTCH1 (n=12, 15%), SF3B1 (n=10, 13%), MYD88 (n=6, 8%), BIRC3 (n=2, 3%), XPO1 (n=6, 8%), FBXW7 (n=4, 5%). Thirty patients (38%) did not have a mutation in one of these genes. Mutation distribution is demonstrated in Figure 1. Five of 7 mutations in TP53 occurred in the DNA-binding core domains (exons 5-8) and one mutation in the homo-oligomerization domain and one in exon 11. The majority of NOTCH1 mutations were identified in the PEST domains (11 of 12) and all but one mutation in SF3B1 were identified in the PP2A repeats 5-9. Correlation with patient characteristics, IGHV mutational sequencing and clinical outcome data is proceeding. [Display omitted] ConclusionWe demonstrate a high proportion of genomic mutations in fit patients age 65 years and older undergoing first-line therapy for CLL. By comparison with data from Italian CLL trials, UK LRF CLL4 and the CLL8 study, our study demonstrates similar mutational frequencies in previously described recurrently mutated genes. These mutations are assessable by massive parallel sequencing and may potentially offer useful predictive information in patients requiring therapy for CLL. AcknowledgmentsSupported by grants from CLL Global Research Foundation and Cancer Institute of NSW / Northern Translational Cancer Research Unit. Disclosures:Mulligan:Roche: Consultancy, Honoraria, Membership on an entity's Board of Directors or advisory committees, Research Funding; Sanofi: Consultancy, Honoraria, Membership on an entity's Board of Directors or advisory committees, Research Funding.

Abstract OT3-18-02: A comparative study of two detection platforms of circulating tumour cells in patients with metastatic breast cancer

Abstract Background: The enumeration of circulating tumour cells (CTCs) using the FDA-cleared CellSearch system has repeatedly been proven to bear prognostic significance in patients with localized and metastatic breast cancer (MBC) (Bidard et al. Lancet Oncol 2014; Janni et al. CCR 2016). In addition, CTC count has been suggested to be a valuable alternative for physician-based decision making in choosing between hormone therapy and chemotherapy as frontline treatment for patients with estrogen receptor-positive, HER2-negative MBC (Bidard et al. SABCS 2018). Challenges for the implementation of CTCs as a biomarker or a source of liquid biopsy in clinical practice include that most isolation methods only detect a limited number of CTCs in a blood sample in a subset of patients, even in those with clinically proven metastatic disease, and the limited availability of comparative data on technical variability between different CTC detection assays. Methods: In this study, we compare CTC detection rates in paired 7.5 ml blood samples from patients with MBC starting a new line of treatment using two commercially available CTC detection assays. All patients are recruited at GZA Hospitals, campus Sint-Augustinus, Antwerp, Belgium. The first CTC detection system is the CellSearch system, currently the only FDA-cleared system for the enumeration of CTCs in patients with MBC. This system employs an immunomagnetic CTC enrichment based on the expression of Epithelial Cell Adhesion Molecule (EPCAM) followed by an immunofluorescent CTC detection based on the expression of cytokeratins (CK) in EPCAM+ enriched cells. With this system, only EPCAM/CK double positive cells can be qualified as CTCs. The second system is the Rarecytes system, an automated scanner for immunofluorescently stained cell smears of the entire buffy coat of blood samples. With this system, CTCs will be enriched and detected in a single-step measurement using a cocktail of anti-EPCAM and anti-CK fluorescently labeled antibodies, thereby allowing to detect also single-marker positive CTCs. Based on CTC detection prevalence data in patients with MBC that are available for the CellSearch system and a prespecified hypothesis that with both detection methods a discordant result (relative to a cut-off of >= or <5 CTCs/7.5 ml blood) will be measured in 25% of patients, we aim to include 100 patients with MBC in this benchmarking study in order to be able to detect at least a 15% difference in CTC detection rate in favor of one of both techniques with 80% power at a significance level of 0.05. Concordance/discordance of CTC results between both detection methods will be analyzed both as a dichotomized variable relative to different cut-off values using the McNemar test and Cohen’s kappa test and as a continuous variable using the intraclass correlation coefficient. Expected results and conclusions: Upon submission of the abstract, 30 patients with MBC have been included in this trial after an accrual period of 4 months. Full results are expected mid 2020. Detailed comparison of CTC detection rates between both techniques will not only render analytical data on the performance of the Rarecytes system compared to the CellSearch system - the current gold standard in the field - but might also shed light on the frequency of EPCAM/CK single-positive CTCs in patients with MBC. Conflicts of interest: This study is partially funded by a grant from Rarecyte, Inc. Contact details for collaboration: Dieter Peeters (HistoGeneX NV, Antwerp, Belgium): dieter.peeters@histogenex.com; Andy Buys (Translational Cancer Research Unit, Antwerp, Belgium): andy.buys@gza.be. Citation Format: Dieter Peeters, Andy Buys, Hannah Ledegen, Kathlyn Moehlig, Steffi Oeyen, Dieter Rondas, Alexia De Beukelaer, Steven Van Laere, Liesbet Vervoort, Peter Vermeulen, Christopher Ung, Mark Kockx, Luc Dirix. A comparative study of two detection platforms of circulating tumour cells in patients with metastatic breast cancer [abstract]. In: Proceedings of the 2019 San Antonio Breast Cancer Symposium; 2019 Dec 10-14; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2020;80(4 Suppl):Abstract nr OT3-18-02.

42O - Profiles of genome complexity identify HORMAD1 as a driver of homologous recombination deficiency and platinum therapy response in triple-negative breast cancer

A. Grigoriadis1, D. Weekes1, J.A. Watkins1, P. Gazinska1, J. Frankum2, C. Lord2, A. Ashworth2, J.M. Ford3, M. Telli3, A. Tutt4 Breakthrough Breast Cancer Research Unit, King’s College London Guy’s Hospital, London, UK Cancer Genomics and Therapeutics, Institute of Cancer Research ICR, London, UK Division of Oncology, Stanford University School of Medicine, Stanford, CA, USA Breakthrough Breast Cancer Research Unit, King’s College London School of Medicine, London, UK

421 Molecular profiling for factors predicting sensitivity or resistance to therapy in relapsed child cancer

F. Saletta, C. Wadham, J. Byrne, D. Ziegler, G. McCowage, M. Haber, G. Marshall, M. Norris. The Children Hospital at Westmead, Children Cancer Research Unit, Westmead, Australia; Children’s Cancer Institute, Molecular Diagnostics, Randwick, Australia; Sydney Children’s Hospital, Kids Cancer Centre, Randwick, Australia; The Children Hospital at Westmead, Oncology department, Westmead, Australia; Children’s Cancer Institute, Experimental Therapeutics, Randwick, Australia

Improvements in SlicerRT, the radiation therapy research toolkit for 3D Slicer

Purpose: Recent years have seen growing interest in adaptive radiation therapy (RT), but the existing software tools are not ideal for research use, as they are either expensive and closed proprietary applications or free open-source packages with limited scope, extensibility, reliability, or user support. To address these limitations, we have developed SlicerRT, an open-source toolkit for RT research, providing fast computations and user-friendly interface for researchers. It is a medium for RT researchers to integrate their workflows in, assists clinical translation of experimental approaches, and enables comparative testing.Methods: SlicerRT builds on the widely used medical image visualization and analysis platform, 3D Slicer (www.slicer.org). In addition to the tools that 3D Slicer offers for visualization, registration and segmentation, the toolkit provides functions specifically designed for RT research. Established development processes as well as testing and validation environment ensure software quality. Standardized software development mechanisms of 3D Slicer were applied for documentation, distribution, and user support. The design and architecture of the toolkit was outlined in [1]. SlicerRT has evolved to contain the most widely used features in the field (see www.slicerrt.org), which have been defined through consensus discussions with a large pool of RT researchers.Results: Numerous modules have been developed for RT-specific features such as DICOM-RT data import and export, as well as dose analysis tools including dose volume histogram, dose accumulation, dose comparison, and isodose line and surface generation. The toolkit also includes contour analysis modules for handling various contour representations, contour morphology, and contour comparison. Advanced registration tools are provided by the included Plastimatch library [2]. The main new features developed in the last year are RT plan and RT image import and the subject hierarchy module, which arranges the loaded data to a structure familiar to clinicians, while providing advanced automation features through the underlying extendible mechanism. The toolkit is being used by several research groups to support and evaluate adaptive radiation therapy workflows. SlicerRT is available for download through the Extension Manager component of 3D Slicer 4.3 or later (see instructions on our website).Future work: As SlicerRT has matured to contain most of the planned functions, our focus has shifted from feature development to usability and stability. The subject hierarchy module, and its features such as bulk deformation of studies embodies our efforts toward usability, while the integration of the subject hierarchy and the advanced contour handling mechanism into the 3D Slicer core points towards stability, also benefiting the whole 3D Slicer community. There is ongoing work to evaluate and potentially standardize the way the 2D contours are rasterized into 3D volumes, thus mitigating the great variability between the different solutions for this problem. Current funding ensures continuous development for several years, so SlicerRT is expected to develop further.Acknowledgement: This work was in part funded by Cancer Care Ontario through Applied Cancer Research Unit and Research Chair in Cancer Imaging grants, and the Ontario Consortium for Adaptive Invention in Radiation Oncology (OCAIRO).

Abstract A10: A mutagenesis screen identifies tumor suppressors and kinases as potential driver genes of ovarian cancer

Abstract Background: The goal of this project is to increase our understanding of the molecular aetiology of ovarian cancer by utilizing Sleeping Beauty (SB) insertional mutagenesis. Methods: Breeding colonies of the following genetically engineered mice were established and cross bred: homozygous floxed SB (STOCK Rosa26-LsL-D-SB11;T2/Onc2,TG6113), homozygous floxed Brca1 knock-out (C57BL/6.Brca1tm2Brn) and Tp53 mutant (C57BL/6-Trp53tm1Tyj/J). CRE recombinase packaged into an adenoviral vector (AdCreM2, MicroBix Biosystems Inc, CA) was surgically injected under the ovarian bursal membrane of mature female mice to delete Brca1 and activate SB mutagenesis in the ovarian surface epithelium. Mice were monitored and sacrificed at ethically defined endpoints or a maximum of 15 months post-surgery. Tumors were assessed for SB transposase activity by immunohistochemical staining. DNA was extracted from paraffin embedded sections of ovarian tumors and underwent high-throughput sequencing for T2/Onc2 insertion sites (Illumina, University of Iowa). Genes with insertion sites were interrogated against SEOC data from The Cancer Genome Atlas (TCGA) to identify genes also altered in the human malignancy. Functional annotation clustering was performed using DAVID (Database for Annotation, Visualization and Integrated Discovery, v6.7). Results: Ovarian tumors were observed at low penetrance starting at 30 weeks post-surgery in SBflox/+Tp53mut/+ mice (6%, 3/48) and SBflox/+Brca1flox/flox p53mut/+ mice (8%, 4/50). No ovarian tumors were observed in SBflox/+Brca1flox/flox mice (n=38) or in SBflox/+Brca1flox/+ mice (n=26). Sequencing of the insertion sites identified a number of genes of which 67 were altered in 10 - 30% of cases in the TCGA SEOC dataset (N = 316). This gene-set was enriched for kinases (P = 0.003, False discovery rate (FDR) = 3%) including Fgfr2, Dyrk1a and Gsk3b, and small GTPase regulators (P = 0.01, FDR = 11%) including Smap2, Trio and Dock10. Other genes of interest included tumor suppressor genes Arid1b, Cdh4 and Wwox and the E3 ubiquitin ligase Ube3a. In the TCGA dataset, decreased progression free survival was associated with high expression of GSK3B, TRIO and UBE3A and low expression of WWOX. Conclusions: This screen identified a number of novel potential driver genes of ovarian cancer. In addition genes previously associated with ovarian cancer were also identified, providing proof of principle for this approach. Investigation of these novel genes may lead to further insights into the pathogenesis of ovarian cancer. This work was supported by Cancer Institute NSW, Cancer Council NSW and The Northern Translational Cancer Research Unit, NSW, Australia. Citation Format: Emily K. Colvin, Emily Fuller, Jizhou Cheng, Anthony Gill, Deborah J. Marsh, Viive M. Howell. A mutagenesis screen identifies tumor suppressors and kinases as potential driver genes of ovarian cancer. [abstract]. In: Proceedings of the AACR Special Conference on Advances in Ovarian Cancer Research: From Concept to Clinic; Sep 18-21, 2013; Miami, FL. Philadelphia (PA): AACR; Clin Cancer Res 2013;19(19 Suppl):Abstract nr A10.

Impact of the new EGF receptor and ALK testing guideline on personalized lung cancer medicine.

All advanced-stage lung adenocarcinoma patients should be tested for EGF receptor (EGFR) mutations and for EML4–ALK translocations as a guide for selecting them for tyrosine kinase inhibitor (TKI) therapy. The new EGFR and ALK guideline provides useful recommendations for appropriate testing [1]. Lung tumors are the leading cause of cancerrelated mortality worldwide [2]. Almost 80% of cases are non-small-cell lung cancer (NSCLC) and extensive disease is present at the time of diagnosis in the majority of patients. Systemic treatment with chemotherapy has reached a plateau of 10–20% response rates and 4–5 months of progression-free survival (PFS), and has not improved in more than a decade [3–5]. A first milestone in a new era of targeted therapy for this disease was reached with the discovery of a subset of tumors carrying activating mutations of EGFR, which can be selectively blocked with TKIs such as erlotinib and gefitinib. Numerous clinical studies have demonstrated the efficacy of these TKIs in the treatment of EGFR-mutated lung tumors, with 55–83% response rates and 9–13 month PFS [6,7]. A second milestone was the discovery, in 2007, of the EML4–ALK translocation in 6.7% of Japanese NSCLC patients [8]. The translocation was later found in 4–7% of Chinese [9], European [10] and US patients [11]. When treated with a novel ALK inhibitor, crizotinib, these patients showed response rates of 57% with a PFS greater than 6 months [12]. As a result of these new developments, many laboratories worldwide are currently testing lung tumor samples for EGFR mutations and EML4–ALK fusions. In an effort to standardize testing practices, the College of American Pathologists, the International Association for the Study of Lung Cancer and the Association for Molecular Pathology constituted writing and advisory panels, presided by three cochairs, to

Abstract 330: Utilization of Sleeping Beauty mutagenesis for the identification of potential driver genes of ovarian cancer.

Abstract Background: The goal of this project is to increase our understanding of the molecular aetiology of ovarian cancer by developing a mouse model of this tumor. Inherited mutation of BRCA1 is the best known risk factor for the most common subtype of ovarian cancer, i.e. serous epithelial ovarian cancer (SEOC); however, loss of Brca1 alone is insufficient for SEOC development in the mouse. Somatic mutation of TP53 is the most common molecular change in SEOC, but does not result in SEOC in the mouse, either alone or in combination with loss of Brca1. These findings suggest that additional currently unknown genetic factors are required for ovarian carcinogenesis. We hypothesized that random mutagenesis would activate these genetic factors and utilized Sleeping Beauty (SB) insertional mutagenesis to initiate ovarian tumors in mice that we have sequenced to determine potential genetic drivers of these tumors. Methods: Breeding colonies of the following genetically engineered mice were established and cross bred: homozygous floxed SB (STOCK Rosa26-LsL-D-SB11;T2/Onc2,TG6113), homozygous floxed Brca1 knock-out (C57BL/6.Brca1tm2Brn) and Tp53 mutant (C57BL/6-Trp53tm1Tyj/J). CRE recombinase packaged into an adenoviral vector (AdCreM2, MicroBix Biosystems Inc, CA) was surgically injected under the ovarian bursal membrane of mature female mice to delete Brca1 and activate SB mutagenesis in the ovarian surface epithelium. Mice were monitored and sacrificed at ethically defined endpoints or a maximum of 15 months post-surgery. Tumors were assessed for SB transposase activity by immunohistochemical staining. DNA was extracted from paraffin embedded sections of ovarian tumors and underwent high-throughput sequencing for T2/Onc2 insertion sites (Illumina, University of Iowa). Results: Ovarian tumors were observed at low penetrance starting at 30 weeks post-surgery in SBflox/+Tp53mut/+ mice (6%, 3/48) and SBflox/+Brca1flox/flox p53mut/+ mice (8%, 4/50). No ovarian tumors were observed in SBflox/+Brca1flox/flox mice (n=38) or in SBflox/+Brca1flox/+ mice (n=26). Sequencing of the insertion sites identified a number of genes of interest including kinases and tumor suppressors. Conclusions: While ovarian cancer had low penetrance in these models, a number of potential driver genes were identified which may provide insights into the pathogenesis of these tumors. The low penetrance of tumor from targeted epithelial cells with SB T2/Onc2 is similar to that seen for pancreatic cancer. For this reason, mice with SB T2/Onc3 which is reported to preferentially induce epithelial tumors are currently being monitored for the development of ovarian tumors. This work was supported by Cancer Institute NSW, Cancer Council NSW and The Northern Translational Cancer Research Unit, NSW, Australia. Citation Format: Emily K. Colvin, Emily Fuller, Jizhou Cheng, Anthony Gill, Deborah J. Marsh, Viive M. Howell. Utilization of Sleeping Beauty mutagenesis for the identification of potential driver genes of ovarian cancer. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 330. doi:10.1158/1538-7445.AM2013-330

Targeting of Rac GTPases blocks the spread of intact human breast cancer

High expression of Rac small GTPases in invasive breast ductal carcinoma is associated with poor prognosis, but its therapeutic value in human cancers is not clear. The aim of the current study was to determine the response of human primary breast cancers to Rac-based drug treatments ex vivo. Three-dimensional organotypic cultures were used to assess candidate therapeutic avenues in invasive breast cancers. Uniquely, in these primary cultures, the tumour is not disaggregated, with both epithelial and mesenchymal components maintained within a 3-dimensional matrix of type I collagen. EHT 1864, a small molecule inhibitor of Rac GTPases, prevents spread of breast cancers in this setting, and also reduces proliferation at the invading edge. Rac1+ epithelial cells in breast tumours also contain high levels of the phosphorylated form of the transcription factor STAT3. The small molecule Stattic inhibits activation of STAT3 and induces effects similar to those seen with EHT 1864. Pan-Rac inhibition of proliferation precedes down-regulation of STAT3 activity, defining it as the last step in Rac activation during human breast cancer invasion. Our data highlights the potential use of Rac and STAT3 inhibition in treatment of invasive human breast cancer and the benefit of studying novel cancer treatments using 3-dimensional primary tumour tissue explant cultures.

Governance approval for multisite, non‐interventional research: what can Harmonisation of Multi‐Centre Ethical Review learn from the New South Wales experience?

In 2007, New South Wales Health mandated the separation of ethical and scientific review from research governance at all New South Wales public health sites based on their distinction in the National Health and Medical Research Council National Statement. This separation allowed for single-site ethical review of multicentre studies. To investigate the time taken for governance approval of multicentre studies through the site-specific approval (SSA) process. A retrospective audit of the SSA process for five non-interventional studies proposed by a university cancer research unit. The median total governance approval time for all submissions (n= 28) was 12 weeks (range 2.5-64); median time from starting the SSA to submission was 8 weeks (range 1-48) and median time for governance approval was 5 weeks (range 0.3-40). Approval times were shorter for public compared to private institutions. Reasons for delays in finalising submissions for approval were the absence of institutional governance officers, lack of clarity regarding signatories, the need to identify a principal investigator employed by the institution, and lack of recognition of ethical approval by private institutions. The need to develop legal agreements between the university and hospital was the main reason for lengthy delays in obtaining approval. The advantages of a harmonised single ethical review process were undermined by the coexistence of a fragmented, complex and lengthy governance approval process. This experience has implications for the success of the national Harmonisation of Multi-Centre Ethical Review (HoMER) model. A harmonised and fully supported national approach to research governance should be developed contemporaneously with HoMER.

Breast tumour stroma is a prognostic indicator and target for therapy

The development of the breast is exquisitely sensitive to interactions between the epithelium and stroma. Experimental evidence indicates that a reduction in signalling between any of the stromal cell types (fibroblasts, macrophages, endothelial cells and adipocytes) results in reduced or absent breast development [1], although all interactions appear to be orchestrated by the epithelial cell oestrogen receptor alpha [2]. The epithelial-stromal interactions that occur in tumours are less well characterised but there is no doubt there is expansion of the stroma as well as of the epithelium during tumour development [3,4]. Recent data indicate that the prognosis after breast cancer diagnosis relates to stromal type, and experimental and clinical studies directed at modifying the stroma (for example, angiogenesis inhibitors) suggest that the stroma is a target for therapy that is worthy of further exploration Studies of separately microdissected breast stroma and epithelium from normal lobules compared with ductal carcinoma in situ (DCIS) and invasive cancer indicate that extensive changes in gene expression in both the epithelial and stromal compartments occur during cancer development. These data strongly support the hypothesis that performing microdissections can be less optimal for gene expression profiling studies or to exclude cancers with a prominent stroma. Some array-based studies have had a requirement of more than 50% of cancer cells in the biopsies taken for array profiling; this may result in exclusion of biologically important cancers. Compared with the intralobular stroma of the normal breast lobule, Ma and colleagues reported that 2,338 genes were upregulated and 1,234 genes were downregulated in the stroma of DCIS [5]. A further 76 genes were upregulated and 229 genes were downregulated in the stroma of invasive tumours, indicating that most of the changes had occurred in DCIS suggesting that paracrine and endocrine influences are driving stroma formation rather than cell interactions, since the basement membrane is largely intact in DCIS. In a similar study examining stroma separated from the epithelium, Casey and colleagues demonstrated that the major changes of gene expression were upregulation of genes for the extracellular matrix and proteases in the stroma and downregulation of cytoskeletal proteins such as keratins, tubulins and adhesion molecules leading to increased cell motility in the tumour epithelium [6]. Invasive tumours have been likened to ‘wounds that do not heal’ [7]. In order to establish whether tumours induced gene expression similar to wounds, Chang and colleagues investigated whether they expressed the genes induced by serum in fibroblasts (the equivalent of wounding) [8,9]. The expression of 422 selected genes changed by serum in tumours was associated with a poor prognosis, whereas tumours with no change tended to have a good prognosis. In this study, although the genes were produced in serum-treated fibroblasts, they could have been expressed in epithelial cells of the tumours studied. In order to assess the prognostic and predictive significance of genes strictly of stromal origin, Finak and colleagues isolated stroma from normal lobules and tumours by laser capture microdissection, and derived a 26-gene expression signature that was a poor prognostic indicator irrespective of breast tumour subtype and standard prognostic indicators and that also indicated resistance to standard treatments [10]. The stromal signature, however, has been described to be associated with a basal type of breast cancer in three independent datasets, including the Canadian study [11]. Other gene signatures derived from the whole tumour and searched for potential stromal genes were also able to detect a poor prognosis signature [12] and to detect a stromal signature that indicated failure to respond to neoadjuvant 5-fluorouracil, epirubicin, and cyclophosphamide chemotherapy [13]. More recently two groups have demonstrated downregulation of a protein (caveolin-1) that acts as a scaffold protein in cell surface pits or caveolae (important for * Correspondence: Anthony.Howell@christie.nhs.uk Breakthrough Breast Cancer Research Unit, Paterson Institute for Cancer Research, University of Manchester, The Christie NHS Foundation Trust, University Hospital of South Manchester, Manchester M20 4BX, UK Howell et al. Breast Cancer Research 2009, 11:S16 http://breast-cancer-research.com/content/11/S3/S16

Vincristine pharmacokinetics is related to clinical outcome in children with standard risk acute lymphoblastic leukemia

Vincristine is a key drug in the treatment of childhood and adult acute lymphoblastic leukemia (ALL), and many other childhood malignancies. Despite decades of wide clinical use, no data on the correlation between vincristine pharmacokinetics and long-term clinical outcome have been published. We here report clinical data (median follow-up time 10.5 years, range 7.3-12 years) for 86 children with B-cell precursor ALL, in whom vincristine kinetics were studied on treatment day 1. The median total plasma clearance was 429 and 331 ml/min per m(2) and the area under the plasma concentration-time curve (AUC) was 4.49 and 5.40 mg/l x min in relapse and non-relapse patients, respectively (not significant). In standard risk patients, where treatment depends more heavily on vincristine than in other subgroups, the relative risk (RR) of relapse was significantly increased for patients with clearance values above median (RR 5.2; P = 0.036), or AUC values below median (RR 5.8; P = 0.025). Our data suggest a relationship between the antileukemic effect and the systemic exposure of the drug, which warrants further studies.

The Institute of Cancer Research, Clinical Trials and Statistics Unit (ICR-CTSU)

This section provides current contact details and a summary of recent or ongoing clinical trials being coordinated by The Institute of Cancer Research, Clinical Trials and Statistics Unit (ICR-CTSU). Clinical trials include: The NCRI Adjuvant Breast Cancer (ABC) trial. ISRCTN31514446The UK randomized trial of hormone replacement therapy (HRT) in women with a history of early stage breast cancer. ISRCTN29941643NCRI Standardisation of Breast Radiotherapy (START) trial. ISRCTN59368779TACT: A randomized trial of standard anthracycline-based chemotherapy (fluorouracil, epirubicin and cyclophosphamide (FEC) or epirubicin and CMF (Epi-CMF)) versus FEC followed by sequential docetaxel in women with early breast cancer. ISRCTN 79718493SoFEA: Study of Faslodex with or without concomitant Arimidex versus Exemestane following progression on non-steroidal Aromatase inhibitors. ISRCTN: 44195747TACT2: Trial of accelerated adjuvant chemotherapy with Capecitabine in early breast cancer. ISRCTN68068041FAST trial: Prospective randomized clinical trial testing 5.7 Gy and 6.0 Gy fractions of whole breast radiotherapy in terms of late normal tissue responses and tumour control.IMPORT low trial.IMPORT high trial.Adjuvant chemotherapy in older women (ACTION).

Researchers reflect from the cancer precipice

This paper presents the reflections of two researchers employed at a cancer research unit. We reflect often together about the difficult emotional issues involved in working with cancer patients. To formalize these conversations we audiotaped and transcribed four dialogue sessions during which we asked ourselves the following questions: What are we learning about ourselves as we become immersed in this work? How do we negotiate our positions as both social scientists and as individuals who empathize with research participants about emotional/existential issues? This paper will explore our responses and reflections about some of our emotional connections to the women with whom we work, as well as our limitations and points of disconnect. We conclude that researchers are not dispassionate observers of others’ experiences and could benefit from working in an atmosphere where reflecting and processing painful emotions with colleagues or through writing is fostered and valued.

Statistical problems with 'optimal' thresholds in studies of new prognostic factors in urology.

BJU InternationalVolume 85, Issue 4 p. 392-397 REVIEW Statistical problems with ‘optimal’ thresholds in studies of new prognostic factors in urology P.E. Keegan, P.E. Keegan Departments of Surgery, Cancer Research Unit, University of Newcastle upon Tyne, UKSearch for more papers by this authorJ.N.S. Matthews, J.N.S. Matthews Statistics and theSearch for more papers by this authorJ. Lunec, J. Lunec Cancer Research Unit, University of Newcastle upon Tyne, UKSearch for more papers by this authorD.E. Neal, D.E. Neal Departments of Surgery,Search for more papers by this author P.E. Keegan, P.E. Keegan Departments of Surgery, Cancer Research Unit, University of Newcastle upon Tyne, UKSearch for more papers by this authorJ.N.S. Matthews, J.N.S. Matthews Statistics and theSearch for more papers by this authorJ. Lunec, J. Lunec Cancer Research Unit, University of Newcastle upon Tyne, UKSearch for more papers by this authorD.E. Neal, D.E. Neal Departments of Surgery,Search for more papers by this author First published: 10 December 2003 https://doi.org/10.1046/j.1464-410x.2000.00491.xCitations: 16 Mr Philip E. Keegan, The Department of Surgery, The Medical School, Framlington Place, Newcastle upon Tyne NE2 4HH, UK. Read the full textAboutPDF ToolsRequest permissionExport citationAdd to favoritesTrack citation ShareShare Give accessShare full text accessShare full-text accessPlease review our Terms and Conditions of Use and check box below to share full-text version of article.I have read and accept the Wiley Online Library Terms and Conditions of UseShareable LinkUse the link below to share a full-text version of this article with your friends and colleagues. Learn more.Copy URL Share a linkShare onFacebookTwitterLinkedInRedditWechat Citing Literature Volume85, Issue4March 2000Pages 392-397 RelatedInformation